Asian Pacific Journal of Tropical Medicine

510-513

Authors: M Karthikeyan, MK Deepa

Objective To evaluate the acute toxicity and to investigate the effect of Premna corymbosa ethanolic extract (PCEE) at doses of 200 and 400 mg/kg body weight in acute and chronic models of inflammation in experimental animals. Methods In the acute toxicity study, a single dose of PCEE of 2 000 mg/kg body weight, p.o. was administered and observed for 48 h. In acute models as egg albumin induced paw edema and chronic model as cotton pellet methods was followed. Results In acute models, egg albumin induced paw edema PCEE significantly (P<0.01) inhibited the edema formation. In chronic model, cotton pellet induced granuloma formation in rats PCEE significantly (P<0.01) reduced the granuloma formation with percentage inhibition of 35.17% and 50.38 % respectively. Conclusions The present study establishes the antiinflammatory activity of Premna corymbosa leaves.

523-525

Authors: Shahbudin Saad, Muhammad Taher, Deny Susanti, Haitham Qaralleh, Nurul Afifah Binti Abdul Rahim

Objective To investigate the antimicrobial activities of n-hexane, ethyl acetate and methanol extracts of the leaves of Lumnitzera littorea (L. littorea) against six human pathogenic microbes. Methods The antimicrobial activity was evaluated using disc diffusion and microdilution methods. Results The antimicrobial activities of the crude extracts were increased with increasing the concentration. It is clear that n-hexane extract was the most effective extract. Additionally, Gram positive Bacillus cereus (B. cereus) appear to be the most sensitive strain while Pseudomonas aeruginosa (P. aeruginosa) and the yeast strains (Candida albicans (C. albicans) and Cryptococcus neoformans (C. neoformans)) appear to be resistance to the tested concentrations since no inhibition zone was observed. The inhibition of microbial growth at concentration as low as 0.04 mg/mL indicated the potent antimicrobial activity of L. littorea extracts. Conclusions The obtained results are considered sufficient for further study to isolate the compounds responsible for the activity and suggesting the possibility of finding potent antibacterial agents from L. littorea extracts.

530-534

Authors: Qiang Wu, Qiongyao Fu, Qian Chen, Qunfang Cai, Zhigang Fan, Zhinong Zhan, Lina Niu, Hua Pei, Peng Zhou

Objective To obtain fbpB–esxA fusing gene of Mycobacterium tuberculosis (MTB), express the encoded fusing protein in Escherichia coli (E. coli), identify protein acquired, and predict the structure and function of the protein utilizing methods of bioinformatics. Methods fbpB and esxA gene were amplified from genome of MTB H37Rv by PCR. The fbpB–esxA fusing gene ligated by (Gly4Ser)3 linker was gained by means of Gene Splicing by Overlapping Extension PCR (SOE-PCR), and fusing gene was cloned into expression vector pET-30a. The recombinant plasmid was sequenced and expressed in E. coli BL21(DE3). The protein was identified by Western blot using anti-HIS antibody. Secondary structure and antigenic epitopes of the protein were predicting using tools of bioinformatics. Results The DNA sequences of fbpB–esxA were identical with that published by GenBank. The Ag85B-ESAT-6 fusion protein about 50 kDa comprised 485 amino acids was efficiently produced from expression system in E. coli BL21(DE3) under the induction of IPTG. Bioinformatics analysis showed the protein contained one transmembrane region and fourteen potential antigenic epitopes. Conclusions The Ag85B-ESAT-6 fusion protein is successfully expressed with N-terminal HIS-tag. Gel filtration demonstrated that it exists as insoluble inclusion bodies mainly. The existence of linker doesn't affect immunogenicity of Ag85B and ESAT-6. It will allow for characterization in vitro and establish a foundation of further function research such as vaccine or diagnostic reagent.

541-546

Authors: Ritu Arora, Ganesh S Dhale, Pooja R Patil, Shobha D Chitambar

Objective To conduct a comparative analysis of the VP4 gene sequences of Indian wild type (06361, 0613158, 061060 and 0715880) and cell culture adapted (06361-CA, 0613158-CA, 061060-CA and 0715880-CA) G1P[8] rotavirus strains. Methods Full-length VP4 genes of each of the four wild type G1P[8] rotavirus strains and their cell culture adapted counterparts displaying consistent cytopathic effect were subjected to RT-PCR amplification and nucleotide sequencing. Results All four cell culture adapted G1P[8] rotavirus strains showed nucleotide and amino acid substitutions in the VP4 gene as compared to their wild type strains. The number of substitutions however, varied from 1-64 and 1-13 respectively. The substitutions were distributed in both VP5* and VP8* subunits of VP4 gene respectively of permeabilization and hemagglutinating activity. The presence of unique amino acid substitutions was identified in two of the four wild type (V377G, S387N in 061060 and I644L in 0715880) and all four cell culture adapted (A46V in 0613158-CA, T60R in 06361-CA, L237V, G389V and Q480H in 061060-CA and S615G and T625P in 0715880-CA) strains for the first time in the VP4 gene of P[8] specificity. Amino acid substitutions generated increase in the hydrophilicity in the cell culture adapted rotavirus strains as compared to their corresponding wild type strains. Conclusions Amino acid substitutions detected in the VP4 genes of G1P[8] rotavirus strains from this study together with those from other studies highlight occurrence of only strain and/or host specific substitutions during cell culture adaptation. Further evaluation of such substitutions for their role in attenuation, immunogenicity and conformation is needed for the development of newer rotavirus vaccines.

