Asian Pacific Journal of Tropical Medicine

1105-1110

Authors: Gabriel Alfonso Gutiérrez-Rebolledo, Susan Drier-Jonas, María Adelina Jiménez-Arellanes

Leishmaniasis is considered as an emerging, uncontrolled disease and is endemic in 98 countries. Annually, about 2 million cases of cutaneous and 500 000 cases of visceral-type leishmaniasis are recorded and 60 000 persons died from the disease. In Mexico, cutaneous leishmaniasis is known as chiclero's ulcer and is reported in 22 states, it is considered as a health problem. For its treatment, pentavalent antimonial drugs are administered. These drugs cause severe side effects, are costly. Drug-resistant cases have been reported and have been developing for over 70 years. One alternative to the drugs that are currently available is to find active molecules in medicinal plants. Dihydrocorynantheine, corynantheine and corynantheidine are active against Leishmania major, while harmane, pleiocarpin, buchtienin, luteolin and quercetin are active against Leishmania donovani. In Mexico, about 20 medicinal plants have been evaluated against Leishmania mexicana, among which the most active are Tridax procumbens, Lonchocarpus xuul and Pentalinon andrieuxii. From these plants, active compounds with IC50 ≤ 30 μg/mL or μM have been isolated, such as 3(S)-16,17-didehydrofalcarinol or Oxylipin, cholestra-4,20,24-trien-3-one or pentalinosterol, 24-methylcholest-4-24(28)-dien-3-one, cholest-4-en-3-one, 6,7-dihydroneridie-none, neridienone, cholest-5,20,24-trien-3β-ol, and isocordoin. Today, only pentalinonsterol has been synthesized and assayed in the visceral leishmaniasis experimental model using BALB/c mice infected with Leishmania donovani. Liposome formulation of this compound administered by intravenous route at 2.5 mg/kg showed a significant reduction of parasite load in mouse liver and spleen.

1117–1122

Authors: Prashamsa Koirala, Su Hui Seong, Hyun Ah Jung, Jae Sue Choi

Objective To discover lead lupane triterpenoid's potential isolated from Pueraria lobata roots against β-site amyloid precursor protein cleaving enzyme 1 (BACE1), which serve as a rate limiting step in amyloid beta (Aβ) production altering the course of Alzheimer's disease. In addition, enzyme kinetics study and molecular docking were conducted to establish the inhibition type and structure activity relationship. Methods A systematic study of 70% ethanolic P. lobata root extract was employed to identify its BACE1 inhibitory potential. Further, BACE1 inhibitory potential of two lupane terpenoids, yielded from ethanolic extract, was assessed. In order to determine their inhibition mode, Lineweaver–Burk plots and Michaelis–Menten model for BACE1 was performed. AutoDock 4.2 program in addition determined the molecular interaction of BACE1 with isolated terpenoids. Results Considering the inhibitory potential of 70% ethanolic extract of P. lobata against BACE1 (IC50 = 80.35 μg/mL), lupeol and lupenone were subsequently isolated and exhibited notable or moderate BACE1 inhibitory activity with IC50 values of 5.12 and 62.98 μmol/L, respectively, as compared to the positive control quercetin (IC50 = 21.28 μmol/L). The enzyme kinetics study enabled us to identify both compounds as competitive inhibitors, where lupeol displayed a very potent inhibition against BACE1 with low inhibition constant (Ki) value of 1.43 μmol/L, signifying greater binding affinity. In order to understand the binding mechanism and structure–activity relationship of two triterpene-based BACE1 inhibitors, we employed computer aided docking studies which evidently revealed that hydroxyl group of lupeol formed two hydrogen bonds with the ASP32 (catalytic aspartic residue) and SER35 residues of BACE1 with the binding energy of (−8.2 kcal/mol), while the ketone group of lupenone did not form any hydrogen bonds with BACE1 giving evidence for less binding affinity. These results in turn have predicted the dependence of the inhibitory activity in the presence of hydroxyl group which has provided a new basis for BACE1 blockade. Conclusions Our results have successfully explored the molecular mechanism of lupane triterpenoids via BACE1 inhibition, suggesting that lupeol in particular could be utilized as a useful therapeutic and preventive agent to mitigate Alzheimer's disease.

1134–1139

Authors: Kaouther Majouli, Assia Hamdi, Malek Besbes Hlila

Objective To test the antioxidant, antimicrobial and α-glucosidase inhibitory activities of the roots extracts from Hertia cheirifolia (H. cheirifolia) L. Methods Total phenolics and total flavonoids content of the different extracts were determined by colorimetric methods and reverse phase high-performance liquid chromatography (RP-HPLC) was performed to identify various chemical components. The different extracts were evaluated for antioxidant activities by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis-3-ethylenebenzothiozoline-6-sulfonic acid (ABTS+) and β-carotene bleaching tests and α-glucosidase inhibitory properties. The antimicrobial activity was carried out in vitro by the broth dilution method. Results Trans-cinnamic acid, rutin hydrate, naringin and quercetin were the main compounds of the ethyl acetate extract from H. cheirifolia L. This extract has significant scavenging activity to decrease free radicals especially for DPPH and ABTS radicals. As well as, the ethyl acetate extract exhibited an antimicrobial property against bacterial strains. Bacillus licheniformis and Salmonella enterica were the most sensitive strains with minimum inhibitory concentration values of 0.156 mg/mL. Conclusion The ethyl acetate extract was found to be selectively antioxidant and antimicrobial.

1146–1153

Authors: Hafiz Muhammad Abdur Rahman, Khaled Ahmed, Muhammad Fawad Rasool, Imran Imran

Objective To investigate the pharmacological basis of its uses of Phyla nodiflora (P. nodiflora) for the treatment of anomalies predominantly by smooth muscle containing tissues such as gastrointestinal/vascular/broncho spasm and cardiovascular modulation. Methods The crude hydroalcoholic extract of P. nodiflora (Pn.Cr) and its fractions were evaluated on isolated rabbit jejunum, rat trachea, aorta and atrium. To access the contractile or relaxant effects of testing materials, the tissues were mounted in isolated organ bath and responses were recorded with transducers coupled with data acquisition system. BALB/c mice were challenged with castor oil for the assessment of in-vivo antidiarrheal activity. Normotensive rats were used for in-vivo hypotensive study. Results Hydroalcoholic extract of Pn.Cr at variable concentrations inhibited the natural spontaneous rhythm and 80 mmol/L K+ mediated contractions in isolated segment of jejunum with EC50 values of 3.18 and 1.91 mg/mL respectively. Verapamil, a Ca2+ channel blocker, demonstrated akin pattern in jejunum and therefore possibly suggesting calcium blocking activity. On isolated rat tracheal tissue, Pn.Cr showed relaxation of high-K+ and carbachol mediated contractions (EC50 values: 1.24 and 2.42 mg/mL). Pn.Cr treatment relaxed the rat aortic ring in a cumulative doses with high-K+ and phenylephrine-induced contractions (EC50 values 0.25 and 0.92 mg/mL). Activity based fractionation of Pn.Cr showed that dichloromethane fraction was more potent for relaxing the tissues spasm compared to aqueous fraction. In-vivo experiments, significant protection by Pn.Cr (P < 0.05) was observed in castor oil-induced diarrhea (50–500 mg/kg) whereas hypotensive effect in anesthetized rats was seen at the dose range of 1–10 mg/kg of Pn.Cr (P < 0.05). Conclusion This study suggests the blockage of calcium channel in the smooth muscles as a pharmacological application to make usage of P. nodiflora in the management of diarrhea, asthma and hypotensive effect.

