Asian Pacific Journal of Tropical Biomedicine

796–800

Authors: Daniella de Sousa Mendes Moreira Alves, Rodrigo Gurgel-Gonçalves, Patr´ ıcia Albuquerque, Cesar Augusto Cuba-Cuba, Maria Imaculada Muniz-Junqueira, Selma Aparecida Souza Kuckelhaus

Objective: To evaluate whether the inoculation of contaminated cultures in the peritoneal cavity of mice could implement decontamination of Acanthamoeba cultures. Methods: Suspensions of Acanthamoeba, Acanthamoeba polyphaga ATCC 30461, or Acanthamoeba spp. isolated from soil (UnB13 strain) were inoculated in the peritoneal cavity of Swiss mice (n = 24). After 1, 6, 12, or 24 h of exposure the peritoneal cavity was washed and assessed for the presence of bacteria, fungi, and Acanthamoeba. Results: After 1 h of intraperitoneal inoculation at least 97% of the bacteria and 96% of the fungi (P < 0.05) and 99% of the bacteria (P < 0.05) were successfully eliminated from the ATCC 30461 strain and from the soil isolate UnB13 strain, respectively. This method also allowed the recovery of most trophozoites and cysts from both Acanthamoeba cultures at the end of 24 h. Conclusions: Our data demonstrated that this technique has great potential for decontamination of Acanthamoeba cultures in a short period of time.

806–811

Authors: Xian-Wen Yang, Hai-Xing Jiang, Xiao-Li Huang, Shi-Jia Ma, Shan-Yu Qin

Objective: To assess the role of Th9 and Th17 cells in malignant ascites (MA). Methods: MA from 30 hepatic carcinoma patients and benign ascites from 30 cirrhotic patients were collected. Corresponding peripheral blood samples from these hepatic carcinoma and cirrhotic patients as well as 30 healthy subjects were collected. The frequency of Th9 and Th17 cells was tested by flow cytometry. Serum levels of interleukin (IL)-9 and IL-17 were examined by ELISA. Results: The observed frequency of Th9 and Th17 cells, and the IL-9 and IL-17 serum levels were significantly higher in MA patients than those in cirrhotic patients and healthy control samples (P < 0.05). Moreover, the Th9 cells demonstrated positive correlation with Th17 cells as well as IL-9 in MA patients; however, this positive correlation was not observed in the cirrhotic patients or healthy control samples. The frequency of Th9 and Th17 cells was distinctly higher in MA patients presenting with stage III or IV malignancy and with lymph node or distant metastasis than those in patients in stage I or II and without distant metastasis (P < 0.05). Conclusions: The increased frequency of Th9 and Th17 cells in MA patients suggests that these two T cell subsets play a synergistic role in MA pathogenesis. This study also demonstrated that Th9 and Th17 cells may perform their biological functions in conjunction with IL-9 production.

819–823

Authors: Junsei Taira, Eito Tsuchida, Masatsugu Uehara, Natsuko Ohhama, Wakana Ohmine, Takayuki Ogi

Objective: To find anti-melanogenesis materials used in whitening cosmetics. Methods: The ethanolic leaf extract of Mallotus japonicus (M. japonicus) having an anti-melanogenesis activity was separated by a sephadex LH-20 chromatography. Each fraction was measured for its tyrosinase inhibitory activity together with its polyphenol content using the Folin–Ciocalteu method. The anti-melanogenesis activity of the active fractions was determined by the melanin content in the murine B16F1 melanoma. The active fractions were put together due to similar constituents, and then separated by high performance liquid chromatography using a C-18 ODS column. The major antimelanogenesis compound was identified using 1H and 13C-NMR and liquid chromatography-mass spectrometry. Results: The ethanolic leaf extract of M. japonicus showed an anti-tyrosinase activity with a high polyphenol content, resulting in suppression of melanin production in the B16F1 melanoma. The extract was separated and the active compound was identical as rutin based on the 1H, 13C-NMR and liquid chromatography-mass spectrometry analysis data. In addition, the rutin treatment with cells reduced the melanin content in a concentration dependent manner without any cell toxicity. The leaf extract of M. japonicus containing rutin would be useful in whitening cosmetics for protection from UV-light exposure to be limiting the accumulation of melanin in skin. Conclusions: The leaf extract of M. japonicus and/or rutin isolated from the extract as a key whitening agent would be useful as a whitening cosmetic material for protecting against disorder skin due to melanin accumulation.

829–833

Authors: Samira Asgharzade, Zahra Rabiei, Mahmoud Rafieian-Kopaei

Objective: To evaluate the effect of ethanolic extract of chamomile on balance and motor learning in rats receiving scopolamine and intact rats. Methods: Fourty-two rats were divided into 6 groups (n = 7). Control group received distilled water. Rats in Group 2 were given 1 mg/kg scopolamine. Groups 3 and 4 received chamomile extract 200 mg/kg and 500 mg/kg, respectively, and scopolamine simultaneously for 20 days. Intact groups (Groups 5 and 6) only received chamomile extract 200 mg/kg and 500 mg/kg, respectively. Motor coordination of rats was assessed with rotarod apparatus. Results: According to the obtained results, compared with the control group, scopolamine significantly decreased time spent on rotarod performance (P < 0.001). Compared with scopolamine group, the strength and staying on rotarod apparatus in Group 3 significantly increased (P < 0.05). The results of this research showed that intact groups that received only chamomile extract at doses of 200 mg/kg and 500 mg/kg significantly increased time spent on rotarod, compared with scopolamine group (P < 0.001). Conclusions: The results of this study indicated the high antioxidant property and protective effect of chamomile extract on motor coordination in the groups that received scopolamine.

841–844

Authors: Zong Ning, Yu-Long Bai, Hua Lu, Kang-Lin Mo

Objective: To investigate the prognostic value of plasma C-reactive protein (CRP) level in patients with paraquat poisoning. Methods: This study included 162 patients with paraquat poisoning. The data of plasma paraquat, CRP level and arterial blood gas were analyzed. Cox regression analysis was applied to evaluate the risk factors of prognosis. Receiver operating characteristics curve analysis and area under curve were used to calculate the predictive power of significant variable. Differences in patient survival were determined using the Kaplan–Meier method and a log-rank test. Results: Plasma CRP level was significantly increased in non-survival patients compared with survival patients (P < 0.05), and positively correlated with plasma paraquat level (P < 0.05). Cox regression analysis revealed that plasma CRP level was an independent prognostic marker of mortality within 30 days. The receiver operating characteristics curve analysis indicated that area under curve of plasma CRP level was 0.867 (95% CI: 0.81–0.93), and the cut-off value was 18 mg/L, and patients with CRP level over this value had a poor survival time compared with those with less than this value. Conclusions: These results suggest that plasma CRP level is distinct increased in patients with paraquat poisoning, and the plasma CRP level may be useful for the prediction of prognosis in paraquat poisoning.

