Asian Pacific Journal of Tropical Biomedicine

183–187

Authors: Alhaji Hamisu Maimusa, Abu Hassan Ahmad, Nur Faeza Abu Kassim, Hamdan Ahmad, Hamady Dieng, Junaid Rahim

Objective: To determine abundance, distribution and diversity of potential breeding container habitats of the dengue vectors in public places including schools, restaurants, mosques and parks in southwest areas of Penang Island, Malaysia. Methods: Premises at restaurants, schools, parks and mosques were surveyed simultaneously and inspected visually for container habitats and production of immature mosquitoes from March 2015 to March 2016. Abundance (mean ± SE) of breeding containers between sites was compared using One-way ANOVA. Independent sample ttest was used to compare total number of Aedes albopictus (Ae. albopictus) and Aedes aegypti (Ae. aegypti) surveyed. Results: The surveyed locations yielded a total of 3741 breeding containers and 19537 immature mosquitoes from four areas. Concurrent artificial and natural containers produced 78.4% immature Ae. albopictus and 6.3% Ae. aegypti mosquitoes in wet season, with 14.2% Ae. albopictus and 1.1% Ae. aegypti mosquitoes in dry season. Artificial containers accounted for 98.1% of the total containers recorded, with restaurants being the most productive locations (8012) and schools being the least productive (2234). Conclusions: It was concluded that public places are good sources of potential container habitats of Aedes mosquitoes in Penang Island, Malaysia and Ae. albopictus has exclusively replaced the home-grown Ae. aegypti even in urban areas. Therefore, treatment of artificial containers in such locations is critical in Aedes mosquito control campaigns during dengue outbreaks.

193–197

Authors: Prabhu Raj Joshi, Mahesh Acharya, Rajan Aryal, Kamal Thapa, Trishna Kakshapati, Rathanin Seng, Anjana Singh, Sutthirat Sitthisak

Objective: To investigate the molecular epidemiology and antimicrobial resistance patterns of methicillin-resistant Staphylococcus aureus (MRSA) among healthcare workers and patients. Methods: MRSA isolates were recovered from nasal swabs collected at a tertiary care hospital of Nepal and confirmed on the basis of Gram staining, conventional biochemical tests, and PCR amplification of mecA gene. PCRs were also used for detection of the different resistance genes and staphylococcal cassette chromosome (SCC) mec types. Antibiotic susceptibility patterns of isolates were assessed by disc diffusion method and minimum inhibitory concentrations were determined by E-test. Results: A total of 29 MRSA were isolated from 536 nasal swabs (5.4%) of health care workers and patients at a tertiary care hospital in Nepal. All isolates were susceptible to amikacin, gentamicin, vancomycin (minimal inhibitory concentrations < 2 mg/mL), tigecycline, tetracycline, nitrofurantoin, rifampicin, quinupristin-dalfopristin, and linezolid. Among the 29 MRSA isolates, resistance to erythromycin (72%), ciprofloxacin (75%), co-trimoxazole (62%), clindamycin (10%), and chloramphenicol (10%) was found, and fifteen isolates (51%) exhibited high-level mupirocin resistance (minimal inhibitory concentrations > 1 024 mg/mL). Fourteen isolates were found harboring the mupA gene and one isolate was found carrying the novel mupB gene. High prevalence (68%) of SCCmec I type was found, followed by SCCmec V (13%) and SCCmec III (3%) among all the MRSA isolates. Conclusions: We found the emergence of SCCmec type I with high-level mupirocin resistance among MRSA in Nepal. Data also suggest that MRSA SCCmec type V strain has spread from the community to the hospital.

208–213

Authors: Agung Nugroho, Hesty Heryani, Jae Sue Choi, Hee-Juhn Park

Objective: To characterize the types, contents, and peroxynitrite-scavenging activities of flavonoids in the leaf of Carica papaya (C. papaya). Methods: Chromatographic and spectroscopic techniques along with high performance liquid chromatography quantitative analysis and peroxynitrite-scavenging assay were performed to isolate and quantify flavonoid compounds in the flavonoid-rich fraction (BuOH fraction) derived from MeOH extract of C. papaya leaves and evaluate their peroxynitrite-scavenging activities. Results: Seven flavonoids were isolated from the leaves of C. papaya, including quercetin 3-(2G-rhamnosylrutinoside), kaempferol 3-(2G-rhamnosylrutinoside), quercetin 3- rutinoside, myricetin 3-rhamnoside, kaempferol 3-rutinoside, quercetin, and kaempferol. All of the substances exhibited potent activities on peroxynitrite scavenging (IC50
4.15 mmol/L), which were stronger than the positive control, L-penicillamine (6.90 mmol/L). The content of kaempferol 3-(2G-rhamnosylrutinoside) was significantly higher than other identified compounds (123.18 mg/g BuOH fraction and 7.23 mg/g MeOH extract). Conclusions: The results of the present study demonstrate the potent antioxidant flavonoids of C. papaya leaf, with kaempferol 3-(2G-rhamnosylrutinoside) as the major one.

