Asian Pacific Journal of Tropical Medicine

762-764

Authors: Yingzi Lin, Shixiang Bao, Guanghong Tan, Fengying Huang, Huiqin Huang

Objective To construct and identify recombinant plasmid p UIS3-BLC+. Methods The cDNA of B-lymphocyte chemoattractant (BLC) was amplified from the total RNA of spleen tissues by PCR method, and were inserted into plasmid of Plasmodium berghei with UIS3 knockout by digestion of restrictive endonuclease and T7 ligation. The recombinant plasmids were screened, and then underwent restriction enzymatic digestion and DNA sequencing. Then the confirmed plasmid was further transfected into COS-1 cells by lipofectamine and the BLC expression was tested by RT-PCR and Western blotting. Results The cDNA of BLC gene was correctively amplified by RT-PCR and the recombinant plasmid p UIS3-BLC+ was constructed successfully, which was confirmed by restriction enzymatic digestion and DNA sequencing. RT-PCR and Western blotting also showed the BLC gene expression in COS-1 cells. Conclusions The recombinant plasmid p UIS3-BLC+ has BLC expression in COS-1 cells, and is useful for further study on BLC transgene and UIS3 gene knockout in Plasmodium berghei.

769-774

Authors: Subramanion Jo Thy Lachumy, Sreenivasan Sasidharan, Vello Sumathy, Zakaria Zuraini

Objective To elucidate its pharmacological activities and medicinal potential of extract of Etlingera elatior (E. elatior). Methods Phytochemical screening of the flower extract was done to determine the phytochemical in the extract. The pharmacological study included the determination of antimicrobial activity and minimum inhibitory concentration (MIC) of metabolic flower extract. The antimicrobial activity of the extract was tested against medically important bacterial, yeast and fungal strains. Apart from that, the methanolic extract of E. elatior flower was further tested in vivo toxicity using the brine shrimp lethality test. Moreover, the flower extract was qualitatively screened for their free radical scavenging activity by 2, 2-diphenyl-1-picrylhydrazyl radical (DPPH) assay. Results The extract was effective on tested microorganisms and MIC values were in the range of 1.563-50.000 mg/mL. The brine shrimp lethality test exhibited no significant toxicity (LC50 = 2.52 mg/mL) against Artemia salina. The E. elatior flower extract with high LC50 value signified that this plant is not toxic to humans. While the phytochemical screening of the flower extract revealed the presence of the following compounds: flavonoids, terpenoids, saponin, tannins and carbohydrates whereas, alkaloids, anthraquinone and reducing sugars were absent. The concentration of the flower extract required for 50% inhibition of DPPH radical scavenging effect (IC50) were 9.14 mg/mL and 8.08 mg/mL for butylated hydroxytoluene 8.08 mg/mL. Conclusions These findings indicate that the extract of E. elatior flower possesses pharmacological properties and potential to develop natural products based pharmaceuticals products.

783-787

Authors: Meena Kumari, Eesha BR, Mohanbabu Amberkar, Sarath babu, Rajshekar, Neelesh Kumar

Objective To evaluate the wound healing effect of aqueous extract of Crotalaria verrucosa (C. verrucosa) in rats. Methods Three wound models including incision, excision and dead space wounds were used in this study. The parameters studied were breaking strength in incision models, granulation tissue dry weight, breaking strength and hydroxyproline content in dead space wounds, percentage of wound contraction and period of epithelialization in excision wound model. Results Two doses of the extract with and without dexamethasone showed significant increases in mean hydroxyproline, total protein content and dry weight of granulation tissue but it was higher with dose 800 mg/kg comparing with the control. The dexamethasone treated group showed a significant (P<0.001) reduction in the wound breaking strength when compared to control group in incision type of wound model. Coadministration of C. verrucosa with dexamethasone significantly (P<0.001) increased the breaking strength compared to the dexamethasone treated only group. In excision wound model, the percentage of the wound contraction was significantly (P<0.01) increased by two doses of test extract on all the days except the lower dose which exhibited only on 12 th, 16 th days of drug treatment and it also reversed the dexamethasone suppressed wound contraction. It significantly (P <0.001) reduced the time required for epithelialization and reversed the epithelialization delaying effect of dexamethasone (P<0.001). Conclusions C. verrucosa was found to possess significant wound healing property. This was evident by decrease in the period of epithelialization, increase in the rate of wound contraction, skin breaking strength, and granulation tissue dry weight content. Hence C. verrucosa could be a good wound healing agent.