852–857

Authors: Sri Rahayu Lestari, Muhammad Sasmito Djati, Achmad Rudijanto, Fatchiyah Fatchiyah

Objective: To monitor the physiological characteristics and genes expression of obesity rat model after rambutan peel extract (RPE) treatment. Methods: Twenty-four 12-week-old male rats were divided into 4 groups: normal, obesity, obesity treated with ellagic acid (O-EA) and obesity treated with RPE30 (ORPE30). Physiological characteristics were monitored by measuring body weight, calorie intake, size of adipocyte and level of triglyceride. Peroxisome proliferator activated receptor gamma (PPARg), CCAAT/enhancer-binding proteins a and fatty acid binding protein 4 (FABP4) expression were observed using immunohistochemistry, Western blotting and quantitative RT-PCR methods. Results: Body weight gain of O-EA and O-RPE30 rats were lower than obesity group and size of adipocyte cells were smaller than obesity group (P < 0.05), but when we compared to normal group, those groups had higher body weight gain and larger adipocyte cells. The level of triglycerides, protein expression of PPARg and mRNA level of FABP4 genes were significantly downregulated on O-EA and O-RPE30 compared to obesity group (P < 0.05). Our results indicated that RPE had potential substance as inhibitor of body weight gain, declining of size of adipocyte, level of triglycerides, PPARg expression and mRNA level of FABP4 gene on obesity rat model. Conclusions: RPE have anti-obesity activity by inhibiting body weight gain, declining size of adipocyte, decreasing triglyceride, PPARg expression and mRNA level of FABP4 gene on obesity rat model.

865-869

Authors: Ferry Efendi, Retno Handajani, Nursalam Nursalam

Objective: To explore the potential of local agar of genus Gracilaria, Eucheuma, Gelidium and local brands as an alternative for imported agarose for DNA electrophoresis, and to examine their ability related to separation and migration of DNA fragments in DNA electrophoresis. Methods: Their performance at various concentrations were compared via an experimental study with a specific brand of imported commercial agarose used in molecular biology research. The measured variables were separation and migration during electrophoresis of a DNA fragment. Results: The local agar genus Gracilaria gigas, Gelidium, brand “B” and brand “S” could separate DNA fragments at a concentration between 1% and 2%, with an optimum concentration of 2% w/v, as good as a specific brand of imported commercial agarose. Conclusions: Their performance were very close to that of commercial agarose and can still be improved by further agar purification as well as by pH and sulfur control.

874–877

Authors: Mohamed Afifi, Aaser Mohamed Abdelazim

Objective: To test the ability of both zinc oxide nanoparticles (ZnONPs) and silver nanoparticles (SNPs) to ameliorate the oxidative stress resulted from diabetes in diabetic rats. Methods: Fifty male albino rats were used; ten of them were served as control group and forty, as the experiment group, were injected with streptozotocin at the single intraperitoneal dose of 100 mg/kg. Then, the experiment group was subdivided into, diabetic, diabetic + ZnONPs, diabetic + SNPs and diabetic + insulin groups. The activities and mRNA expression levels of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase were determined in brain tissues. Malondialdehyde, total antioxidant capacity, zinc and silver concentrations were estimated in the brain tissues of all rats. Results: A significant increase in the activities and mRNA expression levels of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase was shown. Malondialdehyde levels were significantly decreased while there was a significant increase in the zinc, silver concentrations and total antioxidant capacity in brain of ZnONPs and SNPs treated rats, compared with diabetic or diabetic + insulin group and their control group. Conclusions: ZnONPs and SNPs can be used to ameliorate the oxidative stress in brain resulted from diabetes mellitus.

880–884

Authors: Sylvester Okhuelegbe Eraga, Mathew Ikhuoria Arhewoh, Rosemary Ngozi Chibuogwu, Magnus Amara Iwuagwu

Objective: To investigate the pharmaceutical equivalence of ten brands of ibuprofen tablets (400 mg) purchased from pharmacies in Benin City, Nigeria. Methods: The drug samples were subjected to uniformity of weight, crushing strength, friability, melting point, disintegration and dissolution tests following acceptable and official protocols. The ibuprofen content was determined using UV and high performance liquid chromatography method. Results: Crushing strength values of the drug samples ranged between 6 and 16 kp while the disintegration times were between 7.43 and 10.40 min (for uncoated tablets) and 3.25–37.32 min (for coated tablets). Friability values were less than 1% and the melting points of recrystallized ibuprofen from the samples ranged from 73.5 to 76.0
C. The amount of ibuprofen released within 1 h ranged between 18% and 102% and two brands failed the content of active ingredient in the UV method of assay while all the brands passed the test using HPLC. Conclusions: Ibuprofen (400 mg) tablets marketed in Benin City, Nigeria vary in pharmaceutical quality.

702–706

Authors: Rosario Galarza-Seeber, Juan David Latorre, Xochitl Hernandez-Velasco, Amanda Drake Wolfenden, Lisa Renee Bielke, Anita Menconi, Billy Marshall Hargis, Guillermo Tellez

Objective: To evaluate the ability of Bacillus spp. as direct-fed microbials (DFM) to biodegrade aflatoxin B1 (AFB1) by using an in vitro digestive model simulating in vivo conditions. Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 mg/mL of AFB1 in modified Czapek-Dox medium. Plates were incubated at 37
Cand observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16S rRNA sequence analyzes for identification. Tolerance to acidic pH, osmotic concentrations of NaCl, bile salts were tested, and antimicrobial sensitivity profiles were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time (3:15 h), supernatants and digesta were collected for highperformance liquid chromatography fluorescence detection analysis by triplicate. Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of fluorescence and area of clearance around each colony in modified Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identified the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions (pH 2.0), tolerant to a high osmotic pressure (NaCl at 6.5%), and were able to tolerate 0.037% bile salts after 24 h of incubation. No significant differences (P > 0.05) were observed in the concentrations of AFB1 in neither the supernatants nor digesta samples evaluated by high-performance liquid chromatography with fluorescence detection between positive control or DFM treated groups. Conclusions: In vitro digestion time was not enough to confirm biodegradation of AFB1. Further studies to evaluate the possible biodegradation effects of the Bacillus-DFM when continuously administered in experimentally contaminated feed with AFB1, are in progress.

714–719

Authors: Haytham Ahmed Zakai, Wajihullah Khan

Objective: To analyse the efficacy of diethylcarbamazine (DEC), tetramisole and chlorpromazine on the longevity and activity of glucose-6-phosphatase and succinate dehydrogenase in the microfilariae recovered from the peripheral circulation of the rats before and after the treatment. Methods: Setaria cervi worms were implanted in white rats via laparotomy and microfilaraemic rats were divided into 4 groups. Groups 1, 2 and 3 were treated with DEC, tetramisole and chlorpromazine respectively, while Group 4 served as infected control. Longevity of microfilariae and differential leucocyte counts were recorded till the disappearance of microfilariae from peripheral blood. Glucose-6-phosphatase and succinate dehydrogenase enzymes were localized in the microfilariae recovered from normal and treated rats. Results: The microfilariae survived for 48 days in untreated rats while survival was reduced to 15, 21 and 27 days after treatment with DEC, tetramisole and chlorpromazine, respectively. Eosinophils and neutrophils increased during 2nd and 3rd weeks, whereas the lymphocytes increased during 4–7 weeks. DEC treatment resulted in slight decrease in the localization of succinate dehydrogenase but not in glucose-6-phosphatase. Tetramisole and chlorpromazine treatment did not show any appreciable change in the localization of both the above enzymes. Conclusions: DEC proved the most effective drug which cleared the microfilaraemia within 15 days and reduced the activity of succinate dehydrogenase to some extent followed by tetramisole and chlorpromazine which took more time for the clearance of microfilariae and had no effect on the localization of both glucose-6-phosphatase and succinate dehydrogenase.

