GJRMI - Volume 2, Issue 6, June 2013 by Dr. Hari Venkatesh.K.Rajaraman

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An International, Peer Reviewed, Open access, Monthly E-Journal

ISSN 2277 â&#x20AC;&#x201C; 4289 www.gjrmi.com Editor-in-chief Dr Hari Venkatesh K Rajaraman

Managing Editor Dr. Shwetha Hari

Administrator & Associate Editor Miss. Shyamala Rupavahini

Advisory Board Prof. Rabinarayan Acharya Dr. Dinesh Katoch Dr. S.N.Murthy Dr. Mathew Dan Mr. Tanay Bose Dr. Nagaraja T. M. Prof. Sanjaya. K. S. Dr. Narappa Reddy

Editorial board Dr. Kumaraswamy Dr. Madhu .K.P Dr. Sushrutha .C.K Dr. Ashok B.K. Dr. Janardhana.V.Hebbar Dr. Vidhya Priya Dharshini. K. R. Mr. R. Giridharan

Honorary Members - Editorial Board Dr Farhad Mirzaei Mr. Harshal Ashok Pawar

INDEX – GJRMI, Vol.2, Iss. 6, June 2013 MEDICINAL PLANTS RESEARCH Botany SUPPRESSION OF LEAF BLIGHT PATHOGEN ALTERNARIA LONGIPES OF MEDICAGO SATIVA BY FLUORESCENT PSEUDOMONAS STRAIN BRL-1 Sen Surjit, Acharya Krishnendu

401–409

Animal Production Technology SENSITIVITY OF BUCK (MALE GOAT) SEMEN MICROBIAL ISOLATES TO THREE PLANT EXTRACTS FROM CAMEROON Salah A Martin, Yongabi K A

410–417

Review Article A BRIEF REVIEW ON GINKGO BILOBA L. (MAIDENHAIR TREE) - A RARE MULTIPURPOSE MEDICINAL PLANT Patel Hemangi, Ingalhalli Rajashekhar

418–427

INDIGENOUS MEDICINE Ayurveda – Kaumarabrithya EFFECT OF BADARA STEM BARK (ZIZIPHUS JUJUBA LAMB.) IN THE MANAGEMENT OF VATAJA KASA IN CHILDREN Chethan Kumar V K, Shailaja U

428–434

Ayurveda – Dravya Guna EFFECT OF PARNAYAVANI (COLEUS AMBOINICUS LOUR.) ON MES INDUCED EPILEPSY IN RATS Sharma Monica, Khemani. N, Singh J

435–440

Ayurveda – Dravya Guna ANTIMICROBIAL POTENTIAL OF THE WILD AND CULTIVATED VARIETY OF ERANDA (RICINUS COMMUNIS LINN.) ROOT Doshi Krunal A, Sagar Dhwani H, Acharya R N

441–447

Ayurveda – Review Article – Kaumarabhritya PROBABLE ETIOPATHOGENESIS (SAMPRAPTI) OF AUTISM IN FRAME OF AYURVEDA IN RELATION TO INTENSE WORLD THEORY Yadav Deepmala, Behera Banshidhar, Kumar Abhimanyu

448–459

Ayurveda – Review Article CRITICAL APPRAISAL SKILL FOR THE ARTICLE PUBLISHED IN AYURVEDIC HEALTH DOMAIN Vaikos C D, Waghchoure Ashok

460–464

Unani – Review Article TRADITIONAL USE OF KAHU (LACTUCA SCARIOLA L.) - A REVIEW Arif Mohammad

465–474

COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF ARAGVADHA (CASSIA FISTULA . L. ), OF THE FAMILY CAESALPINACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA HIMALAYA HERBAL HEALTH CARE’S quarterly INFOLINE endorses & features Global Journal of Research on Medicinal plants & Indigenous medicine (GJRMI) in their recent Vol. 9, Jan-mar 2013 issue. The Editor-in-Chief, GJRMI, on behalf of the Board members & Referees, thank the scientific publications division, The Himalaya Drug Company for the endorsement. Click this Face book Link to read the below material

Research article SUPPRESSION OF LEAF BLIGHT PATHOGEN ALTERNARIA LONGIPES OF MEDICAGO SATIVA BY FLUORESCENT PSEUDOMONAS STRAIN BRL-1 Sen Surjit1, Acharya Krishnendu2* 1, 2

Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata 700 019, INDIA

*Corresponding Author: nataraj_2001us@yahoo.com; Fax : +91 033 24764419; Phone : +91 8013167310

Received: 05/04/2013; Revised: 17/05/2013; Accepted: 26/05/2013

ABSTRACT A leaf blight disease caused by the fungus Alternaria longipes was observed in Medicago sativa Linn. commonly known as ‘Alfalfa’, a tonic plant cultivated at different areas of West Bengal, India. A potential biocontrol agent viz. fluorescent Pseudomonas strain BRL-1 showed both in vitro and in vivo antagonistic activity against the pathogen. In dual culture bioassay as circular and semicircular patterns, the isolate quantitatively inhibits the growth of the pathogen by about 74% and 65%, respectively. Foliar application of a talc-based formulation of the antagonist to field condition revealed that the maximum mean disease index reached to 1.097 and 1.19 in 2010 and 2011, indicating around 77% and 76% reduction in disease severity when compared to non-treated control. Transferring the outcome of this work in the field would benefit the growers by facilitating bioorganic production of this important medicinal plant and finally consumers can get hazard free natural plant product. KEYWORDS: Alfalfa, biocontrol, leaf blight, medicinal plant

Cite this article: Sen. S., Acharya K., (2013), SUPPRESSION OF LEAF BLIGHT PATHOGEN ALTERNARIA LONGIPES OF MEDICAGO SATIVA BY FLUORESCENT PSEUDOMONAS STRAIN BRL-1, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 401–409

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INTRODUCTION Medicago sativa (Alfalfa) is a perennial flowering plant in the family Fabaceae cultivated as an important forage as well as medicinal plant in many countries. It is used as tonic due to presence of high percentage of proteins (60.5%), minerals, enzymes, vitamins etc. Alfalfa is a valuable source of vitamins A and E, fresh is rich in Vitamin C (1.78 mg/g) but it loses 80% of vitamins on drying (Rashmi et al., 1997). The leaves are also rich in vitamin K, which is used medicinally to encourage the clotting of blood (Weiner, 1980). Alfalfa leaves, either fresh or dried, have traditionally been used as a nutritive tonic to stimulate the appetite and promote weight gain (Foster and Duke, 1990). The plant is antiscorbutic, diuretic, oxytocic, haemostatic, nutritive, stimulant (Duke and Ayensu, 1985). The expressed juice is emetic and is anodyne in the treatment of gravel (Duke and Ayensu, 1985). The plant is taken internally for debility in convalescence, anaemia, haemorrhage, menopausal complaints, pre-menstrual tension, fibroids etc (Bown, 1995; Chevallier, 1996). A poultice of the heated leaves has been applied to the ear in the treatment of earache (Moerman, 1998). The plant is grown commercially as a source of chlorophyll and carotene, both of which have proven health benefits (Foster and Duke, 1990). The leaves also contain the anti-oxidant tricin (Foster and Duke, 1990). The root is febrifuge and is also prescribed in cases of highly coloured urine (Duke and Ayensu, 1985). Extracts of the plant has antibacterial activity (Duke and Ayensu, 1985). Paul and Singh, (2002) reported fungal phytopathogens attack medicinal and aromatic plants leading to significant quantitative and qualitative loss. Alternaria leaf blights were very common in medicinal plants cultivated in various districts of West Bengal, India (Maiti et al., 2007a, b, c). Today biocontrol agents are gaining importance in the field of disease management of medicinal plants as they do not have the adverse effects like that of fungicides which eventually contaminate the purity of

plant drugs (Mathivanan et al., 2005). Extensive research is underway globally to exploit the potentiality of pseudomonads, which help to protect crops from phytopathogens and are metabolically and functionally more diverse (Choudhury et al., 2009). A wide range of fluorescent pseudomonads have been reported for having in vitro and in vivo biocontrol potentiality against variety of phytopathogens (Kishore et al,. 2005; Mansoor et al., 2007; Sen et al., 2009; 2012; Maiti et al., 2012). The present study demonstrates the efficiency of a potential biocontrol agent fluorescent Pseudomonas BRL-1 for controlling leaf blight disease in Medicago sativa. MATERIALS AND METHODS Organisms The pathogenic organism was isolated from the diseased leaves of M. sativa as a pure culture on potato dextrose agar medium (PDA), identified as Alternaria longipes (Maiti et al., 2007b). The culture was maintained in the same medium and stored at 4Â°C for further study. The biocontrol agent fluorescent Pseudomonas BRL-1 was obtained from our laboratory culture stock. The antagonist was subcultured and maintained on tryptic soy agar (TSA) medium for subsequent use. Dual Culture Bioassay Fluorescent Pseudomonas BRL-1 from 24 h old culture (107 cells ml-1) was streaked in the peptone (1%) glucose (2%) agar (2%) (PGA) plate as circular / O and semicircular / U pattern. Then mycelial disc (5 mm diameter) of 3 days old culture of A. longipes was subsequently inoculated at the center of O or U shaped region on the PGA plates (Skidmore and Dickinson, 1976). Inoculation only with the pathogen served as control. The plates in triplicate were incubated at 30Â°C for 5 days and diameter of colony growth was measured at every 24 h intervals. Light microscopic (Zeiss AX 10) studies were also performed to detect physical and / or morphological changes of mycelia.

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 401–409

Talc based formulation and survival of fluorescent Pseudomonas BRL-1 Talc-based formulation of the antagonist was prepared using a method developed by Vidyasekaran and Muthamilan (1995) modified. Ten gram carboxy methyl cellulose (CMC) per kg of sterile talc was used as adhesive. The bacterial suspension (8 × 109 CFU ml-1) was mixed with sterile talc (400 ml kg-1) and air dried (approximately to 35% w/w, moisture content). The formulation was stored at 4°C for up to 180 days. The antagonist in the talc-based formulation was monitored in vitro with respect to its shelf-life and viability under this storage condition. Survival of the bacterial population in the formulation was assayed at 30 days intervals using King’s medium B in a dilution plate assay according to Vidyasekaran and Muthamilan, (1995). Survival of fluorescent Pseudomonas BRL-1 on phylloplane Survival and multiplication of fluorescent Pseudomonas BRL-1 on the phylloplane of Medicago was determined following the method of Kishore et al. (2005). The talc-based formulation (4 g l-1) of the antagonist was applied as foliar sprays with an initial cell density of 106 CFU g-1 of fresh leaves. Leaves were collected at 3 days intervals, washed in sterile water, serially diluted and plated (in triplicate) on King’s B agar medium. Colonies of BRL-1 was determined after 48 h of incubation at 30°C utilizing its colony morphology, fluorescence characteristics. The bacterial populations were expressed as log CFU g-1 of leaf. The whole experiment was performed thrice. Field studies The Medicinal Plant Garden, R.K. Mission Ashrama, Narendrapur, India was used for the field experiments during 2010 and 2011 when the environmental conditions were conducive (January to July) for the rapid spread of A. logipes on M. sativa. The trial was conducted as a randomized complete block design with

three replicate plots (3 × 4 m2) and forty plants per plot. Well-rotted farmyard manure was mixed well into the soil before planting the seedlings. Four-week-old disease free seedlings were transplanted to the random blocks on end of January allowing A. longipes leaf blight to develop naturally (Silva et al., 2004). The talc based formulation the antagonist was prepared by dissolving it in water (4 g l-1) allowing it to settle for 1 h, and filtering the solution through muslin cloth. The filtrate was applied as a foliar spray using a low volume sprayer beginning at transplant and repeating every 15 days for up to six months i.e. end of July 2010 and 2011. Plots sprayed with the talc-based carrier without the biocontrol agent was served as control. Twenty-five plants from each plot were rated for disease severity at 15 day intervals starting at transplant for up-to six months using a 0-5 rating scale (Kishore et al., 2005). Statistical analysis The disease severity data were statistical analysed by using analysis of variance (ANOVA) followed by Tukey’s Test to find out significance level at 1% (p<0.01). RESULTS Dual culture of fluorescent Pseudomonas BRL-1 and A. longipes Growth inhibition of A. longipes by strain BRL-1 was observed in dual culture. Mycelial growth was restricted near bacterial streak and continued away from it. The growth inhibition of A. longipes remained proportionate with an increased incubation period of up to 5 days. Quantitatively fluorescent Pseudomonas BRL1 inhibited the growth of A. longipes by 73.76% and 64.66% in circular and semicircular streaks after 120 h of incubation respectively (Fig. 1). After 120 h of confrontation, microscopic examination of the mycelia at the inhibition zone showed markedly damaged hyphae as evidenced by disorganization of the cytoplasm, shriveling, growth deformities, swelling, fragmentation, short branching and lysis (Fig. 2).

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 401â&#x20AC;&#x201C;409

% Inhibition of radial mycelial growth

Fig. 1 Inhibition of A. longipes by fluorescent Pseudomonas BRL-1 under dual plate culture using circular (O) and semicircular (U) method. Each point represents the mean Âą SE (standard error) of three separate experiments, each in triplicate. 80 70 60 50 O Shaped

U Shaped

30 20 10 0 0h

24h

48h

72h

96h

120h

Incubation, h

Fig. 2 [A] Microscopic observation of mycelium of control plate showing normal hyphal structure and spore [B] Microscopic observation of mycelium of treated plate showing hyphal deformities, short branching, and disorganization of hyphal cytoplasmic contents.

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 401–409

Fig. 3 Survival of fluorescent Pseudomonas BRL-1 in talc based formulations stored at 4°C. Each point represents the mean ± SE (standard error) of three separate experiments, each in triplicate.

log cfu/g of talc based formulation

9 8.5 8 7.5 7 6.5 6 5.5 5 0

120

150

180

Storage at 4 C

Fig. 4 Changes in the populations of fluorescent Pseudomonas BRL-1 on M. Sativa phylloplane growing under field condition. Each point represents the mean ± SE (standard error) of three separate experiments, each in triplicate. 7

log CFU / g

2 0

Days after inoculation

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 401–409

Table 1 Field study for the control of leaf blight disease of M. sativa caused by A. longipes in 2010 and 2011. Foliar application of the talc based formulation on date of transplantation and at an interval of 15 days until 180 days. Disease index was rated on a 0–5 scale. Values are mean ± SE of twenty-five randomly selected plants per plot of three individual plot experiments Days after

Disease index

transplantation

2010

2011

Control

Treated

0.159 ± 0.032 0.102 ± 0.016*

0.182 ± 0.009

0.162 ± 0.006*

0.254 ± 0.014 0.183 ± 0.007*

0.358 ± 0.024

0.213 ± 0.009*

0.678 ± 0.021 0.321 ± 0.012*

0.773 ± 0.014

0.424 ± 0.016*

0.947 ± 0.006 0.578 ± 0.014*

1.047 ± 0.019

0.612 ± 0.015*

1.542 ± 0.019 0.634 ± 0.012*

1.96 ± 0.017

0.69 ± 0.033*

2.126 ± 0.015 0.719 ± 0.012*

2.724 ± 0.044

0.794 ± 0.017*

105

3.241 ± 0.014 0.801 ± 0.018*

3.81 ± 0.017

0.912 ± 0.009*

120

3.734 ± 0.008 0.886 ± 0.005*

3.965 ± 0.026

0.98 ± 0.014*

135

4.225 ± 0.033 0.915 ± 0.018*

4.45 ± 0.014

1.03 ± 0.006*

Control

150

4.412±0.011

0.988±0.024*

4.561±0.032

1.087 ± 0.024*

165

4.61 ± 0.009 1.029 ± 0.015*

4.73 ± 0.019

1.12 ± 0.016*

180

4.76 ± 0.012 1.097 ± 0.016*

4.88 ± 0.014

1.19 ± 0.015*

* Data in each row differ significantly with control according to Tukey’s test (P<0.01) Survival of fluorescent Pseudomonas BRL-1 in talc-based formulation and on the M. sativa phylloplane

of leaves at 15th day after treatment (Fig. 4). No colonies of BRL-1 were recovered on the controls.

Survival of the antagonist in talc-based formulation stored at 4°C was monitored for 180 days (Fig. 3). With time, the initial population of BRL-1 (8.7 log CFU g-1) in talc based formulation gradually decreased. Approximately 30% decrease in the colonyforming unit was estimated on 180th day after storage at 4°C. Foliar application of this formulated antagonist showed the initial population (within 1 h after application) to be log 6.4 CFU g-1 of leaves. The initial population steadily declined to log 3.3 CFU g-1

Field studies Talc based formulation of fluorescent Pseudomonas BRL-1 was evaluated in the field on M. sativa for two successive seasons. After the 3rd and 4th spray of the formulation, new symptoms of leaf blights were inhibited in treated plots. The mean disease index after 180 days in control field reached to 4.76 and 4.88 in 2010 and 2011, respectively, where most of all plants were severely affected and more than 60% leaves were defoliated (Table 1). On the

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contrary, the disease index in BRL-1 treated fields reached only 1.097 and 1.19 in 2010 and 2011, indicating around 77% and 76% reduction in disease severity respectively. DISCUSSION Over the past twenty years, the control of plant pathogenic fungi by antagonistic bacteria and fungi has been the topic of numerous studies. The majority of these studies dealt with antagonists controlling soil borne pathogens and to a less significant extent, foliar pathogens. Pseudomonas strains were evaluated for their ability to control Sclerotinia homeocarpa and Bipolaris sorokiniana on the phylloplane of Kentucky bluegrass (Hodges et al., 1994). Pseudsomonas, Bacillus and Stenotrophomonas showed in vitro antifungal activity against Verticillium dahliae var. longisporum (Berg et al., 1998). Rozsnyay et al. (1992) showed that some strains of P. fluorescens and some fungi inhibited canker and dieback diseases of apricot. The strain BRL-1 have been reported as a potential biocontrol agent for controlling both soil borne and aerial pathogen as evidenced through in vivo and field experiments conducted earlier (Sen et al., 2009; 2012). An effort to characterize the mycoparasitic action of BRL-1 in the present study revealed that the bioagent had the potentiality to manage A. longipes both in vitro and in vivo. Recently it has been reported that the same antagonist had the ability to control leaf spot disease of Stevia rebaudiana, caused by Alternaria alternata (Sen et al., 2012). Coagulation of cytoplasmic contents and disorganization of fungal hyphae upon dual-culture attributed by the strain BRL1 mainly by the release of secondary metabolites like siderophore, Indole Acetic Acid (IAA) and lytic enzymes (Sen et al., 2012). Similar type of observations were made earlier by Bano and Musarrat (2002), Kapsalis et al. (2008), Ramyasmruthi et al. (2012) Maiti et al. (2012) who demonstrated that the exposure of different phytopathogenic fungi to

secondary metabolites and lytic enzymes such as chitinase and protease could result in the degradation of the fungal cell wall. IAA reduce spore germination, mycelial dry weight and protein content of the pathogenic fungi and thus prevent significantly any chance for disease induction (Sharaf and Farrag, 2004; Sen et al., 2009). According to several studies, siderophore, chitinase and protease produced by fluorescent pseudomonads are known to inhibit the growth of some fungal pathogens (Lim and Kim 1995; Kapsalis et al. 2008; Maiti et al. 2012). CONCLUSION In the present study, we showed that the strain BRL-1 could live well in talc-based formulation stored at 4Â°C for 6 months. It has been reported that fluorescent pseudomonads could stay alive in certain formulations (Connick 1988; Mugilan et al. 2011). Moreover, the success of any biocontrol formulation not only depends on the ability of the biocontrol agent to survive in the formulation, but also its capacity to survive on the host plant to which it is applied. Bunster et al. (1989) and Lindow (1995) reported foliar application of fluorescent pseudomonads are able to survive well on leaf surfaces by occupying particular surface sites. Our study complements their findings by demonstrating that an average of 53% cell population survived for 15 days after application of talc-based formulation of BRL-1 on M. sativa. The formulation when applied in every 15 days, for two consecutive seasons successfully reduced symptoms of A. longipes leaf blight on M. sativa by 77% and 76% respectively. This ultimately helps the farmers to limit the use of hazardous fungicides and simultaneously saving their crops. Finally, the concerted effort of academic, federal and private sector scientists should lead to the development of effective and consistent biocontrol of aerial plant diseases based on an integrated biological control strategy.