222–226

Authors: Damrongpan Thongwat, Supaporn Lamlertthon, Urat Pimolsri, Nophawan Bunchu

Objective: To evaluate the larvicidal activity of the ethanolic and aqueous extracts of the endocarp and seeds of Dracaena loureiri (D. loureiri) against the dengue mosquito vector, Aedes aegypti. Methods: Bioassays were performed by exposing late third-stage to early fourth-stage larvae of Aedes aegypti to various concentrations of the extracts from D. loureiri. The larval mortality was observed after 24- and 48-h exposure. Results: The larvicidal bioassay in this study demonstrated that the ethanolic endocarp extract was the most effective with the LC50 value of 84.00 mg/L after 24 h exposure and < 50 mg/L after 48 h exposure. Extracts from the other parts of the plant were significantly less effective as a larvicide. Conclusions: The ethanolic endocarp extract of D. loureiri demonstrated effective larvicidal activity. It is an alternative source for developing a novel larvicide for controlling this mosquito species.

234–239

Authors: Nabil Abdel Salam Ahmed Hasona, Mohammed Ahmed Qumani, Turki Ahmed Alghassab, Muath Abdulrahman Alghassab, Abdullah Ali Alghabban

Objective: To assess the ameliorative properties of Iranian Trigonella foenum-graecum L. (T. foenum-graecum) seeds and Punica granatum L. (P. granatum) peel extracts against streptozotocin-induced diabetes in male guinea pigs. Methods: Sixty guinea pigs were divided into six groups (10 guinea pigs per group). Group 1 consisted of normal animals. Groups 2 and 3 were treated with Iranian T. foenum-graecum seeds and P. granatum peel extract alone, respectively. Group 4 was treated with streptozotocin only; whereas Groups 5 and 6 receiving streptozotocin were treated with Iranian T. foenum-graecum seeds and P. granatum peel extract, respectively. All animals were treated for 30 days, and the body weight, blood and liver biochemical parameters were measured. Results: Guinea pigs exposed to streptozotocin showed an alteration in body weight gain, fasting glucose level, kidney function parameters (blood urea nitrogen and creatinine) as well as decreased serum and hepatic total protein level. In addition, it increased the cholesterol and triglyceride level, while decreasing the hepatic glucose-6 phosphate dehydrogenase activity, glycogen, glutathione content and hepatic catalase activity. Oral treatment with T. foenum-graecum seeds and P. granatum peel extracts revealed significant protective properties with respect to body weight gain and other biochemical parameters studied. Conclusions: The Iranian T. foenum-graecum seeds and P. granatum peel extracts are significantly potent in ameliorating diabetic condition induced by streptozotocin and improving various biochemical parameters in serum and liver of guinea pigs.

245–248

Authors: Abdelkrim Berroukche, Mohamed Terras, Abdelrahmane Labani, Hafsa Dellaoui, Wassila Lansari

Objective: To assess cadmium sulfate (CdSO4) and zinc chloride (ZnCl2) antagonist effects on the prostate specific antigen and prostatic cell organization in rats. Methods: The study included 40 adult male rats, divided into four groups: Group 1 (CdSO4), Group 2 (ZnCl2), Group 3 (CdSO4eZnCl2) and Group 4 (control). Animals were treated with CdSO4 and ZnCl2 at the same dose (15 mg/L) during 30 days. Results: It was showed a higher body weight and a lowering serum-prostate specific antigen concentration [(1.8 ± 0.6) ng/mL] in animals treated with CdSO4. CdSO4 induced a cyto-nuclear atypia, proliferative lesions, hyperplasia and precancerous foci in prostate tissue. Toxic effects of ZnCl2 were not recorded in this study. Conclusions: Protective role of zinc was exhibited against toxic effects of cadmium in prostate gland.

253–256

Authors: Yeasmeen Ali, Md. Ashraful Islam, Nazmul Hasan Muzahid, Mohd. Omar Faruk Sikder, Md. Amzad Hossain, Lolo Wal Marzan

Objective: To reveal the presence of methicillin resistant Staphylococcus aureus (S. aureus) (MRSA) in poultry samples and to determine the antibiogram pattern against five antibiotics. Methods: Samples from different poultry farm of Chittagong city, Bangladesh were examined for S. aureus by different biochemical tests and confirmed as MRSA by identifying the presence of mecA gene using PCR. Antibiotic resistance pattern in S. aureus was determined by antibiotic disk diffusion method. Results: In this study, a total of 60 samples (30 from nasal swabs and 30 from cloacal swabs) were used, of which 54 were confirmed as S. aureus by different biochemical tests. Among these, 12 were confirmed as MRSA by detecting mecA gene using PCR. During antibiogram study, both nasal and cloacal samples showed the highest resistance against penicillin-G and the lowest resistance was observed against neomycin. Conclusions: Based on the present study, it can be said that different antibiotics are used extensively in poultry that leads to MRSA and is alarming for human health.

