Asian Pacific Journal of Tropical Medicine

239-242

Authors: Yu Zhang, Li-Ke Yu, Ning Xia

To evaluate the diagnostic value of endostatin (ES), vascular endothelial growth factor (VEGF) and carcinoembryonic antigen (CEA) in both serum and pleural effusion of lung cancer patients. Methods: Levels of ES, VEGF and CEA in 52 malignant pleural effusion due to lung cancer and 50 patients with non-malignant disease were measured by using sandwich enzyme-linked immunosorbent assay and microparticle enzyme immunoassay. Results: The ES, VEGF and CEA levels in pleural effusion and serum, and their ratio(F/S) were higher in lung cancer group than that in benign group, and the differences were statistically significant(P<0.05). The diagnostic efficiency of ES+VEGF for lung cancer was superior to either single detection. The diagnostic efficiency of ES+VEGF+CEA was superior to either ES+VEGF or ES+CEA. Conclusions: The results suggest that ES, VEGF and CEA might be useful in the differentiation between benign and malignant pleural effusion due to lung cancer. In comparison with either single determination of concentration in serum or pleural fluid, the combined detection of two or three markers is of important clinical significance in the diagnosis of lung cancer.

680-685

Authors: S Raju, S Kavimani, V Uma Maheshwara rao, K Sreeramulu Reddy, G Vasanth Kumar

Objective To highlight the nephroprotective activity of ethyl acetate extract of dried flowers of Tecoma stans for its protective effects on gentamicin-induced nephrotoxicity in albino rats. Methods For studying acute toxicity study, single oral dose of 5 000 mg ethyl acetate floral extract/kg body weight was administered to albino rats (five females, five males). Nephrotoxicity was induced in albino rats by intraperitoneal administration of gentamicin 80 mg/kg/day for eight days. Effect of concurrent administration of ethyl acetate floral extract of Tecoma stans at a dose of 100, 200 and 300 mg/kg/day given by oral route was determined using serum creatinine, serum uric acid, blood urea nitrogen and serum urea as indicators of kidney damage. The study groups contained six rats in each group. As nephrotoxicity of gentamicin is known to involve induction of oxidative stress, in vitro antioxidant activity and free radical-scavenging activity of this extract was also evaluated. Results For acute toxicity testing both female and male rats administered with the extract at a dose of 5 000 mg/kg. The results showed no toxicity in terms of general behavior change, mortality, or change in gross appearance of internal organs (LD50 > 5 000 mg/kg). It was observed that the ethyl acetate floral extract of Tecoma stans significantly protected rat kidneys from gentamicin-induced histopathological changes. Gentamicin-induced glomerular congestion, peritubular and blood vessel congestion, epithelial desquamation, accumulation of inflammatory cells and necrosis of the kidney cells were found to be reduced in the groups receiving the ethyl acetate floral extract of Tecoma stans along with gentamicin in a dose dependent manner. The floral extract also reduced the gentamicin-induced increase in serum creatinine, serum uric acid, blood urea nitrogen and serum urea levels (P >0.01). Conclusions The present study indicates a very important role of reactive oxygen species (ROS) and the relation to renal dysfunction and point to the therapeutic potential of Tecoma stans in gentamicin induced nephrotoxicity.

689-693

Authors: Shanti Bhushan Mishra, Amita Verma, Alok Mukerjee, M Vijayakumar

Objective To evaluate the antihyperglycemic activity of leaves of Hyptis suaveolens using streptozotocin model. Methods Hyptis suaveolens extract (HSE) 250 and 500 mg/kg body weight was administered orally to streptozotocin induced diabetes, once daily for 21 days. Results A significant reduction in blood glucose was observed in diabetic animals treated with HSE at different doses when compared with diabetic rats. Levels of triglyceride, total cholesterol, low density lipoprotein, very low density lipoprotein were decreased while administering HSE at different doses, compared with their control values in diabetic animals. Conclusions Our results show that HSE possesses significant antihyperglycemic activity which might be attributed to stimulating effects on glucose utilization and antioxidant enzyme.

698-705

Authors: Chinnaperumal Kamaraj, Abdul Abdul Rahuman, Asokan Bagavan, Gandhi Elango, Abdul Abduz Zahir, Thirunavukkarasu Santhoshkumar

Objective To evaluate the larvicidal and repellent activities of ethyl acetate and methanol extracts of Acacia concinna (A. concinna), Cassia siamea (C. siamea), Coriandrum sativum (C. sativum),Cuminum cyminum (C. cyminum), Lantana camara (L. camara), Nelumbo nucifera (N. nucifera) Phyllanthus amarus (P. amarus), Piper nigrum (P. nigrum) and Trachyspermum ammi (T. ammi) against Anopheles stephensi (An. stephensi) and Culex quinquefasciatus (Cx. quinquefasciatus). Methods The larvicidal activity of medicinal plant extracts were tested against early fourth-instar larvae of malaria and filariasis vectors. The mortality was observed 24 h and 48 h after treatment, data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) to kill 50 and 90 per cent of the treated larvae of the tested species. The repellent efficacy was determined against two mosquito species at five concentrations (31.25, 62.50, 125.00, 250.00, and 500.00 ppm) under the laboratory conditions. Results All plant extracts showed moderate effects after 24 h and 48 h of exposure; however, the highest activity was observed after 24 h in the leaf methanol extract of N. nucifera, seed ethyl acetate and methanol extract of P. nigrum against the larvae of An. stephensi (LC50 = 34.76, 24.54 and 30.20 ppm) and against Cx. quinquefasciatus (LC50 = 37.49, 43.94 and 57.39 ppm), respectively. The toxic effect of leaf methanol extract of C. siamea, seed methanol extract of C. cyminum, leaf ethyl acetate extract of N. nucifera, leaf ethyl acetate and methanol extract of P. amarus and seed methanol extract of T. ammi were showed 100% mortality against An. stephensi and Cx. quinquefasciatus after 48 h exposer. The maximum repellent activity was observed at 500 ppm in methanol extracts of N. nucifera, ethyl acetate and methanol extract of P. nigrum and methanol extract of T. ammi and the mean complete protection time ranged from 30 to 150 min with the different extracts tested. Conclusions These results suggest that the leaf and seed extracts of C. siamea, N. nucifera, P. amarus, P. nigrum and T. ammi have the potential to be used as an ideal ecofriendly approach for the control of the An. stephensi and Cx. quinquefasciatus.

711-716

Authors: Siddharthasankar Banerjee, Someshwar Singha, Subrata Laskar, Goutam Chandra

Objective To investigate the role of leaf extract of Limonia acidissima L. (Rutaceae) as a biocontrol agent against the larval form of Culex quinquefasciatus, and characterization of bioactive component responsible for larvicidal activity. Methods Larval mortality of mosquito species was observed after 24, 48 and 72 hours of exposure to different concentrations of aqueous extract, solvent extract and subsequently bioactive compound. The bioactive compound was subjected to IR and GC-MS analysis. Results Mortality rate at 3% concentration of crude extract were highest (90%) amongst all concentrations tested and subsequently highest (95%) mortality was achieved in chloroform: methanol extract at 100 ppm concentrations. IR and GC-MS analysis of bioactive compound revealed the presence of steroid compound which may act as larvicide. Conclusions The chloroform: methanol extract of mature leaves of Limonia acidissima was found to exhibit considerable mosquito larvicidal activity against Culex quinquefasciatus.