1161–1166

Authors: Fabrizio Bertelloni, Giovanni Tosi, Paola Massi, Laura Fiorentini, Maria Parigi, Domenico Cerri, Valentina Virginia Ebani

Objective To investigate some pathogenic characters of Salmonella enterica strains isolated from poultry. Methods Twenty-three genetically distinct Salmonella enterica strains, of different serovars and pulsotype, were examined for virulence traits. Resistance to gastric acid environment was estimated by measuring the percentage of survived bacterial cells after exposure for 2 h to a synthetic gastric juice. Strains were analyzed with PCR for the presence of the following virulence genes: mgtC and rhuM located on SPI-3, sopB and pipB located on SPI-5, Salmonella virulence plasmid (spv) R (spvR), spvB and spvC located on Salmonella plasmid virulence and sodCI, sopE, and gipA located on prophage. Finally, resistance to 21 antibiotics was tested with Kirby–Bauer method. Results A percentage of 82.60% of strains were resistant to gastric environment after induction and 60.87% of the strains exhibited constitutive resistance too. Nineteen different virulence profiles were detected. The phage related genes sodCI and sopE and the plasmid mediated operon spvR, spvB and spvC (spvRBC) were detected in 82.60%, 47.82% and 52.17% of strains, respectively. Typhimurium and Enteritidis strains showed the highest number of virulence genes. Twenty-one different antibiotic resistance profiles were obtained and two isolates (Typhimurium and Enteritidis) resulted sensible to all the tested molecules. The ampicillin, streptomycin, sulfonamide and tetracycline resistance profile was detected in seven isolates (30.43%). Conclusion Our results show that paratyphoid Salmonella strains with several characters of pathogenicity, that may be cause of severe pathology in animals and humans, are circulating among poultry.

1172–1176

Authors: Tao Zhang, Jia-Yi Liao, Li Yu, Guo-Sheng Liu

Objective To study the influence of glycyrrhetinic acid (GA) on bronchial asthma (BA) smooth muscle proliferation and apoptosis as well as inflammatory factor expression and its molecular mechanism. Methods Male SD guinea pigs were selected and made into asthma models, bronchial asthma smooth muscle cells were cultured and divided into BA group, GA group and GA + LM group that were treated with serum-free RPMI1640 culture medium, serum-free RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid, serum-free RPMI1640 culture medium containing 50 ng/mL glycyrrhetinic acid and 100 ng/mL LM22B-10 respectively; normal guinea pigs were collected and bronchial smooth muscle cells were cultured as control group. The cell proliferation activity as well as the expression of proliferation and apoptosis genes, inflammatory factors and p-ERK1/2 was determined. Results Proliferation activity value and mRNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in BA group were significantly higher than those of control group while mRNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly lower than that of control group (P < 0.05); proliferation activity value and mRNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40, protein expression of p-ERK1/2 of airway smooth muscle cell in GA group were significantly lower than those of BA group (P < 0.05) while the mRNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly higher than those of BA group (P < 0.05); proliferation activity value and mRNA expression of Bcl-2, TNF-α, IL-4, IL-6, YKL-40 of airway smooth muscle cell in GA + LM group were significantly higher than those of GA group (P < 0.05) while mRNA expression levels of Bax, caspase-9 as well as caspase-3 were significantly lower that of GA group (P < 0.05). Conclusion GA can inhibit the proliferation of bronchial smooth muscle cells and reduce the expression of inflammatory factors by inhibiting the phosphorylation of ERK1/2.

1185–1189

Authors: Hua Luo, Xiao-Qiong Wu, Min Zhao, Qiong Wang, Geng-Pan Jiang, Wei-Jun Cai, Ming-Ying Luo

Objective To investigate the distribution and contents of vimentin (Vim) and glial fibrillary acidic protein (GFAP) immunoreactivities in the central nervous system (CNS) of normal newborn, adult and aged rats. Methods In this study, thirty healthy and normal Sprague–Dawley rats were simply classified into three groups: Newborn (7 days aged), adult (5 months aged) and aged (24 months aged) rats. Brains and spinal cord were dissected and cut into frozen sections. The expression of Vim and GFAP in CNS were detected by confocal immunofluorescence. Results In each group, Vim was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was highest in neuron-like cell of newborn rats, while Vim was mainly expressed in cell bodies in adult and aged rats. GFAP was expressed in all the regions of CNS including the hippocampal, cerebral cortex, the third ventricle and spinal cord, and the expression was in astrocytes of aged rats. In the third ventricle, Vim was detected in all groups, and only observed in neuron-like cells of newborn. Meanwhile, the GFAP expression showed no significant differences between adult and aged rats in this region. The co-localization of Vim and GFAP were mainly observed in hippocampus and cerebral cortex of newborn, but this co-localization was found in the third ventricle of the rats in all groups. Conclusion Our data demonstrate for the first time that the expression of Vim and GFAP in the rat's CNS during development. This data may provide a foundation for the further mechanistic studies of these two main intermediate filaments during development of CNS.

1197–1200

Authors: Ling Fan, Li-Feng Chen, Jing Fan

Objective To investigate the electrophysiological changes of autonomic cells in left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with chronic heart failure. Methods Guinea pigs model of iron deficiency anemia complicated with chronic heart failure in 10 guinea pigs of the experimental group was made by feeding a low iron diet, pure water and subcutaneous injection of isoproterenol. The control group consisting of 11 guinea pigs was given normal food, normal water and injected with normal saline. The left ventricular outflow tract model specimen was also prepared. The standard microelectrode technique was used to observe electrophysiological changes of autonomic cells in the outflow tract of left ventricular heart failure complicated with iron deficiency anemia in guinea pig model. The indicators of observation were maximal diastolic potential, action potential amplitude, 0 phase maximal depolarization velocity, 4 phase automatic depolarization velocity, repolarization 50% and 90%, and spontaneous discharge frequency. Results Compared with the control group, 4 phase automatic depolarization velocity, spontaneous discharge frequency and 0 phase maximal depolarization velocity decreased significantly (P < 0.01) and action potential amplitude reduced (P < 0.01) in model group. Moreover, repolarization 50% and 90% increased (P < 0.01). Conclusions There are electrophysiological abnormalities of the left ventricular outflow tract in guinea pigs with iron deficiency anemia complicated with heart failure.

1019–1029

Authors: Rodrigo Siqueira Batista, Andréia Patrícia Gomes, Jorge Luiz Dutra Gazineo, Paulo Sérgio Balbino Miguel, ... Mauro Geller

Meningococcal disease is the acute infection caused by Neisseria meningitidis, which has humans as the only natural host. The disease is widespread around the globe and is known for its epidemical potential and high rates of lethality and morbidity. The highest number of cases of the disease is registered in the semi-arid regions of sub-Saharan Africa. In Brazil, it is endemic with occasional outbreaks, epidemics and sporadic cases occurring throughout the year, especially in the winter. The major epidemics of the disease occurred in Brazil in the 70's caused by serogroups A and C. Serogroups B, C and Y represent the majority of cases in Europe, the Americas and Australia. However, there has been a growing increase in serogroup W in some areas. The pathogen transmission happens for respiratory route (droplets) and clinically can lead to meningitis and sepsis (meningococcemia). The treatment is made with antimicrobial and supportive care. For successful prevention, we have some measures like vaccination, chemoprophylaxis and droplets' precautions. In this review, we have described and clarify clinical features of the disease caused by N. meningitidis regarding its relevance for healthcare professionals.

1037-1042

Authors: Abdul Waheed Khan, Sadia Nawab, Zeeshan Nasim, Abdul Haleem Khan, Syed Ishfaq Ahmad, Fazli Zahir, Israr Ud Din

Infection due to hepatitis C virus (HCV) is a major cause of fibrosis and hepatocellular carcinoma in Pakistan. In the current review, pattern of HCV genotypes and subtypes in Khyber Pakhtunkhwa province was ascertained in light of the available literature. After thorough analysis, genotype 3 (58.27%) was determined to be the leading HCV genotype, followed by genotypes 2 (12.39%), 1 (9.54%) and 4 (0.86%). The proportions of genotypes 5 and 6 were recorded as 0.09% and 0.22% respectively. Subtype wise, 3a accounted for 48.67%, followed by subtype 2a (10.91%), 3b (9.43%), 1a (5.84%), 1b (3.66%), 2b (1.45%) and genotype 4 with its undefined subtypes contributed a portion of 0.86%. The cumulative share of subtypes 1c, 2c, 3c, 5a and 6a was less than 1%. In 11.51% cases, the subtype was untypeable while in 7.17% cases mixed subtypes were recorded. Gender wise, proportions of most HCV subtypes were marginally higher among males as compared to females. On the basis of studied groups, 3a was pervasive among all groups except in intravenous drug users where 2a was the major HCV subtype. Similarly, based on various geographical locations (provincial divisions), subtype 3a revealed a ubiquitous distribution. Conclusively, HCV 3a persists to be the principal subtype across the province of Khyber Pakhtunkhwa. The considerable number of untypeable subtypes in most studies urges for an improved genotyping system on the basis of local sequence data and practice of sequencing for determination of underlying subtype in untypeable cases. Further, studies on identification of subtypes transmission pattern are imperative for assessment of transmission origin and reinforcement of efficient control strategies. In addition, the current review emphasizes the need of attention toward HCV risk groups and ignored southern side of Khyber Pakhtunkhwa province for better holistic understanding of HCV genotype distribution pattern in the province.