262–264

Authors: Nawfal Rasheed Hussein, Amer Abdullah Balatay, Alpay Turan Sezgin, Rana Saadi Jawad

Objective: To investigate the prevalence of hepatitis C virus (HCV) and its genotypes in Duhok City, Kurdistan Region, Iraq. Methods: A cross sectional study was conducted to investigate the prevalence of HCV and its genotype. A total of 2109 subjects, who attended the hospital for complaints other than hepatitis, were recruited in this study. Results: First, anti-HCV antibody positivity was examined by ELISA. About 5.2% (111/ 2109) of our samples were tested positive for anti-HCV antibodies. To confirm the positivity, RT-PCR was performed. Amongst all samples, 2.8% (60/2109) was positive by RT-PCR. Then, we genotyped all the RT-PCR positive samples, and it was found that 50.0% (30/60) of our samples were typed as HCV genotype 4, 43.3% (26/60) as genotype 1 and 6.7% (4/60) as genotype 3. Conclusions: The prevalence of HCV was higher than that was reported previously and genotype 4 was the most prevalent. Further population based study is required to investigate the prevalence of HCV.

96–102

Authors: Bamidele Abiodun Iwalokun, Senapon Olusola Iwalokun, Bernice Enobong Udoh, Modinat Balogun

Objective: To assess the occurrence and pattern of Toll-like receptor 4 (TLR4) cosegregated genotypes among children with Plasmodium falciparum malaria in Nigeria. Methods: In this case-control study, a total of 79 Plasmodium falciparum infected children aged 2–7 years and 105 age-matched uninfected controls of Yoruba descents in Lagos were studied. The extracted DNA samples were used for TLR4 genotyping at codons 299 (Asp > Gly) and 399 (Thr > Ile) by PCR-restriction fragment length polymorphism. Malaria infection was diagnosed by blood smear microscopy and infected children were stratified into asymptomatic, uncomplicated and severe malaria sub-groups. Malnutrition was determined by measuring the mid upper arm circumference and anemia was defined as hemoglobin < 11 g/dL. Results: The proportions of children with acute malnutrition and severe anemia were 12.0% and 3.2%, respectively. Parasitemia and malnutrition were not correlated and four distinct patterns of TLR4 genotypes were found in the study population: Asp299Asp/ Thr399Thr (90.2%), Asp299Gly/Thr399Thr (4.3%), Gly299Gly/Thr399Thr (3.8%) and Asp299Gly/Thr399Ile (1.6%). These genotypes did not differ significantly (P > 0.05) in frequency between infected and non-infected children. However, low and high occurrences of the TLR4 Asp299Asp/Thr399Thr and Asp299Gly/Thr399Thr genotypes were observed in the severe malaria subgroup. Conclusions: This study reveals a protective role for TLR4 Asp299Gly/Thr399Ile and Asp299Asp/Thr399Thr genotypes against severe malaria in Nigerian children.

107–109

Authors: Agustin Iskandar, Karomah Sriwedari, Indah Adhita Wulanda, Muhammad Rasjad Indra, Hartojo, Novi Khila Firani, Ery Olivianto

Objective: To know the difference between chemerin and adipocyte fatty acid-binding protein (AFABP) levels in obese individuals with positive Toxoplasma gondii (T. gondii) immunoglobulin G (IgG) compared with negative T. gondii IgG. Methods: This study is a cross-sectional study by using consecutive sampling methods conducted from January to April 2013. The subjects were 57 obese individuals who were divided into obese group of positive and negative T. gondii IgG. The level of chemerin, AFABP and T. gondii IgG was done by ELISA. The data were analyzed by independent t test. Results: The results showed that the level of chemerin of positive T. gondii IgG group was significantly higher than the negative T. gondii IgG group [(70.0 ± 16.5) vs. (64.4 ± 16.1) pg/mL; P = 0.003], but there was not significant AFABP difference between seropositive and negative IgG groups [(83.6 ± 41.9) vs. (74.2 ± 36.7) pg/mL; P = 0.598]. Conclusions: It can be concluded that the level of chemerin of seropositive T. gondii IgG was higher than that in the negative T. gondii IgG group.

116–120

Authors: Ruben Merino-Guzm ´ an ´ , Juan David Latorre, Ramiro Delgado, Xochitl Hernandez-Velasco, Amanda Desha Wolfende, Kyle Dean Teague, Lucas Elzie Graham, Brittany Danielle Mahaffey, Mikayla Francis Ann Baxter, Billy Marshall Hargis, Guillermo Tellez

Objective: To standardize an ELISA protocol to quantify total immunoglobulin A (IgA) from different biological samples. Methods: Two independent experiments were conducted. In Experiment 1, total IgA levels were quantified from the lachrymal fluid, tracheal swab, and cloacal swab at various time points from Days 30 to 89 in white Leghorn chickens. Experiment 2 was conducted to evaluate the effect of 50 or 500 ppb of aflatoxin B1 (AFB1) on total IgA quantified in samples from the lachrymal fluid, tracheal swab, gut content and cloacal swab in broiler chickens at 21 days of age. Results: Lachrymal fluid contained the highest level of IgA; however, the sampling procedure was time consuming and stressful to the bird, and the sample volume depends largely on the size of the chicken. Cloacal swabs also contained a high concentration of IgA; this sampling procedure was faster than lachrymal fluid sampling and was not affected by the age of the bird. Tracheal sampling was more difficult than cloacal sampling; the age of the bird limited the sampling, and the IgA concentration was the lowest detected at all sampling ages. 500 ppb of AFB1 significantly reduced total IgA concentration in the gut content compared with control or 50 ppb of AFB1 treated groups. Interestingly, a significant reduction in total IgA was also observed in those chickens that received 50 ppb of AFB1 in gut rinse when compared with cloacal swabs. Conclusions: The results of this study suggest that cloacal swab is an easy and reliable way to evaluate mucosal IgA concentration in both Leghorn and broiler chickens.

