Asian Pacific Journal of Tropical Disease

7-12

Authors: Anuj Modi, Vimal Kumar

Luffa echinata Roxb. (Cucurbitaceae) is a spreading climbing herb of tremendous medicinal importance, distributed throughout Pakistan, India, Bangladesh and Northern Tropical Africa. Traditionally various parts of the plant are being used for the treatment of different ailments such as jaundice, intestinal colic, enlargement of liver and spleen, leprosy, diabetes, bronchitis, nephritis, rheumatism, cirrhosis, dropsy, anthelmintic, stomach ache, snake bite, dog bite, fever, diarrohea and hemorrhoid disorder. The plant also possesses antioxidant, analgesic, anti-inflammatory, antidepressant, anxiolytic, antiepileptic, hepatoprotective, antibacterial, antifungal, antiulcer and anticancer activity. Research has been carried out using different techniques to support most of these claims. This review is an attempt to compile an up-to-date data on its ethanomedicinal, phytochemical and pharmacological perspective.

27-32

Authors: Kalpesh Chhotalal Ashara, Jalpa S. Paun, Moinuddin M. Soniwala, Jayant R. Chavada, Nitin Merubhai Mori

Topical delivery systems for drugs make localized administration of the drug anywhere in the body through ophthalmic, vaginal, skin and rectal routes. Topical formulations encompass a wide variety of formulations intended for cosmetic or dermatological application, to healthy as well as diseased skin. Drugs may be administered for localized or systemic effect. Topical preparations can be formulated with varying physico-chemical properties, as solid, semisolid or liquid. Microemulsion of drug is prepared and incorporated into emulgel, having novel topical drug delivery system as dual release control system. Micro-emulsions are micronized; thermodynamically stable systems having low interfacial tension prepared by adding co-surfactant have several features like enhanced permeability, good thermodynemic stability and prolong release. Emulgel prolongs the drug release, increases patient compliance and stability of emulsion. The prepared emulgel is evaluated for various parameteres like pH, viscosity, globule size; spreadability etc. whereas micro-emulsion is evaluated for various parameters like viscosity, pH, zeta-potential etc.

41-47

Authors: Seyyed Shamsadin Athari, Seyyed Moehyadin Athari

Asthma is a syndrome of variable airflow obstruction. It is characterized pathologically by bronchial inflammation and remodeling changes. Eosinophil infiltrate in asthma and a relationship between the degree of eosinophil infiltration in airways and severity of asthma has been suggested. Eosinophil has antigen-presenting cells and main role in allergic asthma. Platelets in inflammatory response is very important. It has also been shown that enzymes released by activated platelets play a direct role in the chronic inflammatory events that lead to airway remodeling in asthma. Dendritic cells (DCs) acquire antigen in the airways and then migrate to the draining lymph node where the cells mature and initiate T cell responses. Allergen challenge induces simultaneous increases in the number of DCs in the lungs. Because DCs are crucial in mounting immune responses during ongoing inflammation in the lung and balance of the allergic immune response.

54-63

Authors: Shakti Rath, Rabindra N. Padhy

Objective: To assess antibacterial activities of leaf and bark extracts of Holarrhena antidysenterica (H. antidysenterica), used by an Indian aborigine for ailments of human gastrointestinal tract, against eight extended spectrum β-lactamase producing multidrug resistant enteropathogens. Methods: Antibacterial activities of eight solvent-extracts of the plant were monitored by the agar-well diffusion method on lawns of all bacteria. Further, minimum inhibitory concentrations and minimum bactericidal concentrations of the best three solvent extracts were determined by the micro-broth dilution method. Preliminary phytochemical analysis of the active leaf and bark extracts were carried out. Results: It was found that Enterobacter aerogenes was resistant to 14 of 16 antibiotics, likewise, Escherichia coli to 13, Klebsiella sp. to 14, Salmonella paratyphi to 7, Salmonella typhi to 15, Shigella dysenteriae and Shigella sonnei to 14, Vibrio cholerae to 4 of 16 antibiotics. It was found that plant-extracts with petroleum ether and n-hexane had the least antibacterial activity. Extracts of leaves with chloroform, methanol, and water registered moderate antibacterial activity, whereas bark-extracts with ethyl acetate, acetone, and ethanol had a comparatively higher antibacterial activity on all these strains. Maximum sizes of zone of inhibition due to leaf extracts with ethyl acetate, acetone, and ethanol, and on the other hand, bark extracts with ethyl acetate, acetone and methanol were recorded against these bacteria; minimum inhibitory concentration and minimum bactericidal concentration values of specifically these extracts were determined. Phytochemical analysis of the methanolic bark extract of H. antidysenterica confirmed the presence of alkaloids, terpenoids, reducing sugars, tannins, and flavonoids. Conclusions: Data analysis revealed that leaves and bark of H. antidysenterica could serve as complementary/supplementary drugs along with suitable antibiotics to control the marauding multidrug resistant enteropathogens.

71-76

Authors: Madhukiran Parvathaneni, Ganga Rao Battu, Alexander I Gray, Pitchaiah Gummalla

Objective: To investigate the in vitro and in vivo anticancer activities of hypophyllanthin and phyllanthin isolated from Phyllanthus amarus Schum & Thonn against breast cancer. Methods: In vitro anticancer activity was evaluated against two cell lines (MCF-7 and MDAMB-231) using MTT assay. In vivo anticancer activity was tested using Sprague-Dawley rats with N-methyl-N-nitrosourea induced mammary cancer. Results: In vitro studies demonstrated a dose-dependent inhibitory effect on cell growth with IC50 values of (35.18±1.48) µg/mL (hypophyllanthin) and (32.51±0.95) µg/mL (phyllanthin) for MCF-7; (38.74 ±1.24) (hypophyllanthin) and (32.2±1.17) (phyllanthin) for MDA-MB-231 breast cancer cell lines. Tumor weights per group at doses of 5 and 10 mg/kg/day for hypophyllanthin (12.82 and 12.06 g) and phyllanthin (11.95 and 8.87 g) treated groups were significantly (P<0.001) lower than untreated N-methyl-N-nitrosourea group (35.85). Conclusions: Results of the present research work indicated that the isolated lignan compounds, hypophyllanthin and phyllanthin showed significant anticancer activities against breast cancer, in vitro and in vivo.

81-86

Authors: Prashant Tiwari, Dheeraj Ahirwae, Anish Chandy, Bharti Ahirwar

Objective: To screen Selaginella lepidophylla (S. lepidophylla) that are used in traditional medicine for their claimed hepatoprotective properties. Methods: Alcoholic and aqueous extracts of S. lepidophylla were evaluated for their hepatoprotective activity using CCl4 and paracetamol induced acute hepatic injury model. Results: Treatment with CCl4 and paracetamol significantly increased liver weight and volume compared to the normal group. Pretreatment with silymarin alcoholic and aqueous extracts significantly prevent increase in liver weight and volume. Conclusions: From the present experimental study it can be concluded that alcoholic and aqueous extract of S. lepidophylla exhibited significant hepatoprotective activity against CCl4 and paracetamol induced hepatotoxicity in rats, as result showed in physical, biochemical and histopathological parameters.