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 401–409

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Mansoor FK, Sultana V, Haque SE (2007). Enhancement of biocontrol of Pseudomonas aeruginosa and Paecilomyces lilaceinus against root rot of mungbean by a medicinal plant Launaea nudicaulis L. Pak J Bot. 39: 2113–2119. Mathivanan N, Prabavathy VR, Vijayanandaraj VR (2005). Application of talc formulations of Pseudomonas fluorescens Migula and Trichoderma viride Pers. ex S.F. Gray decrease the sheath blight disease and enhance the plant growth and yield in rice. Phytopathology. 153: 697–701. Moerman D (1998). Native American Ethnobotany: Timber Press: Oregon. Mugilan I, Gayathri P, Elumalai EK, Elango R (2011). Studies on Improve Survivability and Shelf Life of Carrier Using Liquid Inoculation of Pseudomonas striata. Int J Pharma Biol Arch. 2: 1271–1275. Paul YS, Singh A (2002). Diseases of Medicinal Plants V. In: Sinha P, Grovil JN, Singh VK. (eds.) Recent Progress in Medicinal Plants Diseases and their Management: Sci Tech Publishing LLC U S A. pp.95-104. Ramyasmruthi S, Pallavi O, Pallavi S, Tilak K, Srividya S (2012). Chitinolytic and secondary metabolite producing Pseudomonas fluorescens isolated from Solanaceae rhizosphere effective against broad spectrum fungal phytopathogens. Asian J Plant Sci Res. 2: 16–24. Rashmi R, Sarkar M, Aditya V (1997). Cultivation af Alfalfa (Medicago sativa L). Ancient Sci Life. 17: 117–119 Source of Support: Nil

Rozsnyay ZD, Hevesi M, Klement Z, Vajna L (1992). Biological control against canker and dieback diseases of apricots. Acta Phytopathol Entomol Hung. 27: 551–556. Sen S, Biswas G, Basu SK, Acharya K (2012). Management of leaf spot disease of Stevia rebaudiana Bertoni with antagonistic bacteria. Aust J Crop Sci. 6: 350–356. Sen S, Rai M, Acharya R, Dasgupta S, Saha A, Acharya K (2009). Biological control of pathogens causing the Cymbidium Pseudobulb rot complex using fluorescent Pseudomonas strain BRL-1. J Plant Pathol. 91: 617–621. Sharaf EF, Farrag AA (2004). Induced resistance in tomato plants by IAA against Fusarium oxysporum f.sp lycopersici. Pol J Microbiol. 53: 111– 116. Silva HS, Rumetro RS, Cakker R, Fillio JL, Pukera ES, Mizubuti G, Mucnteer A (2004). Induction of Systemic Resistance by Bacillus cereus against tomato foliar diseases under field conditions. J Phytopathol. 152: 371– 375. Skidmore AM, Dickinson CH (1970). Colony interaction and hyphal interference between Septoria nodorum and phyloplane fungi. Trans Bri Mycol Soc. 66: 57–64. Vidyasekaran P, Muthamilan M (1995). Development of Formulation of Pseudomonas fluorescens for Control of Chickpea Wilt. Plant Dis. 79: 782–786. Weiner MA (1980). Earth Medicine, Earth Food: Ballantine Books. Conflict of Interest: None Declared

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Research article SENSITIVITY OF BUCK (MALE GOAT) SEMEN MICROBIAL ISOLATES TO THREE PLANT EXTRACTS FROM CAMEROON Salah A Martin1*, Yongabi K A2 1

Department of Animal Production Technology, College of Technology, University of Bamenda, Cameroon, P.O Box 39 Bamenda 2 Phyto-biotechnology Research Foundation, P.O. Box 9221, Bamenda-Cameroon *Corresponding Author: E-mail: salahmartin@yahoo.fr

Received: 07/04/2013; Revised: 31/05/2013; Accepted: 07/06/2013

ABSTRACT A study was conducted to investigate the prevalent microorganisms in Buck semen in Bouchi. Ten semen collections from three Bucks (BI, BII, BIII) of the Red Sokoto Breed were cultured aerobically on nutrient agar and MacConkey agar (oxoid), and blood agar (oxoid) for bacteria on potato destrose agar (oxoid) for fungi. Two bacterial species Staphylococcus aureus and Bacillus cereus together with mould Aspergillus niger were used. Aspergillus fumingatus and Torulopsis spp were isolated. The corresponding pH values and nitric contents were indicative of abnormality and infection respectively. The isolates from the samples were tested against three different herbs (Carica papaya L., Vernonia amygdalina Delile., and Jatropha curcas L. A cold ethyl acetate extract of the leaf and stem bark of the herbs showed appreciable inhibition on all the isolates except Jatropha curcas leaf that never showed an effect

KEY WORDS: Artificial insemination, semen, buck herbs, Carica papaya L., Vernonia amygdalina Delile., and Jatropha curcas L. and Antimicrobials

Cite this article: Salah. A. Martin, Yongabi. K. A. (2013), SENSITIVITY OF BUCK (MALE GOAT) SEMEN MICROBIAL ISOLATES TO THREE PLANT EXTRACTS FROM CAMEROON, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 410–417

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INTRODUCTION Artificial insemination is a very important practice by animal scientists, and remains true in Nigeria. During this practice, semen collected is normally preserved in specially compounded diluents pending insemination in the practice of artificial insemination in the country. Animal scientist often includes antibiotics and sometimes antimycotics in the diluents before insemination. This is done to protect the destruction of the semen content before insemination, but unfortunately; the inclusion of antibiotics has often been carried out without knowledge of the kind of bacteria present in the semen during ejaculation (Jimenez et al., 2012). Above all, some insemination do not bother to include antimycotics even then their pathogenic role in the semen with regard to the release of toxin (aflatoxin and mycotoxins is crucial to the reduction of the semen viability. Besides, some bacteria in the semen may develop resistance due to genetic adaptations (Colleuau et al., 2011, Nordstoga et al., 2011) to some of the antibiotics normally used in diluents. Studies in Cameroonian medicinal plants research is a very active domain in our Universities (Salah et al., 2012, Ndip et al., 2007) especially in the domain if antimicrobial studies ( Asob et al., 2011, Noumedem et al., 2013). The occurrence of microorganisms and their pathogenic role in certain cases have been exclusively reviewed (Akiynanju et al., 1986) by blood and radotitls and Zgnorniak Nowosienka et al., 1984). Pathogens and other microflora have been shown to have adverse effects on the fertility of semen by the production of toxins, putrefaction of diluents, components of biological origin, and the utilization of metabolic substrate, lamming (Tuncer et al., 2013). In the practice of artificial insemination, the possible pathogenic effects within the female reproductive tract as well as semen viability during storage of these microbes have been controlled considerably by the use of antibiotics and antimycotics in diluents (Butswat and Choji, 1974). However,

certain constraints have militated against the adoption of artificial insemination in this zone (Audu, 1989) and the farmers have had to depend on natural mating which exposes the female to contamination from males carrying pathogenic organisms within their urino-genital tract. The scarcity of veterinary services, the high cost of drugs particularly antibiotics and antimycotics for inclusion in the diluents as well as for treating animals, and the lack of knowledge for the microbial spectrum of semen are some of the factors that have the space of the adoption of the option of artificial insemination. This study investigates the prevalent microorganisms in Buck semen of the Red Sokoto breed and exploits indigenous means of controlling them with medicinal herbs. MATERIAL AND METHODS Semen Collection Semen was collected from July to November 2006 using the artificial vagina (AV) method after thorough feeding and abstinence from three months. The AV was normally washed with soap and hot water after each collection and disinfected with 70% alcohol after drying (Butswat and Choji, 1994) Isolation and Microorganisms

Identification

Bacteria A loopful of the test semen was aseptically streaked on nutrient agar MacConkey agar and blood agar plates and incubated aerobically at 37oC for 24 h. This was repeated for ten collections with the pH and nitrite values determined, using diagnostic urinalysis test trip (Combi-9 test strip) according to standard medical diagnostic method. Bacterial colonies were carefully picked and purified by repeated sub-cultures on nutrient agar and blood agar plates and their morphologies were studied. Pure cultures were preserved on nutrient agar slants and used for gram staining and other

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biochemical test (Chesborough, 2008), and the isolates were identified according to methods of Buchana and Gibbons (1974) and Harringan and McCance (1976) Fungi A loopful of the test semen was aseptically inoculated onto potato dextrose agar plates (Oxoid Ldt) in triplicates and incubated at 35oC for 5 days. This was repeated for ten collections for the three bucks. Following incubation, the growth were examined using an hand lens (10x) before picking out specimens in lactophenol cotton blue on a microscope slide and examined according to the methods of Harrigance and McCance (1976) and latter identified according to methods of Barnett and Hunter (1972). Preliminary Screening of herbs antimicrobial Property on Isolates

with

Samples Collection Herbs were collected from villages around Bouchi; Yelwa Tudu, Kagadamia, Doka and Dumba in Gwallameji. Those described by Audu were adopted from for the selection of plants (2). Leaves were used and samples were deposited in the herbarium of the institution. Processing of Plant Materials Plant parts were dried under shade for two weeks and then carefully packed in large Khaki enveloped and dried in an oven at 37oC for 12 h before milling. Fine powders were obtained through sieving. Crude extracts of the plant powder were made by adopting a modified existing method of Akinyanju et al., 1986 and Audu 1987. Cold solvent extraction was used. Ten grams each of the fine powder were weighed and soaked in 50 ml of ethyl acetate in the ratio 1:5 weight/volume and allowed to stand and for 48 hours at 25oC. Gravity filtration was done using filter paper (Whatman No. 13) and evaporated to dryness in the sun.

Antimicrobial Screening Test on Isolates The bacterial isolates were cultured in peptone water for 18 hrs. 0.3 ml of each of the bacterial suspension was mixed with 15 ml of nutrient agar in sterile petri-plates and allowed to solidify. A sterile steel borer was used to punch wells into the agar and each well was filled with 0.1 ml extract (at 2% concentration) and with 0.1 ml of distilled water as control. This was incubated at 37Â°C for 24 hours and the diameter of the zone of inhibition in measured to the nearest mm using vernier caliper according to methods of Akinyanju et al., (1986). Spores of the isolated fungi were harvested from stock and seeded onto dextrose agar plates with 0%, 2%, 4% and 16% concentrations of the herbal extracts. Spore germination and growth rate were monitored for a week. RESULTS Bacteria isolated from the semen samples for the three bucks were Staphylococcus aureus and Bacillus cereus, while Aspergillus niger (Var tieghem, 1867), Aspergillus fumigates (Fresenius, 1863), and Toruopsis spp were moulds of yeast respectively (Tables I and II). All the semen from the bucks were acidic and buck I revealed higher acidity (pH = 5.1) The nitrite values for the three bucks were high; with buck I revealed a higher value (Table III). The diameter of zone of inhibition of ethylacetate extract of herbs on bacteria isolate from buck semen is shown in Table IV. The susceptibility of the bacterial isolates to the ethyl acetate extract of the herbs (Carica papaya, Vernonia amygdalina, and Japtropha curcas) and the antimicrobial activity of the combined ethyl acetate extract of the individual herbs on both bacterial and fungal isolates is shown on Table V.

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TABLE I: Bacterial Isolates from the three buck seminal fluid in triplicate REPLICATE

III

BUCK I

S. a. , B.c.

BUCK II

S. a.,

S. a.

BUCK III

S. a., B.c

S. a. , B.c

S. a., B.c

S. a. – Straphylococcus aureus B.c. – Bacillus cereus.

TABLE II: Mould and Yeast isolates from the three-Buck seminal fluid in triplicate. REPLICATE

III

BUCK I

T. s.

BUCK II

A. f

BUCK III

A. n.

T. s. = Torulopsis spp A. f. = Aspergillus fumingatus A. n. = Aspergillus niger.

TABLE III: The Biochemical content of the buck semen REPLICATE

BUCK I

BUCK II

BUCK III

Number of samples collected

5.1

6.1

Nitrite

+++

N/B +++ = Highly positive ++ = Positive

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TABLE IV: The diameter of zone of inhibition of ethyl acetate extract of herbs on bacteria isolate from buck semen Average diameter of Zone of Inhibition 9 mm for 3 replicates at 2% concentration in mm Herbs Staphylococcus Bacillus cereus 1.0 1.0 Carica papaya Paw-paw (Leaf) Carica papaya Paw-paw (stem bark)

3.0

Vernonia amygdalina Bitter leaf (stem bark)

3.0

Vernonia amygdalina Bitter leaf (leaf) Jatropha curcas Physic nut (stem bark)

3.0

4.5

4.0

3.0

Jatropha curcas Physic nut (leaf) Mixture of all extracts 1:1

Resistant

3.5

6.5

Table V: The combined Effects of Ethyl acetate Plant Extracts on both Bacterial and Fungal Isolates Concentration (%)

Aspergillus niger

No growth from 1st – No growth from 1st – No growth from 1st – 7th day 7th day 7th day

Aspergillus fumingatus

No growth from 1st – No growth from 1st – No growth from 1st – 7th day 7th day 7th day

Control

Profuse growth with Profuse growth with Profuse growth with sporulation sporulation sporulation

Discussion The presence of nitrite in the semen samples and above all the higher nitrite nature of the studied semen (pH = 5.1 and 6.1) not only confirms an abnormality, but the degree of abnormality in the semen. It has been reported that pH values of Friesian bull 7.0 is needed for excellent fertility (Jimenez et al., 2012).

16%

The pH values of the semen (pH = 5.1 and 6.1) tally with the observation of Arthur an Allen (1972) that fresh semen samples are slightly acidic and that excessively high values occur with inflammation of the accessory glands. In a study on bacterial contamination of boar semen, (Dede, 1981), Staphylococci were isolated along with ten other genera of bacteria in the tropical environment. In this present

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study, Bacillus cereus, Staphylococcus aureus as bacterial isolates while Aspergillus niger (Vartieghem, 1867), Aspergillus fumingatus (Fresenius, 1863) and yeast, a Toralopsis spp as fungal isolates. This study, however, slightly differs from the result of ZgnorniakNowosielska (1984) who isolated only saprophytic flora from horse semen sample in Poland. Before restraining the buck for the research period, the animal had been feeding wildly and perhaps might have gotten Bacillus cereus through this means or even through the feed used during the period of this study, as there were no quality controls on the feed. The mould, Aspergillus spp, might also have been gotten through this means or may have been in the animals for some time. Bacillus cereus is widely distributed in nature and lives as saprophytes in the soil, dust, water, and on vegetation and cereals as reported by Cheesborough (2008). It is a major pathogen in cattle, sheep and goat and the bacilli are excreted in the faeces, urine and saliva of infected animals as reported by Cheesborough (2008). Aspergillus niger (Vartieghem, 1867) and Aspergillus fumigates (Fresenius, 1863) are ubiquitous saprophytes in soil, on plants, men and animals, while the yeast, Torulopsis spp, appear on foodstuffs and genitals of all the animals (Noumdem et al., 2013). Fungi have been noted to exert more effects on the hormonal activity within the reproductive system of animals. For instance, Aspergillus genus was known to be associated with the secretion of a toxic fungal metabolite, aflatoxin. Moreso ingestion of aflatoxin-

contaminated feed could lead to widespread reproductive abnormality in male chicken, including a reduction in circulating levels of testosterone (Clarke and Ottinger, 1987). From antimicrobial screening test, papaw leaf and stem bark (Carica papaya, Bitter leaf and stem bark (Vernonia amygdalina) and Jatropha curcas stem bark showed appreciable inhibitory property against the bacteria isolates in vitro as against no inhibition on the water control. Similarly, the extract of herbs combined in equal ratios showed a much more appreciable diameter of zone of inhibition than the individual extracts for the bacterial isolates and completely inhibited the growth of the fungal spores at all test concentrations except at 0% concentration for the first three days. This study shows that the knowledge of micro organism before insemination and their sensitivities to antibiotics and antimycotics in compounding diluents is very vital and above all proves the possibility of indigenous knowledge of herbs in controlling these pathogens, so as to ease and enhance artificial insemination practice in this environment. However, more studies should be carried out to map out the specific bioactive ingredients and ways of including the herbs directly in animal feeds or otherwise. ACKNOWLEDGEMENT The Authors are grateful to Dr. M. O. Agho, Director, FEPA/ZER Centre (A. T. B. U. Bouchi), for his assistance.

REFERENCES Akiynyanju, J. A., Owoyale J. A. and Okamla E. O., (1986) Antimicrobial effect of leaf extracts of Acalypha torta. The state of medicinal Plant Research I Nigeria edited by Abayomi Sofowora, R 47â&#x20AC;&#x201C;251.

Assob, Jules CN, Kamga, Henri LF, Nsagha, Dickson S, Njunda, Anna L , Nde, Peter F, Asongalem, Emmanuel A., Njouendou, Abdel J., Penlap, Veronique B, (2011) Antimicrobial and toxicological activities of five

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medicinal plant species from Cameroon Traditional Medicine . BMC Complementary and Alternative Medicine, 11 (1), p.70 Audu J. A. (1989) Medicinal Herbs and their uses in Bouchi State Nigeria field 54, 157–168. Barnett. H. L. and Hunter, B. B., (1972) Illustrated Genera of imperfecti 3rd Edn. Burgress publishing Minneapolis. Buchanan, R. E. and Gibbons, N. E., (1974) Bergey’s Manual of terminative Bacteriology, William and Willims, Baltimore, USA. Butswat, I. S.,and Choji, G. E. (1974) Constraints to adoption of artificial insemination techniques in livestock production in Bauchi Local government Area of Nigeria. J. Animal production, 22:28–31. Chesborough Monica (2008): Laboratory Manual for Countries.

Medical Tropical

Clarke, R. N. Doer J. A. and Ottinger, M. A. (1987) Age related changes in testicular development and productive endocrinology associated with aflotaxicosis in the male chicken. Boil reproduction 36: 117–124. Colleau, Jean-Jacques, Clément, Virginie, Martin, Pierre, Palhière, Isabelle, (2011) Optimized diffusion of buck semen for saving genetic variability in selected dairy goat populations . BMC Genetics, 12 (1), p.25. Colleau, Jean-Jacques., Clément, Virginie., Martin, Pierre., Palhière, Isabelle, (2011) Optimized diffusion of buck semen for saving genetic variability in selected dairy goat populations. BMC genetics, 12, p.25.

Dede, T. L. (1981): Bacteriological Studies of Boar Semen in the tropical contamination of boar semen in the tropical environment. Nigeria J. of Agricultural Science 3: 105–107. Harrigan, W. B. and McCance, M. F., (1976): Laboratory Methods I food and diary microbiology. Academic Press London, U.K. Jiménez-Rabadán, P., Morrell, J M., Johannisson, A Ramón, M., GarcíaÁlvarez, O., Maroto-Morales, A., Alvaro-García, P J., Soler, A J, (2012). Single layer centrifugation (SLC) improves sperm quality of cryopreserved BlancaCeltibérica buck semen. Animal reproduction science, 136 (1-2), p.47– 54. Ndip, Roland N., Malange Tarkang, Alertia E., Mbullah, Susan M., Luma, Henry N., Malongue, Agnes, Ndip, Lucy M, Nyongbela, Kennedy, Efange, Simon M N, (2007) In vitro anti-Helicobacter pylori activity of extracts of selected medicinal plants from North West Cameroon. J. of ethnopharmacology, 114 (3), p.452– 457. Nordstoga, A B., Söderquist, L., Adnøy, T., Paulenz, H, (2011) . Fertility results after vaginal deposition of frozenthawed buck semen diluted with two different extenders using one- or twostep procedures. .Reproduction in domestic animals = Zuchthygiene, 46 (1), p.82–86. Noumedem, Jaurès AK. Mihasan, Marius, Lacmata, Stephen T., Stefan, Marius., Kuiate, Jules R / Kuete, Victor, (2013) Antibacterial activities of the methanol extracts of ten Cameroonian vegetables against Gram-negative multidrug-resistant bacteria BMC Complementary and Alternative Medicine, 13 (1), p.26.

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Salah A. Martin and Njunda L. Anna (2012). Effects of Ruellia praetermissa extracts on erythropoisis in pregnant women. Global J Res. Med. Plants & Indigen. Med., Volume 1(8): pp 309–314.

Safa, (2013) Effects of different doses of trehalose supplementation in egg yolk extender in frozen–thawed Angora buck semen Small Ruminant Research, 136 (1–2), p.47–54.

Tuncer, Pürhan Barbaros, Taşdemir, Umut, Büyükleblebici, Serhat, Özgürtaş, Taner, Coşkun, Erdem, Erol, Halil, Aydın, Fevzi Nuri, Gürcan, İsmail

Zgorniak Nowosielska, I; Biclanski, W. and Kosinial I., (1984). Myocoplasma in Stallion Semen. Animal Repro. Science 7: 343–350.

Source of Support: Nil

Conflict of Interest: None Declared

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Review article A BRIEF REVIEW ON GINKGO BILOBA L. (MAIDENHAIR TREE) A RARE MULTIPURPOSE MEDICINAL PLANT Patel Hemangi1, Ingalhalli Rajashekhar2* 1

C.G Bhakta Institute of Biotechnology, Uka Tarsadia University, Mahuva Road, Bardoli-394601, Surat (Dist. Surat), Gujarat, India. 2 Assistant Professor, C.G Bhakta Institute of Biotechnology, Uka Tarsadia University, Mahuva Road, Bardoli-394601, Surat (Dist. Surat), Gujarat, India. *Corresponding Author: Email: rajashekhar@utu.ac.in; rajshekhar.blr@gmail.com; Mob: +919448371037

Received: 20/04/2013; Revised: 23/05/2013; Accepted: 27/05/2013

ABSTRACT Ginkgo biloba, a native of China, was probably a member of the mixed-mesophytic forest community that once covered the hill country. In developing countries a large proportion of the population relied heavily on traditional practitioners and medicinal plants to meet health care needs. Growing interest in the field of Complementary and Alternative medicine places new obligations on the clinicians. To assist in this process, a comprehensive review of Ginkgo biloba, the well studied botanical medicine, was provided. Studies so far indicated the efficacy of Ginkgo biloba in tinnitus, schizophrenia, psychotic organic brain syndrome, vertigo of undetermined origin. Also double blind research showed Ginkgo biloba’s efficacy in conditions including cerebro-vascular insufficiency, memory impairment, Alzheimer’s disease, multi-infarct dementia, resistant depression, peripheral arterial insufficiency, venous insufficiency and asthma was summarized. KEY WORDS: Ginkgo biloba, ginkgolides, dementia, Ginkgo extract

Cite this article: Patel. H., Ingalhalli. R. (2013), A BRIEF REVIEW ON GINKGO BILOBA L. (MAIDENHAIR TREE) - A RARE MULTIPURPOSE MEDICINAL PLANT, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 418–427

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INTRODUCTION In developing countries a large proportion of the population relies heavily on traditional practitioners and medicinal plants to meet health care needs (Gartoulla, 1993). Ginkgo biloba tree is commonly known as maidenhair tree. Since it has no close living relatives, it is also called a living fossil. Ginkgo biloba is cultivated nowadays in many parts of the world as an ornamental plant and commercially grown for its leaves which contain several medicinal properties (DeFeudis, 1998). The use of Ginkgo leaf extract for pharmaceutical purposes was originally developed in Germany in 1965, and the first commercially available Ginkgo leaf extract was registered for human use in 1974 in France, under the code-name "EGb 761" (DeFeudis, 1998). Scientists from various countries have been conducting clinical trials to establish the efficacy of leaf extract in treating tinnitus, vertigo, Alzheimer’s disease, dementia, anxiety, cerebral dysfunction (Ref; Table 1–5). Clinical trials have also been counducted to find out the efficacy of leaf extract to treat various other diseases (Ref; Table 6). The Ginkgo leaf extract EGb 761 has shown to have low acute and chronic toxicities (Woerdenbag and De Smet, 2000). Ginkgo biloba has proved its efficacy in treating a considerable number of diseases and still provides ample opportunities to future researchers to unravel the hidden medicinal properties. Considering the critically endangered status of Ginkgo biloba in the wild, it has become inevitable to cultivate it in different parts for its utilization. Habit Ginkgo biloba is a dioecious tree, windpollinated, animal-dispersed gymnosperm with evolutionary affinities to both cycads and the conifers which has been used in traditional Chinese medicine for about 5000 years (Saquires, 1999; Norstog et al., 2004). Ginkgos are large trees, normally reaching a height of 20–35 m (66–115 feet), the tree has an angular crown and long, somewhat erratic branches, and is usually deep rooted and resistant to wind and snow damage. Young trees are often tall

and slender, and sparsely branched; the crown becomes broader as the tree ages (Royer et al., 2003). The leaves are unique among seed plants, being fan-shaped with veins radiating out into the leaf blade, sometimes bifurcating (splitting), but never anastamosing to form a network. Two veins enter the leaf blade at the base and fork repeatedly in two; this is known as dichotomous venation. The leaves are usually 5–10 cm (2–4 in), but sometimes up to 15 cm (6 in) long. The old popular name "maidenhair tree" is because the leaves resemble some of the pinnae of the maidenhair fern, Adiantum capillus-veneris (Wang, 1961). Distribution and status As a wild species, Ginkgo biloba is native to China and was probably a member of the mixed-mesophytic forest community that once covered the hill country bordering the Yangtze River valley (He et al., 1997). The species is highly adaptable and grows well in most regions with a distinct seasonality and moderate rainfall, including areas with warmtemperate, cold-temperate, or Mediterranean climates (Shen et al., 2004). For centuries, it was thought to be extinct in the wild, but is now known to grow in at least two small areas in Zhejiang province in eastern China, in the Tian Mu Shan Reserve. However, recent studies indicate high genetic uniformity among Ginkgo trees from these areas, arguing against a natural origin of these populations and suggesting the Ginkgo trees in these areas may have been planted and preserved by Chinese monks over a period of about 1,000 years (Tang et al., 2012). The greater genetic diversity has been found in Southwestern China populations, in mountains surrounding eastern Qinghai-Tibet Plateau (Tang et al., 2012; Fu et al., 1999). Where it occurs in the wild, it is found infrequently in deciduous forests and valleys on acidic loess (i.e. fine, silty soil) with good drainage. The soil it inhabits is typically in the pH range of 5.0 to 5.5 (Barrett, 2004). Medicinal importance Extract of Ginkgo leaves contain bioflavonoids (amentoflavone, bilobetin,

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ginkgetin), flavonoids (quercetin, isorhamnetins, kaempferol), proanthocyanidins, Ginkgolides A,B,C, bilobalide (Barrett, 2004; Kraft and Hobbs, 2004; Deng and Zito, 2003; Evans, 2002; McKenna et al., 2001; Bruneton 2000; Samuelsson, 1999). Most Ginkgo extracts available in the European market are standardized to 24% flavones glycosides and 6% terpene lactones (Mahady et al., 2001; Ahlemeyer and Krieglstein, 1998; Cott, 1995). Although some Ginkgo preparations have also been applied parenterally, the great are ingested as tablets, capsules or extracts (Koltringer et al., 1989). Leaf extract of Ginkgo is made from dried Ginkgo leaves and has a standardized

content of 22–27% flavonol glycosides and 5– 7% terpene trilactones. This extract is taken internally for the treatment of cerebral and peripheral vascular diseases, as well as to alleviate some of the ailments associated with ageing, including dizziness, ringing in the ears, and short-term memory deterioration (DeFeudis, 1998; Juretzek, 1997). Ginkgo supplements are usually taken in the range of 40–200 mg per day. Clinical trials to date have utilized the standardized products EGb761 and to a lesser extent LI 1370 (Bluementhal et al., 2000). Details of clinical trials using Ginkgo biloba are given in Tables 1 to 6.

