Asian Pacific Journal of Tropical Biomedicine

4–9

Authors: Bagus Hermansyah, Loeki Enggar Fitri, Teguh Wahju Sardjono, Agustina Tri Endharti, Samsul Arifin, Niniek Budiarti, Didi Candradikusuma, Erma Sulistyaningsih, Nicole Berens-Riha

Objective: To investigate clinically severe malaria patients with Plasmodium falciparum (P. falciparum), Plasmodium vivax (P. vivax) and mixed species infections. Methods: This study was conducted at Dr. Saiful Anwar General Hospital, Malang, Indonesia, from December 2011 to May 2013. Twenty nine patients (mean age of 41 years, 22% female), who suffered from severe malaria according to World Health Organization criteria (major and minor) and other criteria based on previous studies, were selected by consecutive sampling. Blood samples were obtained at admission from peripheral blood for microscopic diagnostic, nested PCR and laboratory examination of blood chemistry. Laboratory results were compared between the groups and correlated to each other. Results: From 29 samples, eight (28%) were diagnosed as P. falciparum mono-infection, 12 (41%) as P. vivax mono-infection and nine (31%) as mixed infections, confirmed by PCR. Cerebral malaria occurred in P. falciparum or mixed species infection only. Parasitaemia was highest in P. falciparum mono-infection. Mean haemoglobin was signifi- cantly lower in P. falciparum than P. vivax infection (P = 0.01). Mean thrombocyte count (77 138/mL) was low in all groups. Mean urea, creatinine, total and direct bilirubin were significantly higher in P. falciparum mono-infection compared to other groups, whereas aspartate aminotransferase and alanine aminotransferase showed no significant differences. Parasitaemia was positively correlated with an increase in urea, creatinine, bilirubin and leucocytosis in all species. Conclusions: Both Plasmodium species can solely or in combination cause severe malaria. Mixed infection was generally more benign than P. falciparum mono-infection and seemed to have some protective effects.

14–19

Authors: Flor Yohana Flores Hernandez, Sanghamitra Khandual, Inocencia Guadalupe Ramírez Lopez

Objective: To evaluate the cytotoxic effects of Spirulina platensis extracts on acute leukemia Kasumi-1 and chronic leukemia K-562 cancer cell lines. Methods: Various concentrations of Spirulina platensis extracts (0.25–50.00 mg/mL) obtained with different solvents were used to treat cell lines for 72 h. For cytotoxic effect studies, cell viability test with trypan blue solution, MTT assay and microscopic cytomorphological assessment were done. Results: Spirulina extract obtained with 70% ethanol showed significant cytotoxicity in K562 and Kasumi-1 cell lines. With trypan blue solution, IC50 values were found to be 4.64 mg/mL for K-562 and 3.68 mg/mL for Kusumi-1 cell lines. Spirulina aqueous extract also showed cytotoxicity with trypan blue method, at a slightly higher dose; where IC50 values were 12.68 mg/mL for K-562 and 2.13 mg/mL for Kusumi-1 cell lines. The IC50 values were found 0.40 mg/mL for K-562 and 0.31 mg/mL for Kusumi-1 cell lines for the 70% ethanol extract according to the MTT assay. Spirulina extract obtained with water also showed cytotoxicity but the dose was a little higher where IC50 values were 15.77 mg/mL for K-562 and 9.44 mg/mL for Kusumi-1 cell lines. The effect of cytotoxicity with ethanol extract is quite comparable with that observed for cyclophosphamide, which is a chemical used as anticancer agent. Conclusions: The cytotoxicity exhibited by Spirulina extract to cancer cell lines might be due to the presence of phytopigments (carotenoids, chlorophyll, phycocyanin) as well as polysaccharides that were reported previously as constituents of the extract. So crude extracts of Spirulina can be used as a source to develop anticancer drugs.

