Asian Pacific Journal of Tropical Biomedicine

1-6

Authors: MD Nazneen Bobby, Wesely EG, Johnson M

Objective: To establish alkaloids profile for the medicinally important plant Albizia lebbeck (A. lebbeck) using high performance thin layer chromatography (HPTLC). Methods: Preliminary phytochemical screening was carried out by Harborne method. HPTLC studies were followed by Harborne and Wagner et al method. Ethyl acetate-methanol-water (100:13.5:10) was employed as mobile phase for alkaloids. The developed plate was sprayed with Dragendorff’s reagent followed by 10% sodium nitrite reagent as spray reagent and dried at 100 ℃ in hot air oven for 3 min. The plate was photo-documented at UV 366 nm and daylight using photo-documentation chamber. Results: Alkaloids, steroids, terpenoids, flavonoids, saponins and glycosides present in the petroleum ether, ethyl acetate and methanolic extracts of A. lebbeck leaves. The petroleum ether extracts of A. lebbeck leaves displayed the presence of 10 types of alkaloids with 10 different Rf values ranging from 0.02 to 0.85. The ethyl acetate extract of A. lebbeck leaves illustrated the presence of 5 different types of alkaloids with 5 different Rf values ranging from 0.09 to 0.84. The methanolic extract of A. lebbeck leaves demonstrated the presence of 4 different types of alkaloids with 4 different Rf values with range from 0.02 to 0.79. Maximum number (10) of alkaloids has been observed in petroleum ether followed by ethyl acetate (5) extracts of the leaves of A. lebbeck. Conclusions: In the present study we observed various alkaloids profile of the medicinally important plant using HPTLC. This profile can be used for the identification of the medicinally important plants from the adulterant. Further, separation and characterization of the bioactive compound (principles) from the plants is to be evaluated and reported in near future.

12-14

Authors: Sharad Srivastava, Adarsh Pratap Singh, Ajay Kumar Singh Rawat

Objective: To develop a simple precise and novel method for the identification and quantification of bioactive molecules ferulic acid in Lycopodium clavatum. Methods: Bioactive molecules ferulic acid in Lycopodium clavatum was indentified and estimated by the assay combined separation and quantitative estimation of the analyte on silica gel 60F254 HPTLC plates with visualization under UV and scanning at 320 nm. Results: HPTLC studies showed the separation and determination of ferulic acid (0.443%) in Lycopodium clavatum. The result of recovery ranged in 95%-97% for ferulic acid. Conclusions: This method is simple, sensitive, economic, novel and first time reported from this species. Therefore, ferulic acid might be a useful chemotaxonomic marker for the genus Lycopodium and other lycopods.

21-26

Authors: M. Johnson

Objective: To analyze the similarity and variation among different populations of Ocimum sanctum (O. sanctum) by isoperoxidase, isoesterase, acid phosphatase and alkaline phosphatase profile. Methods: The isolation, separations and staining of the enzymes isoperoxidase, isoesterase, acid phosphatase and alkaline phosphatase were performed as described by Sadasivam and Manickam. Results: In isoperoxidase system, the band PRX4 1 (0.36) was present only in the accession collected from Salem, similar to that band PRX4 2 showed its presence only in the accession collected from Kolli Hills and PRX5 1 (0.42) was expressed only in Wyanad accession. The accession collected from Tirunelveli showed its unique banding profile in region ACP2 3 with 0.18 molecular weight-Rf values in the acid phospahatase system. In esterase system, EST5 1 and 8 1 were observed only in the accession collected from Tenkasi. In the alkaline phosphatase enzyme system, only one region (AKP6) of activity with single band (AKP6 1 -0.51) was observed. In this enzyme system, all the selected accession showed their expression. Highest percentage (0.666 7) of similarity was observed between the accessions collected from Tirunelveli and Kolli Hills, Nagercoil and Kolli Hills and Salem and Andaman while the highest percentage (0.800 0) of variation was observed between the accessions collected from Salem and Andaman. The isozyme profile based cladogram showed two clusters, cluster 1 (C1) included the accessions collected from Tenkasi, Tirunelveli, Nagercoil and Kolli Hills and they showed highest percentage of divergence with the other three accessions. The cluster 2 (C2) included the accessions collected from Wyanad, Salem and Andaman. Conclusions: Unique banding profiles of esterase, peroxidase, acid phosphatase and alkaline phospahatase were observed in O. sanctum, which represent the fingerprint of medicinally important plant. Such finger printing is useful in differentiating the species and act as biochemical markers for these species in plant systematic studies in the near future.

30-33

Authors: Solomon Jeeva, Johnson Marimuthu @ Antonisamy, Cosman Domettila, Babu Anantham, Mony Mahesh

Objective: To explore the phytochemical constituents of Ulva reticulata (U. reticulata) and Sargassum wightii (S. wightii). Methods: The preliminary phytochemical screening was performed by Harborne method. Results: The results of the phytochemical screening revealed the presence of steroids, phenolic groups, saponins, tannin, flavonoids, carbohydrates, coumarins, and xantoproteins in the extracts of U. reticulata and steroids, phenolic groups, saponins, tannin, flavonoids, carbohydrates, carboxylic acid, coumarins, and xantoproteins were detected in the extracts of S. wightii. Conclusions: The solvent extracts of U. reticulata and S. wightii show a number of metabolites presence, further work will emphasize the isolation and characterization of active principles responsible for bio-efficacy and bioactivity.

40-45

Authors: Muraleedharannair Jalajakumari Mithraja, Johnson Marimuthu @ Antonisamy, Mony Mahesh， Zachariah Miller Paul, Solomon Jeeva

Objective: To examine the phyto-constituents of Adiantum caudataum (A. caudataum), Adiantum latifolium (A. latifolium), Adiantum lunulatum (A. lunulatum), Christella dentata (C. dentate) and Christella parasitica (C. parasitica), to provide chemical marker and find inter- specific variation between the medicinally important genuses. Methods: The dried and powered leaves materials (50 g) were extracted successively with 250 mL of petroleum ether, ethyl acetate, methanol, chloroform, acetone, benzene and water by using Soxhlet extractor for 8 h at a temperature not exceeding the boiling point of the solvent. The aqueous extracts were filtered using Whattman filter paper (No.1) and then concentrated in vacuum at 40°C using Rotary evaporator. The residues obtained were stored in a freezer -70 °C until further tests. Phytochemical screening of the extracts was carried out according to the standard methods. Results: A total of five plants and 30 extracts were examined for the phytochemical screening. The crude extracts of A. caudataum, A. latifolium, A. lunulatum, C. dentata and C. parasitica showed varied degree of phyto-constituents with reference to solvents of the plant extracts. A total of three plants and 18 extracts were examined for the phytochemical screening A. caudataum, A. latifolium, A. lunulatum. The steroid is totally absent in the tested extracts of A. lunulatum. Similar to that caoboxylic acid also absent in A. caudatum and Coumarin showed its presence only in A. caudatum. A total of two plants and twelve extracts were examined for the phytochemical properties of Christella. Of which phenol is present in 11 extracts tested. Steroid is present only in one extract of C. parasitica and coumarin is present only in C. parasitica five extracts. Conclusions: The present study result provids a simple and cheap tool (Chemical marker) for the characterization and identification of the medicinally important taxa. With the help the phytochemical screenings we can find inter- specific variation between the medicinally important genuses and easily we can differentiate the Adiantum and Christella genuses.

