Asian Pacific Journal of Tropical Medicine

925-929

Authors: Yongxiang Zhao, Jing Tang, Qin Yao, Yuan Zhou, Huange Zhao, Xiaoyun Zeng, Jiaqi Shi, Guorong Luo, Xiaoxun Xie, Sufang Zhou, Zuguo Liu, Xiaoling Lu, Donghai Lin, Jianming Liu

Objective To investigate the effect of fusion proteins expressed by the fused gene of porcine α 1, 3 galactosyltransferase (α1, 3 GT) and enhanced green fluorescent protein (EGFP) on the green fluorescence intensity of EGFP. Methods The fragment containing α 1, 3GT was firstly recovered after the pcDNA3.1- α 1, 3GT recombinant vector were digested with Bam HI and Eco RI, and then, the resultant fragment was ligated to the pEGFP-N1 vector which was also digested with the same enzymes. The new recombinant eukaryotic expression pEGFP/α 1, 3GT vector was obtained and sequenced. The pEGFP/α 1, 3GT was used to transfect human lung carcinoma cells A549 and HEKC 293FT, and the expression of EGFP was quantitatively analyzed by fluorescent microscope and flow cytometry. Results The positive percentage of A549 was 80.5%, and that of 293 FT was 86.5% 48 hours after the two cell lines both were transfected by pEGFP-N1. The positive percentage of A549 was 75.8%, and that of 293 FT was 81.2% 48 hours after the two cell lines were transfected by pEGFP/α 1, 3GT. The mean fluorescence intensities of A549 transfected with pEGFP-N1 and pEGFP/α 1, 3GT were 1.21 and 0.956, respectively when compared with that of A549 without transfection. Meanwhile, the those of the 293FT that were transfected with pEGFP-N1 and pEGFP/α 1, 3GT were 7.66 and 1.00, respectively when compared with that of 293FT cells without transfection. Conclusions These results suggested that the expression of EGFP gene fused with porcine α 1, 3GT gene was partly inhibited.

934-938

Authors: Mauro Prato, Valentina Gallo, Elena Valente, Amina Khadjavi, Giorgia Mandili, Giuliana Giribaldi

Objective To investigate the effect of malarial pigment (hemozoin, HZ) on expression of heat shock proteins (HSPs) and cell viability in human monocytes by using a stable cell line (THP-1 cells). Methods THP-1 cells were fed with native HZ or treated with pro-apoptotic molecule gliotoxin for 9 h. Thereafter, the protein expression of HSP-27 and HSP-70 was evaluated by western blotting. Alternatively, HZ-fed cells were cultured up to 72 h and cell viability parameters (survival, apoptosis and necrosis rates) were measured by flow cytometric analysis. Results HZ increased basal protein levels of HSP-27 without altering those of HSP-70 in THP-1 cells, and promoted long-term cell survival without inducing apoptosis. As expected, gliotoxin inhibited HSP-27 protein expression and promoted long-term cell apoptosis. Conclusions Present data show that HZ prevents cell apoptosis and enhances the expression of anti-apoptotic HSP-27 in THP-1 cells, confirming the previous evidences obtained from HZ-fed immunopurified monocytes. Since the use of a stable cell line is pivotal to perform HSP-27 silencing experiments, monocytic THP-1 cells could be a good candidate line for such an approach, which is heavily required to clarify the role of HSP-27 in survival of impaired HZ-fed monocytes during falciparum malaria.

943-947

Authors: Ukwe Chinwe V, Ekwunife Obinna I, Epueke Ebele A, Ubaka Chukwuemeka M

Objective To determine the suppressive and curative activity of aqueous leaf extract of Ageratum conyzoides (A. conyzoides) in combination with chloroquine and artesunate, respectively against Plasmodium berghei infection in mice. Methods Using malaria (Plasmodium berghei) infected albino mice of both sexes, aqueous extracts of A. conyzoides in combination with chloroquine and artesunate were tested for antimalarial activity, respectively. Four-day suppressive test and Rane's curative test were carried out. Results Suppressive tests showed significant dose dependent reduction in parasitemia level produced by the extract-chloroquine and extract-artesunate combinations. Suppressive activities of both extract-drug combinations were greater than the individual drugs alone. Extract-chloroquine (100:5) produced the highest suppressive effect (98% suppression). Curative tests showed absolute survival in two extract-drug combinations. Two extract-drug combinations produced higher curative effects than the individual drugs alone. The highest dose combinations of extract-chloroquine (100:5) and extract-artesunate (100:5) produced absolute parasitemia clearance (cure) in the infected mice. Conclusions The study indicated that aqueous extract of A. conyzoides had the ability to potentiate the antimalarial activity of chloroquine and artesunate against induced plasmodiasis in mice. It contributes a lot in the malaria endemic and poverty stricken tropics.