126–133

Authors: Sumaiya Nahid, Kishor Mazumder, Zillur Rahman, Saiful Islam, Md. Humayun Rashid, Philip Grey Kerr

Objective: To evaluate the effect of methanolic extract of Syzygium cumini (L.) Skeels (S. cumini) seeds on the major organs in an animal model of diabetes through biochemical and histopathological studies. Methods: The methanolic extracts of S. cumini seeds (100 and 200 mg/kg body weight) were administered to alloxan-induced diabetic rats daily, with fasting blood glucose levels being measured by glucometry at one-day interval for a duration of two weeks. Biochemical assays to evaluate changes in the functions of the heart, liver, pancreas and kidney were carried out. Histopathological changes in the diabetic rat organs (pancreas, liver, heart, kidney and spleen) were also observed after the 14 days of treatment with the extracts. Results: Oral administration of methanolic extracts of S. cumini seeds (100 and 200 mg/ kg body weight), with gliclazide as a positive control (25 mg/kg), showed beneficial effects including lowering blood glucose levels (P < 0.001), improved heart and liver functions, and hyperlipidemia due to diabetes. At 200 mg/kg, the extracts reversed cardiac and liver damage caused by alloxan. Conclusions: In addition to the anti-hyperglycemic activity of methanolic extracts of S. cumini seeds, the extracts demonstrates potential to minimize cardiac and hepatic complications.

139–143

Authors: William Patrick Cruiz Buhian, Raquel Orejudos Rubio, Juliana Janet Martin-Puzon

Objective: To determine the thin-layer chromatography (TLC) fingerprint profiles and to evaluate the in vitro antioxidant activity of ethanol extracts of Muntingia calabura (M. calabura) leaves and stems. Methods: The leaves and stems were extracted using ethanol as solvent. The TLC separation of the phytochemical constituents of the leaf and ethanol extracts was carried out in ethyl acetate: n-hexane and chloroform: ethyl acetate mobile phase systems. Distinct spots were visualized under visible light, UV 254 nm, UV 366 nm and after spraying with vanillin-sulfuric acid. The 2,2-diphenyl-1-picrylhydrazyl free-radical scavenging assay was used to evaluate the antioxidant activity of the extracts. Results: Both the leaf and stem ethanol extracts at 4 mg/mL exhibited 2,2-diphenyl-1- picrylhydrazyl inhibition of more than 90%, relative to gallic acid. The results of TLC showed that the degree of resolution between the constituent spots was comparable between the two mobile phase systems using the different visualization wavelengths. Under the 254 nm visualization, few spots were observed in leaf and stem extracts. Visualization at 366 nm yielded the greatest number of observable spots of various colors in both leaf and stem extracts. More spots were visualized upon post-derivatization with vanillinsulfuric acid in the TLC chromatograms using chloroform: ethyl acetate mobile phase, compared to those in ethyl acetate: n-hexane mobile phase. Conclusions: M. calabura exhibited very high antioxidant activity in its leaves and stems ethanol extracts, both of which are used in traditional medicine. The TLC results demonstrated the presence of diverse secondary metabolites in the leaf and stem ethanol extracts, indicating that the antioxidant activity, including other bioactivities may be attributed to these phytochemical constituents. This paper has reported for the first time the TLC fingerprinting of M. calabura using visible light, UV 254 nm, UV 366 and postderivatization with vanillin-spray to visualize separate spots on TLC plates

147–150

Authors: Abdulrahman Khazim Al-Asmari, Md Tanwir Athar, Ahmed Abdullah Al-Faraidy, Mohammed Salim Almuhaiza

Objective: To focus on the analysis of chemical constituents of the Thymus vulgaris L. (locally known as “Zaitra” or “Za'atar”; Family: Lamiaceae) which is available in the market of Saudi Arabia. Methods: The Zaitra oil was analyzed by gas chromatography-mass spectrometry. The mass spectra were compared with the standard spectra available in National Institute Standard and Technique library. Results: The results indicated that the Thymus oil is composed of many chemical compounds including a pinene, thymol and caryophyllene which are biologically active and also used in various diseases. Conclusions: It can be concluded that the Thymus vulgaris due to presence of many bioactive compounds can be used as a new potential source of medicine for the treatment of various types of illness.

157–160

Authors: Marianne Haddad, Salam Zein, Hawraa Shahrour, Kamar Hamadeh, Nadine Karaki, Hussein Kanaan

Objective: To extract and identify the chemical composition of the polysaccharide isolated from the Eucalyptus cultivated in Lebanon and to evaluate its antioxidant activity. Materials: The water-soluble polysaccharide was isolated from Eucalyptus leaves, and its structure was identified by Fourier transform infrared spectroscopy, proton nuclear magnetic resonance, and carbon-13 nuclear magnetic resonance. The antioxidant activity of the active ingredient was screened for its radical scavenging ability using 2,2-diphenyl- 1-picryl-hydrazyl radical (DPPH) test. Results: The results of the DPPH test have shown that fucoidan, the polysaccharide isolated from Eucalyptus, exhibited almost the same antioxidant activity against DPPH$ as the ascorbic acid did at 100 mg/mL. Conclusions: This natural molecule extracted from a medicinal plant has a promising antioxidant activity and could be used in pharmaceutical and medical applications.