722-726

Authors: Bragadeeswaran Subramanian, Thangaraj Sangappellai, Rajiv Chandra Rajak, Balaji Diraviam

Objective To explore the biomedical and pharmacological activity of Paracondactylis indicus (P. indicus), Paracondactylis sinensis (P. sinensis), Heteractis magnifica (H. magnifica) and Stichodactyla haddoni (S. haddoni). Methods The live sea anemones were kept inside the glass bowl along with some amount of distilled water in an ice container for 15 min. During stress condition, nematocysts released from the tentacles were collected and centrifuged at 5 000 rpm for 15 min. The supernatant were collected in separate cleaned beakers for lyophilisation. Results The protein content of crude extracts was 15.2, 28.7, 18.2 and 35.4 μg/mL. In hemolytic assay, the P. indicus was sensitive (16.842 HT/mg) on chicken blood but P. sinensis was less sensitive (1.114 HT/mg) on chicken and goat blood. Whereas H. magnifica and S. haddoni showed hemolysis (0.879, 0.903 HT/mg and 56.263, 0.451 HT/mg) in chicken and goat blood. In antimicrobial assay, the methanol extract of P. indicus showed maximum inhibition zone of 9.7 mm against S. typhii and P. sinensis showed 9.8 mm against K. pneumonia in methanol and ethanol extracts. Whereas the H. magnifica and S. haddoni showed maximum of 10 mm against S. typhii, K. pneumonia in methanol and ethanol extracts. Conclusions The high toxic sea anemones may also contain some biologically active agents which has haemolytic, analgesic and anti-infilamatory activity.

735-738

Authors: S Sukumaran, S Kiruba, M Mahesh, SR Nisha, Paul Z Miller, CP Ben, S Jeeva

Objective To investigate the preliminary phytochemistry and antibacterial activity of the flower extract of Peltophorum pterocarpum . Methods Phytochemical analysis was done by using the standard methods given by Harbone. The methanolic flower extract were tested against Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus cereus, Salmonella typhi, Serratia marsecens, Acinetobacter baumannii, Enterobacter sp., Proteus mirabilis, Enterococcus faecalis and Streptococcus pyogenes by the agar disc diffusion method. Results Preliminary phytochemical screening of flower extract showed the presence of phenolic compounds, flavonoids, saponins, steroids, tannins, xanthoproteins, carboxylic acids, coumarins and carbohydrates. The flower extract of Peltophorum pterocarpum showed significant activity against four gram positive (Staphylococcus aureus, Bacillus cereus, Enterococcus faecalis and Streptococcus pyogenes) and three gram negative bacteria (Proteus mirabilis, Acinetobacter baumannii and Serratia marsecens), out of 12 pathogenic bacteria studied. Conclusions The findings of the present study confirm the presence of significant antibacterial activity against human pathogens in the flowers of Peltophorum pterocarpum.

743-747

Authors: Chidambaram Kumarapppan, Albert Jaswanth, Karpagam Kumarasunderi

Objective To validate traditional claims of usefulness of the Indian plants in management of poisonous snakebite and evaluate the antivenom properties displayed by the alcoholic extracts of Andrographis paniculata (A. paniculata), Crateva magna (C. magna), Gloriosa superba (G. superba) and Hydrocotyle javanica (H. javanica). Methods These plants were collected, identified and the extracts were prepared by using conventional Soxhlet ethanol extraction technique. The venom neutralization activity was accessed in mice (20-25g) and number of mortalities was observed against clinically important snake (Naja nigricollis) venom. Present study also deals with in vitro membrane stabilizing activity of these plants against hyposaline induced human red blood corpuscles (HRBC). Results Extracts of H. javanica and G. superba gave 80 % and 90 % protection to mice treated with minimum lethal dose of venom (LD99). These two plants showed significant neutralization effect against the venoms of Naja nigricollis venom. H. javanica and G. superba (25-100 mg/mL) produced significant changes of membrane stabilization of human red blood cells (HRBC) exposed to hyposaline-induced haemolysis. Conclusions We conclude that probably due to presence of various phytochemicals plays an important role in the anti-venom potential of these Indian medicinal plants against Naja nigricollis venom. The above observations confirmed that A. paniculata, C. magna, G. superba and H. javanica plant extracts possess potent snake venom neutralizing capacity and could potentially be used as an adjuvants for antivenin therapy in case of snakebite envenomation, especially against the local effects of cobra venoms.

756

Authors: Mauro Prato

Malarial pigment does not induce MMP-2 and TIMP-2 protein release by human monocytes

589-593

Authors: M Ahmed, MN Singh, AK Bera, S Bandyopadhyay, D Bhattacharya

Gastrointestinal (GI) parasitism is the most serious constraint throughout the world in small ruminants which causes significant production loss in animals. GI parasites are major contributor to reduce productivity in terms of meat, milk and wool in animals. Control of GI parasite is done primarily by anthelmintic treatment where choice and schedule of treatment is done after identification and quantitation of individual parasite. Identification of GI parasites is done through microscopic method by identifying specific morphological characteristics of egg and larva (L3). Since most of parasite eggs are having similar morphological characteristics, identification up to species level through microscopy is not possible in most of cases. To address this issue, molecular techniques are the viable alternative for identification of species as well as molecular level differences within a species (isolates) of parasites. Different DNA based molecular techniques viz. PCR, AFLP, RAPD, RFLP, PCR-SSCP, real time PCR, DNA microarray etc. have been used for identification and to assess the genetic diversity among parasite population. For identification of species, the characteristic sequence of genomic DNA of different species should differ to allow the delineation of species, but at the same time, no/minor variation within the species should exist. In contrast, for purpose of identifying population variants (strains/isolates), a considerable degree of variation in the sequence should exist within a species. Various target regions, including nuclear ribosomal DNA (rDNA), mitochondrial DNA (mtDNA) or repetitive DNA elements (microsatellite loci), which show considerable variation in the number of repeats within individuals have been employed to achieve the identification of parasites species or strain.

597-599

Authors: Apichat Vitta, Raxsina polseela, Seangchai Nateeworanart, Muncharee Tattiyapong

Objective To survey the Angiostrongylus cantonensis (A. cantonensis) or the rat lungworm in a rat, definitive host, and in a giant African land snail (Achatina fulica), the intermediate host, in Phitsanulok, Thailand. Methods Rats and giant African land snails were captured from Tha Pho sub-district, Phitsanulok, Thailand. Rats were killed and examined for adult A. cantonensis. The artificial digestion method following Baermann technique were used for isolation third stage larvae of A. cantonensis. Results Sixty-two rats were captured and they were identified as Rattus argentiventer, Rattus rattus (R. rattus), Bandicota savilei, and Bandicota indica but only one animal (R. rattus) of 62 rats (1.61%) was positive with adult worm of A. cantonensis. The third stage larvae of A. cantonensis were examined on 307 Angiostrongylus fulica snails. It was found that the overall infection rate was 12.38% (38 infected out of 307 Achatina snails). Conclusions This study demonstrates that A. cantonensis is available in the natural hosts of Phitsanulok. This suggests that the transmissions of this parasite to human may occur in this region.

604-609

Authors: Teimour Hazratian, Yavar Rassi, Mohammad Ali Oshaghi, Mohammad Reza Yaghoobi-Ershadi, Esmael Fallah, Mohammad Reza Shirzadi, Sina Rafizadeh

Objective To investigate species composition, density, accumulated degree-day and diversity of sand flies during April to October 2010 in Azarshahr district, a new focus of visceral leishmaniasis in north western Iran. Methods Sand flies were collected using sticky traps biweekly and were stored in 96% ethanol. All specimens were mounted in Puri's medium for species identification using valid keys of sandflies. The density was calculated by the formula: number of specimens/m2 of sticky traps and number of specimens/number of traps. Degree-day was calculated as follows: (Maximum temperature + Minimum temperature)/2—Minimum threshold. Diversity indices of the collected sand flies within different villages were estimated by the Shannon-weaver formula ( image). Results Totally 5 557 specimens comprising 16 Species (14 Phlebotomus, and 2 Sergentomyia) were indentified. The activity of the species extended from April to October. Common sand-flies in resting places were Phlebotomus papatasi, Phlebotomus sergenti and Phlebotomus mongolensis. The monthly average density was 37.6, 41.1, 40.23, 30.38 and 30.67 for Almalodash, Jaragil, Segaiesh, Amirdizaj and Germezgol villages, respectively. Accumulated degree-day from early January to late May was approximately 289 degree days. The minimum threshold temperature for calculating of accumulated degree-day was 17.32°. According on the Shannon-weaver (H′), diversity of sand flies within area study were estimated as 0.917, 1.867, 1.339, 1.673, and 1.562 in Almalodash, Jaragil, Segaiesh, Amirdizaj and Germezgol villages, respectively. Conclusions This study is the first detailed research in terms of species composition, density, accumulated degree-day and diversity of sand flies in an endemic focus of visceral leishamaniasis in Azarshahr district. The population dynamics of sand flies in Azarshahr district were greatly affected by climatic factors. According to this study the highest activity of the collected sand fly species occurs at the teritary week of August. It could help health authorities to predicate period of maximum risk of visceral leishamaniasis transmission and implement control program.