791-793

Authors: Sarmistha Saha, Gagan Goswami

Objective To investigate the gastroprotective activity of hydroalcoholic extract leaves of Ficus religiosa (F. religiosa) in different experimental models of gastric ulcer in rats. Methods The hydroalcoholic extract leaves of F. religiosa were studied at two dose levels (250 and 500 mg/kg, oral) in rats against absolute ethanol (0.2 mL oral), aspirin (200 mg/kg) and pyloric ligation induced gastric ulcer. Ranitidine (50 mg/kg, oral) was used as a standard drug. Mean ulcer indices and oxidative stress were measured. Phytochemical tests and acute toxicity tests were also carried out. Results Administration of F. religiosa to rats significantly decreased the ulcer index value when compared with the control treated group. Ranitidine (50 mg/kg, oral) also produced a significant decrease the ulcer index value when compared with the control treated group. Phytochemical analysis revealed the presence of tannins, sterols, saponins, flavonoids, carbohydrates and proteins. Conclusions The results suggest that the leaves of the F. religiosa possess significant anti ulcer activity.

797-799

Authors: D Devina Merin, S Prakash, B Valentine Bhimba

Objective To explore the biosynthesis of silver nanoparticles synthesized by marine microalgae. Methods Marine microalgae was collected from Central Marine Fisheries Research Institute (CMFRI, tuticorin) and cultured in the lab. Silver nanoparticles synthesis were observed in normal and microwave irradiated microalgae and screened against human pathogens for the presence of antimicrobials. Results The presence of silver nanoparticle was confirmed by UV-Visible spectroscopy at 420 nm by the presence of plasmon peak. Further confirmation was done by scanning electron microscope (SEM). Conclusions These results not only provide a base for further research but are useful for drug development in the present and future.

804-806

Authors: Hosseini Vasoukolaei Nasibeh, Telmadarraiy Zakkyeh, Vatandoost Hassan, Yaghoobi Ershadi Mohammad Reza, Hosseini Vasoukolaei Morteza, Oshaghi Mohammad Ali

Objective To determine the tick species parasitizing domestic ruminants in Ghaemshahr county in Mazandaran, a Caspian province in the north of Iran. Methods About 361 sheep, 54 goats and 10 cattle of 18 herds in several villages in Ghaemshahr were inspected for tick infestation. Separated ticks were preserved in 70% alcohol and identified. Results About 323 ticks (207 female, 116 male) were collected, the occurrence of ticks on sheep, goats and cattle were 28.3%, 22.2% and 20.0% respectively. The mean number of ticks on each animal was low (3-5 ticks per animal). Rhipicephalus sanguineus, Rhipicephalus bursa, Ixodes ricinus, Boophilus annulatus, Haemaphysalis punctata and Haemaphysalis numidiana were the tick species we found. Rhipicephalus sanguineus were the most abundant species in the study area. The largest number of ticks were generally present from April to July, mostly in animal ears and tails. Ixodes, Boophilus and Haemaphysalis occurred in mountainous areas of Ghaemshahr, whereas Rhipicephalus were present in both mountains and plains of the study area. Conclusions The result of this study is a survey of tick species from domestic animals in Iran and implication of possible prevention measures for diseases transmitted by ticks.

812-814

Authors: Chintana Chirathaworn, Yong Poovorawan, Viboonsak Vuthitanachot

Objective To determine the levels of IL-18 and IL-18 binding protein (BP) in patients with dengue virus infection. Methods Acute and convalescent sera were collected from each patient. Control group was sera from blood donors. The levels of both IL-18 and IL-18BP were measured by ELISA assays. Results It was shown that IL-18 and IL-18BP levels were significantly higher in patients when compared with controls. In addition, the level of IL-18BP was lower in convalescent than in acute sera. Conclusions These data suggest that both IL-18 and IL-18BP production was induced following dengue virus infection. Investigating the regulation of IL-18 by its natural regulator could lead to further understanding of the immune response or immunopathogenesis following dengue virus infection.