166–172

Authors: Zahra Rabiei

Epilepsy is a disorder in brain in which clusters of nerve cells, or neurons, occasionally signal abnormally and cause strange emotions, sensations, and behavior, or sometimes muscle spasms, convulsions, and loss of consciousness. Neurotransmitters in central nervous system greatly affect and play a very important part in neuronal excitability. Traditional treatments are still a component of health care system in many communities despite the fact that well-established alternatives are available. In this review article, we addressed epilepsy and its treatments with emphasis on medical plants and introduction of antiepileptic plants and their action mechanisms. Relevant articles published since 2010 were retrieved using the search terms including epileptic seizure, anticonvulsant, medicinal plants, and oxidative stress. Most plants/herbal preparations that are ethnomedically used to treat epilepsy or those which have been tested for anticonvulsant activity were reported. Overall, the results of the published articles show that the symptoms of epilepsy seizure can be inhibited or treated by active ingredients derived from medicinal plants.

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.

995–1000

Authors: Aly Younes, Hanaa El-Sherief, Fathia Gawish, Marwa Mahmoud

Objective: To evaluate the predatory potential of the Odonata nymph on freshwater snails that serve as intermediate hosts for Schistosoma species (Bulinus truncatus and Biomphalaria alexandrina). Methods: Observations on the searching, attacking and devouring of the two snail types with series of laboratory-based predation experiments, whose aims were to determine daily predation rate, differential predation, prey preference considering small-, mediumand large-sized snails were conducted. Results: Laboratory evaluation revealed that, the Odonata nymph could kill and consume the two intermediate hosts. The number of snails consumed differed according to the snail type, size and density. The times taken for searching and handling times were dependent on the snail size, type and satiation of the predator. The predation rate varied also with respect to snail type, size and density. This study also evaluated that Odonata nymphs consumed more Bulinus truncatus than Biomphalaria alexandrina per unit time, and that there may be a preference for smaller than larger snails. Conclusions: According to our observation, the predator, Hemianax ephippiger nymph may be a suitable biocontrol agent in connection with Schistosoma intermediate hosts.

1008–1014

Authors: Panan Ratthawongjirakul, Vorraruthai Thongkerd, Wanpen Chaicumpa

Objective: To investigate the impact of the fliD gene on the biofilm formation of Helicobacter pylori (H. pylori). Methods: H. pylori fliD mutant was constructed using inverse PCR mutagenesis. The mobility of the bacteria and its adhesion ability to human epithelial cells were assessed using a motility assay and a fluorescein isothiocyanate staining adhesion assay, respectively. The formation of biofilm was evaluated using a pellicle assay and a crystal violet staining assay. The cyto-architecture of the biofilm was documented with scanning electron microscopy. Results: It was found that there was no significant difference in the levels of bacterial adhesion and the biofilm formation between the wild-type ATCC 43504 and the fliD mutant. Apart from a poor motility, the fliD mutant had a slightly delayed formation of its biofilm and an incomplete cyto-architecture of its biofilm. The bacterial cells residing in the biofilm of the fliD mutant showed a loose accumulation with less apparent crosslinking fibrils. Most of the mutant cells had truncated flagella. Conclusions: This study provides the preliminary evidences that fliD potentially regulates biofilm formation and is required for the motility of H. pylori. Further studies need to be performed in order to develop fliD as a novel target for vaccine or antimicrobial agent in future.

1022–1028

Authors: Doroth
ee Dinangayi Tshilanda, Philippe Bila Babady, Damase Nguwo Vele Onyamboko1, Christian Muamba Tshilolo Tshiongo, Damien Sha-Tshibey Tshibangu, Koto-te-Nyiwa Ngbolua, Philippe Vuka Tsalu, Pius Tshimankinda Mpiana