728–732

Authors: Farida Hussan, Rina Haryani Osman Basah, Mohd Rafizul Mohd Yusof, Nur Aqilah Kamaruddin, Faizah Othman

Objective: To determine the effect of Plantago major (P. major) extract on the liver injury following acetaminophen (APAP) toxicity. Methods: The male Sprague Dawley rats (n = 38) were randomly divided into normal control (n = 6) and experiment (n = 32) groups. The latter was subdivided into four groups and induced with APAP (1000 mg/kg) per oral, followed by P. major extract and N-acetylcysteine orally to the respective groups for six days. Results: On the seventh day, the serum bilirubin, liver enzymes and tissue malondialdehyde were increased in APAP groups whereas the total protein in serum, tissue superoxide dismutase and glutathione levels were reduced. The plant extract treatment reduced the histological deteriorations such as aggregation of hepatocellular cords, formation of binucleated cells and vacuolisation of the cells with scanty cytoplasm. It also revealed significant reduction of malondialdehyde and increased level of superoxide dismutase and glutathione. The findings in the extract treated groups were comparable to the group treated with N-acetylcysteine. Conclusions: In conclusion, P. major can enhance innate antioxidant activity and ameliorate the APAP-induced liver injury.

738–743

Authors: Hataipan Phienwej, Ih-si Swasdichira, Surattana Amnuoypol, Prasit Pavasant, Piyamas Sumrejkanchanakij

Objective: To investigate the effect of Tinospora crispa (T. crispa) extract on matrix metalloproteinase-13 (MMP-13) expression and cell migration. Methods: The cytotoxicity of T. crispa extract was examined by 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay on head and neck squamous cell carcinoma (HNSCC) cell lines. The effect on expression of MMP-13 was analysed by RT-PCR and ELISA. The migration was assessed by wound healing assay. Results: MMP-13 mRNA was highly expressed in the metastatic human HNSCC cell lines, HN22 and HSC-3. T. crispa extract at a concentration of 100.0 mg/mL caused about 50% reduction of cell survival. T. crispa extract at a non-toxic concentration of 12.5, 25.0 and 50.0 mg/mL significantly suppressed MMP-13 mRNA expression and secreted MMP- 13 in both HN22 and HSC-3. The expression of tissue inhibitors of metalloproteinase-2 (TIMP-2) by HSC-3 cells was attenuated by 25.0 and 50.0 mg/mL of T. crispa extract. Addition of the extract to cells in a wound healing assay showed inhibition of cell migration by HN22 cells. Conclusions: These data suggest that T. crispa could be considered as a potential therapeutic drug to prevent metastasis of HNSCC.

748–755

Authors: Anastasia Wheni Indrianingsih, Sanro Tachibana, Rizna Triana Dewi, Kazutaka Itoh

Objective: To isolate and investigate antioxidant and a-glucosidase inhibitor compounds in the leaves of Quercus gilva Blume (Q. gilva). Methods: Dry leaves of Q. gilva were extracted with methanol and the methanolic extract was further separated by silica gel column chromatography using several solvents with increasing polarity. The antioxidant activities of the isolated compounds were evaluated using various in vitro assays: 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, hydrogen peroxide radical scavenging activity, b-carotene bleaching assay, and reducing power assay. The a-glucosidase inhibitory assay was conducted against aglucosidase from Saccharomyces cerevisiae. Results: Three compounds were isolated and their structures were identified as catechin (1), epicatechin (2), and tiliroside (3) using an instrumental analysis. Compound 2 had higher antioxidant activity with inhibitory concentrations (IC50) of (22.55 ± 2.23) mmol/L than that of quercetin, which was used as the standard, with an IC50 of (28.08 ± 2.39) mmol/L, followed by compound 1 with IC50 of (40.86 ± 3.45) mmol/L. On the other hand, compound 3 had the lowest antioxidant activity with an IC50 of (160.24 ± 8.15) mmol/L. However, compound 3 had the highest a-glucosidase inhibitory activity with an IC50 of (28.36 ± 0.11) mmol/L, followed by compounds 1 and 2 with (168.60 ± 5.15) and (920.60 ± 10.10) mmol/L, respectively. Conclusions: The results obtained for the antioxidant activities and a-glucosidase inhibitory activities in a methanolic extract from the leaves of Q. gilva confirmed the potential of this plant as a source of natural antioxidants and antidiabetic medicine.

761–767

Authors: Hai Nguyen Thanh, Hue Pham Thi Minh, Tuan Anh Le, Huong Duong Thi Ly, Tung Nguyen Huu, Loi Vu Duc, Thu Dang Kim, Tung Bui Thanh

Objective: To investigated the protective potential of ethanol extracts of Scutellaria baicalensis (S. baicalensis) against lipopolysaccharide (LPS)-induced liver injury. Methods: Dried roots of S. baicalensis were extracted with ethanol and concentrated to yield a dry residue. Mice were administered 200 mg/kg of the ethanol extracts orally once daily for one week. Animals were subsequently administered a single dose of LPS (5 mg/ kg of body weight, intraperitoneal injection). Both protein and mRNA levels of cytokines, such as tumor necrosis factor alpha, interleukin-1b, and interleukin-6 in liver tissues were evaluated by ELISA assay and quantitative PCR. Cyclooxygenase-2, inducible nitric oxide synthase, and nuclear factor-kB protein levels in liver tissues were analyzed by western blotting. Results: Liver injury induced by LPS significantly increased necrosis factor alpha, interleukin-1b, interleukin-6, cyclooxygenase-2, inducible nitric oxide synthase, and nuclear factor-kB in liver tissues. Treatment with ethanol extracts of S. baicalensis prevented all of these observed changes associated with LPS-induced injury in liver mice. Conclusions: Our study showed that S. baicalensis is potentially protective against LPSinduced liver injury in mice.

773–777

Authors: Mustafa Kemal Çelen, Murat Sayan, Tuba Dal, Celal Ayaz, Alicem Tekin, Tuncer O¨ zekinci, Suda Tekin Koruk, Tunga Barcin, Recep Tekin, Mehmet Sinan Dal, Sevgi Kalkanlı

Objective: To detect the subtype characterization and drug-resistant mutations in HIV-1 strains after the refugee movement from Syria to Turkey between 2011 and 2014 in south east border lines. Methods: A total of 65 patients were included in this study, of which 57 (88%) patients were antiretroviral therapy-naive patients. HIV-1 RNA was detected and quantified by real-time PCR assay. HIV-1 subtypes and circulating recombinant forms (CRFs) were identified by phylogenetic analysis (neighbor-joining method), and drug-resistant mutations were analyzed. Results: Three major HIV groups were indicated. Two of these groups were located in subtype B. The other group showed heterogeneity. Subtype B (48/65, 73.8%), followed by CRFs (12/65, 18.5%) was the most common strain. Subtype of CRFs consisted of CRF01_AE (9/65, 13.8%) and CRF02_AG (3/65, 4.6%). Subtype C (1/65, 1.5%), sub-subtypes A1 (2/65, 3.1%) and F1 (2/65, 3.1%) were also detected with low prevalence. The rate of overall primary antiretroviral resistance was 4.9% (3/61). Drug-resistant rate for non-nucleoside reverse transcriptase inhibitors was 4.9%. The thymidine analogue mutation rate was 13.1% (8/61). Conclusions: HIV molecular epidemiology studies are necessary to determine transmission patterns and spread. Subtype B and CRF01_AE, CRF02_AG are the most prevalent strains in the south-east of Turkey. However, subtype C, sub-subtypes A1 and F1 are of low prevalence but persist in the south-east of Turkey. In the near future, changing of HIV epidemiology will be possible in Turkey due to migration movement in border lines and resistance testing will play an important role in HIV management.