818-820

Authors: Vishwanath Sathyanarayanan, Ragini Bekur, Abdul Razak, Joydeep Chakraborty

Objective To study disseminated cryptococcal infection in a tertiary care hospital in Southern India. Methods The clinical profile of 12 disseminated cryptococcosis patients with the age group of 28-52 years was retrospectively analyzed. Results 7(58.3%) presented with fever < 30 days and 3(25%) > 30 days whereas 2(16.7%) did not have fever. All the 12(100%) had headache, 2(16.7%) had altered sensorium, one (8%) seizure. 5(41.7%) had diarrhea and vomiting. 6(50%) had oral candidiasis, and anemia. 9(75%) had elevated erythrocyte sedimentation rate (ESR). 6(50%) had neck stiffness. Cerebrospinal fluid (CSF) pressure was elevated in all 12(100%) patients. Blood culture positive for Cryptococcus neoformans(C. neoformans) in 11(91.7%) and CSF culture positive in all 12 (100%), one (8%) had urine culture positive. India ink preparation was positive in 10(83.3%). CD4 count was less than 50/microl in 4 (33.3%), between 50-100 in 6(50%) and 2(16.7%) in the range of 100-200. 6(50%) were treated with parenteral amphotericin B (0.7 mg/kg/d) during intensive phase followed by oral fluconazole 400 mg/d for 8 weeks then maintenance oral fluconazole 200 mg/d. 5(41.6%) were treated with fluconazole alone. 8(66.7%) improved and 4(33.3%) patients died. Among those who succumbed to the illness, 2(16.7%) received amphotericin and fluconazole, 2(16.7%) patients received fluconazole alone. Conclusions Disseminated cryptococcosis can cause considerable mortality in HIV patients and immunocompromised non- HIV individuals. At times, its presentation closely mimics that of Tuberculosis. Early diagnosis and appropriate treatment should be started as early as possible.

828-832

Authors: Ezeani Michael Chukwudi, Onyenekwe CC, Wachukwu CK, Anyiam DCD, Meludu SC, Ukibe RN, Ifeanyichukwu M, Onochie A, Anahalu I, Okafor UU

Objective To investigate the prevalence of microbial antigenic components of circulating immune complexes amongst grades of CD4 T lymphocyte counts in HIV sero positive and sero-negative participants. Methods Polyethelene glycol (PEG-600) and buffering methods of precipitation and dissociation of immune complexes was used to generate immune solution from sera of 100 HIV sero-positive and 100 HIV sero-negative participants. These were categorized into 3 grades based on CD4 count: > 500 cell/mm3, 200-499 cell/mm3 and <200 cell/mm3. The immune solutions were assayed using membrane based immunoassay and antibody titration, along side its unprocessed serum for detection of various microbial antigens and or antibodies. CD4 T cell counts were estimated using Patec Cyflow SL-3 Germany. Results Antigenic component of immune complexes of various infectious agents was detected in 99 and 70 HIV sero-positive and HIV sero-negative participants, respectively. In group A, there were 10 HIV positive participants, including 4 (40.0%) had circulating immune complexes(CIC s) due to Salmonella species only; 1(10.0%) due to Salmonella-Plasmodium falciparum (P. falciparum), Salmonella-P. falciparum-HCV and P. falciparum antigens, respectively. In group B, 45(45.4%) HIV sero-positive participants with CIC s had CD4 T lymphocyte count between 200-499 cells/mm3. Out of these, 20(44.4%) had CIC s due to Salmonella species only; 9(20%) due to Salmonella-P. falciparum. In group C, there were 44(44.4%) HIV sero-positive participants, including 3(6.8%) due to Salmonella species only; 24(54.4%) due to Salmonella-P. falciparum; 2(4.5%) due to P. falciparum only. Conclusions In HIV sero-positive participants, presence of heterogeneity of Salmonella species-P. falciparum antigens was highly incriminated in CD4 count depletion but not homogeneity of malaria parasites antigens. Malaria parasites antigens only were incriminated in CD4+ count depletion amongst HIV sero-negative participants. Before taking any decision on the management of HIV-1-positive individuals, their malaria and Salmonella paratyphi status should be assessed, but not malaria status alone.

835-837

Authors: Vishwanath Sathyanarayanan, Bekur Ragini, Abdul Razak, M Mukhya prana Prabhu

Neurobrucellosis is a rare form of localized brucellosis usually with no systemic manifestations. We report a rare case of brucellosis presenting as meningoencephalitis associated with hydrocephalus. This patient had a lymphocytic predominant CSF and was initially treated with empirical anti tubercular therapy and steroids. A week later, when his CSF culture grew Brucella species, the treatment was changed to a combination of streptomycin, doxycycline and rifampicin and the patient improved with this therapy. This case illustrates the need to consider neurobrucellosis as a close differential diagnosis of neurotuberculosis in endemic areas when the patient presents with meningo encephalitis with lymphocytic CSF