50-56

Authors: J.K. Patra, S. Gouda, S.K. Sahoo, H.N. Thatoi

Objective: To separate the bioactive compounds from crude methanol extract of leaf of Excoecaria agallocha (E. agallocha), a mangrove plant from Bhitarkanika, Odisha, India. Methods: Chromatographic separation was carried out by paper, thin layer chromatography (TLC) and column chromatographic techniques. Bioautography of the column fractionated extract and TLC chromatogram were evaluated in vitro for antibacterial and DPPH free radical scavenging activity. The HPLC and 1 H NMR spectroscopy of the extract was also carried out to detect the presence of compounds and possible functional groups in the crude plant extract. Results: Evaluation of solvent system for chromatographic separation revealed that n-butanol: acetic acid: water in the ratio of 4:1:1 was the most appropriate for the separation of phenolic compounds in the crude extract. The antibacterial bioautography screening of the TLC fractionated extract of E. agallocha showed positive activity against Staphylococcus aureus (MTCC-1141) and Pseudomonas aeruginosa (MTCC 1034). Evaluation of antioxidant properties of TLC fraction of E. agallocha showed 23.36% scavenging activity using DPPH assay at 40 µL concentration. HPLC chromatogram of the plant extract produced seven peaks for the leaf and two peaks for the stem/bark with the highest peaks at 2.563 and 2.493 minutes of retention time, respectively. The 1 H NMR spectroscopic analysis of the plant extract revealed the presence of acyclic aliphatic and α- mono substituted aliphatic group of compounds in the sample. Conclusions: Thus, it is apparent from the present study that methanol crude extract of E. agallocha is rich in aliphatic group of compounds which can be purified and characterized for possible pharmaceutical utilization.

64-66

Authors: Selvamony Sukumaran, Mony Mahesh, Solomon Jeeva

Objective: To identify the phytoconstituents present in the fronds of Tectaria zeylanica. Methods: Screening of phytochemicals was done by using the standard methods given by Harborne. Results: Phytochemical screening of the fronds of Tectaria zeylanica confirmed the presence of various derivatives such as phenols, saponins, steroids, tannins, xanthoproteins, coumarins and carbohydrates. Conclusions: The present observations suggest a possibility to establish high yielding plantlets by in vitro culture for the production of medicinally important bioactive compounds against human pathogenic microorganisms.

74-78

Authors: R. Subramanian, S. Gayathri, C. Rathnavel, V. Raj

Objective: To determine sodium (Na), iron (Fe), magnesium (Mg), manganese (Mn), lead (Pb), zinc (Zn), cadmium (Cd) and copper (Cu) in some medicinal plants regularly used in cooking in Indian curries. Methods: All the nutrients were determined by using atomic absorption spectrophotometer (AAS). Results: The average concentrations detected were ranged from 150.22 to 521.98, 782.42 to 813.84, 15.75 to 532.72, 85.72 to 101.50, 0.478 to 9.890, 0.684 to 2.751, and 11.51 to 94.05, 6.94 to 49.76 mg/kg for Na, Fe, Mg, Mn, Pb, Zn, Cd and Cu, respectively. Conclusions: The mineral and metal contents in the samples were found at different levels. Therefore, these medicinal plants are rich in some essential minerals, especially Fe and Mg which are essential for human health.

83-89

Authors: PN Ansil, SP Prabha, A Nitha, PJ Wills, V Jazaira, MS Latha

Objective: To evaluate the curative effect of the methanolic extract of Amorphophallus campanulatus (A. campanulatus) tuber (ACME) against thioacetamide induced oxidative stress in rats. Methods: The curative potential of two different doses of ACME (125 and 250 mg/kg; p.o.) was evaluated against thioacetamide (TAA) induced oxidative stress in male Wistar rats. Single dose of TAA (100 mg/kg; s.c.) was administered to the rats in all groups except the normal control. In treatment groups, rats were administered with silymarin-the standard drug (100 mg/kg; p.o.) and ACME 2, 24 and 48 h after TAA intoxication. Hepatotoxicity was assessed by quantifying the serum levels of AST, ALT, ALP and LDH. The antioxidant potential of ACME were also evaluated by the estimation of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST), reduced glutathione (GSH) and lipid peroxidation (Thiobarbituric Acid Reactive Substances-TBARS) in hepatic and renal tissues. In addition, the liver sections were also evaluated for its histopathologic changes. Results: ACME significantly (P<0.05) reversed the elevation of serum AST, ALT, ALP, LDH and tissue malondialdehyde levels. Hepatic and renal GSH, GST, GR, GPx and catalase levels were remarkably increased by the treatment with the extract. Quantification of histopathological changes also supported the dose dependent curative effects of ACME. Conclusion: This study demonstrates the antioxidant and hepatoprotective activity of ACME and thus scientifically supports the use of this tuber in traditional medicine for the treatment of liver disorders.

99-103

Authors: Alak Kanti Dutta, Partha Sarathi Gope, Subrata Banik, Sukh Makhnoon, Muhammad Ali Siddiquee, Yearul Kabir

Objective: To study total phenolic content and antioxidant properties of 80% methanol extracts of ten high yielding rice varieties, five each from two different seasons namely aman and boro of Bangladesh. Methods: Total phenolic content was measured by Folin-Ciocalteau method while 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, hydroxyl ion scavenging, ferric reducing antioxidant power (Ferric reducing antioxidant power), and total antioxidant capacity (TAC) by ammonium molybdate, were used to analyze their Antioxidant properties. Results: Rice variaty BR5 of aman and BRRI dhan28 of boro season comparatively showed higher TPC and Antioxidant properties than the other rice varieties. BR22 of aman season showed the highest hydroxyl ion scavenging activity although it displayed the lowest TPC. Except for hydroxyl ion scavenging activity, aman rice varieties displayed comparatively higher total phenolic content and antioxidant property than the boro rice varieties. Conclusions: The results of the present study implies that the selected rice varieties possess moderate antioxidant capacity and therefore, can be considered as health supplements and nutraceuticals foods as rice is the staple food of Bangladesh.

112-115

Authors: S. Sundarammal, R.Thirugnanasampandan, M. Tamil Selvi

Objective: Orthosiphon thymiflorus (Roth) Sleesen (O. thymiflorus) is an aromatic plant widely distributed in the Western Ghats of South India. The present study investigates the chemical composition and antioxidant activities of O. thymiflorus essential oil isolated from the leaves. Methods: The essential oil was hydrodistilled from the leaves by using Clevenger apparatus. Isolated essential oil was chemically analyzed by gas chromatography (GC) and gas chromatography- mass spectrometry (GC-MS). In vitro antioxidant activity of the isolated essential oil was determined. Results: Chemical composition analysis of essential oil reported to the presence of thirty three compounds. The major compound was 2-isopropyl-5-methyl-9-methylene-bicyclo-1-decene (4.4.0) (42.62%) followed by carotol (16.48%), α -cadinol (6.14%) and δ-3-carene (5.15%). In vitro antioxidant activity of the isolated oil showed dose dependent free radical scavenging activity against DPPH (IC50 437.12±0.02 µg/mL) and hydroxyl radicals (IC50317.88依0.02µg/mL). Metal chelating and prevention of deoxyribose degradation activities of the oil was observed with the IC50 of 339.78±0.01µg/mL and 158.33±0.02µg/mL respectively. Antioxidant activity of the oil was low when compared with the positive controls such as BHT and ascorbic acid. Conclusion: The present study shows various chemical components present in the O. thymiflorus essential oil and with it’s significant antioxidant activity thus has great potential to be used as natural food supplement.