614-618

Authors: AN Kumar, K Murugan, P Madhiyazhagan, K Prabhu

Objective Midge egg masses are reported to support non-pathogenic strains of the cholera pathogen, Vibrio cholera (V. cholera). Mosquito born diseases have been reported to cause millions of death worldwide. The present research reveals the toxicity effect of spinosad and neem seed kernel extract (NSKE) against different larval stages of Anopheles stephensi (An. stephensi) and Chironomus circumdatus (Ch. circumdatus). Methods The neem seeds were collected from Marudamalai hills, Bharathiar University, Coimbatore, India. Neem seed kernels were powdered, extracted and diluted for different concentrations (2 ppm to 10 ppm). Spinosad was purchased from Kalpatharu pesticide Limited, Coimbatore, Tamil Nadu, India and thoroughly mixed with distilled water to prepare various concentrations (0.01 to 0.08 ppm) and used for bioassay. Results The results depict that spinosad is more toxic in lower concentrations when compared to NSKE and mosquitoes are more susceptible than chironomids. Lethal concentrations were evaluated using the observed mortality. The lowest LC50 value obtained from spinosad against An. stephensi and Ch. circumdatus were 0.002 05 ppm and 0.008 91 ppm. This study investigated on effect of Spinosad and NSKE on the biology of mosquito. The immature stages of both species were susceptible to Spinosad and NSKE. Spinosad and NSKE in individual as well as combined treatment provided additional days in development for mosquitoes. Conclusions The results conclude that Spinosad and NSKE are potential larvicides against An. stephensi and Ch.

624-631

Authors: Subramaniam Ramachandran, Aiyalu Rajasekaran, KT Manisenthil Kumar

Objective To investigate antidiabetic, antihyperlipidemic and antioxidant activity of methanol extract of Tectona grandis (T. grandis) flowers (METGF) in streptozotocin (STZ) induced diabetic rats to supports its traditional use. Methods Acute toxicity study of METGF was carried out in rat to determine its dose for the antidiabetic study. Oral glucose tolerance test (OGTT) was performed to evaluate METGF effect on elevated blood glucose level. Diabetes was induced in rats by administration of STZ (60 mg/kg, ip.) and it was confirmed 72 h after induction. METGF was orally given to the diabetic rats up to 28 days and blood glucose level were estimated each week. On 28 day of the experiment, diabetic rats were sacrificed after the blood collection for the biochemical parameters analysis and liver, kidney was collected to determine antioxidants levels. Results In acute toxicity, METGF did not show toxicity and death up to a dose 2 000 mg/kg in rats. Administration of METGF 100 and 200 mg/kg significantly (P<0.001) reduced blood glucose levels in OGTT and STZ-induced diabetic rats. Both doses of METGF treatment significantly (P<0.001, P<0.01 and P<0.05) increased body weight, serum insulin, haemoglobin (Hb) and total protein levels in diabetic rats. Also, MEGTF treatment reduced elevated glycosylated haemoglobin (HbA1c) and other biochemical parameters levels significantly (P<0.001) in diabetic rats. Altered lipid profiles and antioxidants levels were reversed to near normal in diabetic rats treated with METGF. Conclusions These results concluded that METGF possesses antidiabetic, antihyperglycemic and antioxidant activity which supports its traditional use.

637-641

Authors: M Amzad Hossain, Muhammad Dawood Shah, Mahyar Sakari

Objective To analyse the chemical composition of different extracts of Merremia borneensis (M. borneensis) by gas chromatography-mass spectrometry (GC-MS). Methods The dried leaves powder was extracted with methanol at room temperature by using Soxhlet extractor. Methanol crude extracts of M. borneensis were extrastel with hexane, chloroform, ethyl acetate and butanol. Results Qualitative analyses of various organic crude extracts showed that majority of these are flavonoids, terpeniods, alkaloids and glycosides. Most of the identified compounds by GC-MS are biologically important. Further the M. borneensis leaf possesses certain characteristics that can be ascribed to cultivation on a domestic plantation. Conclusions The suitable extracts for respective compounds can be chosen on the basis of above GC-MS analysis. All the major compounds from different extracts are biologically active molecules. Thus the identification of a good number of compounds from various extracts M. borneensis might have some ecological significance.

645-648

Authors: Krishnan Kannathasan, Annadurai Senthilkumar, Venugopalan Venkatesalu

Objective To study the antibacterial activity of the leaf methanol extracts of five different species of Vitex namely, Vitex altissima (V. altissima), Vitex diversifolia (V. diversifolia), Vitex negundo (V. negundo), Vitex peduncularis (V. peduncularis) and Vitex trifolia (V. trifolia). Methods Antibacterial assay was carried out by using disc diffusion method, determination of minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) against five strains of Gram-positive and seven strains of Gram-negative human pathogenic bacterial strains. Results The results of antibacterial activity of Vitex species showed that the extracts possessed a broad spectrum of antibacterial activity. The V. peduncularis possessed the highest activity against all the microorganisms screened. It produced a zone of inhibition ranged between (11.000 ± 0.577) and (22.670 ± 0.667) mm and the MIC values were from 62.5 to 1 000.0 μg/mL and the MBC values were from 125.0 to 2 000.0 μg/mL. Conclusions Based on the present study, V. peduncularis is recommended for the isolation of antibacterial molecule responsible for the activity against the tested human pathogenic bacterial strains.

654-657

Authors: S Balakumar, S Rajan, T Thirunalasundari, S Jeeva

Objective To assess antifungal activity of Ocimum sanctum leaves against dermatophytic fungi. Methods Antifungal activity of Ocimum sanctum leaves was measured by 38 A NCCLS method. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of various extracts and fractions of Ocimum sanctum leaves were also determined. Results Ocimum sanctum leaves possessed antifungal activity against clinically isolated dermatophytes at the concentration of 200 μg/mL. MIC and MFC were high with water fraction (200 μg/mL) against dermatophytic fungi used. Conclusions Ocimum sanctum has antifungal activity, and the leaf extracts may be a useful source for dermatophytic infections.

661-665

Authors: Abebe Alemu, Yitayal Shiferaw, Gebeyaw Getnet, Aregaw Yalew, Zelalem Addis

Objective To determine the magnitude of opportunistic and non-opportunistic intestinal parasitic infections among HIV/AIDS patients in Bahir Dar. Methods Cross-sectional study was conducted among HIV/AIDS patients attending Gambi higher clinic from April1-May 30, 2009. Convenient sampling technique was employed to identify the study subjects and hence a total of 248 subjects were included. A pre-tested structured questionnaire was used to collect socio-demographic data of patients. Stool samples were examined by direct saline, iodine wet mount, formol-ether sedimentation concentration and modified Ziehl-Neelsen staining technique. Results Out of 248 enrolled in the study, 171(69.0%) (90 males and 81 females) were infected with one or more intestinal parasites. The highest rate of intestinal parasites were observed among HIV/AIDS patients (80.3%, 151/188), and the infection rate of HIV negative individuals was 33.3% (20/60). Cryptosporidum parvum (43.6%), Isospora belli (15.5%) and Blastocystis hominis (10.5%) were opportunistic parasites that were found only in HIV/AIDS patients. Conclusions Opportunistic parasite infections are common health problem among HIV/AIDS patients in the study area. Therefore, early detection and treatment of these parasites are important to improve the quality of life of HIV/AIDS patients.