Table-1: Selected Clinical Trials Employing Ginkgo on Dementia Reference

Plant part / herbal product

Purpose of study

Ihl R et al., 2010

Ginkgo extract single dose of 240 mg daily 160 mg of ginkgo extract

Treatment of mild to moderate dementia Treatment of dementia

EGb 761 standardized extract

Mazza, M et al., 2006 Kanowski and Hoerr, 2003

Number of Subjects Not available Not available Not available

Results

214

Not effective Effective

Effective Effective

Van Dongen et al., 2003 Le Bars, et al., 2002 Le Bars et al., 2000

EGb 761Standardized extract EGb 761 Standardized extract EGb 761 Standardized extract

Use of ginkgo biloba extract for the treatment of dementia Treatment for dementia Treatment for dementia Treatment for dementia

Van Dongen et al., 2000 Kanowski et al., 1996

EGb 761 standardized extract EGb 761 standardized extract

Treatment for dementia Treatment for dementia

196 156

Not effective Effective

Le Bars et al., 1997 Haase et al., 1996

Ginkgold (EGb 761 tablets) EGb 761standardized extract

Treatment for dementia Treatment for dementia

202

Effective

Hofferberth, 1989

Tebonin forte (EGb 761) Standardized extract

Treatment for dementia

Effective

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Not available 309

Effective

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Table-2: Selected Clinical Trials Employing Ginkgo on Alzheimer's disease Reference

Plant part / herbal product

DeKosky et al, 2008 Stough et al., 2001 Rigney et al., 1999 Kanowski et al., 1996

Twice-daily dose 120 mg extract EGb 761 Standardized extract

Number of subjects

Purpose of study

Alzheimer's disease in Not available adults 75 years or older Memory 61 enhancement Memory Kaveri 31 enhancement Ginkgo extract 120 mg Treatment of Alzheimer 216 daily

Results Not effective Effective Not effective Effective

Table-3: Selected Clinical Trials Employing Ginkgo on Asthma Reference Li et al., 1997 Anonymous, 1989

Plant part / herbal product Concentrated leaf liquid (Chinese product) Ginkgo extract

Purpose of study Treatment for asthma

Number of subjects 61

Inhibitor of platelet Not available activating factor

Results Effective Effective

Table-4: Selected Clinical Trials Employing Ginkgo on Cerebral Insufficiency Reference

Plant part / herbal product

Purpose of study

Number Of subjects

Results

Brautigam et al., 1998

Geriaforce (liquid extract)

Treatment for cerebral insufficiency

197

Effective (improvement of short term visual memory)

Vesper and Hansgen, 1994 Grassel, 1992

Treatment for cerebral insufficiency Rokan ® (EGb Treatment for cerebral 761) insufficiency Kaveri LI 1370 Treatment for cerebral insufficiency Kaveri® LI 1370 Treatment for Cerebral insufficiency LI 1370 Treatment for cerebral (standardized insufficiency liquid extract)

Effective

209

Effective

Bruchert et al., 1991 Schmidt et al., 1991 Eckmann, 1990

Kaveri LI 1370

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Table-5: Selected Clinical Trials Employing Ginkgo on Tinnitus Reference

Plant part / herbal product

Purpose of study

Number of subjects

Results

Raja et al., 2004

Ginkgo biloba Tablets

Not effective

Drew and Davies, 2001 Coles R. 1988

LI 1370 standardized extract Ginkgo biloba extract (40 mg three times a day for 12 weeks)

To evaluate the efficacy of ginkgo for the treatment of tinnitus Treatment for tinnitus Treatment of tinnitus

956

Not effective

Slight improvement but treatment was ineffective

Table-6: Selected Clinical Trials Employing Ginkgo on other diseases Reference Muir et al., 2002 Prasad et al., 2003 Gertsch et al., 2002 Kang et al., 2002

Plant part / herbal product Seredrin (standardized Ginkgo biloba extract) Ginkocer (Ginkgo biloba extract) Ginkgo biloba extract Ginkgo biloba extract

Chen et al., 2003

Ginkgo biloba exocarp polysaccharides (GBEP) capsule preparation taken orally

Brochet et al., 1995

Intravenoous application of ginkgolide B Ginkgo biloba extract 6o mg per day for 12–18 months Ginkgo biloba extract

Sikora ,1989

Haguenauer, 1986 Lagrue et al.,1986

Ginkgo biloba extract

Purpose of study

Number of subjects Not available

Results

Effective

Treatment for acute mountain sickness Treatment for Antidepressant induced sexual dysfunction To observe the clinical efficacy of (GBEP capsule preparation in treating upper digestive tract malignant tumors of middle and late stage Treatment for exacerbations of multiple sclerosis Treatment of impotence

Effective (as pretreatment) Not effective

Improvement in penile arterial blood flow

Treatment of idiopathic vertigo Treatment of cyclic edema

Asymptomatic in 47% patients Out of ten 3 women showed complete elimination of edema and six were improved

Treatment for Raynaud’s syndrome Treatment for vitiligo

104

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Effective

The GBEP Capsule preparation had positive therapeutic effects on upper digestive tract malignant tumors of middle and late stages Not effective

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The Special Ginkgo biloba leaf extracts are generally well tolerated with low incidence of side effects. Less than two percent of the patients develop side effects and no serious side effects have been noted in any trials. The side effects include gastrointestinal complaints, headache, sleep disturbances, dizziness and allergic skin reactions. The ginkgo leaf extract EGb 761 has been shown to have low acute and chronic toxicities (Woerdenbag and De Smet, 2000). Cultivation Ginkgo has long been cultivated in China; some planted trees at temples are believed to be over 1,500 years old. In China, Ginkgo is cultivated across a broad range of moisture, temperature, and topographic gradients between 25° and 42°N latitude, where minimum winter temperatures can reach –32°C and maximum summer temperatures 42°C (He et al., 1997). The first record of Europeans encountering it is in 1690 in Japanese temple gardens, where the tree was seen by the German botanist Engelbert Kaempfer. Because of its status in Buddhism and Confucianism, the Ginkgo is also widely planted in Korea and parts of Japan; in both areas, some naturalization has occurred, with ginkgos seeding into natural forests. In some areas, most intentionally planted Ginkgos are male cultivars grafted onto plants propagated from seed, because the male trees will not produce the malodorous seeds. The popular cultivar 'Autumn Gold' is a clone of a male plant. Ginkgos adapt well to the urban environment, tolerating pollution and confined soil spaces (Gilman, 2008). They rarely suffer disease problems, even in urban conditions, and are attacked by few

insects (Boland et al., 2002; Minnesota, 2008). For this reason, and for their general beauty, ginkgos are excellent urban and shade trees, and are widely planted along many streets. Ginkgos are also popular subjects for growing as penjing and bonsai; they can be kept artificially small and tended over centuries. Furthermore, the trees are easy to propagate from seed. In cultivation in the United Kingdom G. biloba has gained the Royal Horticultural Society's Award of Garden Merit. The Ginkgo leaf is the symbol of the Urasenke school of Japanese tea ceremony. The tree is the national tree of China, and is the official tree of the Japanese capital of Tokyo, and the symbol of Tokyo is a ginkgo leaf (Minnesota, 2008). CONCLUSION With the growing patient demand for alternative, complementary, natural, or integrated approaches in treating disease, it is increasingly important for clinicians to develop referral relationships with well-trained Complementary and Alternative Medicine providers. Ginkgo biloba extract (GBE), one of the most commonly used and best-researched phytomedicines, has documented efficacy for many conditions. Use of GBE in cerebrovascular insufficiency, memory impairment in the elderly, Alzheimer’s disease, multi-infarct dementia, resistant depression, peripheral artery insufficiency, venous insufficiency and asthma is well supported by multiple studies. GBE for tinnitus, schizophrenia, psychotic organic brain syndrome, vertigo of undetermined origin, and PMS, although less supported, still deserves serious consideration because of GBE’s high tolerability, and the limited or complete lack of efficacy with conventional treatments for these conditions.

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Barrett M. (2004) Bilobalide, ginkgolides and flavonoids in Ginkgo biloba L. extract and pharmaceutical Handbook of Clinically Tested Herbal Remedies, 2 Vols. New York: Haworth Herbal Press; pp. 547–672. Bluementhal M. (2000) Ginkgo biloba leaf extract Herbal Medicine Expand Commission E Monographs, American Botany Council, Austin, Integrative Medicine Communications, Boston, US; 160–169. Boland, Timothy, Laura E. Coit, Marty Hair (2002) Michigan Gardener's Guide. Cool Springs Press. ISBN 1-930604-203. Brochet B, Guinot P, Orgogozo JM. (1995) Double blind placebo controlled multicentre study of ginkgolide B in treatment of acute exacerbations of multiple sclerosis. The Ginkgolide Study Group in multiple sclerosis. J Neurol Neurosurg Psychiatry. 58(3):360–362. Bruneton, J. (2000) Pharmacognosy, Phytochemistry, Medicinal Plants. 3rd ed. Paris: Intercept; 2000. Chen HS, Zhai F, Chu YF. (2003) Clinical study on treatment of patients with upper digestive tract malignant tumors of middle and late stage with Ginkgo biloba exocarp polysaccharides capsule preparation [Article in Chinese] 1(3):189–91. Coles R. (1988) Trial of an extract of Ginkgo biloba (EGB) for tinnitus and hearing loss Clin Otolaryngol 13:501–504. Cott J. (1995) NCDEU update. Natural product formulations available in Europe for psychotropic indications. Psychopharmacol Bull. 31(4):745–751. DeFeudis, F.V. (1998) Ginkgo biloba Extract (EGb 761): From Chemistry to Clinic, Ullstein Medical, Weisbaden.Foster.

DeKosky, Steven T.; Williamson, Jeff D.; Fitzpatrick, Annette L.; Kronmal, R. A.; Ives, D. G.; Saxton, J. A.; Lopez, O. L.; Burke, G. (2008). "Ginkgo biloba for Prevention of Dementia". The Journal of the American Medical Association 300 (19): 2253–2262. Deng F, Zito SW. (2003) Development and validation of a gas chromatographicmass spectrometric method for simultaneous identification and quantification of marker compounds including bilobalide, ginkgolides and flavonoids in Ginkgo biloba L. extract and pharmaceutical preparations. J Chromatogr A. 986(1):121–127. Drew S, Davies E. (2001) Effectiveness of Ginkgo biloba in treating tinnitus: double blind, placebo controlled trial. BMJ. 322(7278):73. Eckmann F. (1990) Cerebral insufficiency treatment with Ginkgo-biloba extract. Time of onset of effect in a doubleblind study with 60 inpatients. [Article in German] Fortschr Med 108:557–560. Evans WC. (2002) Pharmacognosy 17 th ed. Philadelphia: W.B. Saunders. Fu Liguo; Li, Nan; Mill, Robert R. (1999). "Ginkgo biloba". In Wu, Z. Y.; Raven, P.H.; Hong, D.Y. Flora of China 4. Beijing: Science Press; St. Louis: Missouri Botanical Garden Press. p. 8. Retrieved 31 March 2008. Gartoulla RP. (1993) Anthropology of contemporary substitute medications in Nepal. Doctoral thesis, North Bengal University, Darjeeling, India. Gertsch JH, Seto TB, Mor J, Onopa J. (2002) Ginkgo biloba for the prevention of severe acute mountain sickness (AMS) starting one day before rapid ascent. High Alt Med Biol. 3(1):29–37.

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Gilman, Edward F. and Dennis G. Watson (1993). "Ginkgo biloba 'Autumn Gold'" (PDF). US Forest Service. Grassel E. (1992) Effect of Ginkgo-biloba extract on mental performance. Doubleblind study using computerized measurement conditions in patients with cerebral insufficiency. [Article in German] Fortschr Med. 110(5):73–76. Haase

J, Halama P, Horr R. (1996) Effectiveness of brief infusions with Ginkgo biloba Special Extract EGb 761 in dementia of the vascular and Alzheimer type. [Article in German] Z Gerontol Geriatr.29(4):302–309.

Haguenauer JP. (1986) Treatment of disturbances of equilibrium with Ginkgo biloba extract. A multicentre, double blind, drug of press Med 15:1569–1572. He, S. A., Y. Gu & Z. J. Pang. (1997) Resources and prospects of Ginkgo biloba in China. Pp. 373–38. Hofferberth B. (1989) The effect of Ginkgo biloba extract on neurophysiological and psychometric measurement results in patients with psychotic organic brain syndrome. A double-blind study against placebo [Article in German]. Arzneimittelforschung. 39(8):918–922. Ihl

R, Bachinskaya N, Korczyn AD, Vakhapova V, Tribanek M, Hoerr R, Napryeyenko O (2010) "Efficacy and safety of a once-daily formulation of Ginkgo biloba extract EGb 761 in dementia with neuropsychiatric features: a randomized controlled trial". Int J Geriatr Psychiatry.

Juretzek, W. (1997) Recent advances in Ginkgo biloba extract (Egb 761). In T. Hori, R.W. Ridge, W. Tulecke, P. Del Tredici, J. Trernouillaux-Guiller, and H. Tobe (eds.), Ginkgo biloba - A Global Treasure. Springer-Verlag, Tokyo, pp. 341–358.

Kang BJ, Lee SJ, Kim MD, Cho MJ. (2002) A placebo-controlled, double-blind trial of Ginkgo biloba for antidepressantinduced sexual dysfunction. Hum Psychopharmacol. 17(6):279–284. Kanowski S, Herrmann WM, Stephan K. (1996) Proof of efficacy of the Ginkgo biloba special extract EGb 761 in outpatients suffering from mild to moderate primary degenerative dementia of the Alzheimer type or multi-infarct dementia. Pharmacopsychiatry. 29(2):47–56. Kanowski S, Hoerr R. (2003) Ginkgo biloba extract EGb 761 in dementia: intent-totreat analyses of a 24- week, multicenter, double-blind, placebocontrolled, randomized trial. Pharmacopsychiatry. 36(6):297–303. Koltringer P, Eber O, Klima G. (1989) Microcirculation in parenteral Ginkgo biloba extract therapy. [Article in German] Wien Klin Wochenschr. 101(6):198–200. Kraft K, Hobbs C. (2004) Pocket Guide to Herbal Medicine. New York: Thieme; pp. 71–73. Lagrue G. (1986) Idiopathic Cyclic edema. Role of capillary hyper permeability and its correction by Ginkgo biloba extract presse Med 15:1550–1553. Le Bars PL, Kastelan J. (2000) Efficacy and safety of a Ginkgo biloba extract.Public Health Nutr. 3(4A):495–9. Le Bars PL, Katz MM, Berman N. (1997) A placebo-controlled, double-blind, randomized trial of an extract of Ginkgo biloba for dementia. North American EGb Study Group. JAMA. 278(16):1327–1332.

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Le Bars PL, Velasco FM, Ferguson JM, (2002) Influence of the severity of cognitive impairment on the effect of the Gnkgo biloba extract EGb 761 in Alzheimer's disease. Neuropsychobiology. 45(1):19–26. Li MH, Zhang HL, Yang BY. (1997) Effects of ginkgo leave concentrated oral liquor in treating asthma [Article in Chinese]. 17(4):216–218. Mahady G, Fong H, Farnsworth N. (2001) Botanical Dietary Supplements: Quality, Safety and Efficacy. The Netherlands: Swets and Zeitlinger. Mazza, M.; Capuano, A.; Bria, P.; Mazza, S. (2006). " Ginkgo biloba and donepezil: a comparison in the treatment of Alzheimer's dementia in a randomized placebo-controlled double-blind study". European Journal of Neurology 13 (9): 981–5. McKenna DJ, Jones K, Hughes K. (2001) Efficacy, safety, and use of Ginkgo biloba in clinical and preclinical applications. Altern Ther Health Med. (5):70-86, 88–90. Muir AH, Robb R, McLaren M. (2002) The use of Ginkgo biloba in Raynaud's disease: a double-blind placebo-controlled trial. Vasc Med. 7(4):265–267. Norstog, K. J., E. M. Gifford & D. W. Stevenson. (2004) Comparative development of the spermatozoids of cycads and Ginkgo biloba. Bot. Rev. 70: 5–15. Parsad D, Pandhi R, Juneja A. (2003) Effectiveness of oral Ginkgo biloba in treating limited, slowly spreading vitiligo. Clin Exp Dermatol. 28(3):285– 287.

Raja PV, Blumenthal JA, Doraiswamy PM. (2004) Cognitive deficits following coronary artery bypass grafting: prevalence, prognosis, and therapeutic strategies. CNS Spectr. 9(10):763–72. Rigney U, Kimber S, Hindmarch I. (1999) The effects of acute doses of standardized Ginkgo biloba extract on memory and psychomotor performance in volunteers. Phytother Res. 13(5):408– 415. Royer, D. L., L. J. Hickey, S. L. Wing. (2003) Ecological conservatism in the “living fossil” Ginkgo. Paleobiology 29: 84– 104. Samuelsson G. (1999) Drugs of Natural Origin 4 th ed. Stockholm: Swedish Pharmaceutical Society. Saquires R. (1999) Ginkgo biloba. Australian traditional medicine society (ATOMS) 5: 9–14. Shen, L; Chen, X-Y; Zhang, X; Li, Y-Y; Fu, CX; Qiu, Y-X (2004) "Genetic variation of Ginkgo biloba L. (Ginkgoaceae) based on cpDNA PCR-RFLPs: inference of glacial refugia". Heredity 94 (4): 396–401. Sikora R. (1989) Ginkgo biloba extract in the therapy of erectile dysfunction.J Urol.141:188A. Stough C, Clarke J, Lloyd J, Nathan PJ. (2001) Neuropsychological changes after 30day Ginkgo biloba administration in healthy participants. Int J Neuropsychopharmacol. 2001; 4(2):131–134. Tang,

CQ, (2012). "Evidence for the persistence of wild Ginkgo biloba (Ginkgoaceae) populations in the Dalou Mountains, southwestern China". American Journal of Botany 99 (8).

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University of Minnesota. Retrieved 29 March (2008) "Examples of Plants with Insect and Disease Tolerance". SULIS Sustainable Urban Landscape Information Series. Van Dongen M, van Rossum E, Kessels A. (2003) Ginkgo for elderly people with dementia and ageassociated memory impairment: a randomized clinical trial. J Clin Epidemiol. 56(4):367–376. Van Dongen MC, van Rossum E, Kessels AG. (2000) The efficacy of ginkgo for elderly people with dementia and ageassociated memory impairment: new

Source of Support: Nil

results of a randomized clinical trial. J Am Geriatr Soc. 48(10):1183–1194. Von Wedel H., Calero L., Walger M. (1995) Soft-laser/Ginkgo therapy in chronic tinnitus. A placebocontrolled study. Adv Otorhinolaryngol. 49:105–108. Wang, C.W. (1961) The Forests of China, Maria Moors Cabot Found., publ. 5. Harvard Univ., Cambridge, Mass. Woerdenbag H J and De Smet P A G M. (2000) Adverse effects and toxicity of Ginkgo biloba extract Med Aromat plants Ind Profiles; 12: 443–451.