92-96

Authors: Thota Venkata Chaitanya Kumar, Tollamadugu Naga Venkata Krishna Vara Prasad2* , K Adilaxmamma, Mekapogu Alpharaj, Yagireddy Muralidhar, Pagadala Eswara Prasad

Objective: To develop a reliable, eco-friendly and easy process for the synthesis of silver nanoparticles using aloin, the active principle of medicinal plant ‘Aloe vera’ and to evaluate antimicrobial activity against Staphylococcus aureus (S. aureus), a causative organism of most of the diseases in livestock and to standardize the level of safety of synthesized silver nanoparticles. Methods: Characterization using UV-vis spectrophotometry, DLS technique, FT-IR and SEM. Tube dilution method was carried out to evaluate the MIC of the compound against S. aureus. MTT assay was performed to evaluate the level of safety of nanoparticles. Results: UV-vis absorption spectrum showed a maximum absorption around 200 nm for aloin mediated silver nanoparticles (ANS). The size of the particles as measured by DLS technique was 67.8 nm. The results of FT-IR analysis indicated the involvement of hydroxyl, carboxyl, amine and nitrile groups in the synthesis and stabilization of aloin mediated silver nanoparticles. SEM images showed that ANS with cubical, rectangular, triangular and spherical morphology and measured sizes of the agglomerated nanoparticles are in a range of 287.5 to 293.2 nm, however the average size of an individual particle is estimated to be approximately 70 nm. The compound (ANS) showed a MIC of 21.8 ng/mL against S. aureus and showed an in vitro spleenocyte viability of more than 80% at the highest concentration of 87.5 mg/L per well. Conclusions: Aloin consists of functional groups which reduced Ag+ ions to Ago ions and helped in synthesis of silver nanoparticles. The synthesis process has further enhanced the antimicrobial activity of nanosilver. The compound is also proved to be safe at the level many times higher than the MIC.

102-110

Authors: Giji Sadhasivam, Arumugam Muthuvel, Ramya Rajasekaran, Abirami Pachaiyappan, Balasubramanian Thangavel

Objective: To study the biochemical and biomedical properties of predatory gastropod Conus betulinus venom. Methods: Molecular weight of the crude venom extract was determined by using SDS-PAGE. Toxicity studies were carried out using haemolytic and brine shrimp lethality assays. Fibrin plate assay and substrate SDS-PAGE were used to determine the effect of sample on fibrin(ogen) olytic and proteolytic activity. The FTIR characterisation and chemical fingerprinting of amino acid were done with HPTLC. Enzymatic activities like phospholipase and hyaluronidase were measured spectrophotometrically and calculated in units. Anticancer screening was carried out by MTT assay. Results: Studies on this deadly venom revealed six different molecular weight proteins of medical significance ranging between 20.0 kDa and 97.4 kDa. The protein content was estimated as 0.9 mg/ mL. Haemolytic activity in erythrocytes was recorded and LC50 (Artemia) at 31.5 µg/mL. Further the venom showed considerable enzymatic properties like gelatinolytic, caesinolytic, fibrinolytic and fibrinogenolytic activities. The hyaluronidase and phospholipase activities were recorded at meagre range. The venom exhibited significant activity against HeLa cell lines. Moreover the evolution of venom is the crucial nature of Conus peptides in their challenging ecosystem. Periodical study on these peptides will unveil more peptides of biomedical use. Conclusions: Although quite a lot of works have dealt with paralytic effects of Conus venom, it still remains as an unexplored cocktail with promising molecules for drug development.

116-120

Authors: Abdul Viqar Khan, Qamar Uddin Ahmed, Mohd Wasi Khan, Athar Ali Khan

Objective: To conduct ethnopharmacobotanical field explorations in rural areas of five districts of Uttar Pradesh, India with regard to the folk herbal formulations associated with the management of poisons and poisonous bites. Methods: Local traditional healers known as “Vaidya” and “Hakeems” in the study area were interviewed to gather ethnopharmacobotanical information using a questionnaire attending various medical practices. Results: Information on 49 herbal formulations prepared from 39 plant species belonging to 28 plant families in the treatment of poisons and poisonous bites is presented in this scientific communication. Conclusion: Present communication revealed that study area is rich in its ethnopharmacobotanical knowledge. The plant species discussed here also encompasses new reports on Chenopodium album, Solanum xanthocarpum, Solanum melongena, Sesamum indicum, Calotropis procera, Coriandrum sativum, Cynodon dactylon, Brassica campestris, Triticum aestivum, Vitis vinifera, Sorgum vulgare and Nerium indicum. This study further concludes that there lies a lot of potential in the Indian herbal repository which should be explored systematically and later subjected to thorough study under the light of latest available scientific research methodologies for the drug standardization and pharmaco-toxicological studies with a view to making cheaper and safer drugs for the benefit of humanity periodically encountered with poisons and poisonous bites.

129-133

Authors: Bindu Madhavi Boddupalli, Ravinder Nath Anisetti, Ramalingam Ramani, Nagulu Malothu

Objective: To investigate the formulation of gastroretentive microspheres of omeprazole along with piperine and estimate the pharmacokinetic parameters in comparison with omeprazole alone. Methods: In our present investigation, gastroretentive microspheres of omeprazole were prepared with the inclusion of piperine. Pharmacokinetic parameters like Cmax, Tmax and area under curve were estimated by administering the prepared microspheres to rabbits and the results were compared with omeprazole alone. Results: There was a significant increase in area under curve from 3.441±1.093 mg·h/mL to 14.422 ±0.708 mg·h/mL along with an increase in Cmax. Conclusions: This clearly shows the increased absorption and decreased metabolism of omeprazole when administered along with piperine as gastroretentive microspheres.

140-147

Authors: Anne Williams Augustine, Akilavalli Narasimhan, Mangalapriya Vishwanathan, Balasubramanian Karundevi

Objective: To evaluate the antidiabetic effect of the aerial part of Andrographis paniculata (A. paniculata) powder (500 mg/kg body weight) in high fat and sucrose-induced type-2 diabetic rat model. Methods: The fasting blood glucose, oral glucose tolerance test, serum insulin, lipid profile, mRNA and protein levels of insulin signaling molecules, 14C-2 deoxy glucose uptake and 14C glucose oxidation in liver were checked. Results: In the type-2 diabetes-induced group, the fasting blood glucose, oral glucose tolerance, serum insulin, lipid profile, glucose uptake and oxidation, Akt and glucose transporter 2 mRNA, insulin receptor and glucose transporter 2 protein (both cytosolic and plasma membrane) and phosphorylation of insulin receptor substrate 1 and Akt were impaired. A. paniculata was able to successfully reinstate this impairment. In addition to this, A. paniculata did not cause a hypoglycemic condition in normal rat, affirming its activity in hyperglycemic state alone. Conclusions: A. paniculata possesses significant antidiabetic and antihyperlipidemic activities in high fat and sucrose-induced type-2 diabetic rat and the molecular actions at the level of insulin signaling molecules in liver reinforce it.

