Effects of Combretum hereroense and Canthium mundianum water extracts on production and expression of interleukin-4

Background: Combretum hereroense and Canthium mundianum are two plants commonly used by traditional healers in the Northern region of Limpopo, South Africa for the treatment of diarrhea and inflammation. In the present study, the effects of their water extracts on the production and expression of interleukin-4 by peripheral blood mononuclear cells (PBMC’S) from HIV positive and negative individuals was evaluated.Materials and methods: Blood samples were collected from both HIV positive and HIV negative volunteers and were used for the purification of Peripheral blood mononuclear cells (PBMC). The PBMCs were cultured together with the water extracts after activation with phytohemagglutinin (PHA) for three days. Solid-phase sandwich ELISA (MABTECH) kit was used to detect IL-4 on un-stimulated and stimulated PBMC’S with phytohemaglutinin (PHA) and plant extracts, followed by the isolation of RNA using RNAesy Qiagen mini kit from the cells. Reverse transcriptase real time PCR was used to evaluate IL-4 gene expression by the cells.Results: Combretum hereroense showed higher production of IL-4 at three different concentrations and a significant expression of mRNA with 4-fold amplification increase at 300μg/ml and 2-fold amplification increase at 20μg/ml. Canthium mundianum also showed increased production of IL-4 at 300μg/ml, but inhibited its production at 20μg/ml. Both extracts showed no expression at 50μg/ml. The response of the PBMCs from HIV negative individuals was more pronounced than that of HIV positive individuals who mostly increased production of IL4 at smaller concentrations unlike their HIV negative counterparts. Although in vitro studies do not necessarily predict in vivo outcomes, the plant extracts modulated the immune system by enhancing the production and expression of IL-4 in both HIV- and HIV+ individuals at different concentrations.Conclusions: For the first time we have shown that the immunomodulatory effect of medicinal plants may depend on the clinical status of the individual. The present study revealed that the effect of the water extracts from the two plants on IL-4 expression and production is dependent on the microbiological state of the individual and is dose dependent. Further studies are needed to identify the active components in the extracts and also characterize the patients further for a better understanding of the mechanisms of action of these extracts.Keywords: interleukin-4, peripheral mononuclear cells, HIV, ELISA, gene expressio

EFFECTS OF COMBRETUM HEREROENSE AND CANTHIUM MUNDIANUM water EXTRACTS ON PRODUCTION AND EXPRESSION OF INTERLEUKIN-4

Background: Combretum hereroense and Canthium mundianum are two plants commonly used by traditional healers in the Northern region of Limpopo, South Africa for the treatment of diarrhea and inflammation. In the present study, the effects of their water extracts on the production and expression of interleukin-4 by peripheral blood mononuclear cells (PBMC’S) from HIV positive and negative individuals was evaluated. Materials and methods: Blood samples were collected from both HIV positive and HIV negative volunteers and were used for the purification of Peripheral blood mononuclear cells (PBMC). The PBMCs were cultured together with the water extracts after activation with phytohemagglutinin (PHA) for three days. Solid-phase sandwich ELISA (MABTECH) kit was used to detect IL-4 on un-stimulated and stimulated PBMC’S with phytohemaglutinin (PHA) and plant extracts, followed by the isolation of RNA using RNAesy Qiagen mini kit from the cells. Reverse transcriptase real time PCR was used to evaluate IL-4 gene expression by the cells. Results: Combretum hereroense showed higher production of IL-4 at three different concentrations and a significant expression of mRNA with 4-fold amplification increase at 300μg/ml and 2-fold amplification increase at 20μg/ml. Canthium mundianum also showed increased production of IL-4 at 300μg/ml, but inhibited its production at 20μg/ml. Both extracts showed no expression at 50μg/ml. The response of the PBMCs from HIV negative individuals was more pronounced than that of HIV positive individuals who mostly increased production of IL4 at smaller concentrations unlike their HIV negative counterparts. Although in vitro studies do not necessarily predict in vivo outcomes, the plant extracts modulated the immune system by enhancing the production and expression of IL-4 in both HIV- and HIV+ individuals at different concentrations. Conclusions: For the first time we have shown that the immunomodulatory effect of medicinal plants may depend on the clinical status of the individual. The present study revealed that the effect of the water extracts from the two plants on IL-4 expression and production is dependent on the microbiological state of the individual and is dose dependent. Further studies are needed to identify the active components in the extracts and also characterize the patients further for a better understanding of the mechanisms of action of these extracts