120-126

Authors: Prabhjit Kaur, Manish Kumar, Bikram Singh, Subodh Kumar, Satwinderjeet Kaur

Objective: To evaluate antigenotoxic, antioxidant and COX-2 inhibitory potential of ‘GG-1’ fraction isolated by Medium Pressure Liquid Chromatography (MPLC) from ethyl acetate extract of Glycyrrhiza glabra (G. glabra) L rhizomes. Methods: The antigenotoxic activity was tested in human blood lymphocytes using comet assay and SOS chromotest using PQ37 strain of Escherichia coli (E. coli) against the diagnostic mutagens viz. 4-NQO and H2O2. Antioxidant activity was determined by DPPH radical scavenging assay. It was also investigated for the COX-2 inhibitory activity using Cayman COX (ovine) inhibitory screening assay. Results: The spectroscopic data of ‘GG-1’ fraction revealed it to be 7-hydroxycoumarin (Umbelliferone). In SOS chromotest, at the concentration of 616.75 µM, umbelliferone exhibited moderate response by reducing the induction factor of hydrogen peroxide by 68.99% (IC50 223.44 µM) and that of 4NQO by 59.71% (IC50 280.74 µM). In comet assay, exhibited a good activity by inhibiting the genotoxicity of both hydrogen peroxide and 4NQO by 61.64% (IC50 330.02 µM) and 50.66% (IC50 577.83 µM) respectively. The umbelliferone exhibited free radical scavenging potential of 43.88% at 616.75 µM. Umbelliferone possessed 95.68% inhibition of COX-2 at 10 µM concentration with IC50<1. Conclusions: The antigenotoxic, antioxidant and COX-2 inhibitory properties showed by the umbelliferone suggests that it may have several applications in nutraceuticals and human health care.

131-135

Authors: Muraleedharannair Jalajakumari Mithraja, Varaprasadham Irudayaraj, Solomon Kiruba, Solomon Jeeva

Objective: To make scientific validation of the ethnomedicinal fern Drynaria quercifolia (L.) J. Smith in relation to urinary disorders. Methods: Antibacterial study was carried out on clinically isolated Urinary Tract Infecting (UTI) bacteria by disc diffusion method. Results: Among the six extracts tested against eight different UTI bacteria, acetone extract was effective against Enterococcus faecalis and Streptococcus pyogenes, while ethanol extract was effective against Pseudomonas aeruginosa. Preliminary phytochemical screening shows the presence of phytoconstituents viz. phenols, tannin, alkaloids, proteins, xanthoproteins, carboxylic acid, coumarins and saponins. Conclusions: Based on the present study, along with previous studies, the ethnomedicinal use of Drynaria quercifolia for the treatment of urinary infection has been scientifically validated.

140-146

Authors: Xavier Devanya Rosaline, Shanmugavel Sakthivelkumar, Kuppu Rajendran, Sundaram Janarthanan

Objective: To screen the antibacterial efficacy of various solvent extracts of marine algae such as Sargassum wightii (S. wightii), Chaetomorpha linum (C. linum) and Padina gymnospora (P. gymnospora) against some selected gram-positive and gram-negative human pathogenic bacteria. Methods: Crude extracts were prepared from the selected marine algae using different solvents namely, hexane, ethyl acetate, acetone and methanol and were tested for their antibacterial activity against human pathogenic bacteria using disc diffusion method. Minimum inhibitory concentration (MIC) was also performed for selected solvent extracts for all the bacterial species. A suitable positive control was also maintained. Results: Among the three marine algae screened P. gymnospora and S. wightii were found to be more active than C. linum. It was observed that the acetone extracts of all the three marine algae showed higher inhibitory activity for the selected bacterial species than other solvent extracts. The results revealed that the crude acetone extracts seem to be a good source material in identifying the effective pure antibacterial compound(s) in all the three marine algae and particularly, S. wightii. Conclusions: The present study showed that the acetone extracts of marine algae such as S. wightii, C. linum and P. gymnospora exhibited good antimicrobial activity. But the acetone extracts of S. wightii possessed highest antibacterial activity than others and so it could be useful in seeking active principles against human pathogenic bacteria.

151-154

Authors: Solomon Jeeva, Johnson Marimuthu @ Antonisamy

Objective: To investigate the antibacterial activity and phytochemical properties of Begonia floccifera Bedd. (B. floccifera) methanolic flower extracts against the selected pathogens. Methods: 20 g of fresh flowers were soaked in 100 mL methanol for 48 h at room temperature. After 48 h, the extracts were filtered using Whatman No. 41 filter paper. The filtrates were collected, made up to known volume and stored in refrigerator at 4 曟. The methanolic extracts were used for phytochemical and antibacterial studies. The preliminary phytochemical screening was performed according to the modified Harborne method. The methanolic flower extracts of B. floccifera Bedd. was tested against Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Salmonella typhi, Serratia marcescens, Proteus mirabilis, Enterococcus faecalis and Streptococcus pyogenes by the disc diffusion method. Results: The results of the phytochemical screening revealed that phenol, tannins, xanthoproteins, steroids, tannins, steroids, phytosterols, triterpenoids, sapogenins, coumarins and carbohydrates presence in the methanolic extracts of B. floccifera. The antibacterial activity has been observed in the methanolic extracts of B. floccifera against the tested bacteria with varied activity. The maximum zone of inhibition was 28 mm for Bacillus cereus, 25 mm for Staphylococcus aureus, 15 mm for Escherichia coli, 13 mm for Proteus mirabilis, 7 mm for Klebsiella pneumonia. Conclusions: The antimicrobial activity of B. floccifera methanolic flower extracts are comparable and their potential as alternative in the treatment of infectious by these microorganisms.

159-161

Authors: Sunil Christudas, Kulathivel TM, Agastian P

Objective: To investigate the phytochemical screening and antibacterial activity of pet. ether, chloroform and ethanolic extracts of Tridax procumbens (T. procumbens). Methods: The antibacterial activity of leaves of T. procumbens was evaluated by agar well diffusion method against three selected gram positive and gram negative bacterial species. Results: The presence of carbohydrates, proteins, tannins, steroids, alkaloids and purines in the different leaf extracts were established by phytochemical analysis. The chloroform extract was more effective against bacteria and the activity was comparable with that of standard drug ampicillin. Conclusions: The results in the present study suggest that T. procumbens leaves can be used for treating diseases caused by the tested organisms.