153-157

Authors: Hamzah Rabiat Unekwu, Jigam Ali Audu, Makun Hussaini Makun, Egwim Evans Chid

Objective: To elucidate the phytochemical content and antioxidant activity of selected wild edible Nigerian mushroom species. Methods: Phytochemical screening was carried out using standard methods while 1,1-Diphenyl picryl hydrazyl (DPPH) radical and reductive power assays were used to evaluate the in vitro antioxidant properties of the selected edible Nigerian mushroom species. Results: The result obtained revealed the presence of alkaloids, cardiac glycosides, saponins, flavonoids, terpenes, steroids, tannins and phenols in the selected mushrooms extracts. The extract of Pleutorus ostearus showed a significantly (P<0.05) higher total phenol and flavonoid content of (248.80±7.63) mg/g and (42.63±0.63) mg/g respectively compared to other mushroom extracts. Cantherale cibarus had the most significant (P<0.05) amount of alkaloids [(135.57±0.27) mg/g] and saponins [(150.41±0.50) mg/g] when compared to other extracts while the tannin content [(170.56±0.74)] mg/g was highest in the mushroom Temitomyces robustus. All mushroom extracts scavenged DPPH radical in a dose dependent manner. However, Lactarus deliciousus had the highest DPPH scavenging activity compared to the other mushroom extracts. Pleutorus ostearus and Lactarus deliciousus had better reductive power than other mushroom extracts concentrations used. Conclusions: The mushroom species analysed have been shown to be good sources of antioxidants and other phytoconstituents, thus it can be used in the management of oxidative stress induced diseases.

162-166

Authors: Sundaram Prasanna Kumar, Sundaram Ravikumar

Objective: To explore the antiplasmodial potential of marine sponge Clathria vulpina (C. vulpina) against chloroquine sensitive Plasmodium falciparum (P. falciparum). Methods: The marine sponge C. vulpina was collected from Thondi coast, authenticated and subjected for extraction by soaking in ethanol:water mixture (3:1 ratio). The percentage of extract was calculated. Filter sterilized extracts (100, 50, 25, 12.5, 6.25, 3.125 μg/mL) were screened for antiplasmodial activity against chloroquine sensitive P. falciparum. The extract was also tested for its hemolytic activity. Results: The percentage yield of extract of C. vulpina was found to be 4.8%. The crude extract of C. vulpina showed excellent antiplasmodial activity (IC50=14.75 μg/mL) which was highly comparable to the positive control chloroquine (IC50=7 μg/mL). Statistical analysis reveals that the significant antiplasmodial activity (P<0.05) was observed between the concentrations and the time of exposure. The chemical injury to erythrocytes was also carried out, which showed that there were no morphological changes in erythrocytes by the ethanolic extracts of sponges after 48 h of incubation. The extract showed slight hemolytic activity which almost equal to chloroquine at 100 μg/mL concentration (1.023%). Conclusions: The marine sponge C. vulpina can be used as a putative antiplasmodial drug after completing successful clinical trials.

173-180

Authors: Kalimuthu Kovendan, Shanmugam Perumal Shanthakumar, Cheruparambath Praseeja, Palanisamy Mahesh Kumar, Kadarkarai Murugan, Savariar Vincent

Objective: To evaluate the mosquito larvicidal and pupicidal activity of the ethanolic extracts from Morinda citrifolia (M. citrifolia) plant and entomopathogenic fungi Metarhizium anisopliae (M. anisopliae) against malaria vector, Anopheles stephensi (An. stephensi). Methods: M. citrifolia leaves were collected in and around Alleppy districts, Kerala, India. M. citrifolia leaf was washed with tap water and shade dried at room temperature. An electrical blender powdered the dried plant leaves. A total of 500 g leaf powder was macerated with 1.5 L of ethanol sequentially for a period of 72 h and filtered. The crude plant extracts were evaporated to dryness in rotary vacuum evaporator. The larvicidal and pupicidal activity was assayed at various concentrations ranging from 100 to 500 mg/L under the laboratory conditions. The LC50 and LC90 values of the M. citrifolia leaf extract and M. anisopliae fungi were determined by Probit analysis. Results: The plant extract showed larvicidal and pupicidal effects after 24 and 48 h of exposure; all larval instars and pupae have considerably moderate mortality; however, the highest larval and pupal mortality appeared in combined treatment at 24 and 48 h. The LC50 and LC90 values of M. citrifolia and M. anisopliae and their combined treatment against the first to fourth instars larvae and pupae of the malaria vector were assessed. M. citrifolia had values of LC50=202.47, 95.75, 57.52, 18.30 and 97.78 mg/L; LC90=384.37, 482.91, 631.22, 757.55 and 944.96 mg/L at 48 h. M. anisopliae had values of LC50=1.40, 3.99, 5.56, 8.77 and 11.49%; LC90=13.84, 17.62, 22.20, 25.71 and 30.78% at 48 h; Combined treatment had values of LC50=3.71, 16.73, 29.71, 40.60 and 138.10 mg/L; LC90=122.29, 150.15, 156.90, 211.99 and 806.67 mg/L at 48 h, respectively. Conclusions: The plant and the fungi are promising larvicidal and pupicidal agents against malaria vector, An. stephensi. This is a new eco-friendly approach for the control of vector. Therefore, this study provides first report on the combined treatment of this plant extract and fungi from India.

186-193

Authors: Zinia T. Nujum, Leela Itty Amma KR, Jeesha C. Haran, Krishnapilla Vijayakumar, Sreelal Thekkumkara Prabhakaran, Sajna Arif Noushad

Objective: To determine the prevalence of any of the clinical manifestations of lymphatic filariasis, parasitological and entomological indices in Thiruvananthapuram district, Kerala, India, prior to launching mass drug administration (MDA) in the district in 2005. Methods: A cross sectional survey was conducted in 7 endemic wards of the district, in a sample of 2 472 individuals above the age of one year. The study consisted of data collection using questionnaire, night blood smear examination and mosquito collection followed by identification and dissection. Results: The prevalence of any of the clinical manifestations of filariasis in this endemic area was 3% (73/2 472) (95% CI between 2.3% to 3.7%). The microfilaria rate was found to be 0.38%. Culex quinquefasciatus formed the predominant mosquito species. The vector infection rate was 1.4% and infectivity rate was 0.47%. Half of the population had satisfactory knowledge regarding the disease. In almost an equal number, the knowledge was poor and only about 10% had good knowledge. Most of the people had not heard about the MDA program. Conclusions: The prevalence of filariasis even in an area considered to be endemic in Kerala, was low. Microfilaraemia was much lower than the clinical manifestations. However, the potential risk of transmission of disease continues. For such areas which had a pre-MDA mf rate less than 1%, there needs to be more stringent criteria for evaluating the effectiveness of the programme, doing transmission assessment surveys and stopping MDA.

199-203

Authors: Prasad Govindrao Jamkhande, Vikas Annarao Suryawanshi, Amruta Sureshrao Wattamwar, Sonal Ramrao Barde

Objective: To investigate the anthelmintic activity of Borassus flabellifer (B. flabellifer) Linn. leaves against Indian adult earth worms (Pheretima posthuma). Methods: The B. flabellifer leaves and Indian adult earth worms (Pheretima posthuma) were collected and authenticated. Earth worms were grouped and treated with extract at 10, 20 and 50 mg/mL concentration, albendazole (10 mg/mL) as a standard and normal saline. The paralysis time and mortality time was considered as indicator of anthelmintic efficacy. Results: All the extracts showed concentration dependent activity but significant activity was observed at 50 mg/mL. At concentration 50 mg/mL extract pertained better activity with paralysis time (13.3 min) and death times (17.92 min) when compared to standard albendazole. Conclusions: The study findings reveal that the methanolic extract of B. flabellifer leaves has effective anthelmintic activity against Indian adult earth worms.