PREVALENCE OF TOXOPLASMA GONDII IGG AND IGM AND ASSOCIATED RISK FACTORS AMONG HIV-POSITIVE AND HIV-NEGATIVE PATIENTS IN VHEMBE DISTRICT OF SOUTH AFRICA

Background: Toxoplasma gondii is a zoonotic parasite that has arisen as an important opportunistic infection that causes morbidity and mortality especially in HIV positive patients. This study was carried out to determine the sero-prevalence of T. gondii (IgG and IgM) and the associated risk factors among HIV positive and negative patients in Northern South Africa. Materials and Methods: The study was conducted in the Vhembe District in Limpopo province from April 2012 to January 2013. A well-structured questionnaire was used to collect socio-demographic information and possible risk factor information on toxoplasmosis from participants. A total of 161 blood samples of both HIV positive and negative patients visiting the local clinics in the Vhembe district were collected. Serum samples were tested for IgG and IgM against T. gondii using commercially available ELISA protocol. Results: The prevalence of T. gondii IgG was 31.7% while that of T. gondii IgM was 4.9%. The prevalence of T. gondii IgG was higher in HIV positive patients (38%) compared to 16.7% among HIV negative patients (p=0.001). Toxoplasma gondii IgG antibodies were more common in patients who were not taking ARV’s (46.2%) compared to those who were taking ARV’s (35.2%) (P25 years)

Advances on Dientamoeba fragilis Infections

Dientamoeba fragilis is an enteric protozoan parasite that remains neglected, probably due to the misconception that it is uncommon and non-pathogenic. As more information became available and antimicrobial agents were developed with activity against this parasite, it became clear that D. fragilis is responsible of an active infection, associated with symptoms such as abdominal pain and diarrhea. The clinical presentation of dientamoebiasis varies from asymptomatic carriage to symptoms ranging from altered bowel motions, abdominal discomfort, nausea and diarrhea with associated eosinophilia reported in up to 50% of paediatric and 10% of adult patients. Moreover, controversy exists over the protective role of the parasite in priming the immune system in a beneficial way such as in selecting beneficial bacteria, keeping potential harmful microbial intruders at bay or producing metabolites beneficial to the host. Thus, a number of ambiguities and obscurities surrounding D. fragilis infections exist. Moreover, the means by which this parasite is transmitted has not been fully defined. The diagnostic recognition of this parasite in fecal examinations requires specific processing and expertise; thus, it is possible that many infections with D. fragilis may go undiagnosed. A number of studies conducted on small numbers of case reports have demonstrated parasite clearance, as well as resolution of clinical symptoms following treatment with various antiparasitic compounds such as paromomycin, hydroxyquinolines and the 5-nitroimidazoles, including metronidazole and tinidazole. In addition there is very little in vitro susceptibility data available for the organism making some current treatment options questionable. This chapter reviews the scientific literature relating to Dientamoeba\u27s life cycle, prevalence, diagnosis and pathogenicity

Inhibitory and Bactericidal Potential of Crude Acetone Extracts of Combretum molle (Combretaceae) on Drug-resistant Strains of Helicobacter pylori

Infection with Helicobacter pylori is strongly associated with a number of gastroduodenal pathologies. Antimicrobial resistance to commonly-used drugs has generated a considerable interest in the search for novel therapeutic compounds from medicinal plants. As an ongoing effort of this search, the susceptibility of 32 clinical strains of H. pylori and a reference strain—NCTC 11638—was evaluated against five solvent extracts of Combretum molle, a plant widely used for the treatment of gastric ulcers and other stomach-related morbidities in South Africa. The extracts were screened for activity by the agar-well diffusion method, and the most active one of them was tested against the same strains by micro-broth dilution and time kill assays. Metronidazole and amoxicillin were included in these experiments as positive control antibiotics. The solvent extracts all demonstrated anti-H. pylori activity with zone diameters of inhibition between 0 and 38 mm. The most potent anti-H. pylori activity was demonstrated by the acetone extract, to which 87.5% of the clinical strains were susceptible. The minimum inhibitory concentration (MIC90) values for this extract ranged from 1.25 to 5.0 mg/mL while those for amoxicillin and metronidazole ranged from 0.001 to 0.94 mg/mL and from 0.004 to 5.0 mg/mL respectively. The acetone extract was highly bactericidal at a concentration of 2.5 and 5.0 mg/mL, with complete elimination of the test organisms in 24 hours. Its inhibitory activity was better than that of metronidazole (p<0.05) as opposed to amoxicillin (p<0.05). The results demonstrate that C. molle may contain therapeutically-useful compounds against H. pylori, which are mostly concentrated in the acetone extract