550-555

Authors: Arshi Malik, Sarah Afaq, Mohammad Shahid, Kafil Akhtar, Abdullah Assiri

Objective To evaluate the effect of allagic acid treatment on the cell viability of human prostate cancer cells. Methods Ellagic acid (10-100 mol/L) treatment (48 h) of human prostate carcinoma PC3 cells was found to result in a dose-dependent inhibition of cell growth and apoptosis of PC3 cells as assessed by MTT assay, western blotting, flow cytometry and confocal microscopy. Results We observed that ellagic acid treatment of PC3 cells resulted in a dose dependent inhibition of cell growth/cell viability. This ellagic acid caused cell growth inhibition was found to be accompanied by induction of apoptosis, as assessed by the cleavage of poly (ADP-ribose) polymerase (PARP) and morphological changes. Further, induction of apoptosis accompanied a decrease in the levels of antiapoptotic protein Bcl-2 and increase in proapoptotic protein Bax, thus shifting the Bax: Bcl-2 ratio in favor of apoptosis. Ellagic acid treatment of PC3 cells was also found to result in significant activation of caspases, as shown by the dose dependent decrease in the protein expression of procaspase−3, −6, −8 and −9. This ellagic acid-mediated induction of apoptosis was significantly (80%-90%) inhibited by the caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp (OMe)-fluoromethylketone (Z-VAD-FMK). Thus these data suggested an essential role of caspases in ellagic acid-mediated apoptosis of PC3 cells. Conclusions It is tempting to suggest that consumption of tropical pigmented fruits and vegetables could be an effective strategy to combat prostate cancer.

560-563

Authors: Kanyanan Kritsiriwuthinan, Warunee Ngrenngarmlert

Objective To determine the prevalence of malaria infections among foreign migrant workers in Thailand. Methods Giemsa-stained thin and thick blood films were prepared from blood samples of 294 foreign migrant workers recruited in the study. Microscopic examination of these blood films was performed for malaria detection. Results Blood film examination revealed 1.36% malaria infections in these 294 subjects. All positive cases were male Myanmar workers in which their blood films only ring stage of Plasmodium spp. was found at low parasite density (mean= 144 parasites/μ L of blood). The prevalence of malaria infections was not significantly different among foreign migrant workers classified by age, gender, and resident province (P>0.05). Thin blood films of these workers also showed 78.91% hypochromic erythrocytes and 61.9% relative Eosinophilia. Conclusions These findings indicate a high risk of malaria transmission. Therefore active malaria surveillance by using molecular methods with more sensitive and specific than microscopy should be considered for malaria control in foreign migrant workers.

568-572

Authors: Lijuan Zhang, Feng Cui, Lingling Wang, Lingling Zhang, Jingshan Zhang, Shiwen Wang, Shuxia Yang

Objective To investigate the situation of anaplasmosis in Yiyuan county, Shandong Province. Methods A total of 26 blood samples from febrile patients suspected of anaplasmosis, 48 blood samples from healthy farmers, 8 from dogs, and 10 from goats and 170 ticks were collected in the same area during 2005-2007, and detected by serological and molecular methods. Results Eight confirmed cases and 6 probable cases were determined using serologic and molecular methods. The seroprevalence of Anaplasma phagocytophilum (A. phagocytophilum) was 26.7% in healthy cases. Nine out of 10 sheep samples and 7 out of 8 dog samples reacted positively to the A. phagocytophilum antigen. PCR amplification and sequencing of the 16SrRNA of A. phagocytophilum gene showed that some samples from patients, goats and ticks were 100% identical. The seroprevalence of Rickettsia typhi was 22.9%, Orientia tsutsugamushi 6.3%, Rickettsia sibirica 27.1%, Coxiella burnetii 18.8%, Bartonella henselae 31.3%, and Borrelia burgdorferi 41.6%. Conclusions It is important to make differential diagnosis of febrile patients and to apply treatment with specific antibiotics. It is needed to enforce essential prevention and control measures including tick control and to improve sanitation conditions.

581-586

Authors: Anish Chandy, Alok Singh Thakur, Mukesh Pratap Singh, Ashish Manigauha

Filariasis is result of parasitic infection caused by three specific kinds of round worm. Lymphatic filariasis is found in under developed region of South America, Central Africa, pacific and Caribbian. It has been found for centuries, with main symptoms as elephant like swelling of the arms, legs and genitals. It is estimate that 120 millions peoples in the world have lymphatic filariasis. The spread of diseases and the challenge encountered in its management are discussed along with a review on drugs against filariasis in this article. Detail on clinical effect of drugs on the infection, safety profile, status in clinical practices and drug resistances are also covered.