Conflict of Interest: None Declared

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Research article EFFECT OF BADARA STEM BARK (ZIZIPHUS JUJUBA LAMB.) IN THE MANAGEMENT OF VATAJA KASA IN CHILDREN Chethan Kumar V K1, Shailaja U2 1

PhD Scholar & Associate Professor, Dept of Kaumarabhritya, S.D.M.College of Ayurveda, Udupi, Karanataka. India 2 Professor & Head, Dept. of Kaumarabhritya, S.D.M.College of Ayurveda & Hospital, Hassan, Karnataka, India *Corresponding Author: Email: drchethankumar@gmail.com; Mob: +919986550745

Received: 07/04/2013; Revised: 21/05/2013; Accepted: 24/05/2013

ABSTRACT Kasa (Cough) is one of the most frequently encountered problems in the Balyavastha (Pediatric age). Recurrent attacks makes the school going child suffer and may have its adverse effects on the studies of the child. The present clinical study was carried out to evaluate the effect of Syrup Badara in Vataja kasa (Dry cough) in children. The present study was conducted on children presenting with Vataja kasa in between the age group of 2–10 years. The clinical trial was conducted on 73 patients. They were divided into two groups; each group consisted of minimum 30 patients. The divided Groups A and B were given syrup Badara and placebo concentrated sugar solution respectively. The results were analyzed statistically. The analysis suggested that Syrup Badara is effective in reducing the signs and symptoms of Vataja kasa. KEY WORDS: Kasa, Cough, Shushka kasa, Badara, Ziziphus jujuba

Cite this article: Chethan K V K, Shailaja U(2013), EFFECT OF BADARA STEM BARK (ZIZIPHUS JUJUBA LAMB.) IN THE MANAGEMENT OF VATAJA KASA IN CHILDREN, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 428–434

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INTRODUCTION Respiratory system is one system which is in continuous contact with the external environment since birth until one’s lifetime, so it is most vulnerable to infections and considered as the prime victim of hyper sensitization in most of the circumstances (Atkins D, 2008). Thus Respiratory Tract Infections (RTI) accounts to about more than 50% of patients attending pediatric OPD in developing and even developed countries worldwide. (Eggenberger K, 1993). Pediatric age group is more vulnerable to respiratory tract infections because of anatomical and physiological peculiarities (i.e. small narrow airways (Drake Lee A B, 1987), hypertrophied lymphoid tissues (Hibber J, 1987), underdeveloped paranasal sinuses (Diane E P, 2008), mucous hyper secretion and peculiarities of Eustachian tube (Maw A R, 1987) and social factors such as attending school, improper food and eating habits. As cough is the most frequent symptom of respiratory diseases (Innes J A, 2006) majority of the patients present recurrent cough as the manifestation of recurrent respiratory disease. In Ayurvedic point of view descriptions of disease Kasa clearly correlate with cough. Moreover the patho-physiology (Samprapti) of Kasa almost exactly correlates the mechanism of cough reflex (Trikamji Yadavji, 2009). Early intervention is necessary in case of Kasa(cough) as it is a potential Nidanarthakara Vyadhi (Diseases as causative factors for other diseases)to produce Kshaya(consumption) (Trikamji Yadavji, 2009). Also it is important to treat any Balaroga (Pediatric disorder)at the earliest as it may hamper the proper Vriddhi (Growth and development) of a child which is clearly described by Acharya Charaka, that Avighata(Absence of inhibiting factors) as shareera vriddhikara bhava (factors responsible for growth of body) (Trikamji Yadavji, 2009). Thus in the present work Kasa (Cough) was taken as the subject of intervention.

The Shamana (Palliative treatment) line of treatment that includes oral administration of medicine is of utmost importance as the administration of syrup is very easy and also effective compared to Shodhana (Purificatory treatment) in children. Many research works have been carried out in relation to the Shamana treatment as directed in Ayurveda and their therapeutic effect is proved. After critical analysis of the drug ie. Stem bark of Badara had been selected as the trial drug to evaluate & establish its efficacy in combating the signs & symptoms of Vataja Kasa (Dry cough). METHODOLOGY Present study was carried out for the scientific understanding of this trial drug stem bark of Badara (Ziziphus jujuba Lam) in the management of Vataja Kasa. The present study was conducted after the approval of the Institutional Ethics committee with reference IEC no – SDM/IEC/09/2008-2009. The study was conducted on children of Vataja Kasa in between the age group of 2–10 years. Patients were divided into 2 groups. Group A and B were treated with Syp. Badara and Syp. Sugar Solution respectively. For preparing the syrup, the following methods were followed at SDP Remedies and Research Centre, Puttur D.K., Karnataka.    

 

After thorough cleaning of stem bark of Badara, it was powdered into a coarse form. One part of the powder of the drug was soaked with 8 parts of water and kept for one night. Next day the contents were boiled and reduced to ¼ th part and filtered. In the obtained mixture 2 parts of sugar was dissolved and re boiled to obtain one thread consistency of the syrup and cooled to room temperature. Lastly it was filtered and sealed in sterilized 100ml bottles. For the purpose of controlled study a Placebo in the form of concentrated sugar syrup of the same consistency was prepared.

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Stem bark of Badara was collected from Kasargod District and authenticated by Smt.Vidya, Botanist, SDP Remedies and Research Centre, Puttur D.K., Karnataka.

Study design For diagnostic purpose the signs and symptoms mentioned below were taken for the study.       

The observations of the cases were recorded according to the research proforma. Criteria for selection of patients Patients with signs and symptoms of Vataja Kasa – viz., Shushka Kasa (Non productive cough), Shirashoola (Head ache), Swarabheda (Hoarseness of voice), Parshwa Shoola (Pain in the flanks), Ura Shoola (Chest pain), Shushka vaktra (Dryness of mouth), Kantodhvamsanam (Throat irritation) were selected randomly irrespective of their sex, religion etc. from Kaumarabhritya O.P.D of S.D.M. Ayurvedic Hospital, Hassan, Karnataka, India Inclusion criteria    

Patients presenting with Vataja Kasa (Dry Cough). Any of two or more symptoms described in the context of Vataja Kasa (Dry Cough). Patients of Vataja Kasa (Dry Cough) with chronicity of less than 15 days duration. Patients irrespective of sex, religion, socioeconomic status and between the age group of 2–10yrs are taken.

Exclusion criteria  

Shushka Kasa (Non productive cough) Shirashoola (Head ache) Swarabheda (Hoarseness of voice) Parshwa Shoola (Pain in the flanks) Ura Shoola (Chest pain) Shushka vaktra (Dryness of mouth) Kantodhvamsanam (Throat irritation)

Dose, duration and mode of administration: Duration of the treatment was 10 days. Group - A - Syp. Badara 5 ml for 5–10 years of age group & 2.5 ml for 2–5 years of age group in QID (6th hourly) after food. Group – B - Syp.Sugar Solution 5 ml for 5–10 years age group & 2.5 ml for 2–5 years of age group in QID (6th hourly) after food. Criteria for assessment Children suffering from Vataja Kasa (Dry Cough) were assessed and evaluated on the basis of the following parameters.     

KasaVega (Bouts of cough) Duration of each bout Anidra (Sleeplessness) Throat congestion Absolute Eosinophil count AEC

Other than Vataja Kasa (Dry cough). Kasa as anubhandha lakshana (Complication) in other systemic diseases.

TABLE 1: Registration data Groups Group A (Syp.Badara) Group B (Syp.Sugar solution) Total

No of patients Total Registered Drop out 31 1

Completed 30

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Table no 3 indicates that Syrup Sugar solution provided relief in some of the clinical symptoms of Vataja Kasa. It provided 23.33% relief in Shushka Kasa (Non productive cough), 89.47% relief in Shirashoola (Head ache), 13.33% in Swarabheda (Hoarseness of voice), 17.39% in Parshwa Shoola (Pain in the flanks), 16.67% in Shushka vaktra (Dryness of mouth) 19.44% in Kantodhvamsanam (Throat irritation), 25% in Kasa Vega (Bouts of cough), 24.72% in duration of each bout of cough, 25.93% relief in Anidra (Sleeplessness). All the changes were statistically significant except in Parshwa Shoola.

RESULTS & DISCUSSION: Table no 2 indicates that Syrup Badara provided relief in all the clinical symptoms of Vataja Kasa (Dry cough). It provided 56.11% relief in Shushka Kasa (Non productive cough), 82.69% relief in Shirashoola (Head ache), 66.67% in Swarabheda (Hoarseness of voice),86.36% in Parshwa Shoola (Pain in the flanks), 91.07% in Ura Shoola(Chest pain), 63.33% in Shushka vaktra (Dryness of mouth), 66.67% in Kantodhvamsanam (Throat irritation), 57.22% in Kasa Vega (Bouts of cough), 56.67% in duration of each bout of cough, 80% relief in Anidra (Sleeplessness). All the changes were statistically significant.

TABLE 2: Effect on symptoms, signs and hematological parameters of Vataja Kasa in Group A Features

0.90

% of relief 56.11

Shushka Kasa-(Non productive cough) 30

1.93

0.03

31.0

<0.001

Shirashoola- (Head ache) Swarabheda- (Hoarseness of voice) Parshwa Shoola- (Pain in the flanks) Ura Shoola- (Chest pain) Shushka vaktra(Dryness of mouth) Kantodhvamsanam(Throat irritation) KasaVega (Bouts of cough)

26 30 22 28 30 30 30

1.0 1.60 0.90 1.03 1.70 1.63 1.93

0.2 0.6 0.17 0.13 0.7 0.53 0.87

82.69 66.67 86.36 91.07 63.33 66.67 57.22

0.09 0.07 0.10 0.06 0.05 0.05 0.05

9.05 14.75 7.71 16.16 20.86 20.86 23.03

<0.001 <0.001 <0.001 <0.001 <0.001 <0.001 <0.001

Duration of each bout Anidra (Sleeplessness) Rhonchi Throat congestion Total WBC Neutrophil (%) Lymphocyte (%) Eosinophil (%) Monocyte (%)

30 1.93 0.90 30 1.57 0.40 8 0.40 0.13 30 1.1 0.23 30 8912.2 8861.7 30 67.3 66.13 30 26.2 28.27 30 6.2 5.6 9 0.3 0

56.67 80 75.0 81.67 0.51 ↓ 1.58 ↓ 9.94 ↑ 8.79 ↓ 100 ↓

0.03 31.00 <0.001 0.07 16.86 <0.001 0.08 3.25 <0.01 0.06 13.73 <0.001 15.49 3.26 <0.01 0.49 2.37 <0.05 0.55 3.77 <0.01 0.10 5.83 <0.001 0.9 3.53 <0.01

AEC ESR (mm/1st hr)

30 30

519.4 10.47

479.2 8.67

6.94 ↓ 18.99 ↓

14.29 2.81 <0.01 0.14 12.95 <0.001

Hb (gm %)

13.37

13.42

0.42 ↑

0.04

1.19

>0.05

Note: ↓decrease , ↑increase ; N = number of frequencies, BT = Before Treatment AT = After Treatment; % = Percentage SE = Standard Error; T = Student ‘T’ Test; P = Probability

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TABLE 3: Effect on symptoms, signs and hematological parameters of Vataja Kasa in Group B Features

% of relief

Shushka Kasa-(Non productive cough) 30

2.00

1.53

23.33

0.09 5.04 <0.001

Shirashoola- (Head ache)

0.70

0.20

89.47

0.12 4.01 <0.01

Swarabheda- (Hoarseness of voice)

1.73

1.43

13.33

0.10 3.07 <0.01

Parshwa Shoola- (Pain in the flanks)

0.83

0.77

17.39

0.10 0.70 >0.05

Ura Shoola- (Chest pain)

0.80

0.97

2.17 ↑

0.08 1.98 >0.05

Shushka vaktra (Dryness of mouth)

1.67

1.30

16.67

0.10 3.61

Kantodhvamsanam (Throat irritation)

1.53

1.10

19.44

0.10 4.18 <0.01

KasaVega (Bouts of cough)

2.20

1.63

0.09 6.16 <0.001

Duration of each bout

2.17

1.57

24.72

0.09 6.60 <0.001

Anidra (Sleeplessness)

1.27

0.97

25.93

0.12 2.52 <0.05

Rhonchi

0.53

0.30

0.09 2.54 <0.05

Throat congestion

0.93

1.0

7.14↑

0.05 1.44

>0.05

Total WBC

30 8938.9 8946.77

0.12↑

9.38 0.84

>0.05

Neutrophil (%)

67.93

68.6

1.14↑

0.48 1.40

>0.05

Lymphocyte (%)

25.53

25.10

0.46↓

0.53 0.82

>0.05

Eosinophil (%)

6.37

6.40

0.83↑

0.06 0.57

>0.05

Monocyte (%)

0.17

100

0.07 2.41

<0.05

AEC

30 549.10

550.87

0.75 ↑

2.90 0.61

>0.05

ESR (mm/1st hr)

8.93

9.07

6.11↑

0.29 0.45

>0.05

Hb (gm %)

12.83

12.62

1.63↓

0.04 4.75 <0.001

<0.01

Note: ↓decrease , ↑increase ; N = number of frequencies, BT = Before Treatment AT = After Treatment; % = Percentage; SE = Standard Error; T = Student ‘T’ Test; P = Probability

Table no 4 gives a clear picture of comparison between the two groups. Group A showing highly significant result i.e p <0.001 in all symptoms of Vataja Kasa (Dry cough) except Shirashoola (Headache). Syrup Badara helps in reducing Shirashoola (Headache), Parshwa Shoola (Pain in the flanks) and Ura Shoola (Chest pain) due

to Ushna Veerya (Hot potency) of the drug Badara which acts as Vedana sthapaka (Analgesics). Kasa vega (Bouts of cough), Shushka kasa (Non productive cough), Shushka vaktra (Dryness of mouth) and Swara bheda (Hoarseness of voice) are due to Rooksha Guna (Dry quality) of Vata and the Snigdha guna

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(Unctous quality) of Badara might provide better relief in those symptoms.

of antigen (allergen/ pathogen) and receptors (such as Dendritic Cells, MHCs those are embedded in the mucosa of nose, pharynx, larynx etc. This drug action explains the symptomatic relief from Kasa (Cough).

Kashaya Rasa (Astringent taste) of stem bark of Badara in Syrup Badara might do the mucosal modulation, which cuts off the contact

TABLE 4: Comparative effect on symptoms, signs and hematological parameters of Vataja Kasa in Group A and Group B Measures

Group A N % Relief 56.11 Shushka Kasa-(Non productive cough) 30 26 82.69 Shirashoola- (Head ache) 30 66.67 Swarabheda- (Hoarseness of voice) 22 86.36 Parshwa Shoola- (Pain in the flanks) 28 91.07 Ura Shoola- (Chest pain) 30 63.33 Shushka vaktra(Dryness of mouth) 66.67 Kantodhvamsanam(Throat irritation) 30 30 57.22 KasaVega (Bouts of cough) 30 56.67 Duration of each bout 30 80 Anidra(Sleeplessness) 8 75 Rhonchi 30 81.67 Throat congestion 30 0.51 Total WBC

Group B T p value N % Relief 30 23.33 5.80 < 0.001 19 89.47 0.71 N.S 30 13.33 6.63 < 0.001 23 17.39 7.23 < 0.001 23 2.17 ↑ 16.64 < 0.001 30 16.67 6.08 < 0.001 30 19.44 6.0 < 0.001 30 25.0 6.12 < 0.001 30 24.72 6.23 < 0.001 27 25.93 7.14 <0.001 11 50 1.57 N.S 27 7.14↑ 11.80 <0.001 30 0.12↑ 2.95 < 0.01

Neutrophil (%)

1.58

1.14↑

2.64

< 0.01

Lymphocyte (%) Eosinophil (%) Monocyte (%) AEC ESR (mm/1st hr) Hb (gm %)

30 30 9 30 30 30

9.94↑ 8.79 100 6.94 18.99 0.42↑

30 30 5 30 30 30

0.46 0.83↑ 100 0.75↑ 6.11↑ 1.63↓

3.16 5.27 0 3.08 4.73 4.28

< 0.01 < 0.001 N.S < 0.01 < 0.001 < 0.001

Note: ↓decrease, ↑increase and N.S not significant; N = number of frequencies, % = Percentage; T = Student ‘T’ Test; P = Probability

Marked decrease in ESR and simultaneous decrease in AEC clearly indicates the drug’s action on allergic conditions. Decrease in Total counts and polymorphs indicate the drug’s effectiveness even in infectious conditions. The present study results goes very well with the study conducted by Rakesh Johri K, (1992) which indicates that the Stem bark of

Badara (Ziziphus jujuba Lamb) contains abundant tannins, which exhibits surface action on the pharyngo-laryngeal mucosa and modulate them to decrease exudation, provides a shield against the contact of any antigen (pathogen/allergen), in addition tannins also have local antimicrobial action which destroys the microbes coming in contact.

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CONCLUSION Kasa (Cough) being a common childhood ailment has been given more emphasis by the medical people because it diverts parents psyche from their routine work. All the changes were statistically highly significant i.e p

< 0.001. Syrup Badara also showed statistically highly significant relief in duration of each bout, frequency of bouts, throat congestion, AEC and also show improvement in Hb gram %. The findings suggest that the Syrup Badara is an effective and safe herbal formulation for the Vataja Kasa (Dry cough).

REFERENCES Atkins,D. (2008). Diagnosis of allergic diseases. In: Behrman , RE.(ed.) Nelson’s Textbook of Pediatrics. Philadelphia, Saunders Elsevier. Ch 140, p.938 Diane, EP. (2008). Sinusitis. In : Behrman, RE. (ed.) Nelson’s Textbook of Pediatrics. Philadelphia, Saunders Elsevier. Ch 377, p.1749. Drake Lee, AB. (1987). The catarrhal child. In : Kerr, AG. (ed.) Scott. Brown’s otolaryngology. Butterworths & Co. Ltd, London. p. 280–290 Eggenberger, K. (1993). Respiratory tract infections most frequently seen in pediatric outpatient care. Ars Medicine. 1 May, p. 24–40. Hibbert, J. (1987) Tonsils and Adenoids. In : Kerr, AG. (ed.) Scott. Brown’s

Source of Support: Nil

Otolaryngology. Butterworths & Co. Ltd, London. p. 368–383. Innes, JA. (2006). Respiratory disease. In : Davidson, S S. (ed.) Davidson’s Principle and Practice of Medicines. Churchill Livingstone Elsevier, Philadelphia. p.657 Maw, AR. (1987). Otitis media with effusion. In : Kerr, AG. (ed.) Scott.I, Butterworths & Co. Ltd, London. p. 157–176. Rakesh Johri, K. (1992). Piperine-mediated changes in the permeability of rat intestinal epithelial cells. The status of γ-glutamyl transpeptidase activity, uptake of amino acids and lipid peroxidation. Biochemical Pharmacology. 1 April, p1401–1407. Trikamji Yadavji, (2009) Kasa Chikitsa Adhyaya., Charaka Samhita, Chaukhambha Sanskrit Sansthan, Varanasi. p. 540, p. 227, p. 332

Conflict of Interest: None Declared

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Research article EFFECT OF PARNAYAVANI (COLEUS AMBOINICUS LOUR.) ON MES INDUCED EPILEPSY IN RATS Sharma Monica1*, Khemani Naresh2, Singh Janardhan3 1

Lecturer, Department of Dravya Guna, CBPACS, Najafgarh, New Delhi, India Professor & HOD, Department of Dravya Guna, NIA, Jaipur, India 3 Department of Pharmacology, PGIMS, Rohtak, Haryana, India *Corresponding Author: Email: monika.harmony@gmail.com ; Mob: +91 7838241559 2

Received: 06/04/2013; Revised: 30/05/2013; Accepted: 04/06/2013

ABSTRACT Epilepsy is considered as the most dreadful disease & it’s prevalence has been estimated at 5–10 person per 1000.The present study was designed to investigate the antiepileptic potential of Coleus amboinicus Lour. on Maximal electroshock (MES) induced seizure model in rats. Total 50 Albino rats of wistar strain were used in the study. Animals were divided into 5 groups each having 10 rats. Fresh juice of Parnayavani i.e. Coleus amboinicus (2 ml/rat) was given in one group while the other group receives Aqueous & alcoholic extract of Parnayavani (500 mg/bw p.o.). Phenytoin (25 mg/kg bw i.p.) served as standard drug for comparision where as control group receives distil water as vehicle. In MES model delay in HLE was taken as end point. C. amboinicus leaf juice (CALJ) & alcoholic extract (CAalE) was having highest efficacy as antiepileptic drug in comparison to its aq. extract. in MES model . Thus we observed that leaf juice as well as alcoholic extracts of Coleus amboinicus possess anticonvulsant and neuroprotective activity. KEYWORDS: Epilepsy, MES, Parnayavani, seizures.

Cite this article: Sharma Monica, Khemani Naresh, Singh Janardhan (2013), EFFECT OF PARNAYAVANI (COLEUS AMBOINICUS LOUR.) ON MES INDUCED EPILEPSY IN RATS, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 435–440

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INTRODUCTION

MATERIAL AND METHODS

Humanity is suffering from various psychosomatic disorders like anxiety, depression, epilepsy, dementia, Parkinson’s (Nayak AP, 2010) and Alzheimer’s inspite of great advances in the field of medical sciences. Epilepsy is considered as the most dreadful disease amongst all, which continues to effect human beings since ancient time It’s prevalence has been estimated at 5–10 person per 1000 and it is considered as the second most common neurological disorder next to stroke (Katzung BG, 2007). The contemporary system of medicine has ample of potent antiepileptic remedies but they also have a risk of developing vast array of side effects including chronic toxicity, teratogenicity, adverse effect on cognition and behaviour etc. (Raza MF et al.,2001). Amid of all these, it is the high time to search remedies from the traditional treasure which may be proven as safe & effective antiepileptic agent.

The study was conducted in the P.G. deptt. of Dravyaguna, NIA, Jaipur & deptt. of Pharmacology, Pt. B.D.Sharma Post Graduate Institute of Medical Sciences, Rohtak. Total 50 Rats were taken for the study.

Coleus amboinicus Lour. Synonym C. aromaticus Benth. or Plectranthus amboinicus Lour. belonging to family Lamiaceae is commonly known as Parnayavani in Sanskrit, Patta ajwain in hindi, Karpurvalli in south India & Country borage in English. It is a large, succulent, aromatic herb found throughout in India and is native of South and East Africa. The leaves of this plant are traditionally used for the treatment of severe bronchitis, asthma, diarrhoea, epilepsy, renal & vesicle calculi & fever (Bhakuni DS et al., 1969). It has been reported to exhibit antilithotic, chemopreventive & antioxidant properties (Padma, P.R. et al., 1998). Although Parnayavani have been in folklore use in Epilepsy no scientific investigation is yet to be done for establishing its antiepileptic activity. Therefore the present study aimed to study the effect of aqueous & alcoholic extract and leaf juice of Coleus amboinicus Lour. (CAaqE), (CAalE), (CALJ) which are known to have anti-epileptic property on animal model.

Collection of Plant Material: The leaves of Coleus amboinicus were collected from the medicinal herbal garden of NIA Jaipur where it grows. The Botanical identification was carried out by BSI, Jodhpur letter no.BSI/AZC/A.19014/SE-1/Estt./162 dt.23.6.2010. Leaves were dried under shade, coarsely powdered & were packed separately in air tight containers. Extract Preparation : For each extraction about 200 g powdered drug was packed in Soxhlet apparatus. About 1.5 litres of solvent i.e. ethanol for alcoholic extraction & distilled water for aqueous extraction, was placed in a round bottom flask and a reflux condenser was attached above the soxhlet. The solvent was heated to boil on heating mantle & was subjected to extraction for 12 h. The filtrate was evaporated to dryness by keeping it on a water bath at 50–60˚C. This process is repeated thrice to get the required amount of extracts of Coleus amboinicus (ethanolic & aqueous extract). Preparation of leaf juice: About 500 g of fresh leaves of Coleus amboinicus were cut into small pieces & juice was prepared by crushing them in a mortar & pestle & by adding 30 ml distil water to it. EXPERIMENTAL ANIMALS Adult Albino rats of Wistar strain (weighing b/w 100–180 g) of either sex were used in the study. Rats were procured from the disease free animal house, Haryana Agricultural University (HAU), Hissar. The experimental rats were housed in poly propylene cages under laboratory conditions of 28 ± 2˚C temp with 75% relative humidity and

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photoperiod of 12 h light & dark cycle. The rats were given standard pellet diet supplied by Hafed, India Limited, Rohtak) and water ad libitum, throughout the experiment.