1047–1053

Authors: Claudia V. Moreno-Quirós, Alberto Sánchez-Medina, Maribel Vázquez-Hernández, Ana G. Hernández Reyes, Rosa V. García-Rodríguez

Objective To evaluate the anti-inflammatory, analgesic, antioxidant and acute toxicity of extracts obtained from a successive extraction with solvents of ascending polarity [hexane, hex; chloroform, CHCl3 and ethanol (EtOH)] of Ternstroemia sylvatica Schltdl. & Cham. Methods The antioxidant potential was evaluated by 2,2 diphenyl-1-picrylhydrazyl, the ferric reducing/antioxidant power assays and by determining the total phenolic content. The anti-inflammatory and antinociceptive effects were evaluated using the in vivo croton oil-induced ear edema, phorbol 12-myristate 13-acetate induced ear edema, carrageenan-induced paw edema, acetic acid-induced writhing and formalin murine models. The acute toxicity was tested using the Lorke's method in mice. Results The EtOH extract was the most active for the antioxidant potential tests diphenyl-1-picrylhydrazyl (68.70% inhibition), ferric reducing/antioxidant power [(2 431.30 ± 102.10) mmol Fe2+ and total polyphenols content (215.80 ± 8.50) meqAG/g]. The anti-inflammatory activity was evaluated by topical application of croton oil (2 mg/ear dose) where the EtOH extract showed the strongest activity compared to the control group (45.13% inhibition), whereas in the phorbol 12-myristate 13-acetate model, at the same dose, the CHCl3 extract showed the highest inhibition (42.88%). In the carrageenan induced edema model, the EtOH extract showed a stronger inhibition compared to indomethacin (56.34% and 50.70% at doses of 250 and 500 mg/kg of extract, respectively) during the first hour. Similarly, the same extract showed the highest analgesic activity (30.60% inhibition) in the acetic acid contortion assay, and in the formalin test it showed a greater effect with respect to the control group in both phases. Conclusions Our work confirms the value of Ternstroemia sylvatica as an important anti-inflammatory and analgesic plant, whose mechanism seems to be associated to its antioxidant effects, and supports its uses in the Mexican traditional medicine.

1059–1064

Authors: Ebru Şebnem Yılmaz, Özkan Aslantaş

Objective To investigate the antimicrobial susceptibility of 97 clinical Staphylococcus aureus (S. aureus) strains against 14 antimicrobials and corresponding resistance mechanisms. Methods The antimicrobial susceptibility of the isolates was determined using a disk diffusion method and antimicrobial resistance genes were screened by polymerase chain reaction. Mutations responsible for ciprofloxacin and rifampicin resistance were investigated by polymerase chain reaction and DNA sequencing. Results All isolates were found to be susceptible to vancomycin. Various rates of resistance to penicillin (83.5%), ampicillin (77.3%), erythromycin (63.9%), tetracycline (16.5%), amoxicillin/clavulanic acid (16.5%), ciprofloxacin (15.5%), trimethoprim/sulfamethoxazole (15.5%), oxacillin (13.4%), fusidic acid (12.4%), rifampin (6.2%), clindamycin (6.2%), gentamicin (6.2%) and mupirocin (5.2%) were determined. In addition, different combinations of resistance genes were identified among resistant isolates. Ciprofloxacin resistant isolates had mutations in codon 84 (Ser84Leu) and 106 (Gly106Asp) in the gyrA gene. Mutations in grlA were mostly related to Ser80Phe substitution. Leu466Ser mutation in the rpoB gene was detected in all rifampin resistant isolates. All methicillin resistant S. aureus isolates were SCCmec type V. Conclusions In conclusion, it was determined that the isolates were resistant to different classes of antimicrobials at varying rates and resistance was mediated by different genetic mechanisms. Therefore, continuous monitoring of resistance in S. aureus strains is necessary to control their resistance for clinically important antimicrobials.

1072–1076

Authors: Mojtaba Shahnazi, Abbas Azadmehr, Moazzam Dosti Jondabeh, Reza Hajiaghaee, Reza Norian, Hamidreza Aghaei, Mehrzad Saraei, Mahmood Alipour

Objective To evaluate the possible involvement of programmed cell death strategy in hydatid cyst protoscolices following treatment with Myrtus communis (M. communis) as an herbal medicine. Methods Protoscolices were aseptically collected from sheep liver hydatid cysts. Evaluating the effect of M. communis extract on programmed cell death and increased activity of caspases 3, 8, and 9 in hydatid cyst protoscolices was conducted by treating the protoscolices with different concentration (5, 50, and 100 mg/mL) of M. communis extract at 37 °C and 5% CO2 for 4 h by using the Bradford test and ELISA commercial kits. Results The extract of M. communis at all concentrations led to initiation of programmed cell death in protoscolices and this effect, was only significant at 50 and 100 mg/mL concentrations, compared to the negative control (P < 0.05). Also, the activity of caspases 3, 8, and 9 in hydatid cyst protoscolices, was shown that the extract at all 3 concentrations could only increase the activity of caspases 3 and 9. Moreover, a significant increase in the activity of caspase 3 was only observed at concentrations 50 and 100 mg/mL by 37.00% and 66.19% while a significant increase in the activity of caspase 9 at the same concentrations was observed by 20.89% and 63.67%, respectively (P < 0.05). Conclusions The extract of M. communis at different concentrations could increase the activity of caspases 3 and 9 and caused programmed cell death in hydatid cyst protoscolices however, this effect was significant at high concentrations of the extract.

1080–1083

Authors: Clarissade Albuquerque Botura, Bruno Ambrósioda Rocha, Thiely Balensiefer, Franciele Queiroz Ames, Ciomar Aparecida Bersani-Amado, Roberto Kenji Nakamura Cuman

Objective To evaluate the efficacy of oral indomethacin, ibuprofen, and paracetamol in oral dosage form on patent ductus arteriosus (PDA) in premature neonates with significant clinical and hemodynamic repercussions (CHRs) and to determine the effect of these respective treatments on renal function. Methods A retrospective study of cases of PDA in premature neonates in the Neonatal Intensive Care Unit was conducted. The treatments consisted of indomethacin [0.2 mg/(kg·d), 3-day cycle], ibuprofen [10 mg/(kg·d) followed by 5 mg/(kg·d), 3-day cycle], and paracetamol (15 mg/kg every 6 h, 5-day cycle). The drugs were administered as an oral solution. The following variables were considered: gestational age, newborn weight at birth, Apgar score, diuresis, serum creatinine and urea levels, and serum electrolyte levels (sodium and potassium). Results Treatment with indomethacin presented efficacy of 87.5% in closure of the ductus with a mean outcome period of 3.5 d. In premature neonates with CHRs and contraindications for indomethacin, the initial treatment with either ibuprofen or paracetamol failed to close the ductus. However, when this treatment was followed by indomethacin, closure occurred in 66.7% of the neonates, with an outcome period of 9.66 d. The initial treatment with one cycle of ibuprofen followed by one or two cycles of paracetamol failed to close the ductus. Conclusions Oral indomethacin was effective for closure of the PDA in premature neonates with severe CHRs. Oral paracetamol or ibuprofen for PDA closure in premature neonates with severe CHRs and contraindications for indomethacin was ineffective. However, results in clinical improvements of neonates allowed the subsequent use of indomethacin and successful closure of the ductus. A significant reduction of diuresis occurred in neonates who were treated with indomethacin, either as a first-line treatment or after the failure of ibuprofen or paracetamol.