211-213

Authors: Guruprasada Shetty, B Sanjeev Rai, K Shreedhara Avabratha, Habeeb Ullah Khan

Objective: To determine the prevalence of hyponatremia and its association with severity and species of malaria. Methods: This is a prospective study, done at Father Muller Charitable Hospital Mangalore, in Karnataka, India. Serum sodium and parasite counts were estimated in all positive cases of malaria patients under 15 years admitted in hospital between August 2010 to July 2012. Data regarding all positive cases of malaria under 15 years admitted in hospital between Jan 2010 to June 2011 were obtained. Data was analysed by Chi-square test, Fisher’s exact test and student t test. P value less than 0.05 was taken significant. Results: A total of 60 patients with malaria who met the inclusion criteria were enrolled for study. Plasmodium vivax was the common species causing malaria (53.3%). Hyponatremia was reported in 39 (62%) patients. Of the 39 cases, 2 (3%) patients had severe hyponatremia, 19 (31.67%) patients had moderate hyponatremia, whereas 18 (30%) patients had mild hyponatremia on admission respectively. Mean sodium for falciparum malaria (131.03±4.31) mEq/L cases was significantly lower than that of vivax malaria cases [(134.2±3.3) mEq/L, t=2.20, P=0.029]. Severity of hyponatremia increased with severity of infection. Conclusions: Hyponatremia is common in children with malaria and its severity increases with severity of infection. It’s more common and more severe in falciparum malaria than vivax malaria.

221-225

Authors: P. Satyanarayana, Farid Babu Meka, Rekha Kumari Dulala, Mrinmoy Ghosh, K.R.S. Sambasiva Rao

Objective: To observe the distribution of malaria infection in the tribal area of East Godavari District of Andhra Pradesh, India . Methods: The data for the present study was collected from 6 342 and 765 patients who were admitted at tribal hospital and hill forest camps respectively during the study period. The data was collected from the malaria suspected patients admitted in paediatric and adult critical care wards of a tertiary hospital and in interior hill and forest camps in and around the Rampachodavaram. The detailed medical case sheet proforma were prepared and data has been analyz. It was found that RDT’s can be helpful to screen Plasmodium falciparum (P. falciparum) inendemic areas and remote tribal belt. Results: A total number of 6 342 and 765 patients were admitted at tribal hospital and hill forest camps respectively during the study period. Among 6 342 of individuals tested, 4 387 individuals were reported for malaria positive. Out of the 4 387 slides examined, 59.0% were positive P. falciparum infection, 19.4% slides showed positive Plasmodium vivax infection and 21.5% had mixed infection. The total P. falciparum burden was estimated as 80.5%. In hill forest camps, out of 765 admitted patients, 650 patients who had clinical history showed suggestive of malaria were examined for malaria parasites. Conclusions: The maximum numbers of malaria infection (4 387) were reported from the tribal based hospital. Malaria is responsible for major health concern in this region, particularly in rainy season. P. falciparum was the major parasite type causing malaria, and most of the complications were due to Plasmodium vivax. Compared to rest of the hills and forest areas, where most of the tribal people reside has the heavy load of malaria mainly P. falciparum. One important finding from the present study was the sex-difference observed in the admission rate. The rate of malaria infection was significantly high for male (53.5%), followed by female (46.5%) and children (33%).

233-235

Authors: Purna A. Chander, Haritha Y. Sri, Nishitha B.M. Sravanthi, Uma V. Susmitha

Objective: To study the anthelmintic activity of Barleria buxifolia leaf and to estimate the total flavonoid content. Methods: The aqueous and ethanolic leaf extracts were prepared and these were analyzed for total flavonoid content by aluminium chloride colorimetric method and Pheretima posthuma was used for anthelmintic activity by using the different concentrations (10, 20, 40, 80 and 100 mg/mL). Results: All the investigational extracts showed an anthelmintic activity at concentration of 10 mg/mL. The ethanolic extract of 100 mg/mL has produced an significant effect (P<0.001) when compared to aqueous extract. The total flavonoid content was found to be 5.67 mg QE/100 g. Conclusions: From the above study, the leaf extract has shown a good anthelmintic activity.

240-245

Authors: Shayeghi Mansoreh, Doosti Soghra, Bazrafkan Sara, Hosseini-Vasoukolaei Nasibeh, Vatandoost Hassan, Akhavan Amir Ahmad, Vatandoost Sajad, Arandian Mohamd Hossein

Objective: To determine the fauna of aquatic insects in Zayandeh River of Isfahan carried out in Tehran University of Medical Sciences in 2011. Methods: This study was performed in Isfahan, central of Iran in Zayandeh River. This was a descriptive study. Having fulfilled sampling for several times, we collected nearly 76 samples from different parts of river. Then they were sealed in an individual jars containing some water obtained from their habitat. Next, the insects were put in jars containing 70% ethylic alcohol. Results: A total of 76 matured samples of aquatic insects from the Zayandeh River were obtained. Among them, the order of Hemiptera which were the most prevalent order including two families: gerridae (n=27, 35.52%), and notonectidae (n=11, 14.47%). Other order were found belonging to Odonata from the family of coenagrionidae (n=12, 15.78%), coleoptera from the family of Carabidae (n=15, 19.73%) and prostigmata from the family of Hydrachindae (n=11, 14.47%). This was the first faunestic study carried out in Zayandeh River of Isfahan of Iran. Conclusions: The results are appropriate for future researches to detect more ecological aspects of aquatic arthropods and their role for biological control of vectors which transmit disease to human and animals.

250-254

Authors: Mahdi Amirmohammadi, Shole Khajoenia, Mahmoud Bahmani, Mahmoud Rafieian-Kopaei, Zohre Eftekhari, Mostafa Qorbani

Objective: To evaluate the effects of Salvia officinalis and Artemisia abrotanum extracts against digestive system parasites of mice. Methods: The ethanol extract was prepared and dissolved in distilled water. The mebendazole was used as positive control and distilled water as negative control. After counting eggs per gram feces, infected mice with 16 eggs per gram feces contained two to three parasites of Syphacia obvelata, Aspicoloris terepetra and Hymenolipis nana designated in 4 groups. The first group was given extracts of Artemisia (150 mg/kg), the second group was given Salvia extract (150 mg/kg), the third group was given mebendazole (10 mg/kg) and finally the fourth group was given distilled water (2 mL/kg). Results: The ethanol extracts of Artemisia and Salvia plants reduced the number of parasite eggs per gram of feces. Results showed significant reduction (P-value<0.001) in the number of eggs excreted by Hymenolepis nana, Aspiculuris tetraptera, Syphacia obvelata in mice. Conclusions: These results revealed that antiparasitic effects of Artemisia and Salvia are reasonable and these two plants might be used as antiparasitic natural products.

259-262

Authors: Kabita Mahato, Bibhuti Bhusan Kakoti, Sudarshana Borah, Mritunjay Kumar

Objective: To carry out an exhaustive study with a view to substantiate the therapeutic potential of the plant in terms of its anthelmintic activity against Pheretima posthuma using mebendazole as a reference standard. Methods: Forty five worms were collected and were divided into nine groups and were washed in normal saline before they were released into 10 mL of respective drug solutions using distilled water as vehicle. Different concentrations (25 mg/mL, 50 mg/mL and 100 mg/mL) of the test (MEHI and AEHI) and standard solution were prepared before the commencement of the experiment. Time for paralysis and time for death were recorded for each group. Results: Crude methanolic extract with concentrations of 25 mg/mL, 50 mg/mL, 100 mg/mL produced dose-dependent paralysis. Time of paralysis and death increased with concentration of 25 mg/mL, 50 mg/mL as compared with that of standard drug mebendazole. Methanolic extract of Heliotropium indicum Linn. (H. indicum) gave shorter paralysis and death time at 100 mg/mL as compared to aqueous extract H. indicum Linn. Results are expressed as mean±SEM (P<0.05) of 5 worms in each group. Conclusions: From the investigation, conclusion can be drawn that the methanolic extract of H. indicum (Boraginaceae) showed better activity than aqueous extract of the same to treat intestinal worm infections. In comparison with the standard drug mebendazole, methanolic extract of H. indicum showed significant anthelmintic efficacy.