781–784

Authors: D´ebora Oro, Andreia Heissler, Eliandra Mirlei Rossi, Diane Scapin, Patrıcia da Silva Malheiros, Everton Boff

Objective: To evaluate the antifungal effect of Cinnamomum zeylanicum (C. zeylanicum) and Melaleuca alternifolia essential oils and honey against strains of Candida sp. from HIV-positive patients in order to subsidize new therapeutic strategies for candidiasis. Methods: The study evaluated the antifungal effect of natural antimicrobials against 30 strains of Candida sp. isolated from oral cavities in HIV-infected patients. Then, they were compared to the action of fluconazole and amphotericin B. Antifungal susceptibility was evaluated by the broth macrodilution technique and the minimum inhibitory concentration and the minimum fungicidal concentration were determined. Results: Among all antifungals evaluated in this study, amphotericin B was the one showing the best results; however, all compounds studied here showed inhibitory activities against isolates of Candida sp. Honey (0.0313–64 mg/mL) demonstrated fungistatic activity inhibiting 70% of the isolates. C. zeylanicum essential oil (0.0313–64 mg/ mL) inhibited 93.3% of the Candida strains and Melaleuca alternifolia essential oil (0.0313–64 mg/mL) was able to inhibit 73.3% of them. Conclusions: Therefore, all natural compounds evaluated in this study, especially C. zeylanicum essential oil, may become promising agents for oral candidiasis therapy including in HIV-positive patients.

612–618

Authors: Wan Fatma Zuharah, Chan Jia Ling, Nurfazlina Zulkifly, Nik Fadzly

Objective: To evaluate the lethal concentration, oviposition deterrence and ovicidal activity of acetone extracts of Melanochyla fasciculiflora (M. fasciculiflora) leaf and Gluta renghas (G. renghas) leaf against Aedes albopictus (Ae. albopictus). Methods: To determine the lethal concentration of Anacardiaceae, ten test concentrations of the extracts ranging from 200 to 650 mg/L were selected for larvicidal bioassays and 25 early fourth instar larvae were exposed to the extracts for 24 h. The sub-lethal concentrations used for oviposition deterrence was the value of LC25, LC50 and LC75 from above study which is 235 mg/L, 470 mg/L and 705 mg/L for M. fasciculiflora extract and 187.5 mg/L, 375 mg/L and 562.5 mg/L for G. renghas extract, respectively. Twenty gravid Ae. albopictus were allowed to oviposit in different treated concentrations. For oviciding procedure, a total of 300 eggs of Ae. albopictus were soaked in solution with each treated concentration as mentioned above for 24 h. After 24 h, eggs were sieved and soaked in seasoned water, and hatching rates were calculated. For comparison, only seasoned water was used in control experiment. Results: G. renghas demonstrated lower LC50 value of 372.80 mg/L compared to M. fasciculiflora (467.90 mg/L). The activity index of negative oviposition revealed the deterrent effect and thus, caused a remarkable negative response resulting in oviposition of fewer eggs compared with control (without plant extract). The acetone extract of M. fasciculiflora was more effective than G. renghas extract in displaying oviposition deterrence potential since the latter did not possess the deterring effect within the concentration range tested. However, both plant extracts exhibited excellent oviciding effect as 92.33% of eggs failed to be hatched when treated with 705.0 mg/L of M. fasciculiflora and 86.67% with 562.5 mg/L of G. renghas. The oviposition deterrence and percentage of egg mortality were directly proportional to the concentrations of extracts in both plants tested. Conclusions: These results clearly indicate that the acetone extract of G. renghas could be served as potential larvicide, whereas M. fasciculiflora has better sub-lethal effect for oviposition deterrence and against Ae. albopictus as an oviciding agent.

624–628

Authors: Mohamed Habib Oueslati, Faraj A. Al-Ghamdi, Adel Noubigh

Objective: To isolate and characterize the bioactive secondary metabolites from aerial parts of widespread Chenopodiaceae taxa growing in Saudi Arabia: Salsola villosa Delile. ex Schul. Methods: Antibacterial activities of chloroformic extract, fractions and isolate compounds was evaluated against five bacterial strains (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus and Staphylococcus epidermidis), using a paper disc diffusion method. The purification of compound(s) of chloroform extract was done by chromatographic column of silica gel. The structure elucidation was determined by extensive spectroscopic analysis (1H and 13C nuclear magnetic resonance, correlation spectroscopy, heteronuclear multiple bond correlation, heteronuclear multiple quantum coherence and nuclear overhauser enhancement spectroscopy) and high resolution electrospray ionization mass spectroscopy analysis. Results: Bioactivity guided fractionation of the chloroformic extract led to the isolation of two bioactive compounds: 4-(40-hydroxy-20-methylcyclopent-20-enyloxy)- 4-methylcyclopent-2-enol (1) named salsolanol and 40-[3-(hydroxymethyl)oxiran-2-yl]-3- [(E)-3-hydroxyprop-1-en-1-yl]-6, 20-dimethoxy [1, 10-biphenyl]-2-ol (2) named biphenylsalsinol. The antibacterial effects of the chloroform extracts, fractions and isolated compounds 1 and 2 were also evaluated in this work. Results showed that the compounds 1 and 2 exhibited antibacterial activities against four strains: Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Pseudomonas aeruginosa with diameter of zone of inhibition ranging between (9.33 ± 0.94) to (26.33 ± 0.94) mm. Conclusions: Based on data presented here, two new natural compounds secondary cyclic alcohol 1 and biphenylpropanoid 2 isolated from bioactive chloroformic extract from aerial parts of Salsola villosa can be responsible for its antibacterial activities.

636–644

Authors: Tarik Khouya, Mhamed Ramchoun, Abdelbassat Hmidani, Souliman Amrani, Hicham Harnafi, Mohamed Benlyas, Younes Filali Zegzouti, Chakib Alem

Objective: To evaluate the anti-inflammatory, anticoagulant and antioxidant effects of aqueous extracts of thyme varieties from Moroccan. Methods: The aqueous extracts of tree medicinal plants [Thymus atlanticus (T. atlanticus), Thymus satureioides and Thymus zygis (T. zygis)] were screened for their antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl radical-scavenging, ferric reducing antioxidant power assay, radical scavenging activity method, the inhibition of 2,2'-azobis (2-amidinopropane) dihydrochloride that induces oxidative erythrocyte hemolysis and thiobarbituric acid reactive substances assay. The anti-inflammatory activity of aqueous extracts was evaluated in vivo using croton oil-induced ear edema and carrageenan-induced paw edema in mice and rats, respectively. This extracts were evaluated in vitro for their anticoagulant activity at the different concentrations by partial thromboplastin time and prothrombin time activated. Results: All thyme varieties were found to possess considerable antioxidant activity and potent anti-inflammatory activity in the croton oil-induced edema. Administration of aqueous extracts of two varieties (50 mg/kg) (T. zygis and T. atlanticus) reduced significantly the carrageenan-induced paw edema similar to non-steroidal anti-inflammatory drug (indomethacin, 10 mg/kg). In partial thromboplastin time and prothrombin time tests, T. atlanticus and T. zygis extracts showed the strongest anticoagulant activity. In contrast, Thymus satureioides did not show the anticoagulant activity in these tests. Conclusions: All aqueous extracts possess considerable antioxidant activity and are rich in total polyphenol and flavonoid but they act differently in the process of inflammatory and coagulation studied. This study shows great variability of biological activities in thyme varieties.