Group 1- Served as Control & receives normal saline (10ml/kg) as Vehicle.

The protocol of the study was approved by Institutional Ethical Committee (IAEC, PGIMS, Rohtak) & the experiments were carried out as per ethical guidelines for animal protection and welfare bearing the CPCSEA 438/01/a/cpcsea/dt 17.07.2006 in its resolution No: 9/IAEC/SVU/2006/dt 04.03.2006.

Group 3- Receives aqueous extract of Coleus amboinicus (CAaqE) (500 mg/kg bw p.o.)

Group 2- Receives Fresh leaf juice of Coleus amboinicus (CALJ) (2 ml/rat).

Group 4- Receives alcoholic extract of Coleus amboinicus (CAalE) (500 mg/kg bw p.o.) Group 5- Receives Phenytoin (25 mg/kg bw i.p.) standard drug for comparison.

Chemicals: Phenytoin (Epsolin, Cadilla, India) was used in the study. All other chemicals used were of analytical grade. Selection of Doses: As per OECD (2002) guideline no. 425, the LD 50 of Coleus amboinicus was estimated to be >5000 mg/kg. Hence, 1/10 th of the LD 50, i.e. 500 mg/kg, dose was selected for the study. Fresh leaf juice was administered at a rate of 2 ml/rat (Baskar R. et.al., 1992). Administration of test substance: The test drugs were prepared by dissolving the required extracts in distilled water. The volume of administration was kept at 1 ml/kg bw. A gastric catheter was used for oral drug administration. Phenytoin was dissolved in normal saline before i.p. administration. Induction of Epilepsy Seizures were induced by MES methods. Experimental design (Study Protocol) In the present study Method of Tomar & Everett (1974) & (Cashin et al., 1962) was used. Electric shock seizures were produced by delivering a current of 150 mA through corneal electrode for a period of 0.2 sec. from an electro-convulsiometer. Animals which showed tonic extension of hind limb were selected & given overnight rest. On the next day the animals were divided into 5 groups of 10 animals each.

Animals of Group 2 were pretreated with leaf juice of Parnayavani (Coleus amboinicus) electroshocks were given after 60 min of drug administration. Animals of Group 3 & 4 received aqueous and alcoholic extract of Coleus amboinicus respectively & again electroshocks were given after 60 min of drug administration. Group 1 and 5 were control & standard drug group respectively. Animals were considered protected if the drug prevents or delays the appearance of hind limb extension component of the Seizures. Effect of extracts on different stages of convulsions i.e. hind limb extension, flexion & clonic convulsions were also noted. Recovery & death of Rats were also recorded in each group. Statistical Analysis All the results were expressed as mean Âą standard error of mean (SEM) and analyses of variance (ANOVA) for the data were calculated by using the SPSS (Statistical Package for Social Sciences) software. RESULTS Phytochemical screening The preliminary phytochemical analysis showed presence of alkaloids, carbohydrates, glycosides, proteins, amino acids, flavonoids, quinine, tannins, phenolic compounds and terpenoids.

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Effect of CALJ, CAaqE and CAalE on MES induced epilepsy The duration of tonic HLE in animals treated with vehicle i.e. in control group was 10.1 ± 0.53 seconds. The CAaqE treated group showed significant reduction in duration of

HLE (2.3 ± 0.99) whereas CALJ, CAalE and standard drug Phenytoin treated group exhibit completely abolished HLE phase as compared to control group.The CALJ, CAaqE,CAalE and phenytoin treated group have shown 100%, 60%,100% and 100% protection respectively (Table-1).

Table No. 1. Effect of Coleus amboinicus leaf juice (CALJ), aqueous extract (CAaqE) & alcoholic extracts (CAalE) on MES induced Seizures in rats. Grou p I. II. III. IV. V.

Duration (seconds) in various phases Dose mg/kg of convulsions Mortality % Protecti Tonic HL Clonus on flexion extension 3.6 ± 10.1±0.53 5.6±0.58 2/10 0.34 CALJ 2ml/rat p.o. 4.0±0.49 NIL*** 10.5±0.84** 0/10 100% CAaqE 500 mg/kg 4.1±0.433 2.3±0.99*** 12.7±0.817* 0/10 60% p.o. ** CAalE 500mg/kg 3.2±0.388 NIL*** 14.7±1.69** 0/10 100% p.o. * Phenytoin 25mg/kg 1.9±0.233 NIL*** 2.3±0.335 0/10 100% i.p. * Data represented as mean+ SEM of 10 rats. . Data compared with one way ANOVA P <0.05 non significant **P <0.01 significant *** P <0.0001 highly significant Design of treatment n= 10 Control

DISCUSSION The MES induced seizures test is the most validated experimental method for assessment of antiepileptic drugs effective in generalised tonic clonic seizures (Loscher W, Schmidt D 1988) (Oliveira FA et.al., 2001). The MES model is used to identify compounds which prevents seizures spread (Kupferberg HJ 1989) (Stables JP, Kupferberg HJ 1995) and most of these compounds have the ability to inactivate voltage dependant Na+ channels in a dose dependant fashion. Such compounds suppress sustained repetitive firing in cultured neurons (Mcnamara J, 2001). In the present study CALJ, CAaqE, CAalE significantly inhibited the tonic HLE in MES test . It suggests the presence of antiepileptic compounds and their potential utility in the management of

generalized tonic clonic seizures. Further CALJ and CAalE were equally potent antiepileptic agents in comparison to CAaqE. Hence CALJ, CAaqE, CAalE may be expected to have similar type of mechanism. Carvacrol & Thymol (main active constituents of Coleus amboinicus (Haque IU, 1988), (Pino J et al., 1989) belongs to the class of monoterpenes. Terpinoids, the oxygenated derivatives of terpenes are reported to exhibit antioxidant properties (J. Grabmann, 2005), (Johanna Grassmann et al., 2002). There is convincing evidence that in seizures, level of secondary metebolites of Lipid peroxidation (LPO) i.e. Thio barbituric acid reactive substances (TBARS) was significantly increased & antioxidant drugs facilitate the suppression of LPO enhancement (Bashkatova

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V et al., 2003). Coleus amboinicus is reported to have antioxidant property (Rao BS et al., 2006) (Kumaran A et al., 2006). Thus it might be suggested that the suppression of LPO enhancement may be involved in the mechanism of action of antiepileptic potential.

CONCLUSION Thus we conclude that leaf juice as well as alcoholic extract of Coleus amboinicus possess anticonvulsant and neuroprotective activity and thus can be effectively used for the treatment of epilepticseizures.

REFERENCES Bashkatova V, Narkevich V, Galina Vitskova G and Vanin A (2003) . The influence of anticonvulsant and antioxidant drugs on nitric oxide level and lipid peroxidation in the rat brain during penthylenetetrazole induced epileptiform model seizures.Progress in Neuro - Psychopharmacology & Biological Psychiatry. Vol. 27, (3): p487–92. Baskar R, Varalaksmi P and Amsaveni (1992). Changes in tissue enzymes produced by Coleus aromaticus experimental Urolithiasis. Indian Drugs 29 (6): p254–58 Bhakuni DS, Dhar ML, Dhar MM, Dhawan BN, Mehrotra BN (1969). Screening of Indian plants for Biological activity part II. Indian J Exp Biol 7: p-250–62. Cashin CH,Jackson H. (1962). An apparatus for testing anticonvulsant drugs by electroshock seizure in mice. J Pharmacol ;14: p-445–75. Haque

IU (1988). Analysis of volatile constituents of Coleus aromaticus. Journal of the Chem. Society of Pakistan. 10 (3): p-369–71.

J. Grabmann (2005) .Terpenoids as Plant Antioxidants. Vitamins & Hormone. Vol 72: p-505–535.

Johanna Grassmann, Susanne Hippeli and Erich F. Elstner (2002). Plant’s defence and its benefits for animals and medicine: role of phenolics and terpenoids in avoiding oxygen stress. Plant physiology & biochemistry Vol 40 issue 6-8, p- 471–478. Katzung BG (2007). In “Basic & Clinical Pharmacology”, 10th Ed. Mc-Graw Hill, Boston, Toronto; p-374–94. Kumaran A and Karunakaran Joel R (2006). Antioxidant and free radical scavenging activity of an aqueous extract of Coleus aromaticus. Food Chemistry. 97: p109–14. Kupferberg HJ (1989). Antiepileptic drug development program: a cooperative effort of government and industry. Epilepsia,30 (1); p-S51–S56. Loscher W, Schmidt D (1988). Which animal models should be used in the search for new antiepileptic drugs? A proposalbased on experimental and clinical consideration. Epilepsy Res., 2; p-145–181. Mcnamara J (2001). Drugs effective in the therapy of epilepsy, chapter 21 In: Goodmann and Gilmann’s the pharmacological basis of therapeutics. Hardman GJ et. Al. Mcgrew hill company Inc. Newyork , p-521–47.

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Nayak AP (2010). Parkinsonism in Ayurvedic perspective, A bird’s eye view. Global J. Res.Med.Plants & Indigen. Med. 1 (11): p-629–38. OECD (2002). Acute oral toxicity. Acute oral toxic class method guideline 425 adopted 23.03.1996. In: Eleventh Addendum to the, OECD, guidelines for the testing of chemicals organisation for economical co-operation and development, Paris, June, 2000. Oliveira FA, Almeida RN, Sousa MFV, Barbosa-Filho JM, Diniz SA, Medeiros IA (2001).Anticonvulsant properties of Nsalicyloyltryptamine in mice. Pharmacol Biochem Behav. 68; p-199– 202. Padma,

P.R., Bhuvaneshwari, V and Silambuchelvi, K (1998). The activities of enzymatic antioxidants in selected green leaves. Indian J Nutr Dietet 35; :p-1–3.

Pino J, Rosado A and Borges P (1989).Volatile components in the essential oil of wild oregano (Coleus amboinicus Lour.). Food / Nahrung. 34 (9): p-819–23.

Source of Support: Nil

Rao BS Shanbhoge R, Upadhya D, Iagetia GC, Adiga SK, Kumar P, Guruprasad K, and Gayathri P (2006). Antioxidant, anticlastogenic and radioprotective effect of Coleus aromaticus on Chinese hamster fibroblast cells (V79) exposed to gamma radiation. Mutagenesis. 21 (4): p-237–42. Raza MF, Shaheen MI, Choudhary A, Suria AU, Rahman S, Sombati and Delorenzo RJ (2001). Anticonvulsant activities of the FS-1 Sub-fraction isolated from roots of Delphinium denudatum. Phytother. Res., 15; p- 426–430. Stables JP, Kupferberg HJ (1995). The NIH Anticonvulsant Drug Development (ADD) Program: Preclinical Anticonvulsant Screening project. In: Antiepileptic Drugs, 4th edn. Ed. Levy RH, Mattson RH, Meldrum BS, Raven Press, New York.p- 4–17. Toman JEP,Everett GM (1974). In Evaluation of drug activit Pharmacometrics. Laurence DR & Bachard AL,Eds. Academic press, London; p-287.

Conflict of Interest: None Declared

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Research article ANTIMICROBIAL POTENTIAL OF THE WILD AND CULTIVATED VARIETY OF ERANDA (RICINUS COMMUNIS LINN.) ROOT Doshi Krunal A1*, Sagar Dhwani H2, Acharya R N3 1

Ph D Scholar, Department of Dravyaguna, IPGT&RA, GAU, Jamnagar.Gujarat, India Head, Department of Microbiology, IAPS, GAU, Jamnagar, Gujarat, India 3 Associate professor, Department of Dravyaguna, IPGT & RA, GAU, Jamnagar, Gujarat, India. *Corresponding author: krunaldoshi760@gmail.com; Mob : 09898907572 2

Received: 04/04/2013; Revised: 23/05/2013; Accepted: 28/05/2013

ABSTRACT Methanolic extracts of wild and cultivated variety of Eranda (Ricinus communis) roots were evaluated for their antimicrobial activity against pathogenic microorganisms such as Escherichia coli, Salmonella paratyphi, Staphylococcus aureus, Staphylococcus epidermidis, and Aspergillus niger using agar well diffusion method. Cultivated variety showed additional antimicrobial activities than wild variety. Wild root extract at 0.282 mg/ml showed inhibiting growth of E.coli and S. epidermidis and cultivated root extract at 0.224 mg/ml showed inhibiting growth of E. coli and Salmonella paratyphi. These findings established the potential of the cultivated variety of Eranda root as an effective antimicrobial agent against selected organisms. However, further studies are needed to evaluate active compounds and probable medicinal benefits in humans by clinical trials.

KEYWORDS: Castor oil plant, Eranda root, Ricinus communis, Methanolic extracts,

Cite this article: Doshi K. A., Sagar D. H., Acharya R. N., (2013), ANTIMICROBIAL POTENTIAL OF THE WILD AND CULTIVATED VARIETY OF ERANDA (RICINUS COMMUNIS LINN.) ROOT, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 441–447

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INTRODUCTION For the treatment of infections occurred by bacteria a wide variety of antibiotics are commonly used (Tumah H, 2005). Due to excess use of existing antimicrobial drugs, multiple drug resistance has been developed. Antimicrobial resistance is an alarming condition for mankind, because most of the infectious bacteria have developed multiple drug resistance (Saeed S et al., 2007). In concern to drawbacks of conventional medicine, the use of natural products as an alternate to the conservative treatment in healing and cure of various diseases has been raised in the last few decades (Saeed S and Tariq P, 2007). Ricinus communis Linn (Euphorbiaceae), commonly known as Eranda in Ayurveda (Kirtikar KR & Basu, 1985), is a soft-wooded small tree widespread throughout tropics and warm temperate regions of the world. It is cultivated widely in India, especially in the region of Saurashtra of Gujarat for seed oil. In Ayurveda, the leaf, root and seed oil of this plant have been used for the treatment of inflammation and liver disorders (Kirtikar KR & Basu, 1985), roots are indicated in rheumatism, inflammation, backache, diseases of abdomen, fever etc (API, 1999). It is reported that this plant possesses hepatoprotective (R Yanfg LL et al., 1987 and Visen P et al., 1992), antidiabetic (Shokeen P et al.,2008), laxative (Capasso F, 1994), and antifertility (Sandhy K & Bobby RG, 2003) activities. Methanol extract of root showed Anti- inflammatory and Free radical Scavenging activity (Ilavarasan R et al., 2006). Methanol extracts of root also reported for its antimicrobial activity (Abhishek M. et al., 2012). Nowadays, due to lack of sufficient quantity of wild variety, the cultivated variety is mainly used to full-fill the market demand of its root. T antimicrobial effect of Eranda root extract have been published. But no research has yet been carried out to evaluate the comparative antimicrobial effect of wild and

cultivated root extracts. Hence, both the varieties were compared for their antimicrobial activity. MATERIAL AND METHODS Collection of Drugs Fresh roots of wild (more than six months) and cultivated variety (six months old) were collected after proper identification of the plant as Ricinus communis Linn. (Euphorbiaceae), from the adjacent area of Jamnagar town of Gujarat, India, with the help of a taxonomist and a specimen (no. 1491wild/cultivated) of the two were preserved in the department, for further reference. The obtained roots were shade dried and made into coarse powder with the help of a mechanical grinder (Mess no 20). Preparation of methanol extracts Both the samples were subjected for extraction with methanol (Shukla V.J. and Bhatt U.B., 2001). The concentration of both extracts for antimicrobial activity was 15 mg/ml. Test microorganisms The antibacterial experimental organisms were Escherichia coli, Salmonella paratyphi (Gram negative), Staphylococcus aureus, Staphylococcus epidermidis (Gram positive). The antifungal experimental organism was Aspergilus niger. Both bacterial and fungal strains were provided by by IMTECH, Chandigarh, India and collected from MBRC, Jamnagar. Antimicrobial assay was performed by inoculation of sub cultured pathogenic strains in nutrient broth. The experimental work was carried out at Dept. of Microbiology, IAPS, GAU, Jamnagar. Determination of Minimum Inhibitory Concentration (MIC) MIC of the extracts against the above mentioned organisms were determined by using broth dilution technique. Different sets containing a range of serial dilution of extracts

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Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 6 | June 2013 | 441–447

were prepared for the selected organisms. 3 ml of nutrient broth was filled in each test tube and autoclaved. Test tubes were left to cool down at room temperature. In each set 200 µl of antimicrobial extract was serially diluted in 13 test tubes and one test tube was kept as blank. Test tubes were inoculated with different pathogenic strains except blank. The inoculated sets were kept for incubation at 37ºC for 24 hours. Optical densities of each were measured at 600 nm. (Phillips R and Martyn R., 1999) Antimicrobial Assay Antimicrobial assay was carried out by agar (well) diffusion method (Jahir Alam Khan et al., 2011). Sterile nutrient agar plates (containing 20 ml N-agar) were prepared. 300 μl of the selected pathogenic strains were spreaded on different N-agar plates with the help of L-shape spreader. 5 wells of 9 mm diameter were bore with the help of sterile borer. 150 µl and 200 µl of each wild root and cultivated root extract were taken for the assay. The antibacterial drug Gentamycin (80 µg/ml) and antifungal drug Fuconozocon DT (80 µg/ml) were taken as standards. Plates were incubated at 37ºC for 24 hours and observed for zone of inhibition on next day. All susceptibility tests were performed in triplicates.

RESULT AND DISCUSSION Upon performing minimum inhibitory concentration (MIC) of methanolic extract of wild sample and cultivated sample against the above mentioned two gram positive and gram negative organisms, the minimum concentration of the wild variety sample extract required to inhibit growth of E.coli and S. epidermidis was 0.282 mg/ml, while inhibition of S. aureus and S. paratyphi were not observed in the concentration range selected for the assay. Minimum concentration of cultivated root extract required to inhibit growth of E. coli and Salmonella paratyphi was 0.224 mg/ml while no remarkable inhibition of the other bacteria was observed (Table 1, Graph 1, Table 2 and Graph 2). Upon performing the Agar diffusion method, 150 µl of wild root extract showed inhibition against S. epidermidis (1.1 mm zone) and A. niger (1 mm zone) where as zone measuring 1 mm was obtained against E. coli when 200 µl of the same extract was taken. Similarly 150 µl cultivated variety root extract showed inhibition against A. niger (1 mm zone) and 200 µl of this extract showed zone of inhibition against all the remaining organisms. The zones of inhibition of the extracts were compared to that of the standards where it was found that the zone obtained with standards were larger than both the extracts. (Table 3 and figure 1).

Table 1: MIC of wild root extract on different bacteria Sr no.

concentration of wild root sample extract (µg/ml)

1 2 3 4 5 6 7

0.00 14.10 56.40 84.60 126.90 169.20 282.00

O.D. (Optical Density) at 660 nm on different organism E.coli Salmonella S. S. paratyphi aureus epidermidis 0.20 0.16 0.38 0.30 0.18 0.15 0.37 0.29 0.17 0.14 0.35 0.25 0.16 0.14 0.34 0.17 0.16 0.13 0.30 0.14 0.15 0.12 0.29 0.07 0.12 0.10 0.27 0.05

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Graph 1: Concentration of wild root extract v/s O.D. (Optical Density) at 660 nm on different organisms E.coli Salmonella paratyphi

Optical density at 660 nm

0.4

S. aureus

0.35

S. epidermidis

0.3 0.25 0.2 0.15

0.1 0.05 0

100

200

300

Concentration of wild root extract

Sr no.

1 2 3 4 5 6

Table 2: MIC of cultivated root extract on different bacteria concentration of cultivated root O.D. (Optical Density) at 660 nm on different extract (Âľg/ml) organism E.coli Salmonella S. S. paratyphi aureus epidermidis 0.00 0.2 0.21 0.21 0.18 0.2 11.20 0.18 0.19 0.17 56.00 0.16 0.19 0.18 0.16 100.80 0.15 0.18 0.17 0.15 168.00 0.12 0.16 0.15 0.12 224.00 0.07 0.1 0.11 0.1 Graph 2: Concentration of cultivated root extract v/s O.D. (Optical Density) at 660 nm on different organisms

Optical density at 660 nm

0.25

E.coli Salmonella paratyphi

0.2

S. aureus 0.15

S. epidermidis

0.1

0.05 0

100

200

300

Concentration of cultivated root extract

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Table– 3: Antimicrobial susceptibility assay – wild and cultivated varieties of Eranda root. Sr. No.

1 2 3 4

Name organism

Escherichia coli Salmonella paratyphi Staphylococcus aureus Staphylococcus epidermidis Aspergillus niger

Zone of inhibition by wild variety root extract(mm) 150µl 200 µl (169.2 (282 µg/ml) µg/ml) 0 1.0 0 0

Zone of inhibition by cultivated variety root extract(mm)

Gentamycin (concentr-ation 80 µg/ml) (mm)

Fluconozocon DT (concentration 80 µg/ml) (mm)

150 µl (164 µg/ml) 0 0

200 µl (224 µg/ml) 2.2 2.1

12 10

− −

1.0

−

1.1

2.2

1.2

−

1.0

2.0

1.0

2.0

−

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CONCLUSION The methanol extract of (R. communis L.) both the wild and cultivated root showed considerable antimicrobial effect. The cultivated variety showed inhibition towards the growth of the gram +ve and gram –ve organisms where as wild variety root extract showed effect against the specific organisms S. epidermidis and E. coli. Both the extracts showed similar effect against the selected fungus A. niger. The result of the study was restricted to the selected concentration range of

the extracts. Further antimicrobial assay can be done with a higher range of concentration. ACKNOWLEDGEMENT The authors wish to thank Miss Jigisha Pancholi, Head dept. of Biochemestry, IAPS, GAU, Jamnagar for her valuable suggestions throughout the study. We are thankful to Mr. Manish Vyas, Ph.D. Scholar, Dept. of RS & BK, IPGT & RA, GAU, Jamnagar for his support in arranging the Microbial and Fungal strains.

REFERENCES Abhishek M, Satish K, Sajad Y, Santosh K. Singh, Gbks Prasad, V. K. Dua (2012). Antimicrobial potential of Roots of Ricinus communis against pathogenic microorganisms, Jan-March; Issue – 1 ; vol-2 ; 545–548. Anonymous (1999), The Ayurvdic Pharmacopoeia of India, edition 1st, Govt. of India. Ministry of Health and Family welfare, Department of I.S.M. & H., New Delhi, Part-I, Volume –I. p. 34–35. Capasso F, Mascolo N, Izzo AA, Gaginella TS. (1994) Dissociation of castor oil induced diarrhoea and intestinal mucosal injury in rat: effect of NGnitro-l-arginine methyl ester. Bri J Pharmacol; 113: 1127–30. Ilavarasan R, Mallika M, Venkataraman S. (2006). Anti-inflammatory and free radical scavenging activity of Ricinus communis root extract. J Ethnopharmacol; 103: 478–80. Jahir Alam Khan, Krishna Pratap Yadav (2011), Assessment of Antifungal property of Ricinus communis, JPBMS, 11(02).