268-272

Authors: Palanivel Bharadhirajan, Sambantham Murugan, Alagarsamy Sakthivel, Periyasamy Selvakumar

Objective: To examine the effect of six different isopods from five different food fish species. Methods: The specimens were collected directly from the trawlers landed at Parangipettai coast during the year of 2012 (January-December). Isopods were removed from the body surface and buccal cavities of the fish hosts and immediately preserved in 70% ethanol. Data regarding the prevalence of isopod parasites was calculated and site of attachment. Results: During the study period, 310 fishes were collected and examined for parasites infection, and it includes five different species. Among the examined fishes, 106 fish were infected among the total number of 120 collected parasites. The occurrence of isopod infection in all the infected fishes was 34.19%. The occurrence of Liza parsia was 25%, Thryssa dussumieri was 36%, Sardinella albella was 17.5% and Otolithes cuvieri was 65%. The maximum isopoda parasite infected noticed Otolithes cuvieri and minimum value was recorded in Sardinella albella. Conclusions: This is the new host for the parasite Nerocila phaeopleura in Liza parsia. Thus parasitic attack the growth and normal function of the fishes will suffer and lead to economic loss or the marketability may reduce then. In along the lifespan, the host population was may be affected by the parasitic outbreak.

276-278

Authors: Maharudra S. Rakh, Rajendra V. Pawar, Amol N. Khedkar

Objective: To investigate the anthelmintic potential of various extracts of the rhizomes of Curcuma amada Roxb. Methods: Cyclohexane, ethyl acetate, methanol and aqueous extracts of Curcuma amada Roxb. against Pheretima posthuma, an Indian species of earthworm. Various concentrations (25, 50 and 100 mg/mL) of these extract were evaluated for anthelmintic activity by recording the time required for paralysis and death of worms. Albendazole was used as a reference standard drug. Results: Methanol and aqueous extracts shows most significant anthelmintic activity compared with standard drug. Conclusions: Since methanol and aqueous extracts were tested by different chemical tests. It shows presence of carbohydrates, steroid, alkaloids, glycosides and flavanoids. These phytoconstituents may be responsible for the said activities.

284-289

Authors: Mohammad Mehdi Darvishi, Mohammad Reza Youssefi, Emad Changizi, Parviz Shayan, Rasoul Rostami Lima, Mohammad Taghi Rahimi

Objective: To investigate and survey the biology of Dermacentor marginatus (D. marginatus) under laboratory conditions. Methods: In this investigation, D. marginatus adult ticks were collected from sheep in Semnan province. Then various developmental stages of D. marginatus including larvae, nymphs and adult males and females under laboratory condition were bred and their biology was scrutinized. Results: The requisite time to complete the life cycle of D. marginatus under controlled laboratory conditions for larvae (26 °C, 75% relative humidity) and nymph (26 °C, 95% relative humidity) moulting, was on average 92 d (range 75-104 d), including preoviposition and egg incubation (22.5 d), larvae incubation (20.5 d), nymphal stage (28 d) along with male maturation (21 d). The index of conversion efficiency and the index of reproduction efficiency in females were 0.397 and 8.300, respectively. Conclusions: Although in this investigation, there was no meaningful correlation between preoviposition period and the weight of female ticks which were laid successfully. The significant linear relationship was fully observed between the weight of engorged female of D. marginatus and the number of eggs laid. The mean of preoviposition period from 5.4 d in autumn to 34.2 d in spring increased. The minimum weight of ticks with laying capacity was 69 mg and lighter ticks (21 mg) either did not lay or if they did their eggs did not hatch.

297-301

Authors: Majid Goodarzi, Shahram Nanekarani, Nasir Landy

Objective: To examine the effect of onion (Allium cepa L.) as an antibiotic growth promoter substitute on growth performance, carcass traits, and microflora composition in broiler chickens. Methods: A total of 192 one-day old mixed sex broiler chicks (Ross 308) were weighed and randomly allocated to four treatment groups, each with 4 replicate pens of 12 chicks. The dietary treatments consisted of the basal diet (control), antibiotic (15 mg virginiamycin/kg), and control+10 or 30 g fresh onions bulb/kg diet. Body weights of broilers were determined at Day 1, 21, and 42, feed intake was determined at the same periods, and feed conversion ratio was calculated accordingly. At Day 42, two birds per replicate were slaughtered for determination of carcass and organ weights. The populations of Lactobacilli spp. and Escherichia coli were enumerated in ileum by conventional microbiological techniques using selective agar media. Results: Dietary supplementation of 30 g/kg onion increased final body weight of broilers at 42 d of age compared to that of the other treatments (P<0.05). Birds fed 30 g onion/kg in the diet had the highest feed intake than other treatments at different growth periods (P<0.05). Feed conversion ratio, carcass yield and internal organ weights were not affected by the dietary treatments at Day 42. The Lactobacilli spp. population in birds supplemented with onion at the level of 30 g/ kg significantly was higher than other groups at 42 d of age (P<0.05). The lowest Escherichia coli loads were detected in broilers fed diets containing 15 mg virginiamycin/kg. The Escherichia coli loads significantly decreased in broilers fed diets containing 10 or 30 g onion/kg (P<0.05). Conclusions: In conclusion, the results of the current study indicated that supplementing broiler diet with 30 g onion/kg could induce favorable influences on performance and ileum microflora composition.

307-310

Authors: Abdolhussein Shakurnia, Abdolhussein Qasemzadeh, Manijeh Afra, Mojtaba Oraki Kohshour

Objective: To identify sero-prevalence of brucellosis among blood donors in Ahvaz city, Southwest Iran. Methods: A total number of 1 450 serum samples from blood donation were collected and were screened for the presence of brucella antibody. Rose Bengal Plate Test, Standard Agglutination Test (SAT), and 2-mercaptoethanol (2ME) agglutination were tested in the sample. SAT dilution ≥1/80 and 2ME agglutination ≥1/40 were considered positive. Results: Sero-prevalence of brucellosis among the blood donors was 0.70%, 0.34%, and 0.20% by Rose Bengal Plate Test, SAT, and 2ME respectively. Conclusions: Considering the 1/80 titer of SAT as the criteria of contamination with brucella, routine screening of sero-prevalence of brucella in blood donors is not recommended in this area.