27–31

Authors: Suzanah Abdul Rahman, Nur Amalina Ahmad, Nadia Hanis Abdul Samat, Syazana Zahri, Afif Raihan Abdullah, Kit-Lam Chan

Objective: To evaluate the effects of Eurycoma longifolia (E. longifolia) standardized extract on the oestrous cycle, levels of reproductive hormones and histology of the ovaries of Sprague-Dawley rats. Methods: Female rats were orally treated with E. longifolia standardized extract at the dose levels of 2.5, 5.0, 10.0, 25.0, 50.0 and 100.0 mg/kg of body weight over 5 days. Vaginal smears were monitored daily within the duration and after withdrawal of the treatment before being sacrificed. The body weights of the females were recorded before and after the 5 days treatment. At the end of the experiments, blood samples were collected for determination of testosterone, oestradiol and progesterone levels. Ovaries were removed, weighed and examined for histomorphological changes. Results: The administration of E. longifolia standardized extract did not significantly alter the oestrous cycle of the rats during the 5 days treatment and after withdrawal of the treatments. This was supported by normal testosterone, oestradiol and progesterone levels as well as normal morphology of the ovaries. Conclusions: The data obtained showed that E. longifolia standardized extract did not exhibit any toxic effect on reproductive activities of female rats suggesting potential use in the management of infertility.

37–42

Authors: Fatemeh Maleki, Mitra Zarebavani, Mehdi Mohebali, Mohammad Saaid Dayer, Fateme Hajialiani, Fatemeh Tabatabaie

Objective: To evaluate the efficacy of some medicinal plants and systemic glucantime in a comparative manner against the causative agent of cutaneous leishmaniasis both in vitro and in BALB/c mice. Methods: For in vivo testing, inbred mice were challenged with Leishmania major parasites and the resultant ulcers were treated with extract based-ointments applied topically two times per day for a period of 20 days. A group of 56 mice were randomly divided into 7 subgroups. The control group received the ointment void of extracts, whereas the reference group received glucantime only. The efficacy of treatments was evaluated by measuring ulcer diameter, parasite burden and NO production. Results: Our results indicated that plant extract based-ointments were effective in reducing ulcer size and parasite burden in spleens, but their effects did not differ significantly from that of glucantime. The plant extracts tested in this study were able to increase NO production that helped parasite suppression. Conclusions: Our findings indicate that the tested plant extracts are effective against Leishmania major both during in vitro and in vivo experiments, but further researches are required to recommend a potential plant extract as an alternative drug.

49–52

Authors: Sabiha Chowdhury, Saikat Kumar Poddar, Sarah Zaheen, Farah Ashrafi Noor, Najneen Ahmed2, Sanjana Haque, Abhijit Sukul, Sauda Binte Sunjida, Md. Mahabob Ullah Mazumder, Nusrat Akbar

Objective: To investigate the presence of different phytoconstituents in Mangifera indica (M. indica) peel and evaluate its cytotoxicity to Artemia salina and hypoglycemic potential in Swiss albino mice. Methods: The methanolic extract of M. indica peel was used to determine the presence of phytoconstituents. Brine shrimp lethality bioassay method was followed to determine the cytotoxic potential of plant extract. In the case of hypoglycemic activity, oral administration of extract at 200 and 400 mg/kg and standard glibenclamide at 10 mg/kg was done, followed by determining the percentage of reduction of plasma glucose from the initial level. Results: The methanolic extract of M. indica peel showed the presence of flavonoid, saponin, steroid, tannins, terpenoids, glycosides and alkaloids. In brine shrimp lethality bioassay, the LC50 of the extract and standard vincristine sulfate was found to be 2.04 and 0.41 mg/mL, respectively. After 90 and 150 min, the methanolic extract at 200 and 400 mg/kg showed prominent plasma glucose reduction of 13.95%, 22.48% and 14.16%, 26.18% respectively compared to standard glibenclamide showing 14.90% and 20.67% plasma glucose reduction. Conclusions: This current research affirms prominent cytotoxic and moderate hypoglycemic potential of M. indica peel. Further bioactivity guided isolation of phytoconstituents and investigation on higher animals can lead to development of new drug molecules.