669-671

Authors: Weihong Li, Yu Guo, Haiyan Niu, Song Jin, Li Wang

Objective To explore the application of cervical cancer screening system, TruScreen in detecting atypical squamous cell of undetermined significance (ASCUS) patients. Methods A total of 42 cases were selected, who were diagnosed as ASCUS by thinprep cytologic test (TCT). Area from site 15 to 20, site 21 to 32 were detected by TruScreen. And the result was compared with those of cases which had positive pathological result of cervical biopsy. Results There were 16 cases with abnormal pathological result in ASCUS cases, including 6 cases with cervical intraepithelial neoplasia (CIN) I, 6 cases with CIN II, 3 cases with CIN III and 1 case with infiltrating carcinoma. The consistency between TCT and pathological test was 38.10% (16/42). The positive rate of TruScreen at site 15-20 was 61.91% (26/42). There was significant difference in consistency with pathological test between TCT and TruScreen (x2=4.762, P=0.029). The positive rate of TruScreen at site 21-32 was 66.67% (28/42)(Kappa=0.181, P=0.016). There was significant difference in consistency with pathological test between TCT and TruScreen (x2=9.4919, P=0.002). And no case was missed when site 21-32 of patients with CINII and above were detected by TruScreen. Conclusions TruScreen is effective in detecting ASCUS patients.

505-509

Authors: M Govindarajan, R Sivakumar, A Amsath, S Niraimathi

Objective To determine the larvicidal efficacy of different solvent leaf extract of Ficus benghalensis (F. benghalensis) against Culex tritaeniorhynchus (Cx. tritaeniorhynchus) and Anopheles subpictus (An. subpictus). Methods Twenty five early third instar larvae of Cx. tritaeniorhynchus and An. subpictus were exposed to various concentrations and were assayed in the laboratory by using the protocol of WHO 2005. The larval mortality was observed after 24 h of treatment. Results Among three solvent extracts tested the maximum efficacy was observed in the methanol extract. The LC50 and LC90 values of F. benghalensis against early third instar of Cx. tritaeniorhynchus and An. subpictus were 100.88, 159.76 ppm and 56.66, 85.84 ppm, respectively. No mortality was observed in controls. The chi-square values were significant at P<0.05 level. Conclusions From the results it can be concluded the crude extract of F. benghalensis was an excellent potential for controlling Cx. tritaeniorhynchus and An. subpictus mosquito larvae.

514-522

Authors: Omeje Edwin Ogechukwu, Osadebe Patience Ogoamaka, Nworu Chukwuemeka Sylvester, Akira Kawamura, Peter Proksch

Objective To provide further evidence for the ethnomedicinal use of the Eastern Nigeria mistletoe, Loranthus micranthus (L. micranthus), as an immunostimulant. Methods Solvent fractions from the crude extract of the mistletoe plant was obtained and screened by the cell mediated delayed type hypersensitivity reaction (DTHR) model in mice. Then the immunomodulatory potentials of a major lupane triterpenoid ester isolated from an active hexane fraction of the Eastern Nigeria mistletoe was investigated. Three lupeol-based triterpenoid esters: 7β 15α-dihydroxyl-lup-20(29)-ene-3β-palmitate (I), 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-stearate (II) and 7β, 15α-dihydroxyl-lup-20(29)-ene-3β-decadecanoate (III) were isolated from the plant leaves epiphyting on a local kola nut tree and were characterized. Compound 1 was subjected to cell proliferation studies using C57Bl/6 splenocytes at three dose levels (5, 25 and 100 μg/mL) in presence of controls. Furthermore, the effect of this compound on IL-8 receptor expression was evaluated at three doses (1, 5 and 10 μg/mL) using the real time polymerase chain reaction assay. Results This triterpenoid ester produced some enhancement of the splenocytes at the tested doses but at doses higher than 5 μg/mL caused inhibition of the IL-8 receptor expression. Conclusions The present findings support the ethnomedicinal use of the Eastern Nigeria Mistletoe in the management of diseases affecting the immune system. The triterpenoid(s) have some immunomodulatory abilities on splenocytes and IL-8 receptors and may partly account for the overall immunomodulatory activity of this plant.

526-529

Authors: Vishal Gulecha, Thangavel Sivakuma

Objective To investigate anticancer activity of different fractions of Tephrosia purpurea [TP] (Sharapunkha, Fabaceae) and Ficus religiosa [FR] (Peepal, Moraceae). Methods The fractions of TP and FR were prepared and tested for in vitro anticancer activity using human MCF 7 cell line by trypan blue exclusion method. Results The result showed that among all these fractions of TPI, TPIII, FRI and FRIII showed better anticancer activity compared to other fractions. The IC50 value for TPI (152.4 μM), TPIII (158.71 μM), FRI (160.3 μM) and for FRIII (222.7 μM) was observed. Conclusions The present study shows anticancer potential of TP and FR fractions in MCF 7 cell line.

535-540

Authors: Kamol Suwannakarn, Apiradee Theamboonlers, Yong Poovorawan

Objective To understand the epidemiology of the East, Central and South African (ECSA) genotype of Chikungunya virus (CHIKV) in terms of emerging and re-emerging infections, this study has been aimed at investigating the evolutionary parameters, genomic signatures and molecular tracking of the CHIKV ECSA genotype in South-east Asia and coastal areas of the Indian Ocean between 2006 and 2009 by using phylogenetic analysis and the Bayesian Markov Chain Monte Carlo (BMCMC) evolutionary estimation. Methods Nearly complete genome sequences of 53 CHIKV isolates from all genotypes were subjected to phylogenetic analysis and evolutionary parameter estimation. The amino acids of 67 of ECSA genotype during 2006 to 2009 were compared for finding molecular signature tracking. The ECSA genotype signatures were visualized to find the possible transmission root was projected onto a geographic map. Results Phylogenetic analysis showed the ECSA genotype was divided into 2 groups. The first group comprises viruses from India and Southeast Asian countries. The second group consists of strains typically circulating in Sri Lanka in 2008. The evolutionary parameters of these groups depicted the time of the most recent common ancestor at approximately 7.5 years ago. The genomic signatures revealed the positions of amino acid variation in each group. Conclusions The molecular evolution projected onto a geographical map showed the routes of CHIKV transmission from 2006 to 2009. Molecular tracking will assist in understanding transmission routes, epidemiology and molecular evolution of CHIKV.

547-549

Authors: Xiaoning Sun, Cheng Lan, Yu An, Ye Sun

Objective To investigate the role of Runx3 protein and TGF-β1 in the pathogenesis of irritable bowel syndrome (IBS), as well as the correlation of these two proteins. Methods Colonic tissue was collected from patients with IBS and normal persons. The colonic expression of Runx3 protein and TGF-β1 was detected with immunohistochemistry method. Semi-quantitative analysis was used to evaluate the staining degree of these two proteins. Results Compared with their counterparts, patients with IBS did not show any changes in the colonic expression of Runx3 protein and TGF-β1 (P>0.05). Interestingly, there was a significant correlation between Runx3 protein and TGF-β1 in patients with IBS(P<0.05). Conclusions The role of Runx3 protein and TGF-β1 in the pathogenesis of IBS remains to be further studied.

556-559

Authors: Suchun Yan, Yiqiang Xie, Bide Zhu, Yingguang Han, Wei Guo

Objective To compare effect of different formulation of Dang-Gui-Bu-Xu-Tang(DGBXT) on myelosuppression mouse. Methods HPLC was used to measure active ingredients of two DGBXT formulations. Hemopoesic function of bone marrow was measured by hemopoietic progenitor cell culture and peipheral blood count. And hemopoietic factors in bone marrow were tested by ELISA. Results The content level of astragaloside A in granule formulation was higher than that in decoction and the content ratio of active ingredient was close to 5 : 1. Two DGBXD formulations could significantly improve amount of peripheral blood cells, and bone marrow cells of bone marrow suppression mice (P<0.05). DGBXD granule formulation significantly increased the colony quantity of all progenitor cell lines and amount of G2/M and S phase cells (P<0.05). It also significantly decreased amount of G0/G1 phase cells in the bone marrow and was more effective (P<0.05). Conclusions DGBXT decoction and the granule formulation all can improve the hematopoietic function of bone marrow suppression mouse. They can improve quantities of peripheral blood and nucleated bone marrow cells, and yield of the hematopoietic stem/progenitor cells in vitro colony; balance the expression of cytokine (EPO, TPO and GM-CSF) in bone marrow microenvirement. They can also facilitate hematopoietic stem/progenitor cells to enter the cell cycle. And the effect of granule formulation is more satisfactory.