Objective: To carry out a phyto-chemical characterization of essential oil from Ocimum basilicum L. (O. basilicum) harvested in DR Congo and to assess the antioxidant potential of crude extracts with respect to the polarity for comparison reason. Methods: The phyto-chemical characterization of essential oil produced by hydrodistillation was performed by coupled gas chromatography-mass spectrometer analysis and the antioxidant potential evaluation by in vitro 2,2-diphenyl-1-picrylhydrazyl free radical scavenging activity method. Results: A previously weighed amount of fresh leaves of O. basilicum produced 0.65% of essential oil that led to the identification of a set of 84.44% out of 99.98% as major compounds (> 1.5%). The chemo-type of this essential oil was linalool-methyl chavicol. Chemical components of oil were characterized by oxygenated aromatic hydrocarbons (46.00%) and oxygenated monoterpenes (26.75%). With respect to the amount of components, methyl chavicol also known as estragole (35.72%) constituted the very large quantity afterward linalool (21.25%) and then epi-a-cadinol (8.02%), a-bergamotene (6.56%), eugenol (4.60%), 1,8-cineole (4.04%), germacrene D (2.06%), thymol (1.64%), and (E)- citral (1.55%), respectively. Essential oil exhibited antioxidant potential and IC50 = (1.180 ± 0.015)mg/mL. Non-polar crude extracts yieldswere low compared to the one of polar extracts. Only methanol and ethyl acetate had considerably manifested antioxidant potential with IC50 values equal to (0.025 ± 0.013) mg/mL and (0.085 ± 0.012) mg/mL, respectively. As concerns to IC50 values, essential oil was less active than methanol and ethyl acetate extracts. The methanol crude extract exhibited the highest activity. Non-polar extracts showed insignificant radical scavenging ability that did not allow assessing IC50 values. These results highlighted the occurrence of antioxidant potential compounds in polar media. Conclusions: Essential oil and crude extracts of O. basilicum growing in DR Congo can be advocated as natural sources of antioxidant potential compounds not only in food but also in pharmaceutical industries. The high antioxidant potential of polar crude extracts highlights antioxidant character of its composition particularly butyl stearate and rosmarinic acid we isolated and identified, respectively in the methanol crude extract.

1037–1043

Authors: Kayini Chigayo, Paul Eanas Lesedi Mojapelo, Simon Mnyakeni-Moleele, Jane Masiiwa Misihairabgwi

Objective: To determine suitable phytochemical extraction solvents, screen for phytochemicals, determine the total phenol and flavonoid contents and the antioxidant activities of different solvent extracts of Kirkia wilmsii (K. wilmsii), an ethnomedicine in South Africa. Methods: Extractionswere performed fromdried tubers of the K. wilmsii plant, using several solvents and varying extraction times. Extract yields were determined and suitable extraction solvents were selected. Total phenol and flavonoid contents of the extracts were determined spectrophotometrically using gallic acid and quercetin as standards. The free radical scavenging activity of the extracts was investigated using 1,1-diphenyl-2-picrylhydrazyl radical. Results: Phytochemical screening confirmed the presence of phenolics, flavonoids, terpenoids, tannins, cardenolide deoxy sugars and reducing sugars.Of the 12 solvent extracts used, six gave yields higher than 5%, while the other six gave yields less than 1%. The highest extract yield of 52.9%was obtained using 80%methanol while the lowest yield of 7.3%was obtained using ethanol at 60 min. The 80% methanol, methanol/chloroform/water (12:5:3) and 60% methanol extracts were significantly higher than those of ethanol, methanol and water (P < 0.05). Total phenolic content recorded extracts ranged from (45.32 ± 0.50) to (122.84 ± 0.31) mg gallic acid equivalent per gram. A maximum total flavonoid content of (917.02 ± 0.10) mg quercetin equivalent per gram and a minimum of (206.26 ± 0.10) mg quercetin equivalent per gram were recorded for methanol and water, respectively. The flavonoid content for methanol was significantly higher than all the other extracts (P < 0.05). The scavenging profiles of K. wilmsii extracts were significantly lower (P < 0.05) than that of ascorbic acid and IC50 values ranged from 129.94 mg/mL for methanol to 225.04 mg/mL for water. An IC50 value of 56.52 mg/mL was obtained with ascorbic acid. Conclusions: Ethanol, methanol, methanol/chloroform/water, 80% methanol, 60% methanol and water can be used as suitable phytochemical extraction solvents for K. wilmsii tubers. Total phenolic content and total flavonoid content analysis proved the presence of high levels of phenolic compounds as well as flavonoids. The presence of phenols and flavonoid could be responsible for the radical scavenging activities observed.

1050–1055

Authors: Patcharawan Tanamatayarat

Objective: To evaluate all parts of Protium serratum (Wall. ex Colebr.) Engl. (P. serratum) for their phytochemistry and biological activities including antityrosinase, antioxidant, and brine shrimp lethality. Methods: Nine ethanolic extracts from different P. serratum parts such as twig, whole fruit, pericarp, and root were investigated for their phytochemical screening and biological activities in terms of tyrosinase inhibition, antioxidant against 1,1-diphenyl-2- picrylhydrazyl and hydroxyl radicals, and lethality to brine shrimp larvae. Results: Phytochemical screening also revealed the presence of flavonoids, condensed tannins, alkaloids, triterpenoids, steroids, and sugars in P. serratum. The root extract was the most effective for antityrosinase activity with IC50 of (21.63 ± 0.31) mg/mL, and the leaf extract exhibited the highest antioxidation activity using 1,1-diphenyl-2-picrylhydrazyl and hydroxyl radical scavenging methods, with IC50 of (4.34 ± 0.09) and (119.80 ± 1.01) mg/ mL, respectively. The essential oil extracted from the whole fruit displayed the highest toxicity against brine shrimp, with LC50 of (3.57 ± 1.82) mg/mL. Conclusions: This study indicates that ethanolic extracts from each P. serratum part have differences in phytochemistry and biological activities (antityrosinase, antioxidant, and brine shrimp lethality). Some parts of the plant should be considered in the further study.