650–657

Authors: Atamgba Agbor Asuk, Margaret Akpana Agiang, Kayode Dasofunjo, Amonor James Willie

Objective: To analyse the phytochemical contents of leaf, stem bark and root of Jatropha curcas (J. curcas) in four solvent extracts and their proximate and mineral compositions. Methods: Standard analytical procedures were used for the determination of phytochemicals, proximate and mineral compositions of the leaf, stem bark and root extracts of J. curcas. Results: Results of the analysis showed the presence of polyphenols, flavonoids, alkaloids, cardiac glycosides, coumarins, saponins, terpenoids, steroids, triterpenoid saponins, carotenoids, phlobatannins and tannins in the leaf, stem bark and root of all the solvent extracts. Flavonoids were present in the highest amount in the ethyl acetate extracts of the leaf (7.35% ± 0.02%), stem bark (4.12% ± 0.01%) and root (3.35% ± 0.02%) followed by polyphenols in the methanol extracts of leaf (4.62% ± 0.02%), stem bark (2.77% ± 0.05%) and root (2.49% ± 0.02%). Poly-acetylated compounds were absent in all the solvent extracts of the leaf, stem bark and root. However, some anti-nutritional agents such as oxalates, phytates and cyanates were present in all the solvent extracts of the leaf, stem bark and root except the ethyl acetate. Phytates were high in the aqueous solvent of the leaf (6.12% ± 0.00%) but low in the stem bark (1.00% ± 0.05%) and root (0.89% ± 0.03%). Proximate composition showed appreciable amounts of total carbohydrate (36.33% ± 0.72%), crude protein (26.00% ± 0.47%) and reducing sugars (5.87% ± 0.14%) in the leaf, while crude fat was more in the stem bark (16.70% ± 0.30%). There was corresponding substantial energy in the leaf [(1514.77 ± 20.87) kJ/100 g] and stem bark [(907.00 ± 8.52) kJ/100 g]. Moisture and ash contents of the leaf, stem bark and root were within acceptable limits for the use in drugs formulation. The mineral composition showed substantial amounts of important elements such as Fe, Ca, Na, Mg and Zn. Others were P, K and Se. Conclusions: The outcome of this study suggests that the leaf, stem bark and root of J. curcas have very good medicinal potentials, meet the standard requirements for drug formulation and serve as good sources of energy and nutrients except for the presence of some anti-nutritional elements predominant in the leaf.

663–670

Authors: King Shimumbo Nalubamba, Eugene Chisela Bwalya, Ntombi Basimbi Mudenda, Hetron Mweemba Munangandu, Musso Munyeme, David Squarre

Objective: To determine the gastrointestinal tract helminthic fauna in domestic and wild guineafowl in Zambia. Methods: Post-mortem and laboratory parasitological examinations for helminth identification and enumeration were conducted on 198 guineafowls (148 domestic and 50 wild) from November 2010 to October 2011. Results: All guineafowls were infested with one or more helminths. Eleven helminth species, namely, Raillietina echinobothrida, Raillietina tetragona, Raillietina cesticillus, Ascaridia galli, Allodapa suctoria, Gongylonema ingluvicola, Tetrameres spp., Heterakis spp., Acuaria spiralis, Syngamus trachea, and Streptocara pectinifera were identified with no trematodes recorded. Mean nematode burden between domestic and wild fowl showed no differences having 113.7 [confidence interval (CI) 98.9–128.6] and 108 (CI 76.6–139.5) nematodes respectively. In contrast, female guineafowls had a mean of 151.9 (CI 128.4–177.8) nematodes per host which was significantly more than the males that had a mean of 79.6 (CI 66.8–94.4). However, there were differences in helminth species richness between domestic and wild guineafowls with domestic guineafowls having more species present at a mean of 4.2 (CI 3.91–4.44) than the wild ones at a mean of 3.4 (CI 2.92–3.88) but there were no sex differences. Eight of the eleven helminth species cooccurred in domestic and wild fowl and five of the helminth species had higher prevalence in domestic guineafowls. Conclusions: Syngamus trachea, Streptocara pectinifera and Acuaria spiralis are reported for the first time in domestic poultry in Zambia. This study represents the first comparative study of helminths in domestic and wild guineafowls at an interface area and adds to the knowledge base in a discipline where a dearth currently exists.

676–683

Authors: Essa Ajmi Alodeani, Mohammad Arshad, Mohammad Asrar Izhari

Objective: To deal with the anti-uropathogenic and in silico screening of (E-)-N0- (substituted-benzylidene)-2-(quinolin-8-yloxy)acetohydrazide analogues in order to search the potential anti-uropathogenic agents. Methods: Three (E-)-N0-(substituted-benzylidene)-2-(quinolin-8-yloxy)acetohydrazide analogues were synthesized. Structure elucidation was done using various spectroscopic techniques including infrared radiation, 1hydrogen-nuclear magnetic resonance, carbon- 13 nuclear magnetic resonance, etc. Physicochemical score, bioactivity score and molecular docking studies were carried out using Lipinski's rule of five, Molinspiration (web based software), Autodock 4.2 tools. In vitro anti-uropathogenic activity was carried out against four pathogens named as Staphylococcus aureus (S. aureus), Staphylococcus epidermidis, Proteus mirabilis and Escherichia coli by disc diffusion method and macrodilution test following their morphological and biochemical characterization. Results: The formation of (E-)-N0-(substituted-benzylidene)-2-(quinolin-8-yloxy)acetohydrazide is confirmed from the spectroscopic results. All the compounds were found in compliance with Lipinski's rule of five and exhibited bioactivity score from −0.50 to 0.00. Docking results revealed that compound-1 is forming one hydrogen bond with TYR 576 and two hydrogen bond with GLU 569, while compound-2 is forming one hydrogen bond with ARG 599, and compound-3 forming 0 hydrogen bond. The anti-uropathogenic evaluation exhibited that compound one exhibited better activity against S. aureus, while it was found to possess moderate to good activity against both Gram-positive bacteria and Gram-negative bacteria excluding S. aureus. Conclusions: Our study revealed that compound one exhibited better activity than the standard in case of S. aureus and moderate to good activity against rest of the pathogens. Molecular docking, physicochemical and bioactivity studies strongly supported the experimental results. From the well obtained results it was concluded that compound-1 can lead as potential anti-uropathogenic agents.