320-325

Authors: Jayaraj Fakkirappa Chinchali, Basappa Baswanneppa Kaliwal

Objective: To investigate the histopathological changes of mouse mammary tissue damage in dose and durational pathogenicity of Staphylococcus aureus (S. aureus) inoculums. Methods: The minimal tissue inflammation dose of S. aureus inoculum was determined by inoculating the lactating mice mammary gland to observe the mortality rate at 24, 48, 72 and 96 h of infection for each inoculum dilutions. To study dose pathogenicity, lactating mice were divided into one control and four induced groups of 5 mice each. S. aureus inoculums doses of 0.8×101, 1.4 ×102, 2×102 and 2.8×103 colony forming unit were inoculated to the induced groups II, III, IV and V respectively. Mice were sacrificed and mammary gland tissues were harvested after 48 h of infection. Similarly to study of durational pathogenicity, a dose of 2×102 colony forming unit of S. aureus inoculums was inoculated to the induced groups II, III, IV and V respectively. Mice were sacrificed and mammary gland tissues were harvested after 24, 48, 72 and 96 h of infection. Body and mammary gland were weighed and histopathological responses of the tissue damage were observed for both dose and durational pathogenicity’s of S. aureus inoculums. Results: The observations indicated mastitis symptoms, decrease in body and mammary gland weights. Histopathological evidences revealed massive infiltration of polymorphonuclear neutrophil leukocytes, damage of alveoli and secretory products as the doses and durations of S. aureus inoculum increased compared to the control. Conclusions: The present study revealed that damage in mammary gland tissue of mice increased as the dose and duration of S. aureus inoculums increased.

330-335

Authors: Arun Kumar

Objective: To evaluate paraoxonase activity, antioxidant status and lipid peroxidation in hypertensive participants and to address the hypothesis that oxidative modifications of lipids due to hypertension can cause changes in serum paraoxonase activities. Methods: The serum paraoxonase activities, antioxidants and lipid peroxidation were determined in 130 hypertensive participants and 130 age-sexes matched normotensive healthy volunteers served as control. Serum paraoxonase activities were measured by enzymatic kit. The glutathione peroxidase, superoxide dismutase and catalase activity were determined by standard methods. Malondialdehyde was measured by thiobarbituric acid reaction. Conjugated diene level was measured by Recknagel and Glende method. Serum uric acid, total bilirubin, serum albumin, serum ascorbic acid and lipid profile were analyzed by standard methods. Results: Total cholesterol, triglycerides, low-density lipoprotein cholesterol were significantly higher and high-density lipoprotein cholesterol were significantly lower in hypertensive patients when compared to normotensive healthy controls. The superoxide dismutase, glutathione peroxidase and catalase were significantly lower in hypertensive when compared with normotensive. Similar findings were observed in the levels of albumin, uric acid, bilirubin and ascorbic acid when hypertensives were compared with normotensive. The oxidative stress indicators namely malondialdehyde and conjugated diene were significantly higher and paraoxonase activity were significantly lower in hypertensive. Conclusions: Our study concludes that paraoxonase activities are bound to alter in hypertension which is caused due to interplay of several confounding factors namely oxidative stress, increased oxidized low-density lipoprotein and depletion of antioxidants.

342-346

Authors: Hamid Kassiri, Ali Kassiri, Mehdi Pourpolad-Fard, Masoud Lotfi

Objective: To determine the epidemiology of animal bites in in Islamabad-Gharb County, Western Iran during the years 2004-2008. Methods: This is a case series research that explains biting incidents. In this descriptive study, all the data were gathered from each case by experienced personnel. Pending a 5-year course, by a questionnaire, bitten individuals listed into this research. The essential information of animal bite cases, such as age, gender, job, residence site (rural or urban) and bite place on the body were collected. Data were analyzed using descriptive statistics by SPSS version 18. Results: The results of this study showed that the mean incidence rate of animal bites within 2004-2008 was 2.6/1 000 people. The highest incidence was observed in 2004 (3.3/1 000 people). The total number of animal bites was 2 864 cases. From all who were injured by rabid animals, 96.5% were living in rural areas and 3.5% in cities. The cases were mostly males (72.5%). The most highly affected age group was 10-19 years old individuals (26%). Bites were most frequent among students (29.7%). Feet were the most common bite site (75.8%). Nearly 96.7% were bitten by dogs and 2.4% by cats. The maximum frequencies of animal bites were in the summer (29.7%). Conclusions: The findings indicate that biting incidents are common in Islamabad-Gharb County thus control measures are needed to decrease the prevalence rate of this difficulty.

353-359

Authors: Mirza Ali Mofazzal Jahromi, Seyed Younes Hosseini, Mahmood Bozorgmehr, Kazem Baesi, Kayhan Azadmanesh, Zuhair Mohammad Hassan, Seyed Mohammad Moazzeni

Objective: To explore the effects of induction of Gamma-interferon(IFN-γ) production by adenoviral vectors (AdVs) on dendritic cells (DCs) viability and activity. Methods: AdVs were propagated in human embryonic kidney 293 cell lines and DCs were derived from mouse bone marrow using GM-CSF-IL-4-enriched media. The DCs were infected by either AdVs-green fluorescent protein or AdVs-IFN-γ during a 48 h culture. Phenotypic and functional characteristics of AdV-infected DCs were measured by flowcytometry-based methods. Results: Our results displayed that AdVs-IFN-γ could significantly induce DCs necrosis. Furthermore, AdVs-IFN-γ-infected DCs efficiently expressed the CD86 molecules. Conclusions: According to our finding, AdV-IFN-γ is able to affect murine bone marrowderived DC viability and activity.

366-372

Authors: Subhash Pawar, Sanditi Ram Reddy, Lakshmi Kiran Chelluri, Chelluri Eswara Prasad

Objective: To determine the authenticity of three techniques viz., ELISA, western blot and indirect immune fluorescence assay (IIFA) to establish the connection between Helicobacter pylori (H. pylori) and two obstructive airway diseases, chronic obstructive pulmonary diseases (COPD) and asthma. Methods: Serum samples were collected from 48 patients, 27 with COPD, 21 with asthma diseases and control sera were obtained from 42 healthy volunteer blood donors. Serum samples were analyzed by three sero-based techniques, viz., ELISA, western blot and IIFA. Results: ELISA results revealed no connection between the H. pylori infection and COPD and asthma. Western blot results also did not reveal any relationship between H. pylori and obstructive airway diseases. Antibody pattern also did not support the connection between these two diseases. IIFA tests revealed a positive connectivity and relation between the two diseases. Conclusions: The results of the present investigations reveal an association of H. pylori in COPD and asthma. IIFA is a reliable test and hence it is recommended.

378-384

Authors: Shanmukha Ittagi, Vijay Kumar Merugumolu, Ramachandra Setty Siddamsetty

Objective: To investigate the cardioprotective effect of 70% ethanolic extract of Tecoma stans (T. stans) flowers against isoproterenol-induced myocardial infarction in rat myocardium. Methods: Wister rats were pretreated with 70% ethanolic extract of T. stans flowers (250 and 500 mg/kg) orally for 14 d and then intoxicated with isoproterenol [200 mg/(kg · day), s.c.] for 2 consecutive d. The biochemical markers for myocardial infarction such as alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, creatinine kinase, total cholesterol, triglycerides, low density lipoproteins and high density lipoproteins were determined. In addition the antioxidant status on heart tissue is also evaluated by testing the activities of antioxidant enzymes such as lipid peroxidation, superoxide dismutase, reduced glutathione and catalase. Results: The results indicated that pretreatment with 70% ethanolic extract of T. stans flowers prevented fall in antioxidants and retarded elevation of cardiac damage markers in isoproterenol treated rats, significantly. In addition, these findings were evidently supported by the remarkable protection revealed in the histopathological studies, even GC-MS analysis data also substantiated out investigation. Conclusions: It was concluded that, in addition to poly phenolics, some of the phyto fragments found during GC-MS analysis might also contributed to the cardiac protection offered by the extract.