165-169

Authors: Patel DK, Prasad SK, Sairam K, Hemalatha S

Objective: To evaluate active aldose reductase (AR) inhibitory principles from ethanolic extract and one isolated compound beta sitostirol from Hybanthus enneaspermus (H. enneaspermus) Linn F. Muell. (Violaceae), used in the treatment of various complications including diabetes. Methods: AR inhibitory activities of H. enneaspermus and isolated compound beta sitosterol against rat lens AR enzyme were investigated in vitro using quercetin as a standard drug. Results: There was a significant difference in the AR inhibitory activity, and it was found to be more in H. enneaspermus [IC50 (49.46±2.26 µg/mL)] followed by beta sitosterol [IC50 (82.71±0.42 µg/mL)] but was less compared to standard quercetin [IC50 (3.326±0.11 µg/mL)]. From the value of Vmax, Km and Ki it was found that H. enneaspermus inhibited enzyme in a non competitive manner, whereas beta sitosterol inhibited enzyme in a competitive manner. Conclusions: H. enneaspermus showed significant AR inhibitory activity compared to the isolated compound beta sitosterol, which may be due to its high phenolic and flavonoid content. So this plant can be used to treat diabetic complications in the future, but further evaluation is necessary to know the exact mode of action involved in this process.

175-177

Authors: Sunder singh, Rai AK, Praveen Sharma, Yogesh Barshiliya, Mohini Sihare, Anju Negi

Objective: To evaluate the antidiarrhoeal effect of alcoholic extract of Rotula aquatica by using different experimental models. Methods: Antidiarrhoeal effect was evaluated by castor oil induced diarrhea, charcoal meal test and PGE2 induced diarrhea. Loperamide (2 mg/kg) and atropine (0.1mg/kg) were used as standard drugs. Alcoholic extract was used in 100 and 200 mg/kg dose. Results: In case of castor oil induce diarrhea dose 200 mg/kg showed delay in defecation time (186.980±15.660) min and 100 mg/kg showed delay in defecation time (97.779±4.952) min, while loperamide (2 mg/kg) showed delay in defecation time (221.148±21.919) min. Finally the reading showed that doses 200 mg/kg showed significant antidiarrhoeal effect from 100 mg/kg of alcoholic extract of Rotula aquatica in different model used for antidiarrhoeal activity. Conclusions: This study revealed that the alcoholic extracts has potential pharmacological activity against diarrhea and this may possible explain the use of the plant in traditional medicine.

181-186

Authors: Sheetal S. Chaudhari, Sanjay R. Chaudhari, Machindra J. Chavan

Objective: To evaluate the analgesic, anti-inflammatory and anti-arthritic activity of leaves of Cassia uniflora Mill. Methods: Petroleum ether, ethyl acetate and methanolic extract of Cassia uniflora (100 and 200 mg/kg, body weight) was screened for analgesic (Eddy’s hot plate and acetic acid induced writhing), anti-inflammatory (Carrageenan induced paw edema) and anti-arthritic (Complete Freund’s adjuvant induced arthritis). In complete Freund’s adjuvant arthritis model degree of inflammation was evaluated by hind paw swelling, body weight, and biochemical parameters and supported by radiological analysis. Results: Treatment with extracts of Cassia uniflora showed significant (P<0.05) and dose dependant increase in paw licking time in Eddy’s hot plate method. In writhing test, extracts significantly reduced the number of writhes. A dose dependant and significant inhibition of edema was observed in carrageenan induced paw edema. Petroleum ether extract at a dose of 100 mg/kg body weight showed most potent and significant activity which was supported by the results of body weight, biochemical parameters and radiological analysis in complete Freund’s adjuvant arthritis model. Conclusions: The extract possesses analgesic, anti-inflammatory and anti-arthritic activity which may be mediated through the phytochemical constituents of the plant.

192-196

Authors: Maharudra S Rakh, Amol N Khedkar, Nilesh N Aghav, Sanjay R Chaudhari

Objective: To evaluate the antiallergic and analgesic activities of seed of Momordica dioica Roxb. (M. dioica) Methods: Petroleum ether, ethyl acetate, methanol and aqueous extracts of M. dioica seed were screened for anti-allergic activity (200 mg/kg (i.p.) in mice body wt.) and analgesic (dose dependant 50 mg/kg and 100 mg/kg in mice body wt.). Results: Methanol extract of M. dioica seed shows more significant antiallergic activity (P<0.01) as compare to other extracts in all three models of milk induced leukocytosis, milk induced eosinophilia and differentia leukocytes count in mice. In dose dependant analgesic activity petroleum ether and methanol extracts gives significant hot plate activity (P<0.05, P<0.01) and in acetic acid induced writhing test significantly (P<0.01) reduced the number of writhing. Conclusions: The antiallergic activity may be due to phytochemical constituents such as polar constituents and flavonoids. The extract possesses analgesic activity which may be due to the phytochemical constituents of the plant.

200-202

Authors: Pallara J Wills, Velikkakathu V Asha

Objective: To investigate the acute and subacute toxicity of Lygodium flexuosum (L. flexuosum) extracts to substantiate the tribal or folk claims for its use as a safe hepatoprotective drug. Methods: The water, ethanol and n-hexane extracts of L. flexuosum were tested in vivo in Wistar rats. In acute toxicity model, rats received a single dose (5 g/kg) of water extract, ethanol extract, and n-hexane extract of L. flexuosum and kept under observation for 14 d. In subacute toxicity studies, animals were treated with daily doses (1 g/kg body weight) of water, ethanol and n-hexane extract respectively for 30 d. Results: In acute toxicity, the extracts administration up to a higher dose of 5 g/kg did not result in mortality or any change in behavior and biochemical parameters of the animals. Sub acute toxicity studies in rats showed that treatment with the extracts did not alter the serum biochemical and hematological parameters. Conclusions: The extracts were found to be devoid of any toxicity in acute (5 g/kg) and subacute (1 g/kg) toxicity evaluation in rats.

208-211

Authors: Vinod S Pawar, Shivakumar Hugar

Objective: To investigate the adaptogenic activity of methanolic extract of Trigonella foenum graecum seeds (METFGS) at 100, 250 and 500 mg/kg doses against anoxia stress tolerance in mice and immobilisation stress models. Methods: Withania somnifera (100 mg/kg, p.o.) was selected as reference standard and it showed significant effect in both stress models. The parameters like anoxia stress tolerance time was recorded in anoxia stress and estimation of biochemical marker levels and determination of organs weight were carried out in immobilisation stress model. Results: Concomitant treatment with METFGS at 500 mg/kg showed marked increase in anoxia stress tolerance time. But the lower doses (100 and 250 mg/kg) of METFGS produced mild increase in anoxia tolerance time. There was dose dependant significant reduction in biochemical parameters like serum glucose, cholesterol, triglycerides and BUN levels exhibited by test extract in immobilisation stress model as compared to stressed group. The stress induced increase in liver, adrenal gland weight and decrease in weight of spleen and testes were significantly reversed by the test extract at higher dose. But lower doses of extract caused the protective effect on weight of liver and adrenal gland only. Conclusion: The results from the present study indicate that METFGS possessed significant antistress activity

219-223

Authors: Lambole Vijay, Upendra Kumar

Objective: To investigate aqueous extract of bark of Moringa oleifera for normal wound healing and dexamethasone suppressed wound healing using incision, excision and dead space wound models in Wistar rats. Methods: In incision and dead space the extracts were applied daily topically till the 10th post wounding days while in excision model it was till the complete epithelialization process. Standard group were treated with Povidone iodine ointment topically daily. The breaking strength, percentage of wound contraction, period of epithelialization, dry granulation weight and hydroxyproline content were observed. Results: The aqueous extract of Moringa oleifera significantly increased the wound breaking strength in incision wound model. The aqueous extract treated wounds were found to epithelize faster and the rate of wound contraction was significantly increased as compared to control wounds. Significant increase in granulation breaking strength and hydroxyproline content in dead space wound was observed. The aqueous extract significantly decreased the antihealing activities of dexamethasone in all the wound models. Conclusions: The results indicated that the aqueous extract of Moringa oleifera promotes wound healing significantly and able to overcome the wound healing suppressing action of dexamethasone.