1090–1094

Authors: Jin-Feng Dong, Xiao-Qiang Zheng, Hong-Bing Rui

Objective To observe the effect of taurine on immune function in mice with T-cell lymphoma during chemotherapy. Methods A total of 40 C57BL/6 mice were selected and randomly divided into 4 groups, namely model group, chemotherapy group, taurine group and chemotherapy + taurine group, each containing 10 mice. Hypodermic injection was adopted to inoculate EL-4 cells in order to establish model of T-cell lymphoma. When the tumor achieved the size of 1 cm3, intervention treatments were given to the groups respectively. Mice in model group received 0.2 mL of normal saline which was intraperitoneally injected on Days 1, 8 and 15 with 3 weeks as a cycle; mice in chemotherapy group were administered with 80 mg/kg body weight of gemcitabine which was also intraperitoneally injected on Days 1, 8 and 15 with 3 weeks as a cycle; mice in taurine group were administered with 80 mg/kg body weight of taurine intraperitoneally injected daily for consecutive 8 d; mice in chemotherapy + taurine group were treated in the same manner as the mice in taurine group and chemotherapy group. Five mice were sacrificed at 2 and 3 weeks after intervention respectively, and the tumor tissues were collected and weighted after removal of auxiliary tissue, then the tumor inhibition rate was calculated. The thymus and spleen of mice sacrificed at 3 weeks after intervention were collected and weighted, and thymus and spleen indexes were calculated. Enzyme linked immunosorbent assay was used to detect the serum levels of IL-4, IL-10, IL-12 and IFN-γ in mice of each group. Results The tumor weights in chemotherapy group, taurine group and chemotherapy + taurine group after 2 and 3 weeks of treatment were significantly lower than that in model group (P < 0.05); the tumor weight in chemotherapy + taurine group after 2 and 3 weeks of treatment was significantly lower than that in chemotherapy group (P < 0.05); the tumor inhibition rate in chemotherapy + taurine group was significantly higher than that in chemotherapy group and taurine group (P < 0.05); the thymus and spleen indexes in taurine group and chemotherapy + taurine group were significantly higher than those in chemotherapy group and model group (P < 0.05); the thymus and spleen indexes in chemotherapy group were significantly lower than those in model group (P < 0.05); after 3 weeks of treatment, the serum levels of IL-4, IL-12 and IFN-γ in chemotherapy group, taurine group and chemotherapy + taurine group were significantly lower than those in model group (P < 0.05); the IL-4 level in taurine group and chemotherapy + taurine group was significantly lower than that in chemotherapy group (P < 0.05); the serum level of IL-10 in chemotherapy group and chemotherapy + taurine group was significantly higher than that in model group and taurine group (P < 0.05); the serum level of IFN-γ in taurine group and chemotherapy + taurine group was significantly lower than that in model group and chemotherapy group (P < 0.05); after treatment of 3 weeks, the serum levels of IL-4 and IL-10 in chemotherapy group, taurine group and chemotherapy + taurine group were significantly lower than those in model group (P < 0.05), and IL-12 level was significantly higher than that in model group (P < 0.05); the level of IFN-γ in taurine group and chemotherapy + taurine group was significantly higher than that in model group (P < 0.05), while the level of IFN-γ in chemotherapy group was significantly lower than that in the other 3 groups (P < 0.05). Conclusions Taurine can effectively enhance the immune function of mice with T-cell lymphoma during chemotherapy, reduce the toxicity of chemotherapy.

1101–1103

Authors: Diego Espinoza-Morales, Aldo Lucchetti Rodríguez, Wilmer Silva-Caso, Luis Suarez-Ognio, María J.Pons, Juanadel Valle Mendoza

We present an atypical case of disseminated cutaneous leishmaniasis in the Sihuas district, located in the Andean valleys of Ancash-Peru. A 62-year-old man with no particular medical history presented multiple lesions located on the inferior abdomen, lumbar region and the right anterior thigh. Histological analysis found leishmanial amastigotes in the lesion sample, the Montenegro reaction was positive for Leishmania spp, and the polymerase chain reaction was positive for Leishmania peruviana. In conclusion, the atypical presentation of this disease may be related to the presence of an uncommon parasite strain or host immune deficiencies. The molecular identification of the etiology for disseminated leishmaniasis, will allow a better understanding of the presentation and proper treatment, as well as associated risk factors.

933–939

Authors: Md. Rajdoula Rafe

Plants are used traditionally throughout the globe to treat various diseases. Traditionally used medicinal plants are an essential part of the health sector in Bangladesh due to its abundance of a vast source of ethno-medicine. Rural people from developing country like Bangladesh are greatly dependent on traditional source of medicine. The prevalence of diabetes mellitus is increasing from recent years; therefore various researches are going on to discover better medicine to treat this disease. This study has focused on five plants which are Andrographis paniculata, Ageratum conyzoides, Swertia chirata, Terminalia arjuna, and Azadirachta indica to find out their traditional formulation as anti-diabetic medicine and their pharmacological activity has also been explored through literature search. The available information about traditional anti-diabetic uses of these plants and their pharmacological activities were collected from various electronic sources like Pubmed, SciFinder, Elsevier, Springer, Scopus, Scirus, Science Direct, Google Scholar and Web of Science apart from these locally available books and peer reviewed journal were also used to collect information. This study will help to strengthen the relation between traditional medicine, pharmacology and drug development. A clue may be found from the information provided this review to discover new and better anti-diabetic drugs.

945–951

Authors: Tereza de Jesus Pinheiro Gomes Bandeira, Débora de Souza Collares Maia Castelo-Branco, Marcos Fábio Gadelha Rocha, Rossana de Aguiar Cordeiro, Crister José Ocadaque, Manoel de Araújo NetoPaiva, Raimunda Sâmia Nogueira Brilhante, José Júlio Costa Sidrim

Objective To evaluate the genetic diversity of clinical and environmental isolates of Burkholderia pseudomallei (B. pseudomallei) recovered in Ceará, Brazil, and screen these isolates for the presence of type three secretion system virulence gene. Methods Nineteen B. pseudomallei isolates (9 from clinical cases and 10 from soils) were analyzed. Random amplified polymorphic DNA was performed with primers OPQ-2, OPQ-4 and OPQ-16 to evaluate the genetic diversity, and type three secretion system gene was detected through polymerase chain reaction. Results Random amplified polymorphic DNA showed a genetic relatedness of approximately 50% among the tested B. pseudomallei isolates, which were grouped into two clades, of which the biggest ones comprised 18/19 isolates for primer OPQ-2, and 17/19 isolates for primer OPQ-16. Primer OPQ-4 grouped the isolates into three clades comprising 1/19, 3/19 and 15/19 isolates. Additionally, type three secretion system gene was detected in all tested isolates. Conclusions This is an effort to type B. pseudomallei strains from Ceará, which is important for better understanding this pathogen, contributing for the epidemiological surveillance of melioidosis in this endemic region.

960–966

Authors: Roya Karamian, Mostafa Asadbegy, Siamak Yari

Objective To investigate nephroprotective potential of Meristotropis xanthioides (M. xanthioides) extract against ethanol-induced nephrotoxicity in Wistar rats, and also its total phenolics content, antioxidant and antibacterial activities. Methods Total phenol and flavonoid amounts of the leaf and stem extracts were determined by Folin–Ciocalteu and aluminum chloride reagents, respectively. Antioxidant and antibacterial activities of the extracts were investigated by 2,2-diphenyl-1-picrylhydrazyl radical scavenging and disc diffusion methods, respectively. In addition, protective potential of the leaf extract against ethanol-induced nephrotoxicity was studied by histological and biochemical analyses. Results Obtained results indicated high total phenol [(10.26 ± 0.46) mg GAE/g of dry extract] and flavonoid [(3.63 ± 0.62) mg QE/g of dry extract] amounts in the leaf extract. The leaf and stem extracts possessed stronger antioxidant activity [IC50: (0.119 ± 0.006) mg/mL and IC50: (0.133 ± 0.009 mg/mL)] than that of ascorbic acid [IC50: (0.142 ± 0.002) mg/mL]. Also, the extracts showed good antibacterial activity against the most of bacteria taken in this research, especially Gram-positive ones. Histological examinations revealed tissue injury in the kidney of rats treated with ethanol. Results from biochemical assays showed reduction in total protein content and also in superoxide dismutase activity. In addition, remarkable increased levels (P < 0.05) of H2O2 and malondialdehyde were found in ethanol-treated rats in comparison to control group. However, these injuries were significantly improved in rats treated by M. xanthioides leaf extract. Conclusions Results from present study demonstrates strong pharmaceutical potential of M. xanthioides extract to apply as a new drug supplement.