389-394

Authors: Shirisha Rao, Bibechana Timsina, Varalakshmi Kilingar Nadumane

Objective: To evaluate the potential anticancer activity of Origanum majorana (marjoram) (O. marjorana) on the fibrosarcoma cancer cell line HT-1080 through various in-vitro assays. Methods: Crude extracts were prepared from O. marjorana using ethanol, methanol and water as the solvents following standard protocols. Cytotoxicity was assessed using MTT assay, trypan blue dye exclusion, AO/EB staining and fluorescence microscopical analysis and DNA fragmentation analysis. Results: Ethanol extract has shown significant cytotoxicity (P<0.001) to fibrosarcoma and least toxicity to normal human lymphocytes when compared to the controls. Conclusions: This study shows that O. marjorana ethanol extract has anticancer potentials and can be explored further for active component isolation, identification and characterization.

403-410

Authors: Arbind Kumar Choudhary, Rathinasamy Sheela Devi

Objective: To study whether the oral administration of aspartame (40 mg/kg body weight) for 15 d, 30 d and 90 d have any effect on marker enzymes, some selective liver and kidney function parameter, lipid peroxidation and antioxidant status in serum. To mimic human methanol metabolism, folate deficient animals were used. Method: Animal weight, complete hemogram, marker enzyme in serum, some selected serum profile reflect liver and kidney function, plasma corticosterone level, and in serum, lipid peroxidation, nitric oxide, enzymatic and non-enzymatic antioxidant level was measured . Result: After 15 d of aspartame administration animals showed a significant change in marker enzymes, and antioxidant level. However, after repeated long term administration (30 d and 90 d) showed a significant change in some selected serum profile reflects liver and kidney function, along with marker enzymes, and antioxidant level. Conclusions: This study concludes that oral administration of aspartame (40 mg/kg body weight) causes oxidative stress in Wistar albino rats by altering their oxidant/antioxidant balance.

415-420

Authors: Fatemeh Zare Shahneh, Samira Valiyari, Abbas Azadmehr, Reza Hajiaghaee, Ali Bandehagh, Behzad Baradaran

Objective: To investigate the cytotoxic and apoptotic activity of Ferulago angulata, Echinophora platyloba, Salvia officinalis and Chelidonium majus on leukemia and lymphoma cell lines, nonHodgkin’s B-cell lymphoma (Raji), human leukemic monocyte lymphoma (U937), human acute myelocytic leukemia (KG-1A) cell lines and peripheral blood mononuclear cells. Methods: Cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Cell viability assay was done using trypan blue exclusion experiments and cell death was identified as apoptosis using death detection ELISA. Results: Our results demonstrated that the extracts dose and time dependently suppressed the proliferation of three leukemia and lymphoma tumor cell lines (KG-1A, U937 and Raji) with ascending order of IC50 values, while peripheral blood mononuclear cells were not significantly affected. Nucleosome productions in apoptotic KG-1A, U937, and Raji cells were significantly augmented in a time-dependent manner and paralleled the anti-proliferative activity of the extracts. Conclusions: The extracts were found to time- and dose-dependently inhibit the proliferation of KG-1A, U937, and Raji cells possibly via an apoptosis-dependent pathway.

426-428

Authors: Nidhi Singla, Jagdish Chander, Vishal Guglani, Prabhjot Kaur

Objective: To evaluate the prevalence of congenital cytomegalovirus (CMV) infection in a tertiary health care centre of North India. Methods: In the present study over a period of one year and nine months (January 2011 to September 2012), the tests were put up by μ-capture ELISA for CMV IgM and IgG antibodies on the samples suspected of having congenital CMV infection. Results: During the time period, we came across 10 cases among children, who were suspected with having congenital CMV infection. The case histories of these infants were studied retrospectively. Most of them had hepatitis with deranged liver enzyme levels and hepatosplenomegaly. Conclusions: We conclude that congenital CMV is much more prevalent than we think and therefore, clinicians should be on lookout for the disease.

435-436

Authors: Beuy Joob, Viroj Wiwanitkit

Acute dyspepsia is a common gastrointestinal problem in clinical practice. This disease is due to the poor dietary behavior and there is a scientific evidence that the gastric infection by Helicobacter pylori is the cause of this medical disorder. Here, the authors report on the prevalence of Helicobacter pylori seropositivity among the patients presenting with acute dyspepsia in a primary care center.

438-441

Authors: Amiya Kumar Hati, Indranil Bhattacharjee, Goutam Chandra, Abanindranath Nag, Prabal Chaudhuri

Objective: To find out whether relapse can be differentiated in vivax patients. Methods: Sixty three people suffering from vivax were taken and ultrasonography was done. Results: Among 63 persons, previous history of vivax was obtained in 51. Out of remaining 12, previous history of falciparum was found in 5, four had previous history of uncertain malaria and in three, no previous history of malaria was recorded. The liver was within normal size (≤140 mm) in 36 persons, and increased in size in 27 persons, of which enlarged (141-150 mm) in 14 and highly enlarged (beyond 150 mm) in 13 persons. Out of these 27 persons with enlarged livers 22 suffered from vivax previously. Altogether 18 out of 63 cases considered to be suffering from relapsed vivax cases in this study, which is alarming. Conclusions: This simple, noninvasive procedure might provide a clue as to how relapse vivax cases can be determined.

445-448

Authors: Leila Farzin, Mohammad Esmail Moassesi, Fattaneh Sajadi

Objective: To assess the serum antioxidant trace elements selenium (Se), zinc (Zn) and copper (Cu) in cutaneous leishmaniasis patients. Methods: In this study, serum Se, Zn and Cu was determined by using atomic absorption spectrometry in patients with cutaneous leishmaniasis (n=95). The values were statistically compared between patients and control group (n=100) using One-way analysis of variance (ANOVA). Results: Our results showed that there was a significant difference in the values of Se and Zn between two groups (P<0.000 1 and P<0.01, respectively). Meanwhile, no significant difference in level of Cu was observed between patients and healthy subjects (P>0.05). Se and Zn levels were found to be (4.33±1.06) and (70.23±19.12) µg/dL in cutaneous leishmaniasis cases, and these values were found statistically lower compared to the controls (11.10±2.37) and (119.61±26.18) µg/dL, respectively. Conclusions: The observations that host products are released from stimulated leukocytes and could induce metabolic changes similar to an acute-phase response revealed an endocrine role for the immune system. Characteristic changes in trace-mineral metabolism are an integral part of the acute-phase response. The changes are usually reflected in decreased serum Se and Zn concentrations.