229-231

Authors: Pradeepa K, Krishna V, Venkatesh, Girish Kumar K, Santosh Kumar SR, Joy H Hoskeri, Gnanesh AU

Objective: To evaluate antinociceptive property of the leaf extract of Delonix elata (Linn.). Methods: Antinociceptive activity was evaluated by abdominal writhing and tail flick methods in Swiss albino mice. Acetyl salicylic acid was used as standard drug. Results: A dose dependent significant (P<0.01) antinociceptive activity was observed in three different concentrations 100, 200 and 300 mg/kg. Among them dose at 300 mg/kg, significantly (P<0.01) inhibited (74.94%) the nociception induced by acetic acid but less effective than the standard reference (84.06%). Tailflick test showed that the extract at the doses 300 mg/kg showed efficient results when compared to the other two doses (100 and 200 mg/kg) with increase in the latency to response of tail to the thermal stimulation. Conclusions: The significant antinociceptive activity of the leaf extract is due to the presence of a single active constituent in higher levels or due to the combined effect of more than one phytoconstituent. This investigation supported the ethnomedicinal claims of Delonix elata.

238-242

Authors: Pramod J Hurkadale, Pournima A Shelar, Siddhalingesh G Palled, Yuvaraj D Mandavkar, Ajay S Khedkar

Objective: To study the hepatoprotective activity of methanol and aqueous extracts of Amorphophallus paeoniifolius tubers against paracetamol induced liver damage in rats. Methods: Hepatotoxicity was induced by paracetamol and the biochemical parameters such as serum glutamic pyruvic transaminase (sGPT), serum glutamic oxaloacetic transaminase (sGOT) and serum alkaline phosphatase (sALP), serum bilirubin (SB) and histopathological changes in liver were studied along with silymarin and Liv-52 as standard hepatoprotective agents. Results: The phytochemical investigation of the extracts showed presence of carbohydrates, proteins, steroids and flavonoids. Pre-treatment of the rats with methanol and aqueous extract prior to paracetamol administration caused a significant reduction in the values of sGOT, sGPT, sALP and sB (P<0.01) almost comparable to the silymarin and Liv-52. The hepatoprotective was confirmed by histopathological examination of the liver tissue of control and treated animals. Conclusions: From the results it can be concluded that Amorphophallus paeoniifolius possesses hepatoprotective effect against paracetamol-induced liver damage in rats.

252-256

Authors: Sundaram Ravikumar, Samuel Jacob Inbaneson, Palavesam Suganthi

Objective: To identify the possible antiplasmodial compounds from Carica papaya (C. papaya), Pongamia glabra (P. glabra), Phyllanthus emblica (P. emblica) L, Phyllanthus acidus (P. acidus) L, Pisonia grandis (P. grandis) and Moringa pterygosperma (M. pterygosperma). Methods: The C. papaya, P. glabra, P. emblica L, P. acidus L, P. grandis and M. pterygosperma were collected from Ramanathapuram District, Tamil Nadu, India and the extraction was carried out in ethanol. The filter sterilized extracts (100, 50, 25, 12.5, 6.25 and 3.125 µg/mL) were tested for antiplasmodial activity against Plasmodium falciparum (P. falciparum). The potential extracts were also tested for their phytochemical constituents. Results: Of the selected plants species, the leaf extract of P. emblica L showed excellent antiplasmodial activity (IC50 35.09 µg/mL) followed by leaf extract of M. pterygosperma (IC50 42.36 µg/mL), leaf extract of C. papaya (IC50 46.23 µg/mL) and leaf extract of P. glabra (IC50 48.17 µg/mL). The flower extract of M. pterygosperma showed IC50 values of more than 100 µg/mL. Statistical analysis reveals that, significant antiplasmodial activity (P<0.01) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes was also carried out and it shows that, there were no morphological changes in erythrocytes by the ethanolic extract of all the tested plant extracts. The in vitro antiplasmodial activity might be due to the presence of alkaloids, glycosides, carbohydrates, flavonoids, phenols, saponins, triterpenoids, steroids and tannins in the ethanolic extracts of tested plants. Conclusions: The ethanolic stem extracts of P. emblica L possess lead compounds for the development of antiplasmodial drugs.

260-264

Authors: Utpal Adhikari, Someshwar Singha, Goutam Chandra

Objective: To analyze the mosquito repellency and larvicidal activity of mature leaves of Swietenia mahagoni (S. mahagoni) against Culex quinquefasciatus (Cx. quinquefasciatus) Say in laboratory bioassay.Methods: Various concentrations of crude and petroleum ether extract of S. mahagoni mature leaves were exposed against all instars of Cx. quinquefasciatus larvae and the respective LC50, LC90, Regression equations and R values were calculated. Mosquito repellency was tested with chloroform: methanol (1:1, v/v) extract of mature leaves. Results: Cent percent mortality of 2nd instar mosquito larva was observed at 50 ppm concentration of petroleum ether extract after 72 h of exposure. 1st, 3rd and 4th instar larvae showed 96.66%, 90.00% and 60.00% mortality at 50 ppm concentration of petroleum ether respectively after 72 h of exposure. Preliminary qualitative phytochemical assay revealed the presence of saponins, alkaloids, tannin, flavonoids, and free glycoside bound anthroquinones. Chloroform: methanol extract showed repellency up to 2 h 15 min after application. No mortality was found in non target organisms, such as Gambusia affinis, Tadpole of Bufo and Chironomus larvae. Chloroform: methanol of mature leaves extract of S. mahagoni exhibits 100% repellency upto 2 h 15 min as no mosquito bites up to that time periods in the treated hands. Conclusions: Different solvent extracts of mature leaves of S. mahagoni can be effectively used as a potent ecofriendly biocontrol agent against larval form of Cx. quinquefasciat.

270-275

Authors: Bhavik kumar H Satani, Shri Hari Mishra

Objective: To generate pharmacognostic parameters of leaves of Heterophragma quadriloculare K. Schum since data on pharmacognostic and phytochemical characterization which provide means to authentic raw material or drug for medicinally important formulation are not available. Methods: The present study reports detailed pharmacognostic characters as identification parameters of the leaves which were subjected to morphological, histological, quantitative microscopic and chromatographic studies. Morphological, histological, quantitative microscopic were done using reported methods and chromatographic studies were performed using HPTLC. Results: Diagnostic features for identification of shape of leaves being compound, odd-pinnate, dorsiventral asymmetric at base with unicellular non-covering and glandular trichomes having unicellular stalk and bicellular head, actinocytic stomata and rhombus calcium oxalate crystals. Water soluble and alcohol soluble extractives were found to be (23.6±0.346)% w/w and (10.8±0.115)% w/w, respectively.Conclusions: The determinations from above studies establish the macro- and microscopic parameters which can be utilized for quick identification of the drug both as whole and in the powdered leaf materials and may helpful in laying down pharmacopoeial standards of this important member of traditional medicine.