974–980

Authors: Soukaina El-Guendouz, Noori Al-Waili, Smail Aazza, Youssef Elamine, Soumia Zizi, Thia Al-Waili, Ali Al-Waili, Badiaa Lyoussi

Objective To study the antioxidant properties of Capparis spinosa (C. spinosa) honey and propolis and the effect of combined honey and propolis administration on urine volume and electrolytes in rats. Methods C. spinosa honey [1 000 mg/kg body weight (b.wt)], propolis (100 mg/kg b.wt), honey/propolis mixture (C. spinosa honey 1 000 mg/kg b.wt/ propolis extract 100 mg/kg b.wt ), distilled water (1 mL/kg b.wt) and furosemide (10 mg/kg b.wt) were orally administered to five groups of rats for 21 d. Urine volume, blood and urine sodium, potassium and chloride were measured. The antioxidant activity of propolis and honey was assessed and their total phenols and flavonoids were determined. Results Propolis and C. spinosa honey contain polyphenols including flavonoids and propolis demonstrated higher antioxidant activities than honey. Honey significantly increased urine volume and urine electrolyte excretion. Propolis had no significant effect on urine volume, but co-administration of propolis and honey caused significant diuresis. No major changes were observed in plasma electrolytes with the use of honey, propolis or their combination. Conclusions Honey and propolis have antioxidant activity and contain polyphenols including flavonoids that are more pronounced in propolis. Honey has a significant diuretic activity alone or in combination with propolis. This is the first study comparing the diuretic effect of co-administration of propolis and C. spinosa honey with furosemide.

987–995

Authors: Hesham M. Al-Mekhlafi

Objective To investigate the prevalence and risk factors of Giardia infection among rural communities among rural communities in Yemen. Methods In this cross-sectional study, a total of 605 stool samples were collected and screened for the presence of Giardia duodenalis (G. duodenalis) cysts and/or trophozoites by using three different diagnostic methods: direct smear, formalin-ether sedimentation, and trichrome staining. A pre-tested questionnaire was used to collect information on the demographic, socioeconomic, behavioural and environmental characteristics of the participants. Results Overall, 28.1% (170/605) of the participants were infected by G. duodenalis. The prevalence was significantly higher among male participants compared to female (P = 0.034); however, it was not significant among different age groups (P > 0.05). Univariate and multivariate analyses identified four variables as the significant key risk factors of Giardia infection among the sampled communities. These are, in addition to being of the male gender, using unsafe water sources for drinking water, not washing hands after defecation, presence of other family members infected with Giardia, and close contact with domestic animals. Conclusions The study reveals that Giardia infection is still prevalent among rural communities in Yemen. The provision of clean and safe drinking water, proper sanitation, and health education regarding personal hygiene practices, particularly handwashing, as well as identifying and treating infected family members is imperative and these interventions should be considered in a strategy to control intestinal parasites among these communities in order to curtail the transmission and morbidity caused by G. duodenalis.

1002–1006

Authors: Ibrar Rafique, Muhammad Arif NadeemSaqib, Muhammad ArifMunir, Huma Qureshi, Ijaz-ul-Haq Taseer, Rizwan Iqbal, Waqaruddin Ahmed4, Tasleem Akhtar, Rizwanullah

Objective To determine the asymptomatic dengue infection in adults of Pakistani population. Methods This study was conducted in five major cities (Islamabad, Karachi, Lahore, Multan, and Peshawar) of Pakistan. A total of 5 230 adults aged 18 years and above without a history of dengue fever at any point in their life were enrolled from participating laboratories. Those who were confirmed for dengue previously were excluded. Of the total, 62.6% (n = 3 276) were male with an average age of 34.6 years. Participants were briefed about the objectives of the study, and written consent was obtained to perform dengue IgG test using enzyme linked immunosorbent assay. The brief information related to age, gender and area was also taken on proforma. Results Overall 32.3% (n = 1 691) was having asymptomatic dengue infection which was 67.5% (n = 756) in Karachi followed by 39.1% (n = 391) in Islamabad, 29.9% (n = 316) in Lahore and 21% (n = 228) in Peshawar and none from Multan. More males were affected with asymptomatic dengue infection than females. The asymptomatic dengue infection was significantly higher in different cities; however, there was no significant difference with respect to age groups. Conclusions The asymptomatic dengue infection is higher in cities i.e. Karachi, Islamabad and Lahore which are at risk of developing secondary dengue infections. There is a need of awareness among the public about secondary dengue infection.

1011–1014

Authors: Guang-Xian Wang, Lan-Lan Liu, Li Wen, Yun-Xing Cao, Yu-Chun Pei, Dong Zhang

Objective To identify the morphological parameters that are related to intracranial aneurysms (IAs) rupture using a case-control model. Methods A total of 107 patients with multiple IAs and aneurysmal subarachnoid hemorrhage between August 2011 and February 2017 were enrolled in this study. Characteristics of IAs location, shape, neck width, perpendicular height, depth, maximum size, flow angle, parent vessel diameter (PVD), aspect ratio (AR) and size ratio (SR) were evaluated using CT angiography. Multiple logistic regression analysis was used to identify the independent risk factors associated with IAs rupture. Receiver operating characteristic curve analysis was performed on the final model, and the optimal thresholds were obtained. Results IAs located in the internal carotid artery (ICA) was associated with a negative risk of rupture, whereas AR, SR1 (height/PVD) and SR2 (depth/PVD) were associated with increased risk of rupture. When SR was calculated differently, the odds ratio values of these factors were also different. The receiver operating characteristic curve showed that AR, SR1 and SR2 had cut-off values of 1.01, 1.48 and 1.40, respectively. SR3 (maximum size/PVD) was not associated with IAs rupture. Conclusions IAs located in the ICA are associated with a negative risk of rupture, while high AR (>1.01), SR1 (>1.48) or SR2 (>1.40) are risk factors for multiple IAs rupture.

835–848

Authors: Yahaya Gavamukulya, Fred Wamunyokoli, Hany A. El-Shemy

Annona muricata (A. muricata) is a tropical plant species belonging to family Annonaceae and known for its many medicinal uses. This review focuses on the research history of its traditional uses, phytochemicals, pharmacological activities, toxicological aspects of the extracts and isolated compounds, as well as the in vitro propagation studies with the objective of stimulating further studies on this plant for human consumption and treatment. A. muricata extracts have been identified in tropical regions to traditionally treat diverse conditions ranging from fever to diabetes and cancer. More than 200 chemical compounds have been identified and isolated from this plant, the most important being alkaloids, phenols and acetogenins. Using in vitro studies, its extracts and phytochemicals have been characterized as antioxidant, anti-microbial, anti-inflammatory, insecticidal, larvicidal, and cytotoxic to cancer cells. In vivo studies have revealed anxiolytic, anti-stress, anti-inflammatory, immunomodulatory, antimalarial, antidepressant, gastro protective, wound healing, hepato-protective, hypoglycemic, anticancer and anti-tumoral activities. In silico studies have also been reported. In addition, clinical studies support the hypoglycemic as well as some anticancer activities. Mechanisms of action of some pharmacological activities have been elucidated. However, some phytochemical compounds isolated from A. muricata have shown a neurotoxic effect in vitro and in vivo, and therefore, these crude extracts and isolated compounds need to be further investigated to define the magnitude of the effects, optimal dosage, and mechanisms of action, long-term safety, and potential side effects. Additionally, more clinical studies are necessary to support the therapeutic potential of this plant. Some studies were also found to have successfully regenerated the plant in vitro, but with limited success. The reported toxicity notwithstanding, A. muricata extracts seem to be some of the safest and promising therapeutic agents of the 21st century and beyond that need to be studied further for better medicinal formulations and diseases management.