1065–1075

Authors: Hassan Nasirian

The brown-banded cockroach, Supella longipalpa (Blattaria: Blattellidae) (S. longipalpa), recently has infested the buildings and hospitals in wide areas of Iran, and this review was prepared to identify current knowledge and knowledge gaps about the brown-banded cockroach. Scientific reports and peer-reviewed papers concerning S. longipalpa and relevant topics were collected and synthesized with the objective of learning more about health-related impacts and possible management of S. longipalpa in Iran. Like the German cockroach, the brown-banded cockroach is a known vector for food-borne diseases and drug resistant bacteria, contaminated by infectious disease agents, involved in human intestinal parasites and is the intermediate host of Trichospirura leptostoma and Moniliformis moniliformis. Because its habitat is widespread, distributed throughout different areas of homes and buildings, it is difficult to control. Considering its possible resistance to insecticides, the control situation may be far more complex. For improved control of S. longipalpa an integrated pest management program is needed. Sanitation, indoor insecticide spraying in the initial cockroach control phase and insecticide formulation baits are recommended simultaneously.

909–913

Authors: Masteria Yunovilsa Putra, Tutik Murniasih, Respati Tri Swasono, Joko Tri Wibowo, Annissa Nur Cahya Saputri, Meilia Rahma Widhiana, Irma Shita Arlyza

Objective: To investigate the antioxidant, antibacterial, antimalarial activities and cytotoxicity of the n-hexane, ethyl acetate, n-butanol, and aqueous fractions from a crude extract of Lobophytum sp. Methods: This organism was collected from the Selayar Islands (South Sulawesi). The antioxidant activity was performed by the 1,1-diphenyl-2-picryl hydrazyl radical scavenging method. All fractions from the crude extract of Lobophytum sp. were examined for their cytotoxicity using the brine shrimp lethality bioassay and heme polymerization inhibitory activity assay for antimalarial activity. Results: It was found that the ethyl acetate, n-butanol and aqueous fractions exhibited heme polymerization inhibitory activity with IC50 values of 11.7, 14.3 and 12.0 mg/mL, respectively, while the n-butanol fraction showed moderate antioxidant activity and cytotoxicity with IC50 values of 150.00 and 92.74 mg/mL, respectively. Conclusions: This study provides information on antioxidant, antibacterial and antimalarial activities as well as the cytotoxicity of all fractions from the crude extract of Lobophytum sp. This is a new report of antimalarial substances derived from Lobophytum sp.

924–930

Authors: Yunita Armiyanti, Renam Putra Arifianto, Elisa Nurma Riana, Kartika Senjarini, Widodo Widodo, Loeki Enggar Fitri, Teguh Wahju Sardjono

Objective: To identify antigenic proteins from the salivary glands of female Anopheles maculatus using a proteomic approach to find the biomarker candidate for serological tools. Methods: The identification of antigenic proteins of Anopheles maculatus salivary gland used these techniques: one-dimensional gel electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis), western blot, and liquid chromatography–mass spectrometry. Results: The proteins that have molecular weight (MW) 43 and 34 kDa were the antigenic protein. Computational bioinformatic analysis by Mascot Server revealed seven novel hypothetical proteins (MW: 43 kDa) and two novel hypothetical proteins (MW: 34 kDa). Further analysis (BLASTP, antigenicity, epitope mapping, and specificity analysis) showed that two novel proteins were identified as apolipoprotein D and cathepsin D in Anopheles darlingi. Conclusions: The identified proteins are potential to be developed as a biomarker of mosquito bite's exposure.

937–944

Authors: Idir Moualek, Ghenima Iratni Aiche, Nadjet Mestar Guechaoui, Souad Lahcene, Karim Houali

Objective: To evaluate the antioxidant and anti-inflammatory activities of aqueous extract of Arbutus unedo (A. unedo) leaves. Methods: In this context, the in vitro antioxidant activity was demonstrated by 2,2- diphenyl-1-picrylhydrazyl, hydroxyl radical and H2O2 radical scavenging, ferrous ion chelating, ferric reducing power, total antioxidant capacity and by the protection against peroxidation of b-carotene-linoleic acid in emulsion. The anti-inflammatory activity was evaluated first by studying the membrane of human red blood cells against different hypotonic concentrations of NaCl and against heat, inhibiting the denaturation of albumin. Results: Total phenolic and flavonoid content were found respectively [(207.84 ± 15.03) mg gallic acid equivalent/g, and (13.070 ± 0.096) mg quercetin equivalent/g]. The extract displayed significant scavenging activity of some radicals such as 2,2-diphenyl-1- picrylhydrazyl [IC50 at (7.956 ± 0.278) mg/mL],
OH [IC50 = (1015.74 ± 46.35 mg/ mL)], H2O2 [IC50 = (114.77 ± 16.86) mg/mL] and showed a good antioxidant activity through ferrous ion chelating activity [IC50 = (1014.30 ± 36.21) mg/mL], ferric reducing power [IC50 = (156.55 ± 17.40) mg/mL], total antioxidant capacity [IC50 = (461.67 ± 4.16) mg/mL] and b-carotene-linoleic acid protection against peroxidation [I % = (87.04 ± 1.21)% at 1 000 mg/mL]. Conclusions: A. unedo showed in vitro anti-inflammatory activity by inhibiting the heat induced albumin denaturation and red blood cells membrane stabilization. Our results show that aqueous leaf extract of A. unedo has good antioxidant activity and interesting anti-inflammatory properties. A. unedo aqueous extract can be used to prevent oxidative and inflammatory processes.