284-289

Authors: S. Vijaya Kumar, Dhirendra B. Sanghai, C. Mallikarjuna Rao, C.S. Shreedhara

Objective: To standardize Melia azedarach L. by detailed study of macorscopical, microscopical and physiochemical parameters. Methods: Bark sample and dried powder of Melia azedarach were studied macroscopically and microscopically. Preliminary phytochemical investigation, physiochemical parameters of the bark sample as per WHO recommended guidelines for standardizations were also performed. Results: The detail microscopy revealed the presence of collapsed phloem, non-collapsed phloem, sieve elements, sieve plates, sieve tubes, companion cells, crystals and starch grains, narrow and wide fibres. Physiochemical parameters such as percentage of foreign matters, ash values, loss on drying, swelling index extractive values were determined. Total phenolic and flavonoid content were also determined. Preliminary phytochemical screening showed the presence of carbohydrates, tannins, terpenoids, glycosides, flavonoids and phenolic compounds. Conclusions: The macroscopic, microscopic and physiochemical analysis is useful in standardization of the Melia azedarach bark.

296-304

Authors: Sandhya S , Chandra Sekhar J, Vinod.K.R, Rao KNV, David Banji

Objective: The present research is envisaged on the preclinical studies of a polyherbal phytocomplex hair growth promoting cream incorporated with aqueous extracts of Trichosanthes cucumerina (T. cucumerina) Linn and Abrus precatorius (A. precatoius) Linn on enhancing the hair growth. Methods: Leaves of T. cucumerina and A. precatoius were separately extracted with distilled water and then subjected to preliminary chemical test. It was then made into a phyto-complex and finally formulated into a hair cream. The various evaluation parameters like organoleptic characters, presence of foreign particles/grittiness, determination of pH, irritancy, partition coefficient, drug content, diffusion studies, stability studies, sterility test, SEM, FTIR, solubility and hair growth promoting activity were performed. Results: Microscopic view of phyto-complex showed micelle structure. The formulated phyto-complex hair growth promoting cream had a smooth consistency with slightly acidic pH. FT-IR studies showed that the phytoconstituents were compatible with the excipients used. SEM studies revealed binding nature of the complex. The cream was found to be stable as the globule size, and was not altered on storage. Sterility testing proved the formulation to be free from microbial attack. The preclinical studies showed that the formulated 2% polyherbal phyto-complex hair growth promoting cream was an effective hair growth promoter as the results were analogous to that of minoxidil 2%. It was observed that percentage of hair follicles in the anagen phase increased considerably which predicts that the formulation can be used in alopecia. Conclusions: As the 2% polyherbal phytocomplex hair cream did not show any kind of irritancy or toxicity to the test animals it can be suggested that the prepared cream can be a better alternative for synthetic hair growth promoters.

308-311

Authors: Karthikeyan M, Umarul Mukhthar AK, Megha M, Shadeer Hamza P

Objective: To develop fast dissolving tablets of diclofenac in order to get fast relief from pain. Methods: Fast dissolving tablets of diclofenac potassium were prepared by direct compression method using Indion 214, Indion 234, Indion 244 and croscarmellose as superdisintegrants. Microcrystalline cellulose was used as diluents and mannitol, as sweetening agent. Tablets were evaluated for weight variation; weight variation of all the formulations was observed which were within the acceptable limit for uncoated tablets as per United States Pharmacopoeia. Results: Mechanical strength, the hardness of tablets was determined and was found to be in the range of 6.4 to 7.13 Kg/cm2 . In vitro disintegration time, the formulation containing indion214, indion234 andindion244 showed 34,31and 39 seconds value for in vitro disintegration respectively. Wetting time for all the formulations was found to be (4.425±0.25) to (5.75±2.25) seconds and the cumulative percentage drug release of formulations FD1- FD5. It was observed that in first 10 minutes, only 12.89% of drug was release from formulation FD5 without using superdisintegrants and while it was 88.56% in the case of formulation FD2. At the end of 24 minutes only 85.42 % of drug was released from the control tablet formulation FD5 whereas the cumulative release from other formulations FD1 and FD4 at the end of 12 minutes was 89.52 %and 79.29%, respectively. The results of in vitro disintegration time show that formulation containing Indion 244, showed the disintegration time of 29.66 seconds. Conclusions: Indion 244 was found to have super disintegrant property the addition technique is a best method for preparing fast dissolving tablets for rapid pain mangment.

316-322

Authors: S. Prabhakaran, R. Rajaram, V. Balasubramanian, K. Mathivanan

Objective: To screen the antifouling potential of various extracts from seaweeds (Ulva reticulate, Sargassum wightii, Halimeda macroloba), sea grasseses (Halodule pinifolia, Cymodocea serullata) and mangrove plants (Rhizophora apiculata, Rhizophora mucronata and Avicennia marina) against some marine fouling bacteria. Methods: The different species of seaweeds, seagrasses and mangrove samples were collected, washed, air dried and fine powdered samples were subjected to solvent extraction by cold steep method. The extracts fraction was eluted with the ethanol and subjected to FTIR. The biofilm forming bacteria were scrapped from the marine environment by biofilm formed PVC sheet. Among these ten strains isolated, four isolates (Flavobacterium sp., Bacillus sp., Cytophaga sp., Pseudomonas sp.) were chosen for this study. Results: Among the tested extracts, Avicennia marina limited the growth of Flavobacterium sp. (16 mm) and Bacillus sp. (20 mm) and the extracts of Rhizophora mucronata limited the growth of Flavobacterium sp. (18 mm) and Bacillus sp. (18 mm). While comparing the inhibition activity of all the extracts, mangrove plants extracts had higher inhibiting activity against primary biofilm forming bacteria than seaweeds and seagrasses. The inhibition activity was mainly correlated with the major functional groups [hydroxyl, amino, carbonyl and phosphoryl functionalities, aliphatic (fatty acids), NH2 (amide I & II)] of the extracts. Conclusions: The bioactive fractions from the above results indicates the occurrence of active constituents in the extracts of seaweeds, seagrasses and mangrove. It shows the improved antifouling activity against marine micro-fouling bacteria. These extracts can be used as the possible natural sources for anti-foulant.

329-333

Authors: Govindasamy C, M. Arulpriya, P. Ruban

Objective: To explore the antimicrobial compounds from the rhodophyceae family member algae Gracilaria corticata using 1 H and 13 C NMR spectral analysis. Methods: Antimicrobial activity was done by using the method of Prabhakar et al. Methanol and acetone extracts were tested against the human pathogens. The acetone extract was subjected to column chromatography using acetone as a solvent for purification and also the chemical analysis combined with 1 H and 13 C NMR analysis. Results: Among the two extracts, acetone extract showed higher activity than methanol extract. 1 H and 13 C NMR analysis showed the fraction extracted with acetone had polysaccharide like compounds. Conclusions: Altogether our observations suggest a triggering of antimicrobial activity by this polysaccharide like compound. Further the investigations are now in progress to purify and identify the structure of the compounds.