855–863

Authors: Adel Spotin, Hamid Reza Moslemzadeh, Mahmoud Mahami-Oskouei, Ehsan Ahmadpour, Maryam Niyyati, Seyed Hossein Hejazi, Fatemeh Memari, Jafar Noori

Objective To verify phylogeography and genetic structure of Acanthamoeba populations among the Iranian clinical isolates and natural/artificial environments distributed in various regions of the country. Methods We searched electronic databases including Medline, PubMed, Science Direct, Scopus and Google Scholar from 2005 to 2016. To explore the genetic variability of Acanthamoeba sp, 205 sequences were retrieved from keratitis patients, immunosuppressed cases and environmental sources as of various geographies of Iran. Results T4 genotype was the predominant strain in Iran, and the rare genotypes belonged to T2, T3, T5 (Acanthamoeba lenticulata), T6, T9, T11, T13 and T15 (Acanthamoeba jacobsi). A total of 47 unique haplotypes of T4 were identified. A parsimonious network of the sequence haplotypes demonstrated star-like feature containing haplogroups IR6 (34.1%) and IR7 (31.2%) as the most common haplotypes. In accordance with the analysis of molecular variance, the high value of haplotype diversity (0.612–0.848) of Acanthamoeba T4 represented genetic variability within populations. Neutrality indices of the 18S ribosomal RNA demonstrated negative values in all populations which represented a considerable divergence from neutrality. The majority of genetic diversity belonged to the infected contact lens and dust samples in immunodeficiency and ophthalmology wards, which indicated potential routes for exposure to a pathogenic Acanthamoeba sp. in at-risk individuals. A pairwise fixation index (FST) was from low to high values (0.024 33–0.418 92). The statistically FST points out that T4 is genetically differentiated between north-west, north-south and central-south metapopulations, but not differentiated between west-central, west-south, central-south, and north-central isolates. Conclusions An occurrence of IR6 and IR7 displays that possibly a gene flow of Acanthamoeba T4 occurred after the founder effect or bottleneck experience through ecological changes or host mobility. This is the first systematic review and meta-analysis providing new approaches into gene migration and transmission patterns of Acanthamoeba sp, and targeting at the high-risk individuals/sources among the various regions of Iran.

871–876

Authors: Benjaporn Sornpet, Teerapong Potha, Yingmanee Tragoolpua, Kidsadagon Pringproa

Objective To study the antiviral properties of the five Asian medicinal plants against in vitro infection by the highly pathogenic avian influenza virus (H5N1). Methods Crude extracts of Andrographis paniculata, Curcuma longa (C. longa), Gynostemma pentaphyllum, Kaempferia parviflora (K. parviflora), and Psidium guajava obtained by both water and ethanol extractions were investigated for their cytotoxicity in the Madin–Darby canine kidney cells. Thereafter, they were investigated in vitro for antiviral activity and cytokine response upon H5N1 virus infection. Results The results revealed that both water and ethanol extracts of all the five studied plants showed significant antiviral activity against H5N1 virus. Among these plants, C. longa and K. parviflora showed strong anti-H5N1 activity. Thus, they were selected for further studies on their cytokine response upon virus infection. It was found that ethanol and water crude extracts of C. longa and K. parviflora induced significant upregulation of TNF-α and IFN-β mRNA expressions, suggesting their roles in the inhibition of H5N1 virus replication. Conclusions To the best of the authors' knowledge, this study is among the earliest reports to illustrate the antiviral property of these Asian medicinal plants against the highly pathogenic avian H5N1 influenza virus. The results of this study shed light on alternative therapeutic sources for treatment of H5N1 influenza virus infection in the future.

887–891

Authors: Soraya Sheikhi, Hassan Vatandoost, Mohammad RezaAbai, Mansoreh Shayeghi, Ahmad Raeisi, Morteza Akbari, Fatemeh Nikpoor, Mohammad Sistanizade Aghdam, Akbar Bagheri

Objective To determine the wash resistance of Olyset® Plus using World Health Organization Pesticide Evaluation Scheme standard washing procedure and to assess the value of knock down and mortality rates of Anopheles stephensi at different regimens of long lasting insecticide treated nets washings. Methods The study was conducted at the Bioassay Laboratory of Culicidae Insectary, School of Public Health, Tehran University of Medical Sciences, Iran. The net was made of polyester impregnated with permethrin and piperonyl butoxide at a ratio of 2:1. The washing resistance was assessed using Le Chat® soap and a shaker incubator set at a speed of 155 r/min, 30 °C for 10 min. The cone bioassay test was carried out according to World Health Organization recommended guideline with tolerant field strain of female Anopheles stephensi to pyrethroids. Results The knockdown and mortality rates of female mosquitoes exposed to Olyset® Plus from un-washed nets to 2 washings were 79.7% and 88.8% respectively. Mortality was dropped to zero while active ingredient estimated 0.532 μg/100 cm2 to 0.481 μg/100 cm2 after 15 washings. A positive correlation was seen between residues of permethrin on nets, knockdown rate and mortality rate of female Anopheles stephensi exposed to different regimes of washed Olyset® Plus (r = 0.954, P = 0.001). Conclusions It is recommended that a preliminary survey conducted on resistance level of Anopheles vectors before the distribution of Olyset® Plus in malaria endemic communities.

896–899

Authors: Saied Goodarzi, Hassan Vatandoost, Mohammad Reza Abai, Saeed Tavakoli, Amir Hatamian, Yousef Ajani, Abbas Hadjiakhoondi, Narguess Yassa, Zahra Tofighi

Objective To determine the larvicidal activities of petroleum ether, chloroform, ethyl acetate and methanol fractions of roots and fruits extracts of Astrodaucus persicus from Apiaceae family against malaria vector, Anopheles stephensi (An. stephensi). Methods Twenty five third instar larvae of An. stephensi were exposed to various concentrations (10–160 g/L) of fractions and were assayed according to World Health Organization protocol. The larval mortality was calculated after 24 h treatment. Results Among tested fractions, the highest larvicidal efficacy was observed from ethyl acetate fraction of fruits extract with 50% and 90% mortality values (LC50 and LC90) of 34.49 g/L and 108.61 g/L, respectively. Chloroform fraction of fruits extract was the second larvicidal sample with LC50 of 45.11 g/L and LC90 of 139.36 g/L. Petroleum ether fractions of fruits and roots and methanol fraction of fruits showed moderate toxicity against An. stephensi. Conclusions Astrodaucus persicus is a potential source of valuable and natural larvicidal compounds against malaria vector, An. stephensi and can be used in mosquitoes control programs as an alternative to synthetic insecticides.

907–915

Authors: Tuan Tran Trong, Dieu-HienTruong, Hoang Chinh Nguyen, Dieu-Thai Tran, Huyen-Trang Nguyen Thi, Giap Do Dang, Ho NguyenHuu

Objective To evaluate the impact of plant growth regulators including kinetin (KN), benzyl adenine and naphthalene acetic acid, yeast extract and casein hydrolyzate on biomass accumulation of Vietnamese ginseng Panax vietnamensis (P. vietnamensis) in cell suspension culture. Methods Cell suspension cultures were established from friable calluses derived from leaves and petioles of 3-year-old in-vitro P. vietnamensis plants. The cell suspension cultures were grown in Murashige and Skoog basal media supplemented with various concentrations of KN, benzyl adenine, naphthalene acetic acid, and yeast extract and casein hydrolyzate. Results All tested factors generated an increase in the cell biomass of P. vietnamensis in suspension culture, but the impact of each varies depended on the factor type, concentration, and incubation period. Addition of 2.0 mg/L KN resulted in the largest biomass increase after 24 d, (57.0 ± 0.9) and (3.1 ± 0.1) mg/mL fresh and dry weight, respectively, whereas addition of benzyl adenine or naphthalene acetic acid produced optimum levels of Panax cell biomass at 1.0 and 1.5 mg/L, respectively. Addition of the elicitor yeast extract led to a 1.4–2.4 fold increase in biomass of P. vietnamensis, while addition of casein hydrolyzate enhanced biomass accumulation 1.8–2.6 fold. Conclusions The addition of each factor causes significant changes in biomass accumulation of P. vietnamensis. The largest biomass accumulation is from cultures grown in MS media containing 2.0 mg/L KN for 24 d. The outcome of the present study provides new insights into the optimal suspension culture conditions for studies on the in vitro cell biomass production of P. vietnamensis.