Kirtikar KR & Basu (1985), BD. Indian Medicinal Plants. 2nd ed, Dehradun: International book distributor; pp. 2274–2277. Phillips R and Martyn R. (1999) Annuals and Biennials. London: Macmillan. P. 106. R Yanfg LL, Yen KY, Kiso Y, Kikino H., (1987) Antihepatotoxic actions of formosan plant drugs. J Ethanopharmacology; 19: 103–10. Saeed S and Tariq P (2007), Antimicrobial activities of Emblica officinalis and Coriandrum sativem against Grampositive bacteria and Candida albicans. Pak. J.Bot., 39(3): 913–917. Saeed S, Naim A and Tariq P (2007), A study on prevalence of multi-drug- resistant Gram-negative bacteria. Int. J. Biol. Biotech., 4(1): 71–74. Sandhyakumari K, Bobby RG (2003), Indira M. Antifertility effects of Ricinus communis Linn. on rats. Phytother Res; 17: 508–11.

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Shokeen P, Anand P, Krishna Y M, Tandon V. (2008) Antidiabetic activity of 50% ethanolic extract of Ricinus communis and its purified fractions. Food Chem Toxicol; 46: 3458–66. Shukla V.J. and Bhatt U.B. (2001), Methods of Qualitative Testing of some Ayurvedic Formulations, Gujarat Ayurvedic University, Jamnagar.

Source of Support: Nil

Tumah

H (2005); Fourth generation cephalosporin: In-vitro activity against nosocomial Gram-negative bacilli compared with beta-lactam antibiotics and ciprofloxacin. Chemotherapy, 51(23): 80–85.

Visen P, Shukla B, Patnaik G, Tripathi Kulshreshtha D, Srimal R, Dhawan (1992) Hepatoprotective activity Ricinus communis leaves. Int Pharmacognosy; 30: 241–50.

S, B. of J

Conflict of Interest: None Declared

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Review article PROBABLE ETIOPATHOGENESIS (SAMPRAPTI) OF AUTISM IN FRAME OF AYURVEDA IN RELATION TO INTENSE WORLD THEORY Yadav Deepmala1*, Behera Banshidhar2, Kumar Abhimanyu3 1

Asst.Professor, Dept. of Kaumarbhritya, M.S.M. Institute of Ayurveda, Khanpur kalan, Haryana-131305, India 2 Lecturer, Dept. of Dravyaguna, Gaur Brahman Ayurvedic College, Rohtak, Haryana – 124001, India 3 Director, All India Institute of Ayurveda, Gautampuri, Mathura road, Sarita Vihar, New Delhi-110076, India *Corresponding Author: E-mail: drdeeyashri2011@rediffmail.com; Mob +919414893921, +919414458895

Received: 10/05/2013; Revised: 26/05/2013; Accepted: 30/05/2013

ABSTRACT Autism is one of the pervasive developmental disorders (PDDs). The disease affects the child's interaction skill with the world by involving social responsiveness, communication ability and lack of understanding for the other people showing heterogeneous clinical features of the disease. Hence, to establish the etiopathogenesis (samprapti) of autism will help to interpret its heterogeneity and to frame out its respective management. Hence, an effort has been made to explain the neurobiology of autism on the basis of Ayurvedic fundamentals and „intense world theory‟ of autism. This knowledge of etiopathogenesis may reveal the specific nature of brain dysfunction in autism and may help to understand the development of symptomatology of the disorder and further its precise nature to respond with various treatment strategies described in Ayurveda. KEY WORDS: Autism, PDD, etiopathogenesis, intense world theory

Cite this article: Yadav. D, Behera. B., Kumar. A., (2013), PROBABLE ETIOPATHOGENESIS (SAMPRAPTI) OF AUTISM IN FRAME OF AYURVEDA IN RELATION TO INTENSE WORLD THEORY, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 448–459

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INTRODUCTION According to DSM-IV, (Diagnostic and Statistical Manual of Mental Disorders, 1994) “Autism is a severe developmental disorder characterized by abnormalities in social functioning, language and communication and unusual interests and behavior”. The key features of autism are – Deficient non-verbal behavior such as eye to eye gaze, facial expression, body posture and gestures. Failure to develop peer relationship. Failure to share enjoyments interests or achievements with other people. Impaired development of social language. Echolalia/ idiosyncratic language. Restricted repetitive and stereotyped patterns of behavior, interests and activities. Here, the process of the development of symptomatology of autism is tried to explain on the basis of „Intense World Theory‟ (Markram & Markram, 2010) of autism and fundamental principles of Ayurveda contributing in disease process. AIMS AND OBJECTIVES 

To validate the contribution of components of Samprapti (etiopathogenesis) (i.e. khavaigunya, tridoshas, trigunas etc.) in pathophysiology of autism.



To establish the etiopathogenesis of autism in frame of Ayurveda to understand the development of symptomatology so as to avail the multimodal treatment approaches of Ayurveda in management of autism.

REVIEW OF CONCEPT According to fundamentals of Ayurveda, Samprapti (Etiopathogenesis) of any disease comprises the vitiated dosha-dushya sammurchhana (union). These vitiated components are manifested in full flown disease, when they merge with already existing „khavaigunya‟(Shastri Kaviraj Ambikadutta,

1995) (i.e. structural and functional alteration of body tissues/ system etc.). An etiopathology of autism seems to be evolved with contribution of khavaigunya in greater extent (Flowchart- Step2). Few etiological factors may be consequent as khavaigunya in following wayi. Genetic Factors: Various research studies establish the direct link between genes and their consequence into autism. Some of them are briefly mentioned here (Flowchart -step1)  Over expression of NMDA receptor gene, particularly the receptor subunits NR2A and NR2B as well as the CAM Kinase linked second messenger pathway are observed in autistic brains. (Rinaldi et al., 2007).  Several studies indicate the involvement of glutamatergic systems in autism. Single nucleotide polymorphism (SNPs) in the gene encoding "glutamate6 receptor" (GLUR6 -Jamain et al., 2002) and “glutamate8 receptor” (GLUR8 - Serojee et al., 2003) were reported in autistic brains. Also, several glutamatergic synapse gene mutations on chromosome 22 were also associated with autism (Jamain et al., 2003).  Rett‟s Syndrome (RTT), a trait of autism, is an X-linked dominant progressive neurodevelopmental disorder which exhibits all the three core characteristics of autism. It is caused by mutations in the gene encoding methyl CPG-binding protein (MeCP2). (Amir et al., 1999)  Similarly fragile X-syndrome is another Xlinked disorder which displays the features similar to autism such as abnormal speech pattern, stereotypic movements and abnormal social behavior, particularly shyness and limited eye contact. It is caused by the mutations in the FMR1 gene that encodes for fragile X-mental retardation protein (FMRP).

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Flow chart 1: Probable Etiopathogenesis of Autism

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ii. Epigenetic toxic insults: (Flowchartstep2)  A recent study of autism provides a link between hyper-activation of NMDA receptor and deficiency of hypothalamic inhibitory hormone i.e. digoxin. Lack of digoxin may fail to avoid brain damage due to excitotoxicity (Kurup & Kurup, 2003) and consequent in autism due to toxicity reasoning.  Imbalance of inhibition-excitation activities of neuronal pathways due to altered levels of serotonergic, GABAergic and NMDA neurotransmitters level may result in epigenetic toxic insults. iii. Postnatal environmental (Flowchart-step3)

factors:

Several perinatal environmental factors may cause autism. These factors include obstetrical complications, prematurity, hypoxic-ischemic encephalitis, jaundice etc. MMR Vaccine and certain drug toxicity are causally linked to the development of Autism. Thus, above three etiological factors are responsible for the structural and functional alterations in brain (Khavaigunya). This will further activates the molecular imbalance in cellular and circuit level that sensitizes gene expression pathways to respond excessively to environmental stimulation. (Flowchart-step4) Under normal condition, gene expression pathways would enable for enriched environments to nurture brain development but if these pathways are sensitized then environmental stimulation may cause exaggerated and accelerated development of brain. This will further affect the neuron connectivity in autistic brain. Several MRI studies suggest that in autistic brain long-range connections (essential for complex information processing of higher order functions) between different brain areas are underdeveloped while the short-range connections (essential for primary information processing) are overly developed. (Just et al., 2007; Mottran et al., 2006)

According to Ayurvedic perspective to maintain proper connection between body tissues is a function of pranavayu (i.e. sandhankar karma) (Chaturvedi G. & Shastry K., 1996). Therefore, the hyper-connectivity between short-range synapses and underconnectivity between long-range synapses may be caused by vitiated pranavayu. Similarly to maintain the intactness between these connections or of body tissues is a function of kapha dosha (i.e.sneha, bandha & sthiratva) (Chaturvedi G. & Shastry K., 1996). But the impaired kapha dosha may lack to provide nourishment (Snehana) to brain matter and affects its compactness and stability which ultimately makes brain tissues more vulnerable to endogenous cytotoxicity. Thus, these alterations in neural connectivity leads to cascade of events which comprises the symptomatology of autism in a following wayA. Hyper-connectivity to primary sensory pathway

short-range

Due to hyper-connectivity between short range neurons, the flow of primary sensory information speedily transfers via synapses causing hyper reactivity across different brain regions (Flowchart-step 4, 5, 6). In Ayurvedic context, this process is carried out by enhanced „chalatva guna‟ (quality) of vata dosha (Chaturvedi G. & Shastry K., 1996).This hyper-reactivity contributes central role in pathophysiology of autism, which can be interpreted as below– i.

Locus Coeruleus (LC):

Super-charged micro circuits in primary sensory area may produce enhanced sensitivity to sensory stimulation. This may consequently over sensitize to locus coeruleus for upcoming environmental stimulation and may result into enhanced nor epinephrine secretion, which further leads to over excitation of NE pathway (Flowchart-step7). Over excitatory NE pathway is cause for hyper-perception and hyper-attention. As per a scientific study, higher sustained attention is reported in autism as compare to control group

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i.e. autistic people have the ability to maintain attention to repetitive stimuli over prolonged periods of time (Johnson et al., 2007). This may hamper the capacity of selective attention for relevant sensory stimuli and thereby sensory overload. (Flowchart-step8) In an Ayurvedic context, vitiated pranavayu and kapha dosha may lead to impairment of function of manas (Chaturvedi G. & Shastry K., 1996) and the function of dhee (Chaturvedi G. & Shastry K., 1996) which result into abnormal orientation and engagement of all relevant and irrelevant environmental stimulation, in turn causes for sensory overload. ii.

Neocortex and Amygdala:

In pathophysiology of autism, neocortex particularly prefrontal and somatosensory cortex and amygdala are involved in great extent. At neocortical region, the hyperexcitatory NE pathway are inhibited by increased GABAergic pathway indicating that inhibition may able to recruit a constant matching level of excitation without developing an imbalance (Rinaldi et al., 2008). This will confront the neocortex for excessive processing of primary sensory information. (Flowchartstep9) This can be illustrated by an example of sensory perceptions. Primary processing of environmental stimuli of sensory origin (i.e. visual, auditory, touch etc.) may be carried out excessively by somatosensory cortex while cognitive functions related to simple feature are carried out excessively by prefrontal cortex. Similarly, Amygdala may also become overly reactive for processing emotionally relevant information. It is widely established that the amygdala mediates the formation and storage of fear memories (Le Daux, 2003) and enhances memory formation throughout the other brain regions by acting as an emotional amplifier (Cahill & Mc Gaugh, 1996). Hence in autistic people, dysfunction of amygdala may result into exaggerated and more persistent processing and storage of aversive emotional and fear related memories. This is supported by the theory of imbalance of excitation inhibition

which would result into increased reactivity, due to loss of inhibition at amygdala (Casonova et al., 2003). Thus, the hyper reactive neocortex and amygdala may significantly consequences into excessive perception, attention, learning and emotionality (Flowchart-step10). This process of hyper learning is stored in the form of simple features of touch, sound, light, fear, emotions, language etc. (Flowchart-step11). Hippocampus, Basal ganglia and amygdala are the sites, where memories of these simple features are allocated (Flowchart-step12). Long term potentiation (LTP) is the neuronal mechanism which is widely assumed to underlie memory formation. This LTP mechanism is mediated by glutamatergic neurotransmitters and receptors system particularly NMDA (Nicoll & Malenka, 1999) and alterations in this system may contribute to the above observed hyper-plasticity leading to hyper-memory and hyper-learning. (Flowchartstep13) With excessive learning and memory processes, sensory regions may consolidate into overspecialized modules and lead to hyperpreference processing pathway. (Flowchartstep14) During early development (probably before age 3 years), this may lead to excessive flow of information from sensory areas to the higher integration areas such as association cortices and prefrontal lobe which may cause prematurely accelerated growth of these higher order brain areas as truly observed in autism. This would be the reason of why autistic children have certain unusual talents and older autistics are excellent on task involving long term memory like recall of train time-tables, historical dates, chemical equations or recall of the exact words of songs heard years before (Carper et al., 2002) Ultimately the hyper-preference processing in the sensory domain, may lead to exaggerated selectivity, sensitivity and specialization of simple sensory features. As a consequence-

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 Autistic children would remain with fragmented and amplified perception of bits and pieces of the world.  Autistics may have abnormal and obsessive attention to detail and hence can notice the smallest change in their environment.  Autistics may become hyper-focused in arbitrary subjects of interests and sustain their attention on these subjects for unusually long time periods. On the whole, this hyper-connectivity phenomenon may cause for enhanced perception of sensory fragments; focus on details and deficit in complex and more holistic processing. As per the Ayurvedic aspect, pranavayu promotes and regulates the other biophysical components and sense faculties (by the virtue of prerana karma) to perform their respective functions (Chaturvedi G. & Shastry K., 1996). But as described earlier, vitiated prana vayu may unable to perform its function of synaptic connectivity properly and so as to fail to regulate the functions of other components, rather consequently responsible for impairment of manas, buddhi and other doshas. (Flowchart-step7,8) Due to impaired manas, the functions related to motor control, abstract thinking and thoughtful planning may upset and lead to non-oriented information processing (Chaturvedi G. & Shastry K., 1996) (Flowchart-step9). Due to impaired buddhi, particularly dhriti, the function of selective attention and further retention may disturb leading to excessive storage of primary information (Chaturvedi G. & Shastry K., 1996) (Flowchart-step10). Functions of sadhak pitta simply resemble with the functions of neocortex and amygdala. As cognitive functions represented by sadhak pitta can be represented in the form of buddhi and medha (Arunadutta, 2002) as well as

function of emotional & social cues are represented as bhaya, shaurya (Chaturvedi G. & Shastry K., 1996) etc. But due to derangement of sadhak pitta, the exaggerated processing of sensory, cognitive and emotional cues take place thereby leading to overflow of primary knowledge (Flowchart-step11, 12). Memorization i.e. smriti is a function carried out by udan vayu (Arunadutta, 2002). But due to its derangement and hyper preferential mnemonic pathway, autistic child may show excessive retrieval of working memory as well as long term memory of only primary features (Flowchart-step13). But in older autistics, working memory functions are seen widely upset, although long term memory may remain intact due to repetitive maintenance rehearsal. This may be suggestive of ongoing degradation of Udan vayu due to local background pathology (Flowchartstep14). Concept of hyper-memory can also be explained with the help of an Ayurvedic principle i.e. „Samanyam Vriddhikaranam‟ (Chaturvedi G. & Shastry K., 1996). According to this theory, generic concomitance may augment the same class of characters, in turn may responsible for their overloading. Such an overspecialized hyper-memory may tend to activate even by the mild environmental stimuli of same class of knowledge and further integrate with its processing to develop hyper-preferential pathway.This mechanism is also supported by one of smriti-hetu (Factors responsible for memory) i.e. „sadrishyat smritirupajayate‟ (i.e. similarity between current knowledge and previous experience). (Chaturvedi G. & Shastry K., 1996) (Flowchart -step14). Back on continuing modern pathophysiology of autism, in general, it is observed that the autistic children may seem normal, rather, gifted at the initial period of development, the symptomatology of autism

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initiates after the age of 18 months or before the age of 3 years when child begins to learn more complex task. The symptomatology may be driven by under-connectivity of long range of connections required for complex task. B. Under-connectivity to complex sensory pathway

long

range

As described earlier, long range neuronal connections play a key role in complex information processing (Minshew & Goldstein, 1993). But in autistic brain, under-connectivity may observed between long range columns of different brain regions. Hence there is an increasing impairment in integrating progressively more complex information across different brain regions which in turn results into memory deficits for complex and abstract material (Flowchart- step15). Also the background mechanism of hyperpreferential sensory-mnemonic pathway of short-range columns, also inhibit the higher order information processing. As on exposure to environmental stimuli for complex task (Flowchart -step16), the hyper-preferential pathway become activate and enhances its sensitivity for selective simple features. This may synergistically processed by the consolidated mnemonic inputs (Hypermemory) of previous primary information and results into dominance of the earliest features and avoidance of processing of other features. Such hyper-autonomous and overly selective pathway leads to following symptoms with respect to different brain regions (Flowchart step17). i. Somatosensory step18)

cortex

(Flowchart-

Under-connectivity to long range circuits and hyper-connectivity with microcircuits in neocortex, particularly somatosensory cortex causes exaggerated perception and attention of fragments of sensory world which must be holistically processed at normal circumstances. This may enhance by hyper plasticity component which drives exaggerated memories to further amplify the processing of same stimulus and also drive over generalization of attention to all related forms of the stimulus.

Thereby the positive consequences are exceptional capabilities for primary and specific tasks while the negative consequences are impairment of holistic processing and a limited repertoire of behavioral routines, which may further repeated obsessively i.e. stereotyped behavior, a core characteristic of autism (Flowchart -step19). Similarly in the domains of auditory, visual & touch stimuli, autistic people on positive consequence may exhibit enhanced discrimination capabilities for elementary stimuli and on negative consequence, they may exhibit the diminished global interference for complex stimuli due to impaired pre-pulse inhibition (Foxton et al., 2003 Mottorn et al., 2003). This will lead to hypersensitivity to environmental sound, light and touch stimuli, which is observes as key characteristics of autism (Flowchart -step 20). In an Ayurvedic paradigm, stereotyped behavior and hyper-sensitivity to touch, light and sound are the features demonstrated by impaired vyan vayu (Arunadutta, 2002) (Flowchart -step19, 20). ii. Prefrontal Cortex: (Flowchart -step21) Downside of hyper-connectivity and under connectivity of cognitive regions, particularly PFC, indicates impairment in higher executive functions. Excessive memory in low level sensory and elementary cognitive regions may lead to an early over-specialization of primary feature processing, missed developmental opportunities to acquire a full spectrum of primary processing strategies and to build higher order strategies. This might lead to a fragmented alphabet of feature processing capabilities in the vocabulary of sensory processing and obstruct the development of higher cognitive functions such as abstract thinking and language processing. Thus, this mechanism is responsible for language and speech impairment, which is a characteristic of autism (Flowchart -step22). In an Ayurvedic paradigm, derangement of udan vayu is responsible for speech impairment (Arunadutta, 2002).

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iii. Amygdala: In autism the functional impairment of amygdala is resulted into storage of aversive emotional and fear related memories (Flowchart- step23). Later on, exposure to fear stimuli, this preformed memories may lead to progressive generalization of fear which may consequence into behavior and may account for inappropriate reactions to the environment, sudden & inexplicable anxiety attacks, loss of fitness required for social interaction and phobias. These are also core characteristics of autism. (Flowchart-step24) Impairment in social interactions & communication are other characteristics of autism, i.e. which are also mediated by insufficient activation of amygdala (Pierce et al., 2001). Due to hypo-activation of amygdala, autistic people may become severely unable to “read other people minds” (Flowchart-step25) and empathizing with other people by affecting the following two elements Ability to distinguish between oneself and others and realize that other people have independent minds and may pursue different goals from one-self.  The ability to express an appropriate emotional reaction to the other person‟s mental state, thus to be unable to empathize with others mind. On the whole, the deficits in language & speech, undue fear & social phobia, and inability to read other people's mind may altogether mediate impairment in social interaction and communication. (Flowchartstep26) In Ayurvedic paradigms the features of emotionality is connoted by Sadhak pitta (Chaturvedi G. & Shastry K., 1996) i.e.

bhaya harsha, prasad, krodha, etc. Thus vitiated sadhak pitta may responsible for increased fear and anxiety which lead to social withdrawal and avoidance. iv. Fusiform face area (FFA): Eye contact and watching the facial expressions are one of the first signs of cognitively healthy infants and serve to build the basis for successful navigation through a social environment. But deviant eye gaze is a core characteristic observed in autistic child. This is driven by the impaired cortical region, named as fusiform face area. As suggested by its name, in normal subjects this area is highly reactive to face recognition. But in autistic children, fusiform face area is observed to be hypo-reactive which lead to abnormal face perception and social avoidance. (Kanwishar et al., 2000) (Flowchart-step27) Due to hypo-reactivity of FFA, amygdala may have to confront to response the face and eye recognition stimuli. On exposure, the right amygdala shows greater activation when viewing familiar and unfamiliar faces while left amygdala and left orbito-frontal cortex shows greater activation on viewing emotional faces. Both areas forms part of the emotion circuit of the brain and shows heightened emotional response to these stimuli. Amygdala, simultaneously also makes quick and powerful fear associations with fearful mnemonic-inputs. In consequence to this, Autistic child may spend less time fixating with the eye region resulting into avoidance of eye contact which is a characteristic of autism (Dalton et al., 2005) (Flowchart -Step28 ) In an Ayurvedic paradigm, eye movement is a function of vyan vayu. So deviant eye gaze may also the result of deranged vyan vayu (Arunadutta, 2002).

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Table 1: Probable etiopathological markers of Autism as per Modern in relation to Ayurveda Modern 1. 2.

4. 5.

Genetic factor, epigenetic toxic insults, post natal environmental factor Dysregulation of gene expression path way Hyperconnectivity to short-rang pathway of primary features Underconnectivity to long-range neuronal pathway Information speedly transfer via synapses Increased connectivity of NE pathway

Ayurveda

Reference

Khavaigunya

Shastri Kaviraj Ambikadutta ,1995

Khavaigunya

Shastri Kaviraj Ambikadutta ,1995

Impairment of sandhankar karma of pranavayu

Chaturvedi G. & Shastry K., 1996

Impairment of chalatva quality of Vata dosha Impairment of pranavayu

Hyperperception and hyperattention

Impairment of Dhee

Somatosensory cortex, PFC & amygdala

Excessive storage of all information

Function similar to manas and buddhi Based on principle of “ samanyam vriddhi karanam”

Hyper memory

Chaturvedi G. & Shastry K., 1996 Chaturvedi G. & Shastry K., 1996 Chaturvedi G. & Shastry K., 1996 Chaturvedi G. & Shastry K., 1996 Chaturvedi G. & Shastry K., 1996

Increased Udanvayu

Arunadutta, 2002

10. Hyper preferential pathway

Excessive intake of smritihetu

Chaturvedi G. & Shastry K., 1996

Stereotyped repetitive behavior 11. Hyper sensitivity to light, touch, sound

Impairment of Vyanvayu

Arunadutta, 2002

12. Impairment of language

Impairment of Udan vayu Arunadutta, 2002

13. Fear, anxiety and phobia

Impairment of Sadhak pitta

14.