342-346

Authors: Karunamoorthy Manivannan, Perumal Anantharaman, Thangavel Balasubramanian

Objective: To evaluate the in vitro antioxidant capacity of a Chlorella marina (Butcher, 1952). Methods: Samples were tested for the total phenolic content, antioxidant activity, deoxy ribose radical scavenging activity and reducing power. Results: The methanolic extract was found to have high levels of phenolic (0.647±0.052 mg GAE/g), when compared to diethyl ether and hexane. The diethyl ether extract exhibited higher antioxidant potential as (0.816±0.366 mg AscAE/ g), higher percentage of deoxy ribose radical scavenging activity (0.399±0.004) and reducing power (2.814±0.014). Conclusions: The phenolic compounds were not a major contributor to the antioxidant capacities of these microalgae. This was very different from many other plant species like fruits, vegetables and medicinal plants. The microalgae could contain different antioxidant compounds from other plants.

352-357

Authors: J. Margaret Beula, M. Gnanadesigan, P. Banerjee Rajkumar, S. Ravikumar, M. Anand

Objective: To identify the antiviral, antioxidant and toxicological evalution of the mangrove plant from South East coast of India. Methods: In vitro antiviral analysis such as RT inhibition assay, HBsAg binding assay, HBV DNA polymerase Inhibitory assays were carried out with the leaf extract of Avicennia marina, Avicennia officinalis, Rhizophora mucronata and Rhizophora apiculata mangrove plants. Further, the in vitro antioxidant assays and in vivo toxicological assay was also carried out with the most potent Avicennia marina leaf extract. Results: Of the selected 4 mangrove plants, Avicennia marina leaf extract showed the minimum IC50 values with 403.91, 489.39 and 372.09 µg/mL concentrations against reverse transcriptase inhibition assay, HBsAg binding assay, HBV DNA polymerase Inhibitory assays, respectively. The antioxidant IC50 values were identified as 12.80±0.93, 640.06±34.93, 19.91±3.93 and 142.06±17.93 µg/mL concentrations against SOD, LPO, NO and DPPH assays, respectively. The phytochemical analysis of Avicennia marinaleaf extract showed the presence of reducing sugars, polyophenols, flavonoids and tannin. The LD50 value was 2 500 mg/kg bodyweight and the sub acute toxicity analysis did not show any significant changes in serum and haematological parameters when compared with the control animals. Conclusions: It can be concluded from the present findings that, the leaf extract of Avicennia marina can be further used as potential antiviral drug after completing the clinical trials.

368-372

Authors: RVignesh, M Srinivasan

Objective: To analyze the proximate composition, fatty acids, amino acids and nutritional composition of flour made from Tilapia (Oreochromis mossambicus) head and bones. Methods: Tilapia fish head and bone are the parts with unknown composition. Fish bones and heads were collected, cleaned and dried. The flours were prepared using 14-mesh stainless steel sieve and analysed further. Results: The results in 100 g of Tilapia head flour (THF) were composed of moisture [(5.89±0.01) %] protein [(32.59±0.02) %] total lipids [(0.2014±0.0002)%], and ash [(11.14±0.02) %]. The results in 100 g of Tilapia bone flour (TBF) were: moisture [(4.22±0.02)%], protein [(31.52± 0.02) %], total lipids [(0.8761±0.0002) %], and ash [(0.89±0.01)%]. The fatty acid composition of THF occurring in the highest proportions were alpha linolenic acid [(C18: 3; (2.4390±0.0025)%], stearic acid [(C18:0; (1.9883±0.0025)%], linoleic acid [(C18:2; (1.9860±0.0025)%], palmitic acid [C16:0; (1.1787 ±0.0002) %], oleic acid [(C18:1; (1.5468±0.0002)%] margaric acid acid [(C17:0 (0.4146±0.0002)%] and moroctic acid [(C18:4; (0.1141±0.0002)%]. The fatty acid compositions of TBF occurring in the highest proportions were stearic acid [(C18: 0; (0.7860±0.0025)%], oleic acid [(C18:1; (0.1141±0.0002)%], alpha linolenic acid [(C18: 3; (0.1141±0.0002)%], palmitic acid [(C16: 0; (0.1143±0.0025)%], linoleic acid [(C18:2; (0.0816±0.0002)%], margaric acid acid [(C18:0; (0.0088±0.0003)%] and moroctic acid [(C18:4; (0.0034±0.0003)%]. The major amino acids such as glutamic acid (0.9967%), aspartic acid (1.837%), lysine (1.048%) and leucine (0.807%) were found in THF and in TBF the major amino acids were found as glutamic acid (0.7865%), aspartic acid (0.9686%), lysine (1.342%) and leucine (0.7756%). The THF flour possessed seven essential minerals in milli grams per gram (mg/gm) such as calcium (56.7±0.2 mg), iron (3.098±0.002 mg), potassium (16.78±0.02 mg), magnesium (15.67±0.03 mg), copper (1.414±0.002 mg), sodium (34.67±0.03 mg) and zinc (0.343±0.003 mg). The TBF flour possessed calcium (67.76±0.02 mg), iron (2.41±0.02 mg), potassium (15.67±0.02 mg), magnesium (45.76±0.02mg), copper (0.131±0.001 mg), sodium (29.11±0.01 mg), and zinc (0.2415±0.0002 mg). Conclusions: The results indicate that the flours made from the fish head and bone of tilapia may result as a valuable alternative food in the human diet. This is the first study which reports the nutritive value of tilapia (Oreochromis mossambicus) byproducts such as heads and bones.

377-379

Authors: Vasanthi Rajendran, Sameer Hassan, Jerrine Joseph, Nagamiah Selvakumar, Vanaja Kumar

Objective: To do a comparative homology search for the 32 mycobacteriophages. Methods: This can be estimated as 3 400 proteins from 32 mycobacteriophages assuming each phage has 80-100 genes. The algorithm most widely used for homology detection in comparative genomics is Basic Local Alignment Search Tool (BLAST). Usually a stringent score cutoff is applied to distinguish putative homolog’s from possible false positive hits. As a consequence, some BLAST hits are discarded or put into insignificant hits that are in fact homologous. Assigning function to protein sequences is important. Here, we review the status of sequence (BLAST) based and domain based approaches to proteins that can provide functional insights such as database on Protein Families (PFAM). Results: The findings showed that 31% of proteins showed similarities with homologous protein with known function. Conclusions: In the present study only 31% of mycobacteriophage proteins functions were able to be predicted. Only for 6 proteins, the template structures were available but then they were not directly involved in phage lifecycle. Hence this emphasizes the importance of exploring the structures for mycobacteriophage proteins in order to understand their function and evolutionary significance. Ab initio methods for protein structure prediction can be only alternative for the rest of the proteins but accuracy of prediction is not highly dependable.

383-385

Authors: Jerrine Joseph, Sameer Hassan, Vasanthi Rajendran,Vanaja Kumar

Objective: To functionally annotate the protein sequences for 64 mycobacteriophage genomes using the ACLAME database and to discuss how structured information on mycobacteriophage encoded proteins helps global in silico analysis. Methods: Each of the protein sequences of 64 mycobacteriophage genomes were searched against ACLAME database Version 0.4 using BLAST and were further classified based on function. The default setting of ACLAME 0.4 BLAST was used. Results: Based on ACLAME, assignment of 72 protein functional families was done for the 64 sequenced genomes, and families were compared among and across genomes. Conclusions: The finding of the present study provides scope for designing novel approaches to decipher this huge chunk of unknown proteins by experimental methods.