564-567

Authors: Keerthi Kularatne, Sudhara Budagoda, Kalana Maduwage, Kamal Naser, Rangith Kumarasiri, Senanayake Kularatne

Objective To describe the epidemiology of snake bite in the region and attempt to compare proven Russell's viper with hump-nosed viper bites. Methods All snake bite admissions to the Toxicology Unit of Teaching Hospital Peradeniya over three year from 2006 were included. Results Of the 776 snakebites, 665(86%) were unidentified and non-envenomed. Hump-nosed viper and Russell's viper accounted for 55(7%) and 40(5%) bites respectively, of them, incriminated snakes were found in 36(65%) and 19(48%) cases. The cobra bites-5, krait bites-0. The median ages: Russell's viper bites-41(range 16-66), hump-nosed viper bites-42(range 15-75). The gender incidence, time of bite (>58% daytime) were similar. In hump-nosed viper bite; upper limb involved in 13(36%), happened at home garden in 22(61%), none in paddy fields. In Russell's viper bite; 6(33%) occurred in paddy fields. Dry bites were similar at 5%. In hump-nosed viper bite: local effects 94%, coagulopathy 3%, acute renal failure 3% and one patient died. In Russell's viper bite; local effects 84%, coagulopathy 53%, neurotoxicity 21%. Abdominal pain occurred only in Russell's viper bites 10(53%). Conclusions Overwhelming numbers of unidentified, non-envenomed snakebites are common in the central hills. Some distinctive differences were observed between Russell's viper and hump-nosed viper bites.

573-580

Authors: Shyamapada Mandal, Manisha Deb Mandal, Nishith Kumar Pal

Cholera, caused by the infection of toxigenic Vibrio cholerae (V. cholerae) to humans, is a life threatening diarrheal disease with epidemic and pandemic potential. The V. cholerae, both O1 and O139 serogroups, produce a potent enterotoxin (cholera toxin) responsible for the lethal symptoms of the disease. The O1 serogroup has two biotypes (phenotypes), classical and El Tor; each of which has two major serotypes (based on antigenic responses), Ogawa and Inaba and the extremely rare Hikojima. V. cholerae O1 strains interconvert and switch between the Ogawa and Inaba serotypes. Fluid and electrolyte replacement is the mainstay of treatment of cholera patients; the severe cases require antibiotic treatment to reduce the duration of illness and replacement of fluid intake. The antibiotic therapy currently has faced difficulties due to the rapid emergence and spread of multidrug resistant V. cholerae causing several outbreaks in the globe. Currently, cholera has been becoming endemic in an increasing number of geographical areas, reflecting a failure in implementation of control measures. However, the current safe oral vaccines lower the number of resistant infections and could thus represent an effective intervention measure to control antibiotic resistance in cholera. Overall, the priorities for cholera control remain public health interventions through improved drinking water, sanitation, surveillance and access to health care facilities, and further development of safe, effective and appropriate vaccines. Thus, this review describes the facts and phenomena related to the disease cholera, which is still a great threat mainly to the developing countries, and hence a grave global concern too.

587-588

Authors: Sunita Singh, Sonia Chhabra, Garima Aggarwal, Rajnish Kalra, Amrita Duhan, Rajeev Sen

Cysticercosis is a condition that occurs when man is infected with larvae of Taenia solium. Oral cysticercosis is a rare event, and it represents difficulty in clinical diagnosis. A case of oral cysticercosis in 11 year old girl is presented which complained of painless swelling for 6 months. A FNAC was performed which revealed bluish pink fibrillary material and interspersed nuclei and fragments of wall of larvae. Patient was treated with antihelminthic.

421-425

Authors: Fengying Huang, Qiuping Meng, Guanghong Tan, Yonghao Huang, Hua Wang, Wenli Mei, Haofu Dai

Objective To analysis and identify a bacterium strain isolated from laboratory breeding mouse far away from a hospital. Methods Phenotype of the isolate was investigated by conventional microbiological methods, including Gram–staining, colony morphology, tests for haemolysis, catalase, coagulase, and antimicrobial susceptibility test. The mecA and 16S rRNA genes were amplified by the polymerase chain reaction (PCR) and sequenced. The base sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank database by phylogenetic analysis and multiple sequence alignment. Results The isolate in this study was a gram positive, coagulase negative, and catalase positive coccus. The isolate was resistant to oxacillin, methicillin, penicillin, ampicillin, cefazolin, ciprofloxacin erythromycin, et al. PCR results indicated that the isolate was mecA gene positive and its 16S rRNA was 1 465 bp. Phylogenetic analysis of the resultant 16S rRNA indicated the isolate belonged to genus Saphylococcus, and multiple sequence alignment showed that the isolate was Saphylococcus haemolyticus with only one base difference from the corresponding 16S rRNA deposited in the GenBank. Conclusions 16S rRNA gene sequencing is a suitable technique for non–specialist researchers. Laboratory animals are possible sources of lethal pathogens, and researchers must adapt protective measures when they manipulate animals.

430-435

Authors: Sarita Kumar, Anita Thomas, MKK Pillai

Objective To evaluate the effects of deltamethrin against field-collected adults of Aedes aegypti L. (Ae. aegypti). Methods The adults were selected with 0.025% deltamethrin for 40 successive generations. The selected adults were tested with 4% DDT and the emerging larvae were tested with various insecticides to study the cross-resistance spectrum. The knockdown and irritability studies were carried out in adult mosquitoes to investigate their behavioural response to deltamethrin. Results Forty generations of selection with deltamethrin resulted in only 3.8—fold resistance in the adults of Ae. aegypti. The adults of parent (PS) and selected strains (DAS) exhibited only 0.8-fold cross resistance to 4% DDT. The larvae emerging from the PS and DAS strains did not develop appreciable levels of resistance to various insecticides tested. The knockdown studies revealed KT50 of 14.4 min in PS adults with no signs of recovery even after 24 h. The DAS strains could develop only 1.2 to 1.3-fold knockdown resistance (KDR). The knockdown response of DDT was though 5–6 times slower than deltamethrin but the continued response in deltamethrin-selected adults caused only 1.2-fold KDR. The PS and DAS strains exhibited significant irritability response towards deltamethrin and DDT. The DAS strains showed 5–6 fold increased irritability to deltamethrin as compared to the PS strain. Conclusions The above results suggest the prolonged effective use of deltamethrin against Ae. aegypti as an adulticide.

442-446

Authors: Jude E Okokon, Paul A Nwafor, Ukeme E Andrew

Objective To evaluate antiplasmodial and analgesic activities of ethanolic leaf extract/fractions of Panicum maximum. Methods The crude leaf extract (47–190 mg/kg) and fractions (chloroform, ethyl acqeous and methanol; 96 mg/kg) of Panicum maximum were investigated for antiplasmodial activity against chloroquine sensitive Plasmodium berghei infections in mice and for analgesic activity against chemical and heat-induced pains. The antiplasmodial activity during early and established infections as well as prophylactic were investigated. Artesunate at 5 mg/kg and pyrimethamine at 1.2 mg/kg were used as positive controls. Analgesic activity of the crude extract/fractions was also evaluated against acetic acid, formalin and heat-induced pains. Results The extract and its fractions dose-dependently reduced parasitaemia induced by chloroquine sensitive Plasmodium berghei infection in prophylactic, suppressive and curative models in mice. These reductions were statistically significant (P<0.001). They also improved the mean survival time from 13 to 28 days compared with control (P<0.001). The activities of extract/fractions were incomparable to that of the standard drugs (Artesunate and pyrimethamine). On chemically and thermally-induced pains, the extract inhibited acetic acid and formalin-induced inflammation as well as hot plate-induced pain in mice. These inhibitions were statistically significant (P<0.001) and in a dose-dependent fashion. Conclusions Panicum maximum leaf extract has antiplasmodial and analgesic activities which may in part be mediated through the chemical constituents of the plant.