Inhibitory and Bactericidal Potential of Crude Acetone Extracts of Combretum molle (Combretaceae) on Drug-resistant Strains of Helicobacter pylori

Infection with Helicobacter pylori is strongly associated with a number of gastroduodenal pathologies. Antimicrobial resistance to commonly-used drugs has generated a considerable interest in the search for novel therapeutic compounds from medicinal plants. As an ongoing effort of this search, the susceptibility of 32 clinical strains of H. pylori and a reference strain-NCTC 11638-was evaluated against five solvent extracts of Combretum molle , a plant widely used for the treatment of gastric ulcers and other stomach-related morbidities in South Africa. The extracts were screened for activity by the agar-well diffusion method, and the most active one of them was tested against the same strains by micro-broth dilution and time kill assays. Metronidazole and amoxicillin were included in these experiments as positive control antibiotics. The solvent extracts all demonstrated anti-H. pylori activity with zone diameters of inhibition between 0 and 38 mm. The most potent anti-H. pylori activity was demonstrated by the acetone extract, to which 87.5% of the clinical strains were susceptible. The minimum inhibitory concentration (MIC90) values for this extract ranged from 1.25 to 5.0 mg/mL while those for amoxicillin and metronidazole ranged from 0.001 to 0.94 mg/mL and from 0.004 to 5.0 mg/mL respectively. The acetone extract was highly bactericidal at a concentration of 2.5 and 5.0 mg/mL, with complete elimination of the test organisms in 24 hours. Its inhibitory activity was better than that of metronidazole (p&lt;0.05) as opposed to amoxicillin (p&lt;0.05). The results demonstrate that C. molle may contain therapeutically-useful compounds against H. pylori, which are mostly concentrated in the acetone extract

PREVALENCE OF CRYPTOSPORIDIUM SPP. AMONG ASYMPTOMATIC HEALTHY EXPATRIATE WORKERS IN SHARJAH, UNITED ARAB EMIRATES

Background: Epidemiological data on Cryptosporidium infections in the United Arab Emirates (UAE) is scarce. Therefore, the main objective of this study was to determine the prevalence of Cryptosporidium species among a community of expatriates in Sharjah, UAE working in different sectors, including the food industry, house maids and other domestic occupations. Materials and Methods: One hundred and thirty four stool samples were collected from asymptomatic individuals presenting to the Sharjah Municipality Public Health Clinic (SMPHC) for screening of intestinal parasites for work permission purposes between 2009 and 2011. Demographic information such as age, sex, and country of origin was collected. Genomic DNA extracted from the stool samples were tested for Cryptosporidium species using real-time PCR (qPCR). Results: Twenty-six individuals (19.4%) were positive for Cryptosporidium sp. by PCR. The infection rate was found to be highest in Afghan nationals (33%; 3/9) compared with the rest of the study population; yet, no significant association existed between nationality and infection rate. Moreover, no association was observed between infection rate and gender (χ2 = 2.439; P = 0.118), nor infection rate and age group (χ2 = 1.219; P = 0.544). Conclusion: Infection by Cryptosporidium sp. was common in the study group, and further studies are needed within the native Emirati population before any conclusions can be made about foreigners potentially transmitting the parasite. Furthermore, data provided in this study could help determine its public and veterinary significance particularly in outbreaks in the country

Bacterial Contamination of Vhuswa - A Local Weaning Food and Stored Drinking-water in Impoverished Households in the Venda Region of South Africa

Bacterial contaminants of Vhuswa, a traditional maize-based weaning food, and domestic drinking-water stored in impoverished rural households in Venda of Limpopo province, South Africa, were determined. One hundred and twenty-five samples of Vhuswa fed to children aged less than five years were assessed for Escherichia coli , Campylobacter jejuni , Salmonella , and Shigella . The microbiological quality of 125 drinking-water samples was also evaluated using total coliforms, faecal coliforms, and faecal streptococci as indicators. The frequency of isolation of E. coli, Salmonella, Shigella, and C. jejuni from the Vhuswa samples was 70%, 5%, 5%, and 2% respectively. The geometric mean counts of total coliforms, faecal coliforms, and faecal streptococci in tap-water stored in household containers ranged from 4.9x102 to 5.8x103 cfu 100 mL-1 , 2.6x102 to 3.7x103 cfu 100 mL-1 , and 3.1x103 to 5.8x103 cfu 100 mL-1 respectively, and for stored spring water it was 5.1x103 cfu 100 mL-1 , 3.2x103 cfu 100 mL-1 , and 5.1x103 cfu 100 mL-1 respectively. The frequent contamination of water and food samples in this study has important implications for the health of children from impoverished communities