392-400

Authors: C. Balachandran, V. Duraipandiyan, S. Ignacimuthu

Objective: To isolate active actinomycetes from soil samples of Northern Himalayas and study their culture characterization, protease production and cytotoxic effects on cancer cell line (A549). Methods: Forty six strains of actinomycetes were isolated from the soil collected from Northern Himalayas, India. Isolation of actinomycetes was performed by serial dilution plate technique. Forty six isolated actinomycetes cultures were grown in ISP 2 medium to study the morphology and biochemical characteristics. Isolated strains were studied for protease enzyme production in skim milk agar medium with solubilising capacity. Seven isolates were studied for melanin pigmentation and different NaCl concentration. Effects of environmental conditions on protease enzyme production of seven isolated strains were also studied at different pH, temperature and metal ions (毬-mercaptoethanol, dithiothreitol, iodoacetamide, MgSO4, CaCl2 and EDTA). Seven isolates were also studied for lytic enzyme activity using different bacteria and yeast such as Pseudomonas aeruginosa, Enterococcus feacalis, Escherishia coli, Candida albicans, Bacillus subtilis, Klebsiella pneumonia and Staphylococcus aureus. Results: Isolates ERIA-31 and ERIA-33 produced more protease enzyme activity in modified nutrient agar media compared to other actinomycetes cultures. ERIA-31 and ERIA-33 were tested for cytotoxic effect in human adenocarcinoma cancer cell line (A549). IC50 for ERIA-31 was 57.04 µg/mL and IC50 for ERIA- 33 was 55.07 µg/mL. Conclusions: Actinomycete being a protease producing bacteria has the potential for use in industrial purpose, pharmaceuticals, cytotoxic agent and its proteolytic activity. Isolates of ERIA-31 and ERIA-33 produced significant amount of protease enzymes.

406-410

Authors: Narayanapillai Udhayashree, Duraisamy Senbagam, Balakrishnan Senthilkumar

Objective: To isolate bacteriocin- producing lactic acid bacteria (LAB) and to asses its preservative effects on food products. Methods: Lactobacillus strain was isolated from chicken intestine using MRS broth (Hi-Media, India) at 37 °C for 48 h. The isolate was identified by 16s rRNA sequencing and phylogenetic analysis. Bacteriocin was produced, purified by ammonium sulphate precipitation and dialyzed. Antagonistic characteristics of bacteriocin were studied by agar well diffusion method. Bacteriocin stability was studied in different temperature, pH and enzymes. Biopreservative efficacy was determined for fish and fresh juice. Results: Lactobacillus fermentum(L. fermentum) strain isolated from chicken intestine was capable of producing bacteriocins, inhibiting the widest range of pathogenic bacteria. The bacteriocin was stable at 37 °C and acidic pH and proteinaceous in nature. Their biopreservative effect on refrigerated food products (apple juice and fish) show low count of colonies. Lactobacillus isolate was sequenced which shows high similarity with reference strain L. fermentum CECT 5716. Conclusions: Bacteriocin producing L. fermentum UN01 could be successfully used as biopreservative and to increase the shelf life of the food products.

418-422

Authors: Amrin Ali, Ritika Raj Shrivastava, Nirupama Agrawa

Objective: To observe their specificity, speciation and ecological equivalence by using previous and present records, from freshwater fish from Tanakpur to Sultanpur areas, during 2009 & 2010. Methods: Data is gathered from the published records to compare it with the present records and entered into a computer database. The taxonomy of monogenoideans follows and those of fish follows. The worms were gently scrapped from the gills of fish and counted manually. They were studied mostly live, under a phase contrast microscope (Olympus CX 41 U-DA 4E 03365 Japan) or fixed in 3% formalin, after being relaxed in lukewarm water. Results: These monogenoids have perfect ecological equivalence, as their macro habitat is geologically connected. They exhibit specificity at different taxonomic levels of the parasites and the hosts. Sympatric speciation of parasites and their host is evident in the present study. Conclusions: The monogenoidean fauna, in this region, has its species integrity in the past sixty to sixty five years. From this important body of study, it seems likely that host specificity among monogenoids from the fresh water fish is more widely spread than previously thought. Seasonal variations, probable controlling factor of infection, determine the intensity.

429-434

Authors: S Shanmugam, K Rajendran, K Suresh

Objective: To elucidate the medicinally important herbs and their role in the health cares of the villagers living in Sivagangai district of Tamil Nadu, India. Methods: Interviews and detailed personal discussions were conducted with the herbalist and local people to identify plants and collect the medicinal informations for 12 months (from June 2009 to May 2010). The medicinal important plants were botanically identified. Results: Totally 71 species of 61 genera belonging to 36 families were reported with ethnomedicinal values. Leaves are the mostly used part to prepare medicine. Generally fresh parts are used. Conclusions: Attention should be made on proper exploitation and utilization of these medicinally important plants.

439-445

Authors: Rajendra Yonzone, R. B. Bhujel, Samuel Rai

Objective: To find out the genetic resources, current ecological status and altitude wise distribution of medicinal plants diversity of Darjeeling Himalaya of West Bengal, India. Methods: The intensive field survey was conducted in the entire regions and necessary information were collected from the local inhabitants and literatures. Results: The present study revealed 218 medicinal plants species from 97 families with 195 genera distributed throughout the different altitudinal ranges of entire Darjeeling Himalaya. Of these, 6 plant species were from 3 500-45 00 m altitude, 22 from 1 800-3 500 m, 69 from 1 000-1 800 m and 121 from 130-1 000 m in the study area. There were 24 cultivated medicinal plants and the rest were wild. Conclusions: It is unique for rich, wide genetic diversity resources and distribution of medicinal plants in distinct climatic zones. However, the status of these some plant species are vulnerable, threatened, rare and critically endangered.

449-455

Authors: D. K. Patel, K. Patel, M. Gadewar, V. Tahilyani

Flavonoids are low molecular weight plant phytoconstituents, widely distributed in the leaves, seeds, bark and flowers of plants. It has various pharmacological activities and protects plants against ultraviolet radiation, pathogens, and herbivores. Most of the beneficial health effects of flavonoids are mainly due to its antioxidant properties. Galangin is a flavonol found in honey, Alpinia officinarum, Helichrysum aureonitens and in propolis. It showed anti-mutagenic, anticlastogenic, anti-oxidative, radical scavenging, metabolic enzyme modulating activity and effective against certain types of cancers. The present review described the pharmacological activity and bioanalytical aspects of galangin, which may be benificial to the researchers who wants to explore the hidden potential of galangin.

462-463

Authors: PV Chaitanya Varma, Shrikiran A Hebbar, Ramesh Y Bhat, Suneel C Mundkur, Karthick Annamalai, Sonia Bhatt

Antibodies against the Golgi complex (AGAs) have been reported rarely and are associated with disorders like systemic lupus erythematosus, Sjögren's syndrome and rheumatoid arthritis. We report a case of autoimmune haemolytic anemia with Antigolgi antibodies, the first such case in medical literature.