451-456

Authors: Gollapalle Lakshminaraya Shastry Viswanatha, Shylaja Hanumanthappa, Nandakumar Krishnadas, Srinath Rangappa

Objective To evaluate antidiarrheal activity of the fractions of aqueous extract from stem barks of Thespesia populnea (Malvaceae). Methods From the aqueous extract three fractions namely ethyl acetate fraction (EAF), methanolic fraction (MF) and residue fraction (RF) were made and studied for antidiarrheal activity. Antidiarrheal activity of the fractions were evaluated in castor oil induced diarrhea, prostaglandin E2 (PG-E2) induced diarrhea and charcoal meal test as in vivo models and the most potent fraction was further evaluated with in vitro models to determine the possible antimotility effect. Results In castor oil induced diarrhea model, the RF (10, 25, 50 and 100 mg/kg, po.) and MF (100 mg/kg, po.) has significantly reduced the cumulative wet faecal mass, where as the EAF have not shown any significant antidiarrheal activity, RF was found to be more potent than MF. Based on these results and percentage yield, only RF was evaluated in PG-E2 induced enteropooling and charcoal meal test. RF (10, 25 and 50 mg/kg) had shown significant inhibition of PG-E2 induced secretions (antisecretory) and decreased the movement of charcoal in charcoal meal test indicating its antimotility activity. Furthermore, RF has showed significant inhibition of acetylcholine, histamine and BaCl2 induced contractions on rat colon, guinea pig ileum and rabbit jejunum with EC50 values of 241.7, 303.1 and 286.1 μg/mL, respectively indicating the antimotility effect of RF. The phytochemical analysis of RF showed presence of gums and mucilages and the possible mechanism may be the combination inhibition of elevated prostaglandin biosynthesis and reduced propulsive movement of the intestine. Conclusions RF possesses good antidiarrheal activity comparing with other two fractions and the possible mechanism thought to be associated with combination of antisecretory and antimolity.

462-465

Authors: Murugesan Gnanadesigan, Sundaram Ravikumar, Samuel Jacob Inbaneson

Objective To identify the hepatoprotective and antioxidant activity of Luminetzera racemosa (L. racemosa) bark extract. Methods Wistar albino rats were divided into 6 groups: Group 1 served as control; Group 2 served as hepatotoxin (CCL4 treated) group; Group 3 served as positive control (Silymarin) treated groups; Group 4, 5 and 6 served as (100, 200 and 300 mg/kg bw p.o.) L. racemosa bark extract treated groups. Moreover, in vitro antioxidant indexes, including DPPH, hydroxyl radical scavenging activity (HRSA), NO, ferric reducing antioxidant power (FRAP), lipid hydroperoxide (LPO) and super oxide dismutase (SOD) were also analyzed in the bark extract. Results The results suggested that, the level of serum glutamate oxyloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatise (ALP), bilurubin, cholesterol, sugar and lactate dehydrogenase (LDH) were significantly (P<0.05) increased in hepatotoxin treated rats when compared with the control group. But, the maximum reduction of SGOT [(225.36±13.65) IU/L], SGPT [(96.85±17.36) IU/L], ALP [(315.37±17.16) IU/L], bilirubin [(2.97±0.46) mg/dL], cholesterol [(163.73±17.54) mg/dL], sugar [(127.35±27.35) mg/dL] and LDH [(1 784.00±268.36) IU/L] were observed with 300 mg/kg bw of bark extract treated rats. Histopathological scores showed that, no visible changes were observed with high dose (300 mg/kg bw) of bark extract treated rats except mild fatty changes. The in vitro antioxidant assays showed that, the IC50 values were observed as (44.17±6.87) μg/mL, (42.45±2.81)μg/mL, (62.37±3.98)μg/mL, (54.24±3.09)μg/mL, (87.25±5.90) μg/mL and (71.54±5.42)μg/mL for DPPH, HRSA, NO, FRAP, LPO and SOD radical scavenging activities, respectively. Conclusions The hepatoprotective and antioxidant activities of the bark extract might be to the presence of unique chemical classes such as flavonoids, alkaloids and polyphenols.

470-474

Authors: Nilesh Kumar Jain, Abhay K Singhai

Objective To investigate and compare the hepatoprotective effects of crude ethanolic and aqueous extracts of Phyllanthus acidus (L.) Skeels (P. acidus) leaves on acetaminophen (APAP) and thioacetamide (TAA) induced liver toxicity in wistar rats. Silymarin was the reference hepatoprotective agent. Methods In two different sets of experiments, the P. acidus extracts (200 and 400 mg/kg, body weight) and silymarin (100 mg/kg, body weight) were given orally for 7 days and a single dose of APAP (2 g/kg, per oral) or TAA (100 mg/kg, subcutaneous) were given to rats. The level of serum aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin and total protein were monitored to assess hepatotoxicity and hepatoprotection. Results APAP or TAA administration caused severe hepatic damage in rats as evident from significant rise in serum AST, ALT, ALP, total bilirubin and concurrent depletion in total serum protein. The P. acidus extracts and silymarin prevented the toxic effects of APAP or TAA on the above serum parameters indicating the hepatoprotective action. The aqueous extract was found to be more potent than the corresponding ethanolic extract against both toxicants. The phenolic and flavonoid content (175.02±4.35 and 74.68±1.28, respectively) and 2,2-diphenyl-1-picrylhydrazil (DPPH) [IC50 = (33.2±0.31)μg/mL] scavenging potential was found maximum with aqueous extract as compared to ethanolic extract. Conclusions The results of present study suggests that the aqueous extract of P. acidus leaves has significant hepatoprotective activity on APAP and TAA induced hepatotoxicity, which might be associate with its high phenolic and flavonoid content and antioxidant properties.

478-481

Authors: CH Rasul, MA Bakar, AA Mamun, MS Siraz, RU Zaman

Objective To determine the magnitude and outcome of influenza in southern part of Bangladesh and also to identify intrusion of novel influenza virus. Methods This study was conducted for two years (2008–2009) in outpatient and inpatient department of both paediatrics and medicine discipline of Khulna Medical College Hospital. Nasal and throat swab specimens were collected from each influenza like illness (ILI) case and kept together in a virus transport media and transported to international centre for diarrhoeal disease and research, Bangladesh laboratory. Influenza virus was detected by rRT-PCR including types and subtypes. Results A total of 526 patients were enrolled during the study period with the mean (SD) age of 19 (17) years. Identification of influenza virus was 14% and positive cases were mostly found in under-five children (24%). The vast majority (88%) of influenza cases were found during April to September. Virus detection rate was higher in inpatient department (IPD) (21%) than in outpatient department (12%). The dominant virus type and subtype was influenza A (87%) and H3 (49%), respectively. Type A was significantly higher than type B in causing severe illness compared to ILI (OR=7.04, 95% CI: 2.76–18.7, P<0.01). Headache and chest indrawing was found in significantly higher proportion (P<0.05) in influenza positive IPD cases. Among 31 hospitalized cases majority (77%) recovered completely except two cases that needed referral for additional support. Conclusions Influenza mostly affected under-five children and young adult. The peak season here was late summer and rainy season.

486-491

Authors: C Thenmozhi, R Sankar, V Karuppiah, P Sampathkumar

Objective To isolate marine bacteria, statistically optimize them for maximum asparaginase production. Methods In the present study, statistically based experimental designs were applied to maximize the production of L-asparaginase from bacterial strain of Bacillus cereus (B. cereus) MAB5 (HQ675025) isolated and identified by 16S rDNA sequencing from mangroves rhizosphere sediment. Results Plackett-Barman design was used to identify the interactive effect of the eight variables viz. yeast extract, soyabean meal, glucose, magnesium sulphate, KH2PO4, wood chips, aspargine and sodium chloride. All the variables are denoted as numerical factors and investigated at two widely spaced intervals designated as −1 (low level) and +1 (high level). The effect of individual parameters on L-asparaginase production was calculated. Soyabean meal, aspargine, wood chips and sodium chloride were found to be the significant among eight variables. The maximum amount of L-asparaginase produced (51.54 IU/mL) from the optimized medium containing soyabean meal (6.282 8 g/L), aspargine (5.5 g/L), wood chips (1.383 8 g/L) and NaCl (4.535 4 g/L). Conclusions The study revealed that, it is useful to produce the maximum amount of L-asparaginase from B. cereus MAB5 for the treatment of various infections and diseases.