954-956

Authors: Supratik Chakraborty, Someshwar Singha, Goutam Chandra

Objective To observe the effect of two common organic acids on the larvae of Aedes aegypti (Ae. aegypti) (L), the natural vector of dengue fever/dengue hemorrhage fever, chikugunya and allergic skin reaction especially in children. Methods Two common organic acids (lactic acid and orthophosporic acid of gradually increasing concentration) were used against laboratory reared third instars larvae of Ae. aegypti in order to observe the rate of mortality after 8, 16 and 24 h of post exposure respectively in laboratory. Results Larval mortality rates recorded were in the following sequences: orthophosphoric acid and lactic acid at 1:1 combination >orthophosphoric acid>lactic acid. Conclusions These two organic acids may be used perfectly in combination (1:1) along with other conventional vector control methods to reduce the Ae. aegypti population, especially in those areas where surveillance and supervisory mechanism are poor or insufficient.

961-964

Authors: Shyamapada Mandal, Manisha DebMandal, Nishith Kumar Pal, Krishnendu Saha

Objective To ascertain the potential antibacterial activity of honey against clinical isolates of Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa) and Salmonella enterica serovar Typhi (S. enterica serovar Typhi) by in vitro methods. Methods The partial inhibitory concentration (PIC), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of the autoclaved honey (extracted from Apis indica hive by indigenous method) were determined for S. enterica serovar Typhi (n=8; from blood cultute), E. coli (n=5; from urine culture) and P. aeruginosa (n=5; from pus culture) isolates by in vitro methods. Results The PICs of the honey tested for the isolates ranged 0.50%-1.25 % (v/v) for S. enterica serovar Typhi, 0.75%-1.50% (v/v) for E. coli and 1.00%-1.25 % (v/v) for P. aeruginosa, while the MICs ranged 1.75%–3.00% (v/v), 3.00%-3.50% (v/v) and 3.50% (v/v), respectively. The P. aeruginosa and E. coli isolates had MBC value of 4.00% (v/v); the S. enterica serovar Typhi showed MBCs in between 3.00% and 3.50% (v/v). The bactericidal activity of honey was achieved at concentration 3.00% (v/v) for S. enterica serovar Typhi and E. coli, and at 3.50% (v/v) for P. aeruginosa. Conclusions The excellent antibacterial activity of honey against clinical bacterial isolates indicates the usefulness of honey in clinical practice against bacterial infection.

971-976

Authors: Evan Prince Sabina, Annie Deborah Harris Souriyan, Deborah Jackline, Mahaboob Khan Rasool

Objective To explore the hepatoprotective and antioxidant effects of piperine against acetaminophen-induced hepatotoxicity in mice. Methods In mice, hepatotoxicity was induced by a single dose of acetaminophen (900 mg/kg b.w. i.p.). Piperine (25 mg/kg b.w. i.p.) and standard drug silymarin (25 mg/kg b.w. i.p.) were given to mice, 30 min after the single injection of acetaminophen. After 4 h, the mice were decapitated. Activities of liver marker enzymes [(aspartate transaminase (AST), alanine transaminase (ALT), and alkaline phosphatase (ALP)] and inflammatory mediator tumour necrosis factor-alpha (TNF-α) were estimated in serum, while lipid peroxidation and antioxidant status (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-s-transferase and glutathione) were determined in liver homogenate of control and experimental mice. Results Acetaminophen induction (900 mg/kg b.w. i.p.) significantly increased the levels of liver marker enzymes, TNF-α, and lipid peroxidation, and caused the depletion of antioxidant status. Piperine and silymarin treatment to acetaminophen challenged mice resulted in decreased liver marker enzymes activity, TNF-α and lipid peroxidation levels with increase in antioxidant status. Conclusions The results clearly demonstrate that piperine shows promising hepatoprotective effect as comparable to standard drug silymarin.