59–63

Authors: Wafa Abdullah Al-Megrin

Objective: To investigate the efficacy of pomegranate (Punica granatum) peel extract as an alternative treatment on the white laboratory mice against giardiasis. Methods: Experimental animals were divided into five groups, including Group A: control (infected untreated), Group B: infected and fed with pectin 7 days before infection, Group C: infected and fed with pectin starting from 7th day of infection, Group D: infected and fed with pomegranate peel extract 7 days before infection, and Group E: infected and fed with pomegranate peel extract starting from 7th day of infection. Results: Results from this study revealed that the prevention rate in the experimental groups reached approximately 50% by the 10th day of using pomegranate peel extract. Moreover, stool cyst counts of groups showed a significant reduction in the shedding of cysts approximately 75.6% by day 20 post-infection. ELISA test showed a reduction in Giardia antigen in the stools of the experimental groups which received pomegranate peel extract. The cure rate of these groups was approximately 97.4% by 28th day of infection. Conclusions: Our present findings indicated that the pomegranate peel extract proved to be valuable in prevention and treatment of Giardia lamblia infection. Further studies are required to determine the effective dose of pomegranate peel extract against Giardia lamblia infection.

70–77

Authors: Agustina Tri Endharti, Aswin Djoko Baskoro, Eviana Norahmawati

Objective: To investigate the anti-inflammatory effect of the protein derived from the soluble factor of Heligmosomoides polygyrus (H. polygyrus) excretory-secretory in a colitis model. Methods: Colitis was induced by providing drinking water containing 3% dextran sodium sulfate (DSS) for a week. DSS was administrated in a cycle protocol, each cycle consisted of 7 days of 3% DSS in the drinking water and followed by 7 days of regular water. This study consisted of five treatment groups, including Groups A (control) received untreated water, B (DSS only, without excretory-secretory), and C–E injected (i.p. ) with excretory-secretory protein (H. polygyrus excretory-secretory total, excretorysecretory 28 kDa and excretory-secretory 55 kDa, respectively). Mice received injection every week. The injection of excretory-secretory was started from the 6th weeks and continued until 11 weeks. At the end of 11 weeks of the experiment, mice were sacrificed, colon tissue was removed and then subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, flow cytometry, real-time PCR and histology examination. Results: Mice received H. polygyrus excretory-secretory 55 kDa reduced mono-nuclear cell infiltrations. H. polygyrus excretory-secretory 55 kDa induced the down-regulation of mRNA interferon- g expression. There were significant differences in the expression of mRNA interferon in the colon of mice after the administration of the excretory-secretory 55 kDa protein fraction compared with other groups (P < 0.001), whereas mRNA transforming growth factor- b expression up regulated in the colon of mice after the administration of the excretory-secretory 55 kDa protein fraction compared with total excretory-secretory group (P < 0.05). The treatment of colitis in mice with excretorysecretory 55 kDa protein fractions modulated interleukin-10 (IL-10) expression, whereas excretory-secretory total and excretory-secretory 28 kDa protein fractions insufficient promoted IL-10 expression. Excretory-secretory 55 kDa proteins fraction promoted IL-10 expression via Foxp3-independent pathways. Conclusions: Excretory-secretory 55 kDa protein could reduce inflammation and have potential therapy. H. polygyrus excretory-secretory 55 kDa was the soluble factor that may help in the development of novel treatments to cure colitis.

86–90

Authors: Aline Venticinque Caris, Wanessa Ysis, Valdir de Aquino Lemos, Ricardo Bottura, Ronaldo Vagner Thomatieli dos Santos

Exposure to hypoxia causes damage in several physiological systems, whose tissues are dependent on the O2 supply. Recently, there has been growing attention on the immunosuppressive and inflammatory potential of the hypoxia, including stimulation, nuclear factor kappa B pathway in macrophages and Th2 response from lymphocytes. These changes may result in transient immunosuppression and happen at the same time to worsening of cognition and other psychobiological aspects. Furthermore, exercise and nutrition, especially glutamine supplementation may provide important role, not pharmacological partially reversing the effects of hypoxia. In fact, recent studies show that moderate exercise can improve cognition in people exposed to hypoxia while the exercise associated with glutamine supplementation can reverse the increase in inflammatory markers and the Th1/Th2 balance. This review aims to bring the light of the discussion about nonpharmacological ways to prevent the effects of hypoxia on the connection between the immune system and the central nervous system.