498-504

Authors: Ahmad Ali Hanafi-Bojd, Shahyad Azari-Hamidian, Vatandoost Hassan, Charrahy Zabihollah

Malaria is a main vector-borne public health problem in Iran. The last studies on Iranian mosquitoes show 31 Anopheles species including different sibling species and genotypes, eight of them are reported to play role in malaria transmission. The objective of this study is to provide a reference for malaria vectors of Iran and to map their spatial and temporal distribution in different climatic zones. Shape files of administrative boundaries and climates of Iran were provided by National Cartographic Center. Data on distribution and seasonal activity of malaria vectors were obtained from different sources and a databank in district level was created in Excel 2003, inserted to the shape files and analyzed by ArcGIS 9.2 to provide the maps. Anopheles culicifacies Giles s.l., Anopheles dthali Patton, Anopheles fluviatilis James s.l., Anopheles maculipennis Meigen s.l., Anopheles sacharovi Favre, Anopheles stephensi Liston, and Anopheles superpictus Grassi have been introduced as primary and secondary malaria vectors and Anopheles pulcherrimus Theobald as a suspected vector in Iran. Temporal distribution of anopheline mosquitoes is restricted to April–December in northern Iran, however mosquitoes can be found during the year in southern region. Spatial distribution of malaria vectors is different based on species, thus six of them (except for Anopheles maculipennis s.l. and Anopheles sacharovi) are reported from endemic malarious area in southern and southeastern areas of Iran. The climate of this part is usually warm and humid, which makes it favorable for mosquito rearing and malaria transmission. Correlation between climate conditions and vector distribution can help to predict the potential range of activity for each species and preparedness for malaria epidemics.

337-341

Authors: VU Omoja, AO Anaga, IR Obidike, TE Ihedioha, PU Umeakuana, LI Mhomga, IU Asuzu, SM Anika

Objective To investigate the effects of combination therapy of methanolic leaf extract of Azadirachta indica (A. indica) and diminazene diaceturate (DDA) in the treatment of experimental Trypanosoma brucei brucei (T. brucei brucei) infection in rats. Methods Acute toxicity study of the drug and extract combinations were done. Selection of the best drug and extract combinations was carried out using fifty four rats of both sexes separated into 9 groups. Three dose combinations were derived from selection of the best drug and extract combinations used for the final study viz: 7 mg/kg bw DDA plus 125 mg/kg bw extract (group B), 3.5 mg/kg bw DDA plus 250 mg/kg bw extract (group C), and 1.8 mg/kg bw DDA plus 500 mg/kg bw extract (group D). The final study had in addition to the three groups derived from the dose response study, four other groups viz: uninfected untreated negative control (group F), infected and treated with 3 000 mg/kg bw extract alone (group E), infected and treated with 7 mg/kg bw DDA alone (group A), and infected untreated positive control (group G). The parameters assessed were onset of parasitaemia (OP), level of parasitaemia (LOP), clearance of parasites post treatment (COPPT), relapse infection period (RIP), post infection survival period (PIST). Results There was no significant difference in OP between the groups (P < 0.05). One day post treatment, the mean LOP of groups A, B, and C were found to be significantly lower than that of group D which in turn was lower than that of group E and G respectively. The mean LOP of group E was significantly lower than group G two days post treatment and this trend continued throughout the experimental period. Mean COPPT of group D was significantly longer than that of groups A, C and B. There was no significant difference in the mean COPPT among groups B, C and A. The mean RIP of group D was significantly shorter than group C, and that of group C was significantly shorter than that of group A. There was no relapse of infection in group B. The PIST of group E did not differ significantly from group G. Conclusions This experiment stands to conclude that combination of 125 mg/kg bw extract and 7 mg/kg bw DDA is very effective in the treatment of trypanosomosis, caused by T. brucei. This combination therapy proved to be better than single therapy of DDA.

347-352

Authors: Sunil Kumar, Vipin Kumar, Om Prakash

Objective To investigate antidiabetic, hypolipidemic histopathological analysis of Dillenia indica (D. indica) methanolic leaves (DIME) extract in alloxan induced diabetic rat by administering oral doses (250 and 500 mg/kg body weight). Methods Blood glucose levels were measured using blood glucose test strips with elegance glucometer on weekly intervals till the end of study (i.e. 3 weeks). Other parameters e.g. liver profile, renal profile and total lipid levels were determined in normal and alloxan induced diabetic rats after oral administration of the extract for 21 days. Histopathological changes in diabetic rat organs (pancreas, liver and kidney) were also observed after extract treatment. Results Daily oral administration DIME (250 and 500 mg/kg body weight) and glibenclamide (10 mg/kg) showed beneficial effects on blood glucose level (P < 0.001) as well as improving kidney, liver functions and hyperlipidaemia due to diabetes. The extract treatment also showed to enhanced serum insulin level and body weight of diabetic rats as compared to diabetic control group. Furthermore, the extract has a favorable effect on the histopathological changes of the pancreas, liver and kidney in alloxan induced diabetes. Conclusions D. indica possess antidiabetic property as well improve body weight, liver profile, renal profile and total lipid levels. DIME has also favorable effect to inhibit the histopathological changes of the pancreas and kidney in alloxan induced diabetes.

357-362

Authors: MJ Mendki, AK Sharma, Vijay Veer, OP Agrawal, Shri Prakash, BD Parashar

Objective To characterize the genetic structure of various populations of Culex quinquefasciatus (Cx. quinquefasciatus) from India representing different geoclimatic locations. Methods Inter simple sequence repeat (ISSR) markers were used. A set of 20 primers were screened with the laboratory populations of mosquito species. Finally the IS 40 primer was chosen based on the scorable banding pattern showing 100 percent polymorphism among the various populations. The statistical analysis was done using POPGENE 1.31 software. The consensus tree was generated based on UPGMA modified from NEIGHBOR procedure of PHYLIP Version 3.5. Results The cluster analysis shows the main cluster which is divided into two sub cluster representing all the populations separated as per their phylogeographic and geoclimatic condition. Conclusions The findings will be helpful in understanding the population variation under different ecological conditions and development of effective vector management strategies.

367-370

Authors: Vinod Kumar, Jalaj Kumar Gour, Surabhi Bajpai, Manish Mishra, Rakesh Kumar Singh

Objective To detect leishmanial antigens in pre and post treated urine of visceral leishmaniasis (VL) patients. Methods Urine and serum sample from three VL patients were collected. Ammonium sulphate precipitation and purification of urine sample was done for proteins isolation. SDS PAGE of proteins was done followed by western blotting, with the patient's pre and post treatment serum. Results Eight proteins of molecular weights 17 kDa, 25 kDa, 28 kDa, 42 kDa, 47 kDa, 54 kDa, 60 kDa and 85 kDa were detected in the urine of VL patients before treatment. After treatment with miltefosine, none of the above proteins was detected in urine samples. The western blot analysis with pre treatment serum confirmed the antigenicity of four urinary proteins of molecular weights 25 kDa, 28 kDa, 54 kDa and 60 kDa. The seropositivity with 25 kDa and 28 kDa antigens was negative with serum obtained after the completion of treatment. Conclusions In the context to unavailability of a prognostic tool, urinary leishmanial antigens may offer a better choice and may also be useful as immunoprophylactic candidates.