Lack of communication and social Impairment

15. Lack of eye communication

Chaturvedi G. & Shastry K., 1996 Arunadutta, 2002, Impairment of Udan vayu Chaturvedi G. & Shastry and Sadhak pitta K., 1996 Impairment of Vyana Arunadutta, 2002 vayu

CONCLUSION Etiopathogenesis of autism may predominantly evolve from khavaigunya as consequence of various sahaja (genetic) & agantuja (Epigenetic toxic insults & post natal environmental factor) hetus. Vitiated doshas (physical & mental) may exacerbate the

khavaigunya leading to various core features of autism. Hyperconnectivity & underconnectivity among short range & long range neuronal pathways respectively may evolve from impairment of pranavayu which in turn causes for hyper-perception, hyper-attention & hypermemory (i.e. impairment of functions of manas & buddhi). Impairment of vyana vayu may

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responsible for stereotyped repetitive behavior, hypersensitivity to light, touch & sound and lack of eye communication. Deranged Udanvayu may result into echolalia and language impairment while deranged Sadhak pitta may cause fear, anxiety and phobia. These features, ultimatly contribute to lack of communication & social impairment. On the whole, this Ayurvedic framing of etiopathogenesis of autism may help to understand the contribution of pathological markers in developing symptoms of disease.

This knowledge can be further used in Ayurvedic management of autism by reducing the effect of „khavaigunya‟ through restoring the functions of other pathological markers and normalizing the functions of manas, buddhi (with dhee, dhriti and smriti), vitiated doshas and trigunas, which in turn, may helpful in reducing problematic behavior that would be adaptive at lower rates as well as maintaining this adaptive behavior for longer duration. In addition, it may help in acquiring new skills of leaning and communicating.

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Chaturvedi Gorakhnath, Shastry Kashinath (1996), Charaka Samhita with “Vidyotini” Hindi commentary, Chaukhambha Bharati Academy, Varanasi Part 1, Sharirasthan, Sutrasthan .

Arunadutta (2002). Ashtanga Hridaya with “Sarwanga Sundara” commentaries, Chaukhambha Orientalia, Varanasi, Sutrasthan .

Courchesne, E., Karns, C.M., Davis, H.R., Ziccardi, R., Carper, R.A., Tigue, Z.D., Chisum, H.J., Moses, P., Pierce, K., Lord, C., Lincoln, A.J., Pizzo, S., Schreibman, L., Haas, R.H., Akshoomoff, N.A., Courchesne, R.Y. (2001). Unusual brain growth patterns in early life in patients with autisticdisorder: an MRI study. Neurology, 57, 245–254

Cahill,

L. and McGaugh, J.L. (1996).Modulation of memory storage.Curr.Opin.Neurobiol.,6, 237– 242.

Carper, R.A., Moses, P., Tigue, Z.D. and Courchesne, E. (2002). Cerebral lobes in autism: early hyperplasia and abnormal age effects. Neuroimage, 16, 1038–1051 Casanova, M. F., Buxhoeveden, D. and Gomez, J. (2003). Disruption in the inhibitory architecture of the cell mini column: implications for autism. Neuroscientist, 9, 496–507

Dalton, K.M., Nacewicz, B.M., Johnstone, T., Schaefer, H.S., Gernsbacher, M.A., Goldsmith, H.H., Alexander, A.L. and Davidson, R.J. (2005). Gaze fixation and the neural circuitry of face processing in autism. Nat. Neurosci., 8, 519–526

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DSM-IV. (1994). American Paediatric Association, Diagnostic and Statistical manual of mental disorders, fourth edition, Washington DC.

Kurup, R.K. and Kurup, P.A. (2003). A hypothalamic digoxin-mediated model for autism. Int. J. Neurosci.,113, 1537– 1559

Foxton, J.M., Stewart, M.E., Barnard, L., Rodgers, J., Young, A.H., O‟Brien, G. and Griffiths, T.D. (2003). Absence of auditory “global interference” in autism. Brain, 126, 2703–2709.

LeDoux, J. (2003). The emotional brain, fear, and the amygdala. Cell. Mol. Neurobiol., 23, 727–738.

Jamain, S., Betancur, C., Quach, H., Philippe, A., Fellous, M., Giros, B., Gillberg, C., Leboyer, M., Bourgeron, T. (2002). Linkage and association of the glutamate receptor 6 gene with autism. Mol. Psychiatry, 7, 302–310. Jamain. S., Quach, H., Betancur, C., Rastam, M., Colineaux, C., Gillberg, I.C., Soderstrom, H., Giros, B., Leboyer, M., Cillberg, C., Bourgeron, T. (2003). Mutations of the X-linked genes encoding neuroligins NLGN3 and NLGN4 are associated with autism. Paris Autism Research International Sibpair Study., Nat Genet. 34, 27–29. Johnson, K. A., Robertson, I.H., Kelly, S.P., Silk, T.J., Barry, E., Daibhis, A., Watchorn, A., Keavey, M., Fitzgerald, M., Gallagher, L., Gill, M. and Bellgrove, M.A. (2007). Dissociation in performance of children with ADHD and high-functioning autism on a task of sustained attention. Neuropsychologia., 45, 2234–2245. Just, M.A., Cherkassky, V.L., Keller, T.A., Kana, R.K. and Minshew, N.J. (2007). Functional and anatomical cortical under connectivity in autism: evidence from an FMRI study of an executive function task and corpus callosum morphometry., Cereb. Cortex 17, 951– 961. Kanwisher, N. (2000). Domain specificity in face perception. Nat. Neurosci., 3, 759– 763.

Markram, K. and Markram, H. (2010).The Intense World Theory – A Unifying Theory of the Neurobiology of Autism, Front Hum Neurosci., 4, 224. Minshew, N.J. and Goldstein, G. (1998). Autism as a disorder of complex information processing. Ment. Retard. Dev. Disabil. Res. Rev., 4, 129–136. Mottron, L., Burack, J.A., Iarocci, G., Belleville, S. and Enns, J.T. (2003).Locally oriented perception with intact global processing among adolescents with high-functioning autism: evidence from multiple paradigms. J. Child Psychol. Psychiatry, 44, 904–913 Mottron, L., Dawson, M., Soulieres, I., Hubert, B. and Burack, J. (2006). Enhanced perceptual functioning in autism: an update, and eight principles of autistic perception.J. Autism Dev. Disord., 36, 27–43. Nicoll, R.A. and Malenka, R.C. (1999). Expression mechanisms underlying NMDA receptor-dependent long-term potentiation. Ann. N. Y. Acad. Sci., 868, 515–525. Rinaldi, T., Kulangara, K., Antoniello, K. and Markram, H. (2007). Elevated NMDA receptor levels and enhanced postsynaptic long-term potentiation induced by prenatal exposure to valproic acid. Proc. Natl. Acad. Sci. U.S.A. 104, 13501–13506.

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Rinaldi, T., Perrodin, C. and Markram, H. (2008). Hyper-connectivity and hyperplasticity in the medial prefrontal cortex in the valproic acid animal model of autism. Front. Neural Circuits 2, 4. Serajee, F.J., Zhong, H., Nabi, R. and Huq, A.H.M. (2003).The metabotropic glutamate receptor 8 gene at 7q31:

Source of Support: Nil

partial duplication and association with autism Genet.,40(4), e42.

possible ,J Med

Shastri Kaviraj Ambikadutta (1995), Sushruta Samhita with “Ayurveda Tattva Sandipika” commentary, Chaukhambha Sanskrit Sansthan, Varanasi, part 1 , Sutrasthan.

Conflict of Interest: None Declared

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Review article CRITICAL APPRAISAL SKILL FOR THE ARTICLE PUBLISHED IN AYURVEDIC HEALTH DOMAIN Vaikos C D1*, Waghchoure Ashok2 1

Associate professor and Head of Department, Dept of Rachana Sharir, Govt. Ayurved College, Nanded, Maharashtra, India 2 Assistant professor, Dept of Rachana Sharir, Govt Ayurved College, Nanded, Maharashtra, India *Corresponding Author: Email - vaikosc@gmail.com

Received: 10/05/2013; Revised: 26/05/2013; Accepted: 02/06/2013

ABSTRACT It is the need of all Ayurvedic students, teachers and scientists to have complete knowledge of the process of evaluation and appraisal of research, review or original articles published in peer reviewed valid journals. Most of the students (and more or less teachers also) are ignorant on the appraisal of a published article since this was not part of their designed curriculum. Central council of Indian medicine have designed curriculum of research methodology recently, which includes publication skills. Published articles on Ayurvedic subjects must be evidence based. This article will help to develop an innovative approach on how to teach and learn critical appraisal skills to Ayurvedic students and hence user could decide how much he/she can rely on the results of the article and, therefore, shall or shall not use the results of that article in his/her practice. Research studies, including qualitative studies, form the basis for evidence-based practice among health professionals. However, many practicing Ayurvedic health educators do not feel fully confident to appraise qualitative and quantitative research studies. This review provides guidelines for appraising the strengths and weaknesses of published qualitative research articles and Health educators will be better equipped to evaluate the quality of the evidence through critical appraisals of qualitative research publications. Published research studies, including qualitative studies, provide the evidence for the selection of evidence-based practices in health education and promotion. For health educators, critical reading of research studies can be time consuming and challenging, however they can increase their confidence in appraising research studies by using the guidelines described in this article. KEY WORDS: Evaluation, appraisal, central council of Indian medicine, evidence based medicine, quantitative and qualitative study, Ayurveda

Cite this article: Vaikos C. D., Waghchoure. A. (2013), CRITICAL APPRAISAL SKILL FOR THE ARTICLE PUBLISHED IN AYURVEDIC HEALTH DOMAIN, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 460â&#x20AC;&#x201C;464

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INTRODUCTION Critical appraisal has been defined as The application of rules of evidence to a study to assess the validity of the published research data (wikipaedia.org), completeness of reporting, methods and procedures, conclusions, compliance with ethical standards,” etc. Critical appraisal is the skill that the science of epidemiology offers to scientists and specialists in different disciplines including medicine (Timm DF, Banks DE, McLarty et al., 2012, McAllister D, Wild S et al., 2009, Green BN, Johnson CD, et al., 2007). It is used to evaluate and determine the positive and negative aspects of a published article (Mohsen Rezaeian et al., 2006), in evidencebased healthcare training, to assist in clinical study in decision-making and in evidencebased social care and education provision (wikipaedia.org). Due to the rapid - development, spread and worldwide acceptance of Ayurveda, the expert specialities, students, clinicians, health professionals’, teachers, and research scientists in Ayurvedic field needs to develop efficient skills in critical appraisal of published articles to use or not to use results of that article in support of his/her articles. Ayurveda is an evidence based science, selection of relevant literature, understanding of research findings, elicit patients own preferences etc (Jane M Young, Michael J Solomon, 2009) and use of journal club and letter-to-the-editor writing project (Green BN, Johnson CD, 2007) may create ability in clinician to appraise the research articles. Journal club should be a part of curriculum which will provide exposure to Ayurvedic student to the world of frontline research and safe way to enter in the world of translational research (Tamoghna Biswas, 2011). The quantity of published articles in the databases like Medline, Pubmed, Medknow etc is increasing. According to Medline database more than 12,000 new articles, including papers on nearly 300 randomized controlled trials (RCTs), are added to their database in each week (Glasziou PP et al., 2008) but the question regarding the validity of results and their use in local help is merely doubtful.

Therefore every article should be critically scrutinised to evaluate the robustness of the ﬁndings. Many times the articles on quantitative and qualitative research are questioned by scientific scholars that they lack scientific rigour, credibility, reliability, trustworthiness and validity. In such cases it is necessary to audit the process up to the end product. This will help the user/reader to determine whether the claim made by the researcher is justiﬁable or not (Dorothy Horsbugh, et al., 2003). Qualitative research studies are particularly helpful when looking at context, experiences or meanings related to principals/laws of diagnostics/therapeutic measures or an issue; (for e. g. theory of Panchmahabhootas (five basic constituent element), Prakriti (body constitution), Samanya Vishesha Sidhdhanta, Lokpurusha Sidhdhanta etc). Quantitative research is useful to address the intervention or therapy questions (Jeanfreau, S. & Jack, L. 2010) or drug discovery or safety and toxicity study of herbo-mineral or mineral drugs. Both kinds of research, evidence may be necessary for a complete understanding of how and why an issue occurs (qualitative research questions) and what can be done to address the issue (quantitative research). DISCUSSION Articles on various subjects of Ayurved viz. standardization of Ayurvedic drugs, clinical trial of mineral, herbal and herbomineral drugs mentioned in Ayurvedic texts, conceptual/empirical research on the basic principle like Tridosha Sidhanta, (Theory of three humours of body) Prakriti (theory of body constituent) etc are published in various journals. There are no certain or specific criteria in Ayurvedic texts to assess the strength and weakness of articles published in various national and international peer reviewed journal. Hence criterion based on modern science is considered for the appraisal of articles in Ayurvedic subject. Following discussion with some modification according to the research question may help the Ayurvedic

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research scholar to appraise their own and others articles before publishing. Appraisal scheme (strength and weakness of published qualified articles) How do we appraise? Appraisal can be done using common sense without the help of a statistical expert (Sarah Lawson et al., 2010). Validity and relevancy of various articles can be considered using different types of checklist. Anyone can assess and evaluate the quality of article using questionary of 10 questions in two sets, one is primary question set and other is main question set which will consider â&#x20AC;&#x201C; the results of research study, validity of review study and its help to everyone or local one? (Joan Durrance et al., 2009). These question sets are more or less applicable to evaluate the interventional and observational study. Screening question/Primary question 1. Aims of the research is mentioned by author or not? What Type of Research Question Does the Study Pose? (Jane M Young, Michael J Solomon, et al., 2009). Consider: the goal of the research, its important and its relevance. 2. Adoption of appropriate methodology. Consider: If the research seeks to interpret or illuminate the actions and/or subjective experiences of research participants. Detailed/Main questions 3. Research design is appropriate to address the aims of the research and is the study's research question relevant (Jane M Young, Michael J Solomon et al., 2009). Consider: If the researcher has justified the research design (e.g. have they discussed how they decided which method to use)? 4. Whether the recruitment strategy (as regards to sample size, sample design, inclusion and exclusion criteria etc.) was appropriate to the

aims of the research and did the study methods address and eliminates the key potential sources of bias? (Jane M Young, Michael J Solomon et al., 2009) Consider: if the researcher has explained how and why the participants were selected. If there are any discussions around recruitment (e.g. why some people chose not to take part). 5. Were the data collected in a way that addressed the research issue? Was the Study Performed in Line with the Original Protocol? (Jane M Young, Michael J Solomon et al., 2009) Consider: If the setting for data collection was justified. If it is clear how data were collected (e.g. focus group, semi-structured interview etc.).If the researcher has justified the methods chosen. If the researcher has made the methods explicit (e.g. for interview method, is there an indication of how interviews were conducted, or did they use a topic guide)? If methods were modified during the study. If so, has the researcher explained how and why? If the form of data is clear (e.g. tape recordings, video material, notes etc.) If the researcher has discussed saturation of data. 6. Has the relationship between researcher and participants been adequately considered? Consider: If the researcher critically examined their own role, potential bias and influence during: 1) Formulation of the research questions 2) Data collection, including sample recruitment and choice of location. How the researcher responded to events during the study and whether they considered the implications of any changes in the research design. 7. Have ethical issues been taken into consideration? Consider: If the research was explained in sufficient details to participants. If the researcher has discussed issues raised by the study (e.g. issues about informed consent, confidentiality and how they have handled the effects of the study on the participants during

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and after the study). If approval has been sought from the ethics committee.

considered other ways the research may be used (Joan Durrance, 2009).

8 Was the data analysis and statistical analysis is sufficiently rigorous and correct? Does the Study Test a Stated Hypothesis? (Jane M Young, Michael J Solomon et al., 2009)

Software for appraisals

Consider: if there is an in-depth description of the analysis process. If thematic analysis is used. If so, is it clear how the categories/themes were derived from the data? Whether the researcher explains how the data presented were selected from the original sample to demonstrate the analysis process. If sufficient data are presented to support the findings. To what extent contradictory data are taken into account. Whether the researcher critically examined their own role, potential bias and influence during analysis and selection of data for presentation. 9. Do the Data Justify the Conclusions? Is there a clear statement of findings? Consider: If the findings are explicit. If there is adequate discussion of the evidence (in favour and against the researcherâ&#x20AC;&#x2122;s arguments). If the researcher has discussed the credibility of their findings (e.g. triangulation, respondent validation, more than one analyst). If the findings are discussed in relation to the original research question. 10. How valuable is the research? Does it add anything new? (Jane M Young, Michael J Solomon et al., 2009) Consider: usefulness of research, if the researcher discusses the contribution the study makes to existing knowledge or understanding e.g. do they consider the findings in relation to current practice or policy, or relevant researchbased literature? If they identify new areas where research is necessary. If the researchers have discussed whether or how the findings can be transferred to other populations or

There are software tool which helps to create Critically Appraised Topics, or CATs, (CEBM oxford university) for the key articles you encounter about Therapy, Diagnosis, Prognosis, Aetiology/Harm and Systematic Reviews of Therapy (McAllister D, Wild S et al., 2009). This software may help the Ayurvedic researcher to appraise his or others article before publishing. Though in Ayurvedic fraternity, such software is not available but one can use it by making some modification with the help of software experts based on the concept of Ayurvedic science. CONCLUSION The information in this article can provide deeper understanding and appreciation for published qualitative research. The appraisals of published article using guidelines, questions, and explanations provided in this article are not intended to be all inclusive and may require some modification to answer and assess the research question. No study is perfect, nor does any study answer all questions, It is recommended that qualitative study and its value of evidence must read and assessed critically. It is concluded that rigorous criteria is available to evaluate the quantitative and qualitative research (Popay J., Rogers A. & Williams G. et al., 1998). Software is also available to assist the researcher to evaluate the strength and weakness of their own and published article. This software is designed for modern research which requires some modifications to meet the need of research question. The use of quantitative criteria to evaluate qualitative research may create the impression that the latter is not academically rigorous. Hence the evaluation criteria for qualitative and quantitative research should be different.

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REFERENCES CAT (critical appraisal tool) maker, centre for evidence based medicine, oxford university) http://www.cebm.net/?o=1216

McAllister D, Wild S. (2009) Why should clinicians understand epidemiology? Post Grad Med J 85:31, Med J Aust 189: 84–85

Dorothy Horsbugh, (2003) Evaluation of qualitative research, Journal of Clinical Nursing 12: 307–312

Mohsen Rezaeian, Department of Social Medicine, Rafsanjan Medical School, Rafsanjan University of Medical Sciences,Rafsanjan, How to teach medical students to critically appraise a published article in the public health domain Iran, http://www.jehp.net

Glasziou PP (2008), Information overloads: what's behind it, what's beyond it? Med J Aust 189: 84–85 Green BN, Johnson CD (2007) use of a modified journal club and letters to editors to teach critical appraisal skills. J Allied Health; 36:47–51 Jane M Young, Michael J Solomon, (2009) How to Critically Appraise an Article, Nat Clin Pract Gastroenterol Hepatol;6(2):82–91.

Popay J., Rogers A. & Williams G. (1998) Rationale and standards for the systematic review of qualitative literature in health services research. Qualitative Health Research 8(3), 341– 351. Sarah Lawson, (2010) Critical appraisal of qualitative research

Jeanfreau, S. & Jack, L. (2010) Qualitative research appraisal tool http://www.nccmt.ca/registry/view/eng/ 148.html

Tamoghna Biswas, (2011) Indian Role of journal clubs in undergraduate medical education, Journal of Community Medicine vol 36

Joan Durrance (2009) Creative appraisals skill program http://creativecommons.org/licenses/bync-sa/3.0/

Timm DF, Banks DE, McLarty J. (2012), Critical appraisal process: Step-by-step. South Med J; 105: 144–8. Wikipedia.org http://en.wikipedia.org/wiki/Critical_ap praisal

Source of Support: Nil

Conflict of Interest: None Declared

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Review article TRADITIONAL USE OF KAHU (LACTUCA SCARIOLA L.) - A REVIEW Arif Mohammad1* 1

HSZH Government Unani Medical College & Hospital (Barkatulla University), AYUSH Campus, Nehru Nagar Kolar Bypass Road, Bhopal-462003 *Corresponding Author: E mail: drarifmd@gmail.com

Received: 20/04/2013; Revised: 23/05/2013; Accepted: 27/05/2013

ABSTRACT Lactuca scariola Linn (prickly lettuce) is an important drug in Unani system of medicine. Tukhme kahu (Prickly lettuce seed) has traditionally been used for the treatment of headache, insomnia, nervousness, hypertension, palpitation, fever etc. Recently the discovery of active components from the plant and their biological function in disease control has led to active interest in the plant across the globe. It is easily available and cost effective drug which has drawn interest of many researchers and is screened for various bioactive substances. The present article explores the Unani classical and published scientific literature to compile the traditional and scientific data comprising pharmacognostic description, pharmacological studies, therapeutic uses, and safety profile of Lactuca scariola to date. The drug exhibits varied pharmacological activities such as sedative, hypnotic, diuretic, deobstruent, antipyretic, anti inflammatory, blood purifier, demulcent, refrigerant, anesthetic and antispasmodic, anti cancer, antibacterial, bronchodilator and vasorelaxant. The present review will provide comprehensive information on phytochemical and therapeutic uses with special reference to Unani medicine which will help to tap its unexplored potential with more scientific approach. KEY WORDS: Lactuca scariola, Tukhme Kahu, Unani medicine, Sedative.