453-462

Authors: Prabhakargouda Basanagouda Patil, Shrinivas Venkatesh Kallapur, Vasant Laxmanrao Kallapur, Shankar Narayanrao Holihosur

Objective: To investigate insecticidal properties of organic solvent extracts of Clerodendron inerme (C. inerme) leaves against larval stages of Aedes aegypti (Ae. aegypti) and Culex quinquefasciatus (Cx. quinquefasciatus) mosquito species. Methods: The sundried leaf powder of C. inerme was subjected for extraction using organic solvents viz. methanol, chloroform, petroleum ether and hexane extract, and were tested against third/fourth instar larvae of Ae. aegypti and Cx. quinquefasciatus species in accordance with WHO standard methods. Experiments were conducted in four replicates with control group containing water alone and positive control group containing respective solvent (dimethylsulfoxide/acetone) used for dissolving the extracts. Results: Among the four solvent extracts, hexane extract has effective growth disruptive activity against Ae. aegypti, and showed positive tests for presence of four groups of phytochemical constituents viz. tanin, phytosteriod, terpenoid and cardiac glycoside. The hexane extract was tested against field collected filarial vector Cx. quinquefasciatus larvae for growth disruptive activity. Adult emergence inhibition values for 50 percent suppression (EI50) of the tested population for methanol, chloroform, petroleum ether and hexane extracts treated against third instar larvae of Ae. aegypti were found to be 37.45, 14.79, 2.56 and 1.96 mg/L respectively, while hexane extract treated against Cx. quinquefasciatus was found to be equally effective with EI50 value of 3.74 mg/L. Hexane extract treated against fourth instar larvae of Ae. aegypti and Cx. quinquefasciatus showed EI50 values of 8.07 and 19.55 mg/L respectively in comparison with that of standard insect growth regulator methoprene demonstrating EI50 value of 0.05 mg/L. Besides, the hexane extract was also found to possess toxic effect against non-target organism Gambusia affinis (a bio-control agent), however, the lethal concentration (LC50=172.7 mg/L for 24 h) against Gambusia affinis was much higher in comparison with that of tested concentrations against Ae. aegypti and Cx. quinquefasciatus. Conclusions: We conclude from the present investigation that the hexane extract of C. inerme has positive implications for its consideration in integrated control of dengue/filarial vectors due to their ecological distinctness in comparison with non-target organism and further due to ecofriendly nature of the extract in terms of easy biodegradability.

467-476

Authors: Chandrabhan Seniya, Ghulam Jilani Khan, Richa Misra, Vaibhav Vyas, Shruti Kaushik

Objective: To find the neuraminidase H1N1 inhibition potential of 4-hydroxypanduratin A and its derivatives along with associated binding mechanism through virtual screening and molecular docking. Methods: Initially, the structural templates for neuraminidase proteins were identified from structural database using homology search and performed homology and ab initio modeling to predict native 3D structure using Modeller 9.10 and I-TASSER server, respectively. The reliability of the three-dimensional models was validated using Ramachandran plot. The molecular docking was performed using Autodcok 4.2 and molecular interactions were analyzed through PyMol, Chimera and LigPlot. Results: The neuraminidase protein sequences of ADH29478, ADD85918, AEM62864 (2009) and AFO38701 (2011) from India were modeled and validated. 4-hydroxypanduratin A and its 88 derivatives were docked in to active binding pockets of neuraminidase. The guanidine group of residues Arg152 and Trp179 of ADH29478 (Chennai) and AFO38701 (Gwalior) neuraminidase models present in the hydrophilic domain (-OH and =O groups) was found to have molecular interactions with high binding affinities of -7.40 kcal/mol and -8.66 kcal/mol, respectively to 1-(2,4-dihydroxyphenyl)-3,3-diphenylpropan-1-one (CID: 19875815) than other FDA approved drugs such as oseltamivir, zana-mivir, and peramivir. Conclusions: 1-(2,4-dihydroxyphenyl)-3,3-diphenylpropan-1-one will be a breakthrough for further drug development against swine flu.

481-485

Authors: Minoo Mohraz, Zahra Bayat Jozani, Mohadeseh Behtaj, Shooka Esmaeeli, Seyed Ahmad Seyed Alinaghi, Hossain Jabbari, Asiyeh Ghorbani

Objective: To evaluate the neurological complications among Iranian HIV-positive patients. Methods: This cross-sectional study was conducted among 428 patients diagnosed with HIV infection between 2006 and 2009 at Imam Khomeini hospital, Tehran, Iran. Demographic and clinical variables as well as laboratory tests were extracted and analyzed. Also, another 100 patients refereed to Voluntary Counseling and Testing center of the hospital were visited and evaluated for neurological complications. Results: Among the patients, neurologic manifestations were observed in 34 (7.94%) patients. Twenty three percent of the patients received antiretroviral therapy. Identified causes included brain toxoplasmosis (14.7%), progressive multi-focal leuko encephalopathy (5.9%), HIV encephalopathy (5.9%), TB meningitis (5%) and unknown etiologies (11.8%). Also, among 100 patients who were admitted and visited at the Voluntary Counseling and Testing center, no one was diagnosed for any neurological manifestations. Conclusions: According to our results, toxoplasmosis is the most frequent cause of neurological conditions among Iranian HIV infected patients and should be considered in any HIV/AIDS patient with neurological manifestations.

491-495

Authors: Sunil Kumar, Shalini Sharma, Dinesh Kumar, Karunesh Kumar, Renu Arya

Objective: To confirm the ethnomedicinal claim of Phyllanthus reticulatus Poir as immune enhancer. Methods: The hydroalcoholic extract of fruits and leaves of the plant were evaluated for immunostimulant activity on albino mice at the dose levels of 100 and 200 mg/kg orally. The assessment of immunostimulant activity on specific and non-specific immunity was studied by neutrophil adhesion test, carbon clearance test and cyclophosphamide induced myelosuppression. Cyclophosphamide (30 mg/kg) was used to induce immunosuppresion in mice and levamisole (50 mg/kg) was used as immunostimulating agent. Results: Oral administration of both the doses of fruit and leaf extracts showed significant increase in phagocytic activity (***P<0.001) and the percentage of neutrophil adhesion (**P<0.01). Significant increase in white blood cell (*P<0.05, **P<0.01) count was seen on administration of both the doses of fruits and leaves extracts. Conclusions: The study demonstrates that both the plant parts triggers specific and non-specific responses and thus reveals a promise to hold immunostimulant activity

500-504

Authors: Harresh Adikesavalu, Avijit Patra, Anjan Mondal, Sayani Banerjee, Thangapalam Jawahar Abraham

Objective: To study the association of Aeromonas salmonicida subsp. achromogenes in the haemorrhagic blister of cultured exotic carp Cyprinus carpio in West Bengal, India, its cellular fatty acid composition and antibiotic sensitivity. Methods: The phenotypic characterization and antibiotic sensitivity of bacterial flora of diseased fish were performed. The whole cellular fatty acid composition of Aeromonas salmonicida (A. salmonicida) was determined by MIS Sherlock automatic identification system. Results: Aeromonas hydrophila, A. salmonicida and Pseudomonas alcaligenes were isolated from the haemorrhagic blister. The fatty acid methyl esters pattern confirmed the A. salmonicida strain as A. salmonicida subsp. achromogenes. Unsaturated fatty acid 16:1 w7c/16:1 w6c (39.09%), saturated fatty acid 16:0 (26.84%) and the mono-unsaturated fatty acids 18:1 w7c (8.89%) and 16:1 iso I/14:0 3OH (8.49%) were the most common fatty acids, which accounted for 83.31% of the total fatty acids. A. salmonicida subsp. achromogenes was highly susceptible to broad spectrum antibiotics such as chloramphenicol, ciprofloxacin, co-trimoxazole, gentamycin, nitrofurantoin and oxytetracycline. Conclusions: The finding on the presence of A. salmonicida in carps necessitates molecular level study on establishing the prevalence of this bacterium in Indian aquaculture systems especially on its free-living viable but non-culturable state.