689–693

Authors: Pascalina Chanda-Kapata, William Ngosa, Albertina Ngomah Moraes, Nicole Maddox, Nathan Kapata

Objective: To assess the prevalence of HIV infection, to highlight HIV-testing refusal rates among participants in a population-based tuberculosis survey and to assess the implication for programme implementation. Methods: This cross-sectional study on the characteristics of participants who refused HIV testing was conducted in a national survey in Zambia. All eligible participants were aged above 15 years and included in the analysis. Results: Out of the 44 791 tuberculosis survey participants, 14 164 (31.6%) refused to participate in HIV testing. The unemployed, rural dwellers, married, and those aged 15– 24 years were associated with higher refusal rates. Conclusions: Strategies to improve HIV testing acceptance are necessary. Qualitative research is recommended to understand the reasons for testing refusals so that remedial interventions can be implemented.

510-514

Authors: Sameer Rushdi Organji, Hussein Hasan Abulreesh, Khaled Elbanna, Gamal Ebrahim Haridy Osman, Manal Khider

Objective: To investigate the true incidence of Bacillus cereus (B. cereus) in food and children diarrhea cases. Methods: A total of 110 samples of various dairy products such as raw milk, long life pasteurized milk, yoghurt and infant powdered milk formulas, raw rice, and feces were examined for the presence of B. cereus by selective plating on mannitol-egg-yolkpolymyxin agar. Confirmation of B. cereus was carried out by biochemical tests and PCR. Identification of non-B. cereus isolates was carried out by 16S rDNA sequencing. Antimicrobial susceptibility was done by disk diffusion method. Results: Overall 35 samples (31.8%, n = 110) yielded Bacillus-like growth. Of which 19 samples (54.28%) were positive for B. cereus. All isolates were positive for enterotoxin production. No psychrotolerant B. cereus strains were detected in all samples. All B. cereus isolates were resistant to penicillin G, but susceptible to vancomycin, erythromycin and clindamycin. Conclusions: The results of this study confirm the importance of including B. cereus in disease control and prevention programs, as well as in routine clinical and food quality control laboratories in both Saudi Arabia and Egypt.

525-532

Authors: Demetrio L. Valle Jr., Jeannie I. Andrade, Juliana Janet M. Puzon, Esperanza C. Cabrera, Windell L. Rivera

Objective: To investigate the antibacterial activities of crude ethanol extracts of 12 Philippine medicinal plants. Methods: Crude ethanol extracts from 12 Philippine medicinal plants were evaluated for their antibacterial activity against methicillin-resistant Staphylococcus aureus, vancomycinresistant Enterococcus, extended spectrum β-lactamase-producing, carbapenem-resistant Enterobacteriaceae and metallo-β-lactamase-producing Pseudomonas aeruginosa and Acinetobacter baumannii. Results: The leaf extracts of Psidium guajava, Phyllanthus niruri, Ehretia microphylla and Piper betle (P. betle) showed antibacterial activity against the Gram-positive methicillinresistant Staphylococcus aureus and vancomycin-resistant Enterococcus. P. betle showed the highest antibacterial activity for these bacteria in the disk diffusion (16-33 mm inhibition diameter), minimum inhibitory concentration (19-156 μg/mL) and minimum bactericidal concentration (312 μg/mL) assays. P. betle leaf extracts only showed remarkable antibacterial activity for all the Gram-negative multidrug-resistant bacteria (extended spectrum β-lactamaseproducing, carbapenem-resistant Enterobacteriaceae and metallo-β-lactamase-producing) in the disk diffusion (17-21 mm inhibition diameter), minimum inhibitory concentration (312-625 μg/mL) and minimum bactericidal concentration (312-625 μg/mL) assays. Conclusions: P. betle had the greatest potential value against both Gram-negative and Grampositive multidrug-resistant bacteria. Favorable antagonistic activities were also exhibited by the ethanol extracts of Psidium guajava, Phyllanthus niruri and Ehretia microphylla.

538-543

Authors: Innocent Onyesom, Ejovi Osioma, Precious Chiamaka Okereke

Objective: To assess the effects of hot water leaf extract of Nauclea latifolia (N. latifolia) on antioxidant status, lipid peroxidation values and parasite levels in hepatic and brain tissue of experimental mice (BALB/c) infected with Plasmodium berghei (P. berghei) malaria. Methods: Forty nine mice were divided into seven groups (n = 7) and used for the study. Group A (control) were given 0.2 mL/kg phosphate buffer saline; Group B mice were infected with P. berghei and treated with phosphate buffer saline. Groups C and D mice were also infected but treated with 200 and 300 mg/kg body weight of leaf extract respectively. Groups E and F mice were not infected, but received 200 and 300 mg/kg of leaf extract respectively. Group G mice were infected and treated with chloroquine (5 mg/kg). Liver and brain tissues of mice were prepared for both biochemical assay and microscopic examination. Results: Results showed that P. berghei malaria infection induced oxidative stress in both liver and brain tissues as evidenced by the significant (P < 0.05) decrease in antioxidants: superoxide dismutase, reduced glutathione and catalase. These reductions perhaps caused compromise in membrane integrity as indicated by the significant increase in lipid peroxidation product malondialdhyde. Malaria parasites were also identified in these tissues. However, N. latifolia treatment eliminated the parasites in tissues and protected them from oxidative damage even better than chloroquine treatment did, whose anti-malarial potency also cleared tissue parasites. The measurement of protection by N. latifolia against damage was strengthened by the insignificant micro structural alterations. Conclusions: The bioactive phytochemical(s) in N. latifolia should be structured and the mechanism(s) of its antimalarial tendency should be further investigated.

547-552

Authors: Jicheng Yu, Liu Zhang, Qiu Liu, Xiaohui Qi, Ying Ji, Beom Seok Kim

Objective: To evaluate various types of samples from the different marine environments as sources of actinomycetes from the Yalujiang coastal wetland, North China, and to screen their antimicrobial properties. Further, the identified actinomycetes were characterized based on morphological, biochemical, and physiological characteristics. Methods: Eight different production media were used to isolate actinomycets from different stations of marine soil sediments in Yalujiang coastal wetland and the genotypic positions were established by 16S rDNA. Results: A total of 172 actinomycetal isolates were obtained from 13 samples using five media. The most effective culture media in the isolation of actinobacteria were Gause’s Synthetic agar and Starch-casein agar. Among 172 isolates, 46 isolates (26.74%) showed antibacterial activity, 70.93% belonged to the genus Streptomyces, others were Micromonospora spp. and Rhodococcus spp. Out of the 46 isolates, two cultures were further supported by morphological characterization analysis. Conclusions: This is the first report about actinomycetes isolated from Yalujiang coastal wetland and it seems that the promising isolates from the unusual/unexplored wetland may prove to be an important step in the development of microbial natural product research.