977-981

Authors: Subramanian Kumaran, Balaraman Deivasigamani, Kumarappan Alagappan, Mannikam Sakthivel, Rajamani Karthikeyan

Objective To determine the prevalence and antibiotic resistance of Escherichia coli (E. coli), in seafood obtained from Cuddalore and Parangipettai fish landing centres. Also, to identify the susceptibility of E. coli against predominant seaweeds red alga Kappaphycus alvarezii (K. alvarezii) and brown alga Padina boergessenii (P. boergessenii) extracts as sulfated polysaccharides and polyphenols respectively. Methods A total of 48 samples (Two stations Cuddalore and Parangipettai, Tamil Nadu, India). Sampling area are fish landing centre where fishes caught from sea and estuary, seafood processing plants (packing and ice packed fishes) and local fish markets (fish samples). After isolation totally 80 strains were analyzed for its antimicrobial resistance and sensitivity against commercially 10 antibiotics. The ampicillin resistant E. coli CE21 was identified through molecular techniques as 16S rDNA sequencing. Two seaweeds K. alvarezii and P. boergessenii were screened for antibacterial activity against 12 antibiotic resistant E. coli strains. Results Totally 48 swabbed samples from two different fish handling area were characterized for total bacterial and E. coli count. Mostly, the E. coli strains were isolated from fish local market and seafood processing plants before and after packaging process. In that maximum 56.25% strains were resistant to ampicillin and the minimum 2.5% strains were resistant to chloramphenicol. Therefore, the E. coli CE21 was identified through molecular techniques E. coli (GenBank accession number GU065251), The MIC value for polyphenol extract was slightly less than sulfated polysaccharides. E. coli strain isolated from Parangipettai was considerably increased MIC value that Cuddalore. Conclusions The polyphenol and sulfated polysaccharides showed promising inhibitory response against all antimicrobial resistant E. coli strains and in particular the inhibitory response of ampicillin resistant E. coli.

988-992

Authors: Mohammed Haneefa KP, Shahima Hanan K, Saraswathi R, Guru Prasad Mohanta, Chandini Nayar

Objective To formulate Pothos scandens Linn (P. scandens) leaf extract in to a gel and investigate their burn wound healing activity. Methods Ethanolic extract of dried leaves of P. scandens were subjected to priliminery phytochemical evaluation and wound healing activities studies. Different gel formulations of ethanolic extract of P. scandens (4% w/v) were prepared using polymers carbopol 934 and carbopol 940 by varying their concentration. These formulations were evaluated for physical parameters, drug content, pH, viscosity, extrudability, spreadability, primary skin irritation, pharmacological activity and stability. Results Wound healing studies of ethanolic extract revealed that P. scandens treated animals were found to epithelise in 22 days while the solvent control and the untreated rats epithelise within 35 and 40 days respectively. The formulation with 1.5% w/w carbopol 934 was found to be more promising as it shows better physicochemical characteristics, higher pharmacological activity and stability compared to other formulations. Conclusions P. scandens alcoholic extract shows significant improvement in burn wound contraction and hence this is a promising candidate in burn wound healing.

997-1000

Authors: Kuldeep Kumar, Pankaj Kumar Singh, Juhi Tomar, Swati Baijal

Dengue fever is a mosquito born viral infection, and the complicated form of dengue is dengue hemorrhagic fever (DHF). In the recent decades incidence and distribution of dengue has increased dramatically. Dengue viruses belong to family flaviviridae with four serotypes and are transmitted mainly by mosquito Aedes aegypti. Today almost two-fifth of world's population (2.5 million) is at risk of dengue and no specific antiviral drug or vaccine is available against it. Uncontrolled population growth in Africa and South East Asia has increased number of susceptible hosts in urban and semi urban areas. About 40% of world population resides in the high risk area for dengue transmission. According to latest estimates by WHO, yearly 50 to 100 million infections occur globally, this includes around 500 000 DHF and 22 000 deaths, mostly among children. Only symptomatic treatment in the form of analgesic, antipyretics and body fluid management is provided to the patient. Prevention strategies mainly focus on two approaches, firstly on activities to control vector and secondly on activities to protect human from mosquito bite but there is always concerns regarding their sustainably and effectiveness. Theoretically development of an effective dengue vaccine is feasible and production of an effective and affordable vaccine could be a viable option to save humans from this dreadful disease. Conceptually vaccine production is possible, but it has to be tetravalent, providing immunity against all serotypes. Few candidate vaccines are in advance stage of their development; however international cooperation is needed to make these vaccines available on cheaper rates to the poor and vulnerable countries. Objective of this review is to discuss various aspects related to dengue, its epidemiology, available preventive methods, need for vaccine and challenges in its development.

1004-1005

Authors: Sumeet Bhargava, Rajul Rastogi, Gaurav Jindal, Amrit Kumar Singh, Vaibhav Rastogi

Tuberculous infection manifesting as an isolated intramedullary tuberculoma of the spinal cord is distinctly unusual. We report a case of a 35 year old woman presenting with an insidious onset of myelopathy, where MRI showed characteristic imaging findings suggestive of intramedullary tuberculosis.