950–956

Authors: Poonsit Hiransai, Jitbanjong Tangpong, Chuthamat Kumbuar, Namon Hoonheang, Onrunee Rodpech, Padchara Sangsuk, Urairat Kajklangdon, Waraphorn Inkaow

Objective: To determine the cytotoxicity, reduction in nitric oxide production and antioxidative activity of the aqueous leaf extract from Tithonia diversifolia (T. diversifolia) in an in vitro model. Methods: Leaves of T. diversifolia were collected from natural habitats and extracted with distilled water using the decoction method. The cytotoxic effect of the extract in terms of cell viability was determined using RAW264.7 cells and human peripheral blood mononuclear cells (PBMCs) via the mitochondrial respiration method using the MTT reagent. The effect of the extract on lipopolysaccharide (LPS)-induced nitric oxide production in RAW264.7 cells was measured using the Griess reagent. The chemical antioxidant was evaluated by ABTS- and DPPH-radical scavenging assays. Results: The half-maximal cytotoxic concentration values were 145.87 mg/mL and 73.67 mg/mL for human PBMCs and RAW264.7 cells, respectively. In the presence of phytohemagglutinin-M, the IC50 on PBMCs proliferation was 4.42 mg/mL. The noncytotoxic range of the extracts inhibited LPS-induced nitrite production in RAW264.7 cells with an IC50 value of 11.63 mg/mL. To determine the anti-oxidative properties, the N-acetyl cysteine equivalent antioxidant capacity of the extract was (32.62 ± 1.87) and (20.99 ± 2.79) mg N-acetyl cysteine/g extract, respectively determined by the ABTS-radical and DPPHradical assay. However, the extract did not confer death protection in a hydrogen peroxideinduced RAW264.7 co-culturing model. Conclusions: Our study demonstrated the immunomodulation caused by the aqueous leaf extract of T. diversifolia, resulting from the inhibition of phytohemagglutinin-Minduced PBMCs proliferation and LPS-induced nitric oxide production in RAW264.7 macrophages. Although the anti-oxidative activity was presented in the chemical-based anti-oxidant assay, the extract cannot protect cell death from stress conditions.

962–966

Authors: Muhammad Sulaiman Zubair, Syariful Anam, Subehan Lallo

Objective: To evaluate the cytotoxic activity of wood extracts of Lunasia amara Blanco (L. amara) and to perform further phytochemical standardization. Methods: The wood extracts of L. amara were assessed for cytotoxic activity by in vitro tetrazolium bromide (MTT) method against two human cancer cell lines, cervical cancer cells (HeLa) and breast cancer cells (T47D). Thin layer chromatography, Dragendorf, acetic anhydride-sulfuric acid and ferric chloride were used to detect alkaloids, steroids and polyphenols, respectively. Furthermore, quantitative determination of total alkaloid by ultra fast liquid chromatography-photodiode array detection using lunacrine as a marker compound was performed as well. Results: The ethyl acetate extract exhibited higher cell-growth inhibition than methanol and n-hexane extracts on HeLa and T47D cancer line cells with the IC50 of 71.15 and 79.04 mg/mL, respectively. Total alkaloid in ethyl acetate extract was counted as (10.46 ± 0.28)% (w/w), while lunacrine determined by ultra fast liquid chromatographyphotodiode array detection method was found to be (3.55 ± 0.26)% (w/w). Conclusions: The high total alkaloid and lunacrine concentration on the extract confirm the potential cytotoxic property of ethyl acetate wood extract of L. amara.

973–974

Authors: Silvia Angeletti, Rossella Ottaviani, Arcangela Michela Lanotte, Lucia Pedicino, Marina De Cesaris, Alessandra Lo Presti, Eleonora Cella, Marta Fogolari, Massimo Ciccozzi, Giordano Dicuonzo

No

982–988

Authors: Baha Latif, Azdayanti Muslim

Sarcocystosis is a zoonotic disease caused by a coccidian intracellular protozoan parasite of the genus Sarcocystis. More than 200 Sarcocystis species have been recorded and the parasites are found in mammals, birds and reptiles. They require two hosts to complete their life cycle. In Malaysia, sarcocystosis was reported as a potential emerging food and water-borne disease after a series of large outbreak of human infections. There was not enough attention given before even though it was reported in both humans and animals. The first human case of invasive muscular sarcocystosis among local Malaysian was reported in 1975. Besides, a retrospective autopsy examination on 100 tongues revealed 21% positive cases. On top of that, a sero-epidemiological survey conducted in 243 subjects in West Malaysia showed that 19.7% had Sarcocystis antibodies. The clinical symptoms of muscular sarcocystosis were first described comprehensively in 1999. Meanwhile, many types of animals including livestock were found harbor the sarcocysts in their tissue. The first case of human intestinal sarcocystosis was reported in 2014. This review indicates that human sarcocystosis is currently endemic in Malaysia and parallel to that reported in animals. However, more studies and investigations need to be conducted since the source of human infection remains unknown.