560-565

Authors: Ali Rafati, Mahboobeh Erfanizadeh, Ali Noorafshan, Saied Karbalay-Doust

Objective: To investigate the effects of benzene on rat’s cerebellum structure and behavioral characteristics, including anxiety and motor impairment. Methods: Twenty rats were randomly allocated into two groups orally receiving distilled water and benzene (200 mg/kg/day). A total of 10 rats were used at the beginning of benzene exposure. Two rats died during benzene treatment and 8 rats remained for evaluation of the behavioral test and finally 6 rats underwent histological assessment. At the end of the 4th week, motor function and anxiety were evaluated in rotarod test and elevated plus maze, respectively. Besides, the cerebellum was dissected for structural assessment using stereological methods. Results: Performance of the benzene-treated rats in fixed and accelerating speed rotarod was impaired and their riding time (endurance) was lower compared to the control group (P = 0.02). The benzene-treated rats also spent less time in the open arms and had fewer entrances to the open arms in comparison to the control group, indicating anxiety (P = 0.01). The total volume of the cerebellar hemisphere, its cortex, intracerebellar nuclei, total number of the Purkinje, Bergmann, Golgi, granule, neurons and glial cells of the molecular layer, and neurons and glial cells of the intracerebellar nuclei were reduced by 34%-76% in the benzene-treated rats in comparison to the distilled water group (P = 0.003). The most cell loss was seen in Bergmann glia. Conclusions: The structure of cerebellum altered after benzene treatment. In addition, motor impairment and anxiety could be seen in benzene-treated rats.

571-575

Authors: Ivan Viktorovich Borozda, Mikhail Anatolievich Danilov, Kirill Sergeevich Golokhvast

Objective: To propose an original method of surgical treatment for the acromial extremity of the clavicle rupture (Rockwood II-VI) with modeled Kirschner wire and cortical screw. Methods: Anatomical study and a test method were applied to 43 cadavers of both sexes. During the period between 2000 and 2013, 34 patients of both sexes were operated upon using the new method. In the comparison group (n = 120), the fixation of the acromial extremity of the clavicle rupture was performed with hamate plate, Lee hook and Kirschner wires. Results: Its application allows, according to the evaluation scale of Constant and Murley (1987), 10% more preservation of the function of the shoulder compared with traditional methods of surgical treatment, and shortens the required hospital treatment and temporary disability periods. Conclusions: It is shown that the proposed author’s method combines low invasiveness, minimum dimensions of the construction and low-cost treatment.

581-586

Authors: Rina Girard Kaminsky, Renato Valenzuela Castillo, Coralia Abrego Flores

Objective: To document epidemiologic data, clinical manifestations and treatment in hospitalized children with Trichuris dysenteric syndrome from Honduras during 2010-2012. Methods: Severe trichuriasis cases were identified by routine stool examinations from hospitalized patients (12 years old or less). Relevant epidemiologic, clinical and management data were obtained from review of clinical histories in the ward. Results: Of 122 Trichuris infections diagnosed in 11 528 (1.0%) stool samples for all ages, 81 (66.4%) were identified in the age group 2-12 years old, 21 (25.9%) of which were severe (≥ 100 eggs in 2 mg of feces). Thirteen of those 21 patients collaborated in this study. Patients (9 males and 4 females) were of rural precedence, from large poor or very poor families, chronically parasitized, and between 2 and 12 years old. Dysentery of months duration, severe anemia and stunting were common complaints; clinical characteristics associated with heavy Trichuris infections included egg counts from 232-3 520 eggs per direct smear, hemoglobin from 3.4-10.8 g/dL, eosinophilia up to 43%, severe malnutrition and growth stunting. Orally administered drugs mebendazole, albendazole, metronidazole, nitaxozanide, and piperazine were prescribed at different dosages and duration other than recommended; no cure or egg excretion control was exercised before patient release. A range of 340 to about 10 000 worms were recovered after treatment from 8 patients. Conclusions: This report underlines the need for detailed community studies in trichuriasis morbidity, effective treatment assessment and clinical response in severely malnourished parasitized children.

429-433

Authors: Hamidreza Alizadeh Otaghvar, Mostafa Hosseini, Adnan Tizmaghz, Ghazaal Shabestanipour, Hamid Noori

Metastatic breast cancer is a disease of early breast cancer that usually occurs several years after the early breast cancer. Breast cancer is the most common cancer among Iranian women. According to the new statistics in Iran 6 160 breast cancers are diagnosed in the country each year and 1 063 cases lead to death. In this paper, epidemiology, diagnosis and treatment have been investigated. In this study, case-control clinical trials and open studies with adequate data were collected. Due to the higher risk of age group 40-49 years and the advent of advanced breast cancer in Iranian women, the early diagnosis and determination of the exact size of the tumor before surgery is important in choosing a therapy plan. The decision on the therapy of invasive breast cancer depends on several factors such as cancer stage, tumor size and type, pathological and cytological status of the tumor, the patient’s opinion, the presence or absence of estrogen and progesterone receptors in the cytoplasm of tumor cells and so on.

438-443

Authors: Omar Messaoudi, Mourad Bendahou, Ibrahim Benamar, Djamal-Elddine Abdelwouhid

Objective: To study the antimicrobial activity of actinomycetes isolated from sabkha of Kenadsa and identification of the isolate interesting. Methods: Eighteen strains were isolated, using four culture media from sebkha of Kenadsa (Bechar, Southwestern Algeria). Screening of antimicrobial activity consisted of two steps: in primary screening, antibacterial activity was determined by using the agar plug method against test strains; in secondary screening, better isolate which showed a good activity in the first screening was selected to extract antimicrobial substances. The antimicrobial activities of extracts were evaluated by using Kirby-Bauer disc diffusion method. Partial characterization of antimicrobial products was performed on the basis of chemical revelations, UV-vis spectrometry and infrared spectroscopy. The identification of isolate interesting was performed through morphological, chemical, biochemical and physiological characteristics. Results: All isolates showed antimicrobial activity against at least one microorganism test. One isolate, LAM143cG3, was selected for its broad spectrum and high antimicrobial activity. The isolate LAM143cG3 was identified as Spirillospora sp. The comparison between the species of this genus (Spirillospora rubra and Spirillospora albida) and our isolate indicated the existence of several physiological and biochemical differences which led us to suppose that this was a new member of this genus. Primary characterization of antimicrobial substances produced by the isolate LAM143cG3 indicated the presence of amines and phenols. The UV–vis spectrum suggested a non-polyenic nature of substances secreted by our isolate, while infrared confirmed the presence of amine groups. Conclusions: The result of the present study revealed that sebkha of Kenadsa was rich in rare actinomycetes, that secreted interesting antimicrobial substance.

449-455

Authors: Samah Mamdouh Mohamed Fathy, Ashraf Mohamed Mohamed Essa

Objective: To investigate the effect of intra-peritoneal injection of purified exudates of axenic Spirulina platensis on the mammalian endocrine and nervous systems. Methods: The intra-peritoneal injection of the cyanobacterial exudates in mice was applied. Sex hormonal levels of testosterone and progesterone were measured using radioimmunoassay while the follicular stimulating hormone and luteinizing hormone were evaluated by direct chemiluminescence. In addition, superoxide dismutase, catalase and glutathione peroxidase were monitored in the hippocampus region using spectrophotometric method. The levels of the hippocampal monoamines, dopamine, noradrenaline and serotonin were analyzed by high performance liquid chromatography while the acetyl choline neurotransmitter was measured by colorimetric method using choline/acetylcholine assay kit. Results: A sharp disruption in the sex hormones levels of testosterone, progesterone, follicular stimulating hormone and luteinizing hormone was demonstrated in the serum of the treated mice. At the same time, a significant reduction in the endogenous antioxidant defense enzymes, superoxide dismutase, catalase and glutathione peroxidase was observed in the hippocampus region of the injected mice. Moreover, levels of dopamine, noradrenaline, serotonin and acetyl choline neurotransmitter in the same region were significantly affected as a result of the treatment with Spirulina filtrate. The gas chromatography-mass spectrometer and liquid chromatography mass spectrometry/mass spectrometry analysis showed the presence of some sterol-like compounds in the cyanobacterial filtrate. Conclusions: This study demonstrated the capability of Spirulina to release detrimental bioactive metabolites into their surrounding that can disrupt the mammalian endocrine and nervous systems.