375-378

Authors: Rajendran Darling Anpin Raja, Solomon Jeeva, Juststella Wilfred Prakash, Johnson Marimuthu Antonisamy, Varaprasadham Irudayaraj

Objective To screen the antimicrobial potential of three ethnomedicinal plants Chassalia curviflora Thw. (C. curviflora), Cyclea peltata Hook. F. & Thomson (C. peltata) and Euphorbia hirta L (E. hirta) used in folk medicines in Aarukani hills Kani tribe, Tamil Nadu, India against human bacterial pathogens. Methods Antibacterial efficacy was performed by disc diffusion method against the pathogens viz., Escherichia coli (E. coli) (ATCC 35218), Staphylococcus aureus (S. aureus) (ATCC 6538), Salmonella typhi (S. typhi) (MTCC 733), Proteus vulgaris (P. vulgaris), Proteus mirabilis (P. mirabilis) and Streptococcus pyogenes (S. pyogenes) and incubated for 24 h at 37 °C. Results The maximum degree of antibacterial activity was observed in C. peltata followed by C. curviflora. While E. hirta showed comparatively low degree of antibacterial activity. The methanolic extract of C. peltata showed the antibacterial activity against three pathogens viz., S. pyogenes, P. vulgaris and E. coli with the inhibition zones 12 mm, 10 mm and 9 mm, respectively. hexane extracts of C. peltata also showed the antibacterial activity against two selected pathogens viz., P. vulgaris and P. mirabilis with 15 mm and 12 mm of inhibition zones. All the three different concentrations (0.25, 0.50 & 0.75 mg/mL) of methanolic extract of C. peltata show the inhibitory effect on the three susceptible bacteria S. pyogenes, P. vulgaris and E. coli with the maximum inhibition in the highest concentration (0.75 mg/mL). The methanolic and hexane extracts of C. curviflora exhibited the antibacterial activity against only one bacterium each i.e. P. vulgaris and S. typhi with the maximum zone of inhibition 13 and 11 mm respectively. The methanolic and hexane extracts of E. hirta exhibited the antibacterial activity against only one bacterium i.e. S. pyogenes with the maximum zone of inhibition 13 and 11 mm respectively. Conclusions The present investigation revealed that the C. curviflora, C. peltata and E. hirta are potentially good source of antibacterial agents and demonstrates the importance of such plants in traditional medicines.

386-390

Authors: Abu Arra Basma, Zuraini Zakaria, Lacimanan Yoga Latha, Sreenivasan Sasidharan

Objective To assess antioxidant activities of different parts of Euphorbia hirta (E. hirta), and to search for new sources of safe and inexpensive antioxidants. Methods Samples of leaves, stems, flowers and roots from E. hirta were tested for total phenolic content, and flavonoids content and in vitro antioxidant activity by diphenyl-1-picrylhydrazyl (DPPH) assay and reducing power was measured using cyanoferrate method. Results The leaves extract exhibited a maximum DPPH scavenging activity of (72.96±0.78)% followed by the flowers, roots and stems whose scavenging activities were (52.45±0.66)%, (48.59±0.97)%, and (44.42±0.94)%, respectively. The standard butylated hydroxytoluene (BHT) was (75.13±0.75)%. The IC50 for leaves, flowers, roots, stems and BHT were 0.803, 0.972, 0.989, 1.358 and 0.794 mg/mL, respectively. The reducing power of the leaves extract was comparable with that of ascorbic acid and found to be dose dependent. Leaves extract had the highest total phenolic content [(206.17±1.95) mg GAE/g], followed by flowers, roots and stems extracts which were (117.08±3.10) mg GAE/g, (83.15±1.19) mg GAE/g, and (65.70±1.72) mg GAE/g, respectively. On the other hand, total flavonoids content also from leave had the highest value [(37.970±0.003) mg CEQ/g], followed by flowers, roots and stems extracts which were (35.200±0.002) mg CEQ/g, (24.350±0.006) mg CEQ/g, and (24.120±0.004) mg CEQ/g, respectively. HPTLC bioautography analysis of phenolic and antioxidant substance revealed phenolic compounds. Phytochemical screening of E. hirta leaf extract revealed the presence of reducing sugars, terpenoids, alkaloids, steroids, tannins, flavanoids and phenolic compounds. Conclusions These results suggeste that E. hirta have strong antioxidant potential. Further study is necessary for isolation and characterization of the active antioxidant agents, which can be used to treat various oxidative stress-related diseases.

397-400

Authors: Vishwanath Sathyanarayanan, Abdul Razak, Kavitha Saravu, Shastry Barkur Ananthakrishna, M Mukhyprana Prabhu, KE Vandana

Objective To highlight the spectrum of clinical manifestations, labs, complications, treatment and outcome of brucellosis. Methods Retrospective study was conducted in Kasturba Medical College, Manipal University, Karnataka, India which included 68 confirmed cases of brucellosis from January 2006- April 2010. Diagnosis of brucellosis was made by culturing the sera/body fluids by standard BACTEC method (or) by testing the sera for Brucella agglutinins using the standard agglutination test (SAT). A titer of 1:320 or more was considered as significant. SPSS 16 was used for statistical analysis and Microsoft Excel for graphical representation. Results Of the 68 patients, 46 (68%) were male and 22 (32%) were female patients with age distribution of 9–75 years. Forty four (64.7%) had history of contact with unpasteurized dairy products or infected animals. Symptoms included fever (68, 100%), myalgia (21, 31%), musculoskeletal symptoms (23, 34%), headache (16, 24%), gastrointestinal symptoms (19, 28%) and altered sensorium (3, 4%). Co-morbidities and associations included HIV positivity (2, 3%), type 2 diabetes mellitus (13, 19%), steroid therapy (3, 4%) and HBsAg positivity (8, 12%). Ten (15%) patients had cervical lymphadenopathy, 4(6%) had splenic enlargement, 6 (7%) had hepatomegaly, 19 (28%) had hepatosplenomegaly and 2(3%) got meningeal signs. Anaemia was observed in 39 (57.3%) cases, high erythrocyte sedimentation rate (ESR) was present in 55 (80.8%) cases, leucocytosis in 10(14.7%), leucopenia in 10(14.7%), thrombocytopenia in 23 (33.82%) and thrombocytosis in 2 (2.94%) cases. Conclusions In countries like India, where brucellosis and tuberculosis are endemic; rapid, sensitive and highly specific diagnostic methods are required to make early diagnosis and prevent resistance as there is an overlap in therapy.

404-408

Authors: Adam H Al-Shamiri, Mohammed A Al-Taj, Ameera S Ahmed

Objective To determine the disease prevalence and its relationship with hepatitis B and C viruses among school children in five endemic areas by schistosomiasis. Methods During June 2007 and March 2009, 1 484 school children aged between 5 – 16 years participated in the current study from 32 basic schools in five districts (Al-Dhabab, Hedran, Warazan, Al-Barh and Al-Shmaytin) in Taiz Governorate, Republic of Yemen. Out of school children who participated in the study; 1 406 stool samples, 1 484 urine samples and 214 blood samples were collected and examined. Results Schistosoma mansoni (S. mansoni) was found in all the studied areas except Al-Barh. However, Schistosoma haematobium (S. haematobium) was recorded only in Al-Shmaytin and Al-Barh. Both S. mansoni and S. haematobium were observed in Al-Shmaytin district. The overall prevalence was 20.76% for S. mansoni and 7.41% for S. haematobium. The prevalence rate of infection among males was higher than females, shwing no significant differences. Rate of light, moderate and heavy infections in the case of S. mansoni were 41.78%, 25.34% and 32.87% respectively. Whereas, for S. hematobium it was 50.90% for light infection and 49.09% for heavy infection. Regarding to the prevalence of viral hepatitis among infected school children with schistosomiasis, it could be noticed that hepatitis B virus was higher than the prevalence of hepatitis C virus. But, the presence of HBsAg and anti-HCV was not associated with Schistosoma infection. Conclusions Schistosomiasis infection is an important public health problem in Taiz Governorate, Republic of Yemen. There was a correlation between S. haematobium and hepatitis B, but no association between S. mansoni infections and hepatitis B and C viruses.

412-413

Authors: Jaydeep Sarkar, Charu Mohan, DN Misra, Atul Goel

The spectrum of dengue infection varies from dengue fever to more severe forms such as dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). In DHF bleeding may be overt (external) or internal. At time bleeding can occur at unusual sites. We report one such case of DHF in which bleeding occurred within the tongue manifesting as an intra-lingual hematoma, causing significant upper airway obstruction.

417-420

Authors: Neelam Kaistha, Varsha Gupta, Shailpreet Sidhu, Jagdish Chander

Salmonella are most commonly associated with gastroenteritis and enteric fever in humans. Occasionally, dissemination of bacilli throughout the body results in establishment of localized foci of persistent infection especially in patients with debilitating diseases and immunosuppressive states. Infection at various aberrant sites due to Salmonella has been reported relatively seldom. It has perfected the art of intracellular survival in niches from where they can cause myriad of effects. Six cases with Salmonella infection at unusual sites without any preexisting or underlying disease diagnosed over a period of two years are presented here. Salmonella etiology was not suspected in these patients and the diagnosis was made microbiologically only after culture isolation.