Cite this article: Arif Mohammad (2013), TRADITIONAL USE OF KAHU (LACTUCA SCARIOLA) - A REVIEW, Global J Res. Med. Plants & Indigen. Med., Volume 2(6): 465â&#x20AC;&#x201C;474

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INTRODUCTION: Tukhme Kahu is the seed of Lactuca scariola Linn (Family - Asteraceae) which is commonly known as “Prickly Lettuce” seeds or “Wild Lettuce” seeds. L. virosa is a variety closely related to L. scariola. The species Lactuca sativa is the common or garden variety which is cultivated in many parts of India as a culinary vegetable (Nadkarni, K.M. 1954). The generic name Lactuca and the common name Lettuce derived from the Latin world lactus (milk), a milky fluid that flows from the stems when they break or are cut (Murray, J.A. 1983, Bunney, S.1992). It has been described by different closely related botanical names such as Lactuca sativa, L. capitata, L. virosa, L. bracteata and sativa wall. L. capitata, L. virosa (Nadkarni, K.M. 1954, Anonymous 1962, Watt, G. 1972, Khorey, R.N. et al.1985). Kahu plant and Tukhme Kahu has been used for a long time. According to Decandolle, 1895, it has been cultivated for more than 2000 years. Its medicinal properties were described by Hippocrates (430 BC). It was praised by Aristotle (356 BC). The species were described by Theophrastus (322 BC) and Dioscorides (60 AD). Galen (164 AD) gave the idea of general use. Arab physicians like Altabri (d. 861 CE) in “Firdausul Hikmat”, Razi (865-925 CE) in „Alhawi‟, Ibnesina (980-1031 CE) in Alqanoon and Majusi (d. 1028) in “Kamil-us-Sana” have described the pharmacological actions and uses of Tukhme Kahu in detail under the heading of Khas and Bazrul Khas (Kahu seeds). The common reported pharmacological actions are anxolytic, sedative, antipyretic, diuretic and analgesic which have been identified and proved by scientific studies. Apart from the reported pharmacological actions, Lactuca scariola also have Anti cancer, antibacterial, Spasmolytic, bronchodilator and vaso-relaxant Activities. Keeping in view the increase incidence of cancer, increase resistance of antibiotics and failure of management of hypertension & asthma which are the leading problem of the healthcare system, the present review was conducted which aimed to provide the current and ancient literary knowledge to the researchers of the related fields.

Unani classical literature and ethno medical literature on recent developments in research on Tukhme Kahu (Lactuca scariola) including original articles and papers were taken into study for the report. All the reports of phytochemical, pharmacological and clinical studies from animal and human model system were included in the review. Reported data was analyzed and represented in the form of table for the current review. Vernaculars Tukhme Kahu is popularly known in different dialect and languages as follows. Bazrul Khas (Arabic) Kahu, Salad Beej (Bengali) Prickly Lettuce, Wild Lettuce (English), Thridox (Greek), Guado, Lattuga (Italian), Kahu (Punjabi), Sallattu Virai (Tamil), Kavu Vitula (Telugu), Kahu ke beej (Hindi), In Persian and Urdu, the drug is commonly known as Tukhme Kahu (Nadkarni, K.M. 1954). Morphology Ibne Baitar (1197–1248 H) described in “Aljame-ul-Mufradat-ul-Advia wa Aghzia” with reference to Dioscorides (60 AD), Galen (121–200 AD), Ibne Masoya and Razi (865– 925 AD) as follows:– Kahu (Khas) actually has two varieties based on its occurrence. 1. Bustaani (Baaghi) “Lactuca Sativa” Garden Lettuce. It is a cultivated variety. 2. Barri (Jungali) “Lactuca Scariola” Wild Lettuce. Cultivated variety could be differentiated into two types. (1) This is 1½ meter high, soft, smooth and sweetish stem with pinnate wide leaves, delicately branched which has white flowers. Seeds are small and whitish in color. (2) The second harvested variety is English (Firangi) which is again of two types. Amongst which, one is well breakable and sweetish. Harvested Kahu is used as vegetable (Salad). Wild variety (Barri) of Lactuca Scariola has longer and thinner leaves than cultivated variety, the leaves are dark green and slightly bitter in taste. The latex is sometimes used as

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the substitute of opium which is not as potent as of Khash Khash (Papaver Somniferum) (Ghani, M.N.K. 1921, Ibne-Sina, 2007). Description: It is found as an erect glance sent Annual or biennial, leafy, 60–150 cm high somewhat prickly plant at the Western Himalaya from Marri to Kunawar, at altitude of 6000–11000 feet. It is also found in Western Tibbet at altitudes of 9000–12000 feet. Distributed to Siberia and West ward to the British Islets and canaries .Stem 3–10 cm high, sparingly prickly below, Capitula is 8–15 m long. Achenes have 6–8 mm body elliptical Setose at apex, 5–9 ribbed, grayish beak as long as body. Leaves are Pinnatifide or Lobed; achenes light colored, rarely sinuate, tending to turn edgewise into a vertical position. Flowers are Yellow, Achenes striate dark brown or grayish brown. Seeds are small whitish grey, about 1.0 cm long and 1–2 mm broad (Dymock, W. et al.,1890, Chopra, R.N. et al.,1956, Kirtikar, K.R et al.,1987). Unani description Seeds are whitish or whitish grey, shiny, elongated and smaller in size. They are tasteless or have light bitter taste. According to some Pharmacognostic experts, the seeds are light and soft. Seeds, Oil, milky fluid that flows from the stems (Lactucarium) and Dried Leaves are used as a drug in Unani system of medicine. The Mizaj (Temperament) of Tukhme Kahu is described as Cold 2º & Dry 3º. But there exists a difference of opinion regarding the gradation in mizaj. A majority of the authors described the mizaj as cold in 2nd degree and dry in 3rd degree (Ghani, M.N.K.1921, Kareem, N.A.1765, Gulam, H.1879, Ansari, A.B.H 1885, Haleem, M.A. 1948) while others consider it as cold and dry in 2nd degree (Said Mohd. 1973, Hakeem, M.A. 1953, Ibne-Sina 2007). Phytochemical Studies The plant contains alkaloids 0.02 %, sugar and glycosides 6.5 %, volatile oil in traces; fat 2.2 %, Gums 2.16 %; Organic acids 1.06 %, Carotene 16 mg %, Vitamin B1 22%, Vitamin

C 44 mg %, Vitamin E 32 and Vitamin K 0.2 mg %.The phytochemical investigations of seeds revealed the presence of alkaloids, the bitter substance lettuce, oxalic acid, lactucopicrin (S. R. Baquar, 1989) and sesquiterpene esters (J. Alberto et al., 1992).The alkaloid, lactucin, isolated from the seeds by Dolejs et al., (1958) and Michal et al., (1958), exhibited antipyretic activity (V. S. Agarwal, 1997) and a triterpenoid saponin isolated from stem possesses antibacterial activity (R. N. Yadava et al., 2008). Pharmacological studies The methanolic extract of Lactuca scariola was found to possess spasmogenic, spasmolytic, bronchodilator, and vasorelaxant activities. The spasmogenic activity may be attributed to some cholinergic constituents, whereas spasmolytic effect may be due to Ca++ channel blocking components that may cause relaxation of gastrointestinal, tracheal, and aortic smooth muscles (Khalid hussain janbaz et al., 2013). L. scariola exhibited dose dependent potent analgesic activity. Methanolic extract of L. scariola can produce significant analgesic activity but failed to show antiinflammatory effect (Fayyaz ahmad et al., 1992). The methanol extracts prepared from leaves and stems of L. scariola showed cytotoxic activity against A549, HePG, MCF7 and HCT116 (Eman Elsharkawy1 et al., 2013). A new triterpenoid saponin has been isolated from the seeds of Lactuca scariola. This compound shows antimicrobial activity against various bacteria and fungi (R. N. Yadava et al., 2008). The antioxidant activity of Lactuca scariola (Asteraceae) was investigated by measuring the radical scavenging effect on DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical and found that the methanolic extract of the aerial parts of Lactuca scariola showed strong radical scavenging activity (D. K. Kim, 2001). Lactucin and its derivatives lactucopicrin and 11beta, 13-dihydrolactucin, which are characteristic bitter sesquiterpene lactones of Lactuca virosa were evaluated for analgesic and sedative properties in mice. Lactucopicrin appeared to be the most potent analgesic of the three tested compounds.

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Lactucin and lactucopicrin, but not 11beta, 13dihydrolactucin, also showed sedative properties in the spontaneous locomotor activity test (Wesolowska A. et al., 2006). A Randomized placebo controlled double-blind trial of Tukhme Kahu (seeds of Lactuca scariola Linn.) on mixed anxiety depressive disorder has shown a significant effect in reducing anxiety and depressive symptoms (Ghazala javed, et al., 2009). The oil of Tukhm-e-Kahu (Lactuca scariola seeds) is used in Industries i.e. soap-making, paints and varnish (Anonymous, 1962; Ambasta, S.P. 1986). These studies may provide a scientific basis to validate the traditional use of Tukhme Kahu in the management of some gastrointestinal, respiratory, neurological and vasospastic ailments. The following table (Table -1) shows the Pharmacological studies mentioning the Reported action with references.

AFAL-O-KHAWAS actions)

(Pharmacological

The eminent Unani physician has described the Afal-o-khawas (Pharmacological Actions) in detail. It is commonly used as Musakkin (analgesic), Munawwim (sedative), Mudir baul (diuretic) and refrigerant. The following table (Table -2) shows the Afal-o-khawas (Pharmacological Actions) with Unani and ethno-medical references. Therapeutic uses According to ancient Unani text, Tukhme kahu and its compound formulations i.e. roghan Kahu & roghan laboob saba,Mufarreh shaikur rais etc has been recommended for the treatment of headache, insomnia, nervousness, fever, palpitation, burning micturition etc. The following table (Table -3) shows the therapeutic uses in different ailments with Unani and ethno-medical literature references.

Table- 1: Pharmacological studies with references S.no.

Components used

Reported action

L. sativa L. scariola (whole plant extract)

Lactuca scariola (seeds extract)

Lactuca virosa (isolated biologically)

Lactuca sativa (Seeds oil)

Triterpenoid saponin, isolated from the stems of L.scariola Methanolic extract of theaerial parts of Lactuca scariola Lactucin & lactucopicrin

7. 8. 9.

Methanol scariola Methanolic

extract extract

of L. of

CNS stimulant and depressant action respectively in mice LD50 = 750 mg/kg, IP (Dhawan, B.N., et al., 1977.) Anti-arrhythmic and cardiac function hemodynamic effect LD50 = 79.05 g/kg, IP (Weizhi, W, et al., 1992) Spontaneous locomotor activity and analgesic effect active crude preparation P-1, P-2, P-3 and Lactucin) (Gromek, D et al., 1992) Sedative effects in loco-motor activity, potentiation of the hypnotic effect of barbiturates, analgesic effect of barbiturates,( Said Mohd. 1973) Antibacterial triterpenoid saponin (R. N. Yadava et al., 2008) antioxidant activity (D. K. Kim, 2001)

Analgesic and sedative activities in mice (Wesolowska A. et al., 2006) Spasmolytic, bronchodilator, and vasorelaxant Activities in rabbits (Khalid et al., 2013) potent analgesic activity in mice (Fayyaz ahmad et

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10. 11. 12.

scariola Methanolic extracts of leaves and stems of L. scariola The extract of Tukhm-ekahu

al.,, 1992) Anticancer activity (Eman Elsharkawy1 et al., 2013)

Tukhm-e-kahu scariola)

Anxiolytic & antidepressant (Ghazala javed, et al., 2009)

(Lactuca

Diuretic, sedative (Ansari A.N, 2009)

Table 2: Afal-o-khawas (Pharmacological Actions) with references S.no .

Afal-o-khawas (pharmacologi cal actions)

Reference

Unani Literature

Ethno-Medical Literature

Hypnotic

(Ghani, 1921, Kareem,1765, Gulam, 1879, Ansari, 1885, Haleem, 1948, Ibne-Baitar (1197-1248 AD), Antaki, (1597), Attar (1888), Azam, (1895), Said (1969), Abid, (1907)

Nadkarni 1954, Anonymous 1962, Dymock (1890), Bunney 1992, Watt, G. 1972, Khorey, 1985, Chopra 1956, Said, H.M. 1969, Ambasta 1986.

Sedative

Kareem, 1765; Azam, 1895, Kabir, Nadkarni, 1954; Waren, 1956, 1951, Ibne-Sina 2007 Chopra, 1956; Watt, 1972 Mair,1973; Khory, 1985; Bunney 1992;

Antipyretic

Kabiruddin, H. (1951), Ibne-Sina Anonymous (1962), Khorey, 1985, (2007) Chopra 1956, Kirtikar, 1987, Said, 1969, mbasta, 1986,

Diuretic

Ghani, 1921; Hakeem, 1953; Nadkarni, 1954, Chopra, Fazlullah, 1970. Ibne-Sina 2007. Anonymous, 1962, Watt, Murray, 1983; Khory, 1985

1956; 1972;

Expectorant

Kareem, 1765; Ahmad, Ghani,1921; Hakeem, 1953;

1956;

Blood purifier

Kabir, 1951; Fazlullah 1970

Aphrodisiac by Antaki, 1597; Kareem, 1765; Ansari, decreasing 1885; Attar, 1888; ; Abid, 1907; anxiety level Ghani, 1921; Ibne-e-Sina, 2007,

Anesthetic

Antidote

1887; Nadkarni, 1954; Khory, 1985; Dymock, et al..1890

Kareem, 1765; Gulam, 1879; Ansari, 1885; Ghani, 1921; Haleem, 1948; Hakeem, 1953. Antaki,

1597;

Kareem,

1765;

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Chopra

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Attar,1888; Ghani, 1921; 10.

Anodyne

Nadkarni,1954; Charles. Murry, 1983; Khory, 1985;

11.

Anti inflammatory

12.

Anti bacterial

R. N. Yadava et al.. 2008.

13.

Anti spasmodic

Watt,1972; Chopra,1956;

14.

Cooling

15.

Demulcent

16.

Deobstruent

17.

Diaphoretic

18.

Hair tonic

19.

Purgative

Watt, 1972; Murray, 1983

20.

Soothing

Watt, 1972

21.

Emmenogogue

Antaki, 1597; Ibn-e-Sina, 2007;

Kareem, N.A. (1765); Azam, M.K. Dymock,1890; Nadkarni, (1895) Ibne-Sina (2007) Chopra, 1956; Khory, 1985; -

Nadkarni, 1954; Chopra, Khory, 1985; Kiritikar 1987.

Ghani, 1921; Fazlullah, 1970; -

Azam,1895; 1933

Kabir,1951;

1974;

1954;

1956;

Chopra, 1956; Watt, 1972; Charles, 1974; Murray, 1983

Hakeem

Kareem, N.A. 1765; Ibne-Sina 2007;

Table 3: therapeutic uses with references S.no.

Therapeutic uses

Reference (Unani & Ethno-Medical Literature)

Insomnia

Antaki, 1597; Kareem, 1765; Attar, 1888; Ibne Baitar, 1197-1248 Ad; Azam,1895; Ghani, 1921; Nadkarni, 1954;Anonymous, 1962;

Headache

Fever

Ibne Baitar, 1197â&#x20AC;&#x201C;1248 AD; Antaki, 1597; Kareem, 1765;Gulam, 1879; Ansari, 1885; Ghani, 1921; Haleem, 1948; Hakeem, 1953; Kirtikar & Basu, 1987. Kabir, 1951; Nadkarni, 1954; Said, 1969; Khory, 1985)

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Nervousness

Nadkarni, 1954; Khory, 1985; Ghazala,2009;

Palpitation

Nadkarni, 1954; Khory, 1985.

Asthma

Nadkarni, 1954; Chopra, 1956; Watt, 1972; Murray, 1983;

Chronic bronchitis

(Nadkarni, 1954; Chopra, 1956; Watt, 1972; Kirtikar &

Chest pain

Kareem, 1765;

Acute inflammation

(Antaki, 1597; Ibne-e-Sina, 1930; Nadkarni, 1954; Khory, 1985; Kirtikar & Basu, 1987)

10.

Burning micturition

(Kareem, 1965; Gulam, 1889; Ansari, 1885; Ghani,1921.

11.

Acute Coryza

12.

Dropsy

Murry, 1983;

13.

Nocturnal emission

Antaki, 1597; Ansari, 1885; Attar, 1888; Ghani, 1921; Ibne Sina, 2007; Nadkarni, 1954; Murry, 1983.

14.

Painful ulcer

15.

Pertusis

Nadkarni, 1954; Chopra, 1956; Watt, 1972; Murray, 1983.

16.

Jaundice

Ibne Sina, 2007.

17.

Prevent hair fall

Kabir, 1951; Anonymous, 1962; Kirtikar & Basu, 1987.

18.

Relief of thirst

Azam,1895; Ghani,1921; Hakeem,1953; Ibne Sina, 2007.

19.

Scorpion sting

Antaki,1597; Kareem,1765;Attar,1888;Ghani, 1921; Ibne Sina, 2007

20.

Spermatorrhoea

Antaki, 1597; Ansari, 1885; Attar, 1888; Ghani, 1921; Ibne Sina, 2007; Nadkarni, 1954; Murray, 1983.

21.

Sunstroke

Antaki, 1597, Ibne Sina, 2007

22.

Dysmenorrhoea

Kareem, 1765; Ibne-e-Sina, 2007; Kirtikar & Basu, 1987; Murry, 1983.

Basu,1987; Ibn Baitar, 1197-1248 AD)

cold/ (Kareem, 1765; Ghani, 1921; Hakeem, 1953;

irritable Nadkarni, 1954; Ambasta, 1986; Kirtikar Basu, 1987; Ibne Sina, 2007.

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Substitute: The Majority of authors considered Khashkhash (Poppy seeds, Papaver somniferum) as substitute of Tukhm-e-Kahu (Lactuca scariola seeds). (Kareem, 1765; Ghani, 1921; Hakeem, 1953; Fazulullah, 1970; Ibne Sina, 2007). While others considered Dammul Akhwain (Dracaena cinnabin) as its substitute (Ansari, 1885; Kareem, 1765; Ibne Sina, 2007) Dose: Tukhme Kahu is prescribed in adult dose of 6–12 gms daily orally (Ghani, 1921; Hakeem 1953). Another reported dose is 3–5 gm (Ansari, 1888; Kabir, 1951). Preparation: Tukhme Kahu is used in form of Safoof (powder), Joshanda (decoction), single or in combination with other drugs. It‟s oil used as Zamad (Liniment) topically. Harmful Effects: The herbal drugs may pose harmful affects either in the form of adverse reactions or drugdrug interactions due to numerous phytoconstituents present in each part of a plant. However no serious health risk is noted till date but some authors have reported that continuous use of Tukhme Kahu in high doses can cause atony of body muscles, Dementia, Amnesia, Loss of vision and sexual debility (Antaki, 1597; Kareem, 1765; Abid, 1907; Ghani, 1921; Aziz, 1942; Hakeem, 1953; Ibne Sina, 2007). Correctives:

(Kareem, 1765; Ansari, 1885; Haleem, 1948; Hakeem, 1953, Ibne Sina, 2007). CONCLUSION Tukhme Kahu is an important drug in Unani system of medicine used for ages in the treatment of headache, insomnia, nervousness, hypertension, palpitation, fever, Asthma, Chronic bronchitis, acute cold/Coryza, Scorpion sting etc. Lactuca scariola has its own importance as it has Anti cancer antibacterial, antifungal, Spasmolytic, bronchodilator and vasorelaxant activities. The scientific analysis of Tukhme Kahu proves many of the activities mentioned in Unani classical literature. Further investigations are needed to find out the mechanism of action, active principle(s) and utility of Tukhme Kahu in clinical practice. Though the drug has been found to be safe but the potent curative effects of the drug need to be verified by more controlled and exhaustive clinical trials, especially in the field of cancer and hypertension so that it can be established as a standard drug. ACKNOWLEDGEMENT The author is grateful to Dr Zaki anwar ansari, Principal, HSZH Govt. Unani medical college, Bhopal (MP) for his cooperation in the work. The technical help provided by Mr. Javed ahmed, library attendant, A & U Tibbia college (University of Delhi) Karol Bagh New Delhi-5 and Mrs. Salma, librarian HSZH Govt. Unani medical college, Bhopal (MP) are also acknowledged.

Mastagi (Pistacia lentiscus) and Honey are reported as correctives of Tukhme Kahu

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Ambasta, S.P. (1986): “The Useful Plants of India”, CSIR, New Delhi, 312. Anonymous (1962); The Wealth of India, Vol. VI, CSIR, New Delhi, 12–16.

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Ansari A.N (2009) “Clinical evaluation of a Unani herbal formulation” Hamdard medicus 2009 Vol. 52 No. 3 51–57 Ansari,

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Antaki, D.Z. (1597);Tazkirah-e-Ulil Albad (Arabic)\, Azharia Press Egypt, 87. Attar, Z. (1888);Ikhtiyarat-i-Badiya (Persian), Matba Munshi Nawal Kishore, Lucknow, 156. Azam, M.K. (1895); Muheet-e-Azam (Persian), Vol. I, Matba Nizami, Kanpur, 196. Bunney, S.( 1992); The Illustrated Encyclopaedia of HERB‟S.Chancellor Press, London, 176. Charles, F. Millspaugh (1974); American Medicine, Dover Publications Inc., New York. Chopra, R.N. and Nayer, S.L. (1956); Glossary of Indian Medicinal Plants. SCIR, New Delhi, 148. D. K. Kim, (2001); “Antioxidative components from the aerial parts of Lactuca scariola L,” Archives of Pharmacal Research, vol. 24, no. 5, 427–430. Dhawan, B.N., Patnaik, G.K., and Rastogi, R.P. (1977); “Screening of Indian Plants of Biological Action Part-VI”, Indian J. of Exp. Bio., 15, 208. Dymock, W., Warden, C.J.H. and Hooper, D. (1890); Pharamacographia Indica : Vol. I, II and III; reprinted by the Institute of Health and Tibbi Research, Pakistan, Hamdard : XV (1-22) (edited by M. Saeed in 1972), Hamdard National Foundation, Pakistan, 13–14.242,. Eman Elsharkawy1 and Mona Alshathly (2013);“Anticancer activity of lactuca steriolla growing under dry desert condition of Northern Region in Saudi Arabia” Journal of Natural Sciences Research Vol.3, No.2. Fayyaz Ahmad, Rafeeq A. Khan, Shahid Rasheed (1992); “study of analgesic and anti inflammatory activity from plant

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Kareem, N.A. (1765); Makhazanul-Advia (Urdu), Vol. I, Munshi Nawal Kishore, Kanpur, 313.

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Source of Support: Nil

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Watt, G. (1972); Dictionary of the Economic Products of India, Vol. I, Cosmo Publication, Delhi, 3-4,578-579.Weizhi, W., Hauzhau, W., Zhimin, Q.I., Zhijik, X., and Rixin, L. (1992): “AntiArrhythmic Action of Seeds of Lactuca Sativa and its Effects on Cardiac Function Hemodynamics”, Chinese Traditional and Herbal Drugs, 23(7), 366–368. Wesołowska A, Nikiforuk A, Michalska K, Kisiel W, Chojnacka-Wójcik E, (2006) “analgesic and sedative activities of lactucin and some lactucin-like guaianolides in mice” jethnopharmacol. sep 19;107 (2):254–8.

Conflict of Interest: None Declared

Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||

Call for Papers â&#x20AC;&#x201C; Vol. 2, Issue 8, August 2013 Submit your manuscripts (Research articles, Review articles, Short Communications, Letters to the Editor, Book Reviews) to Global Journal of Research on Medicinal plants & Indigenous medicine â&#x20AC;&#x201C; GJRMI Submit it online through www.gjrmi.com or mail it to submitarticle@gjrmi.com on or before July 10th 2013.

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