460-470

Authors: Sherien Kamal Hassan, Nermin Mohammed El-Sammad, Amria Mamdouh Mousa, Maha Hashim Mohammed, Abd el Razik Hussein Farrag, Amani Nassir Eldin Hashim, Victoria Werner, Ulrike Lindequist, Mahmoud Abd El-Moein Nawwar

Objective: To evaluate the antidiabetic and antioxidant effects of aqueous ethanolic extract of Caesalpinia ferrea (C. ferrea) leaf in normal and streptozotocin (STZ) induced diabetic rats. Methods: Male Sprague-Dawley rats divided into 6 groups of 6 rats each were assigned into diabetic and non-diabetic groups. Diabetes was induced in rats by single intraperitoneal administration of STZ (65 mg/kg body weight). C. ferrea extract at the doses of 250 and 500 mg/kg body weight was orally administered to both diabetic and non-diabetic animals for a period of 30 days. After completion of experimental duration serum, liver and pancreas were used for evaluating biochemical and histopathological changes. Results: Oral administration of C. ferrea leaf extract significantly reduced elevated serum glucose, α-amylase, liver function levels and significantly increased serum insulin, total protein and body weight as well as improved lipid profile due to diabetes. Furthermore, the treatment resulted in a marked increase in glutathione peroxidase, superoxide dismutase, catalase and reduced glutathione, and diminished levels of lipid peroxidation in liver and pancreas of diabetic rats. Histopathological studies demonstrated the reduction in the pancreas and liver damage and confirmed the biochemical findings. Conclusions: From the present study, it can be concluded that the C. ferrea leaf extract effectively improved hyperglycaemia while inhibiting the progression of oxidative stress in STZ-induced diabetic rats. Hence, it can be used in the management of diabetes mellitus.

476-482

Authors: Faridah Yusof, Atheer Awad Mehde, Wesen Adel Mehdi, Hamid Ghazali, Azlina Abd Rahman

Objective: To verify possible associations between adenosine aminohydrolase (ADA) and AMP-aminohydrolase (AMPDA) to E3 SUMO-protein ligase NSE2 (NSMCE2) in patients with renal stones. And to isolate, purify and characterize ADA in patients with renal stones and healthy group. Methods: A total of 60 renal stones patients and 50 control were enrolled in a case-control study. The blood urea, creatinine, uric acid, protein, albumin, ADA and AMPDA were measured by colorimetric tests. The serum NSMCE2 was measured by ELISA. Results: Serum ADA, AMPDA and specific activity of enzymes showed significant decrease (P < 0.05) in patients with renal stones compared to control group, mean levels of sera NSMCE2 and uric acid had a significant increase (P < 0.01 and P < 0.05, respectively) in patients compared to control group. Conclusions: The present study suggests that ADA, AMP deaminase and NSMCE2 can be used as a indicator to monitor the DNA damage and inflammation disorders in the patients with kidney stones.

486-490

Authors: Benjamin Amoani, Elvis Ofori Ameyaw, Du-Bois Asante, Francis Ackah Armah, James Prah, Collins Paa Kwesi Botchey, Johnson Nyarko Boampong

Objective: To investigate the effect of pre-existing Schistosoma haematobium (S. haematobium) infection on malaria disease severity. Methods: The study involved the use of twenty-five imprinting control region mice, fifteen of which were initially infected with S. haematobium. Five of the remaining ten schistouninfected mice together with five schisto-infected mice were infected with Plasmodium berghei (P. berghei) after four weeks (acute stage) of schistosoma infection. The remaining five schisto-uninfected mice together with five schisto-infected mice were also infected with P. berghei after seven weeks (chronic stage) of schistosoma infection. The last five schistoinfected mice were used as control group. They were then monitored for changes in P. berghei parasitaemia on Days 3, 5, 7, 9 and 11 post-infection. Records on their survivability were also taken. Results: The co-infected mice had significantly higher malaria parasitaemia, compared with the mono-infected mice during acute S. haematobium infection. In contrast, the co-infected mice had significantly lower malaria parasitaemia during chronic S. haematobium infection and a higher survival rate. Conclusions: Co-infection of mice with P. berghei during acute S. haematobium infection resulted in rapid P. berghei development and increased malaria parasitaemia. However, the co-infection resulted in slower P. berghei development and decreased malaria parasitaemia with enhanced survivability of the mice during chronic S. haematobium infection. Therefore, pre-existing chronic S. haematobium infection may provide some protection to the host by reducing parasitaemia.

494-500

Authors: Min Li, Douglas Bruce Lowrie, Cheng-Yu Huang, Xiang-Chan Lu,Ying-Chu Zhu, Xing-Hua Wu, Mayila Shayiti, Qiong-Zhen Tan, Hua-Ling Yang, Si-Yuan Chen, Pan Zhao, Sheng-Hua He, Xiu-Rong Wang, Hong-Zhou Lu

Objective: To investigate patients’ perception of service quality at hospitals in nine Chinese cities and propose some measures for improvement. Methods: The ServQual scale method was used in a survey involving patients at out-patient and in-patient facilities in Shanghai, Chongqing, Chengdu, Nanning, Guilin and Laibin of Guangxi, Honghezhou of Yunnan, Wulumuqi of Xinjiang and Zhongshan of Guangdong. The data collected were entered and analyzed using SPSS 20.0. Statistical analyses included descriptive statistics, factor analyses, reliability analyses, product-moment correlations, independent-sample t-tests, One-way ANOVA and regression analyses. Results: The Kaiser-Meyer-Olkin value for the factor analysis of the scale was 0.979. The Cronbach's α for the reliability analysis was 0.978. All the Pearson correlation coefficients were positive and statistically significant. Visitors to out-patient facilities reported more positive perception than visitors to in-patient facilities on tangibles (t = 4.168, P < 0.001) and reliability (t = 1.979, P < 0.05). Patients of 60 years of age and above reported more positive perception than those between 40 and 49 on reliability (F = 3.311, P = 0.010), assurances (F = 2.751, P < 0.05) and empathy (F = 4.009, P = 0.003). For the five dimensions of the scale, patients in Laibin, Guangxi reported the most positive perceived service quality, followed by patients in Shanghai. On the other hand, patients in Chongqing and Nanning and Guilin of Guangxi reported relatively poor perceptions of service quality. Standardized regression coefficients showed statistically significant (P < 0.001) positive values for all ServQual dimensions. Empathy (β = 0.267) and reliability (β = 0.239) most strongly predicted perception of service quality. Conclusions: Chinese patients perceived service quality as satisfactory. Hospitals in various regions of China should enhance their awareness and ability to serve their patients.