921–924

Authors: Wen-Bo Cheng, Dong Li, Qin Yang, Yue-Mei Hou

Objective To investigate the relationship between basal ganglia cerebral infarction and paroxysmal atrial fibrillation (PAF) caused by abnormal vagus nerve tension. Methods A total of 1 483 cases of elder patients with cerebral infarction who received head CT or MRI examination during the period were enrolled, including 830 male and 613 female, with the average age as 78 years. These cases were divided into basal infarction ganglia group (n = 1 045) and non-basal ganglia infarction group (n = 438) according to the anatomic site of cerebral infarction. The differences of the incidence of PAF, left atrial diameter and heart rate variability were compared between the two groups. Results In basal ganglia infarction group, the incidence rate of PAF was significantly higher than that of non-basal ganglia infarction group (P < 0.05). The incidence trend of cerebral infarction in basal ganglia was age-related, in the >79 years basal ganglia cerebral infarction group, the incidence of PAF was significantly higher than that of non-basal ganglia infarction group (P < 0.05). There was no significant difference in the left atrial diameter between the basal ganglia infarction group and non-basal ganglia infarction group. Basal ganglia cerebral infarction patients with high PAF had higher heart rate variability than non-basal ganglia infarction group. Conclusion Elderly patients with basal ganglia infarction have high incidence of PAF. Sympathetic nerve damage in cerebral basal ganglia, increased vagal tension and cardiac vagal tension are the direct causes of PAF. The results indicates that the increased central vagal nerve tension mediated PAF probably is an indication of supplying sympathetic neurotransmitter or cardiac vagal denervation treatment.

929–931

Authors: Henok A. Tegegne, Ivana Koláčková, Renata Karpíšková

Objective To evaluate the diversity and molecular characteristics of livestock associated methicillin-resistant Staphylococcus aureus in livestock animals, food of animal origin and the environment in the Czech Republic. Methods After having been primarily enriched in buffered peptone water, the samples were cultured on Baird–Parker agar. Presumptive colonies were sub-cultured to blood agar and assessed morphologically. Furthermore, presumptive Staphylococcus aureus colonies were confirmed by MALDI-TOF. Multiplex PCR, spa-typing, and MLST have been used to characterize the strains. Each mecA-positive Staphylococcus aureus isolates were examined against 14 different antimicrobials by using disk diffusion method. Results In this study, 13 different spa-types belonging to five sequence types (ST) were detected. Ninety four percent of tested strains belonged to CC/ST398 for which t011, t034, t2123 and t2346 were the vast major spa-types. In addition, non-ST398 clones such as CC1(t127), ST5(t3598), ST8(t064) and ST361(t315) were detected, which are known as human associated clones. Conclusions The diversity of livestock associated methicillin-resistant Staphylococcus aureus has grown, and detecting lineages of human origin in animals and vice-versa becomes more common. Thus, livestock animal and its products will be a potential for the evolvement of methicillin-resistant Staphylococcus aureus in human population. Monitoring of pigs as well as other food-producing animal species and their products is therefore recommended.

723–728

Authors: Valentina Virginia Ebani

Captive reptiles, always more often present in domestic environment as pets, may harbor and excrete a large variety of zoonotic pathogens. Among them, Salmonella is the most well-known agent, whereas there are very scant data about infections by mycobacteria, chlamydiae and leptospirae in cold-blooded animals. However, the investigations that found antibody reactions and/or the bacteria in samples collected from free-ranging and captive reptiles show that herpetofauna may be involved in the epidemiology of these infections. The present review reports the updated knowledge about salmonellosis, mycobacteriosis, chlamydiosis and leptospirosis in reptiles and underlines the risk of infection to which people, mainly children, are exposed.

744–752

Authors: Vijay Kothari, Sakshi Sharma, Divya Padia

Chromobacterium violaceum is a gram-negative bacterium, which has been used widely in microbiology labs involved in quorum sensing (QS) research. Among the QS-regulated traits of this bacterium, violacein production has received the maximum attention. Violacein production in this organism, however is not under sole control of QS machinery, and other QS-regulated traits of this bacterium also need to be investigated in better detail. Though not often involved in human infections, this bacterium is being viewed as an emerging pathogen. This review attempts to highlight the recent research advances on C. violaceum, with respect to violacein biosynthesis, development of various applications of this bacterium and its bioactive metabolite violacein, and its pathogenicity.

760–764

Authors: Yasir Waheed, Sher Zaman Safi, Muzammil Hasan Najmi, Hafsa Aziz, Muhammad Imran

Objective To explore RNA dependent RNA polymerase of Chikungunya virus (CHIKV) and develop T cell based epitopes with high antigenicity and good binding affinity for the human leukocyte antigen (HLA) classes as targets for epitopes based CHIKV vaccine. Methods In this study we downloaded 371 non-structural protein 4 protein sequences of CHIKV belonging to different regions of the world from the US National Institute of Allergy and Infectious Diseases (NIAID) virus pathogen resource database. All the sequences were aligned by using CLUSTALW software and a consensus sequence was developed by using Uni Pro U Gene Software version 1.2.1. Propred I and Propred software were used to predict HLA I and HLA II binding promiscuous epitopes from the consensus sequence of non-structural protein 4 protein. The predicted epitopes were analyzed to determine their antigenicity through Vaxijen server version 2.0. All the HLA I binding epitopes were scanned to determine their immunogenic potential through the Immune Epitope Database (IEDB). All the predicted epitopes of our study were fed to IEDB database to determine whether they had been tested earlier. Results Twenty two HLA class II epitopes and eight HLA class I epitopes were predicted. The promiscuous epitopes WMNMEVKII at position 486–494 and VRRLNAVLL at 331–339 were found to bind with 37 and 36 of the 51 HLA class II alleles respectively. Epitope MANRSRYQS at position 58–66 and epitopes YQSRKVENM at positions 64–72 were predicted to bind with 12 and 9 HLA II alleles with antigenicity scores of 0.754 9 and 1.013 0 respectively. Epitope YSPPINVRL was predicted to bind 18 HLA I alleles and its antigenicity score was 1.425 9 and immunogenicity score was 0.173 83. This epitope is very useful in the preparation of a universal vaccine against CHIKV infection. Conclusions Epitopes reported in this study showed promiscuity, antigenicity as well as good binding affinity for the HLA classes. These epitopes will provide the baseline for development of efficacious vaccine for CHIKV.

773–786

Authors: Omar M.E. Abdel-Salam, Eman R. Youness, Nadia A. Mohammed, Noha N. Yassen, Yasser A. Khadrawy, Safinaz Ebrahim El-Toukhy, Amany A. Sleem

Objective To investigate the effect of NG-nitro-l-arginine methyl ester (l-NAME), a non-selective nitric oxide synthase (NOS) inhibitor, and 7-nitroindazole (7-NI), a selective neuronal NOS inhibitor, on oxidative stress and tissue damage in brain and liver and on DNA damage of peripheral blood lymphocytes in malathion intoxicated rats. Methods Malathion (150 mg/kg) was given intraperitoneally (i.p.) along with l-NAME or 7-NI (10 or 20 mg/kg, i.p.) and rats were euthanized 4 h later. The lipid peroxidation product malondialdehyde (MDA), nitric oxide (nitrite), reduced glutathione (GSH) concentrations and paraoxonase-1 (PON-1) activity were measured in both brain and liver. Moreover, the activities of glutathione peroxidase (GPx) acetylcholinesterase (AChE), and butyrylcholinesterase (BChE), total antioxidant capacity (TAC), glucose concentrations were determined in brain. Liver enzyme determination, Comet assay, histopathological examination of brain and liver sections and inducible nitric oxide synthase (iNOS) immunohistochemistry were also performed. Results (i) Rats treated with only malathion exhibited increased nitric oxide and lipid peroxidation (malondialdehyde) accompanied with a decrease in GSH content, and PON-1 activity in brain and liver. Glutathione peroxidase activity, TAC, glucose concentrations, AChE and BChE activities were decreased in brain. There were also raised liver aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities and increased DNA damage of peripheral blood lymphocytes (Comet assay). Malathion caused marked histopathological changes and increased the expression of iNOS in brain and liver tissues. (ii) In brain of malathion-intoxicated rats, l-NAME or 7-NI resulted in decreased nitrite and MDA contents while increasing TAC and PON1 activity. Reduced GSH and GPx activity showed an increase by l-NAME. AChE activity increased by 20 mg/kg l-NAME and 10 mg/kg 7-NI. AChE activity decreased by the higher dose of 7-NI while either dose of 7-NI resulted in decreased BChE activity. (iii) In liver of malathion-intoxicated rats, decreased MDA content was observed after l-NAME or 7-NI. Nitrite level was unchanged by l-NAME but increased after 7-NI which also resulted in decreased GSH concentration and PON1 activity. Either inhibitor resulted in decreased liver ALT activity. (iv) DNA damage of peripheral blood lymphocytes was markedly inhibited by l-NAME or 7-NI treatment. (v) iNOS expression in brain and liver decreased by l-NAME or 7-NI. (vi) More marked improvement of the histopathological alterations induced by malathion in brain and liver was observed after 7-NI compared with l-NAME. Conclusions In malathion intoxicated rats, the neuronal NOS inhibitor 7-NI and to much less extent l-NAME were able to protect the brain and liver tissue integrity along with improvement in oxidative stress parameters. The decrease in DNA damage of peripheral blood lymphocytes by NOS inhibitors also suggests the involvement of nitric oxide in this process.

792–801

Authors: Hafiza Noreen, Nabil Semmar, Muhammad Farman, James S.O. McCullagh

Objective To evaluate the total phenolic content and compare the antioxidant activity of various solvent extracts and fractions from the aerial parts of Coronopus didymus through various assays. Methods Total phenolic content was determined using the Folin-Ciocalteu assay and the in vitro antioxidant activity of a number of different extracts was investigated in a dose-dependent manner with three different methods: the 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and ferric reducing antioxidant power (FRAP) assays. A flavone was isolated from the most active ethanolic extract with high antioxidant activity using size exclusion chromatography. IC50 values were calculated for the DPPH and ABTS methods. The FRAP activity was assessed in terms of μM Fe (II) equivalent. Results The phenolic content was found to be highest in the ethanol extract (CDA Et; 47.8 mM GAE) and the lowest in the dichloromethane extract (CDA DCM; 3.13 mM GAE). The ethanol extract showed high radical scavenging activity towards DPPH and ABTS radicals with IC50 values of (7.80 × 102) and (4.32 × 102) μg/mL, respectively. The most active ethanol extract had a FRAP value of 1921.7 μM Fe (II) equivalent. The isolated flavone F10C (5,7,4′-trihydroxy-3′-methoxy flavone) was far more effective for scavenging free radicals in the DPPH and ABTS assays with IC50 of 43.8 and 0.08 μg/mL, than the standard trolox, with IC50 values of 97.5 and 21.1 μg/mL, respectively. In addition, the flavone F10C and the standard ascorbic acid had FRAP values of 1621.7 and 16 038.0 μM Fe (II) equivalents, respectively. Conclusions The total phenolic content of extracts in decreasing order is ethanol extract (CDA Et) > acetone extract (CDA ACE) > phenolic extract (CDA MW) > n-hexane extract (CDA nHX)> chloroform extract (CDA CHL) > dichloromethane extract (CDA DCM). The ordering of extracts in terms of antioxidant activity from highest to lowest is CDA Et > CDA MW > CDA DCM > CDA CHL > CDA ACE > CDA nHX in DPPH, ABTS and FRAP assays. A significant relationship is found between antioxidant potential and total phenolic content, suggesting that phenolic compounds are the major contributors to the antioxidant activity of C. didymus.

808–812

Authors: Patel Mahipal, Rajesh Singh Pawar

Asian Pacific Journal of Tropical Medicine Asian Pacific Journal of Tropical Medicine Volume 10, Issue 8, August 2017, Pages 808-812 open access Asian Pacific Journal of Tropical Medicine Original research Nephroprotective effect of Murraya koenigii on cyclophosphamide induced nephrotoxicity in rats Author links open overlay panelPatelMahipalRajesh SinghPawar Show more https://doi.org/10.1016/j.apjtm.2017.08.005Get rights and content Open Access funded by Hainan Medical College Under a Creative Commons license Abstract Objective To evaluate the nephroprotective effect of defatted methanolic extract and aqueous extract of Murraya koenigii (M. koenigii) against cyclophosphamide drug. Methods Nephrotoxicity was induced by cyclophosphamide in 7 days at 150 mg/kg body weight through intraperitoneal route in rat model. Nephroprotective activity of M. koenigii extract (100 mg/kg and 200 mg/kg in intraperitoneal route) was measured, including nephrological source, oxidative stress parameters like superoxide dismutase, glutathione, the lipid peroxide and in vivo assay like blood urea nitrogen, creatinine were determined and analyzed by One way analysis of variance followed by Tukey's test. Results The study result showed that important phytochemicals such as carbohydrates, flavonoids, tannin, alkaloids, glycosides, protein and steroids were found to be present in the extract of M. koenigii. The renal function markers like blood urea nitrogen and creatinine level were found to be decreased significantly by M. koenigii extract treatment. A significant difference was found to be at P < 0.01. Conclusions The present study reveals the protective role of M. koenigii extract against cyclophosphamide induced nephrotoxicity.

819–826

Authors: Behnam Mahdavi, Wan A. Yaacob, Laily B. Din

Objective To report the chemical composition and bioactivity (including antioxidant and antimicrobial activity) of essential oils from the rhizomes, stems, and leaves of Etlingera sayapensis (E. sayapensis) A.D. Poulsen & Ibrahim for the first time. Methods First, the essential oils were obtained using a Clevenger-type apparatus. Then, the essential oils compositions were identified by chromatography methods including GC-FID and GC–MS. For the next step, DPPH radical scavenging activity (RSA), β-carotene bleaching (BCB), and ferrous ion chelating ability (FIC) were chosen to evaluate the essential oils antioxidant activity. Finally, disc diffusion assay and minimum inhibitory concentration method (MIC) was applied to investigate antimicrobial activity of the rhizomes and leaves oils of E. sayapensis against 18 microorganisms. Results All of the oils contained oxygenated monoterpenes (leaves: 74.18%, stems: 75.60%, and rhizome: 54.61%), The essential oil obtained from leaves contained high amount of carvone (21.38%), cis-carveol (13.49%); The rhizomes oil was rich in linalool formate (25.47%), eugenol (11.84%); and the stems oil was dominated by α-terpineol (39.86%), linalool formate (30.55%). The leaves oil represented the highest ability in all of the antioxidant activity tests. For antimicrobial activity, the rhizome oil presented more active when compared to leaves oil against Bacillus subtilis, Bacillus thuringiensis, Staphylococcus aureus, methicillin resistant Staphylococcus aureus (MRSA), Aeromonas hydrophila, Escherichia coli, Enterobacter aerogenes, Proteus mirabilis, Shigella sonnei, Serratia marcescens, Vibrio parahaemolyticus, Candida albicans, and Candida parapsilosis. Conclusions The most components of the essential oils belong to oxygenated monoterpenes. Linalool formate, carvone, and α-terpineol are found as the most abundant compounds in the oils of the different parts of E. sayapensis. The rhizomes oil can prevent the growth of wide spectrum microorganisms; however, the oils are not highly potent in antioxidant assays.

833–834

Authors: Marcel Massoud, Fouad Kerbage, Leony Antoun, Ribal Merhi, Souheil Hallit, Rabih Hallit

Brucellosis is one of the most common zoonotic infections worldwide caused by gram negative bacilli of the genus Brucella. It is transmitted to humans by contact with infected animals or derived food products such as unpasteurized milk. Brucellosis' clinical presentation varies widely from multi-systemic involvement to asymptomatic infection. We present the case of a 52-year-old Lebanese male who was admitted to our hospital with a 3-week history of fever (up to 40 °C), chills, night sweats and abdominal pain. Abdominal CT scan revealed the presence of several mesenteric lymphadenopathies and some retroperitoneal lymphadenopathies. Blood cultures came back positive for Brucella melitensis, and a follow-up CT of the abdomen done after treatment revealed complete resolution of the lymphadenopathies. To our knowledge, this is the first case in the literature of brucellosis presenting as retroperitoneal and mesenteric lymphadenopathies. In endemic areas, the diagnosis of brucellosis should always be raised in front of any long duration fever even in the absence of a typical clinical presentation.