Absence of hybridization with the wild-type and mutant rpoB probes in the Genotype MTBDRplus assay detects 'disputed' rifampicin mutations.

OBJECTIVE: Mycobacterium tuberculosis isolates that fail to hybridize to at least one rpoB wild-type or any mutation probe on the Genotype MTBDRplus strip are assumed to be rifampicin-resistant. However, the precise mutation(s) are unknown. We sought to identify the mutations in isolates with such hybridization patterns and determine if the mutations are associated with resistance to rifampicin. METHODS: In this study, 275 M. tuberculosis isolates were screened with the Genotype MTBDRplus assay to identify isolates with the hybridization pattern. These isolates were sequenced and their minimum inhibitory concentrations (MIC) determined using the Bactec MGIT 960 system. RESULTS: Among the 275 isolates tested, 15 (6%) isolates with the hybridization pattern were identified. Sequencing showed that failure to hybridize to rpoB wild-type probes resulted from the presence of 'disputed' rifampicin mutations, which are mutations not always associated with a rifampicin-resistant phenotype. All, except 3/15, isolates had a rifampicin-resistant phenotype (MIC > 1 µg/mL). One of the three isolates with a rifampicin-susceptible phenotype had the same mutation at position 526 (His526Leu) as another isolate that had a rifampicin-resistant phenotype. CONCLUSION: The recommendation of the Genotype MTBDRplus assay to assume rifampicin resistance based solely on failure to hybridize to rpoB wild-type probe allows the identification of important RIF-resistant isolates. About 20% (3/15) of such isolates could be missed by relying only on the standard MGIT 960 DST assay for drug susceptibility testing.

In vitro antibacterial activity and acute toxicity studies of aqueous-methanol extract of Sida rhombifolia Linn. (Malvaceae).

BACKGROUND: Many bacteria among the Enterobacteria family are involved in infectious diseases and diarrhoea. Most of these bacteria become resistant to the most commonly used synthetic drugs in Cameroon. Natural substances seem to be an alternative to this problem. Thus the aim of this research was to investigate the in vitro antibacterial activity of the methanol and aqueous-methanol extracts of Sida rhombifolia Linn (Malvaceae) against seven pathogenic bacteria involved in diarrhoea. Acute toxicity of the most active extract was determined and major bioactive components were screened. METHODS: The agar disc diffusion and the agar dilution method were used for the determination of inhibition diameters and the Minimum Inhibitory Concentration (MICs) respectively. The acute toxicity study was performed according WHO protocol. RESULTS: The aqueous-methanol extract (1v:4v) was the most active with diameters of inhibition zones ranging from 8.7 - 23.6 mm, however at 200 microg/dic this activity was relatively weak compared to gentamycin. The MICs of the aqueous-methanol extract (1v:4v) varied from 49.40 to 78.30 microg/ml. Salmonella dysenteriae was the most sensitive (49.40 microg/ml). For the acute toxicity study, no deaths of rats were recorded. However, significant increase of some biochemical parameters such as aspartate amino-transferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and creatinine (CRT) were found. The phytochemical analysis of the aqueous methanol extract indicated the presence of tannins, polyphenols, alkaloids, glycosides, flavonoids and saponins CONCLUSION: The results showed that the aqueous-methanol extract of S. rhombifolia exhibited moderate antibacterial activity. Some toxic effects were found when rats received more than 8 g/kg bw of extract.

In vitro cytotoxicity and antioxidant activities of five medicinal plants of Malvaceae family from Cameroon.

The potential antiproliferative and antioxidant activities of extracts from five medicinal plants from Cameroon were evaluated in vitro on HepG-2 cells. The results showed the significant decrease of the viability of the cells in a concentration-dependent manner. According to the IC(50) obtained, the extracts of S. acuta (461.53±0.23) and U. lobata (454.93±0.12) showed significant antiproliferative activity. At fixed concentration (250µgmL(-1)), extracts demonstrated higher antiproliferative activity (67.05%; 65.42%), (52.62%; 56.64%) and (32.98%; 36.85%) respectively during 24, 48 and 72h. Extracts of S. cordifolia and V. album demonstrated significant antiproliferative property after 48h while S. rhombifolia exhibited weak cytotoxicity. The results of the antioxidant properties showed that theses extracts induced significantly increase of SOD, CAT and GsT activity after 48h. Taken together, the results extracts showed that of S. acuta and U. lobata may be a promising alternative to synthetic substances as natural compound with high antiproliferative and antioxidant activities.

Seasonal variation and prevalence of tuberculosis among health seekers in the South Western Cameroon.

OBJECTIVES: To determine the prevalence of tuberculosis (TB) in Fako health District, to assess the effects of seasonal variation on the incidence of TB in the study area and to use sentinel analysis to predict areas of greatest infection. DESIGN: A prospective cross sectional study based on laboratory investigations. SETTING: Fako health District, South Western Cameroon. RESULTS: The prevalence of TB was 23.3%. Tuberculosis was significantly more prevalent in males (12.6%) as compared with females (10.7%) (P = 0.034). TB prevalence was significantly different between age groups, with the highest number of cases recorded in the age group 21-30 (P = 0.002). When the health areas were compared, TB prevalence varied significantly (P = 0.001), with Limbe Town recording the highest number of TB cases. We recorded more TB cases in the wet season compared with the dry season and the difference was statistically significant (P = 0.000). There was a significant drop in the prevalence of TB over the study period (P = 0.000). CONCLUSION: This study is the first to report on the effects of season on the prevalence of TB in Cameroon. These findings will therefore provide additional useful base line data for setting up TB control strategies in Cameroon.

Anti-ulcer actions of the bark methanol extract of Voacanga africana in different experimental ulcer models in rats.

The antiulcerogenic effects of the bark methanol extract of Voacanga africana were studied using various experimental ulcer models in rats. The effects of the extract on the volume of gastric juice, gastric pH, acid output, mucus production and peptic activity were recorded, as well as the preventive action against lesions caused by HCl/ethanol and indomethacin. Oral administration of the extract (500-750 mg/kg) inhibited the formation of gastric lesions induced by HCl/ethanol (40-63% inhibition). The inhibitory effect against HCl/ethanol was significantly (P<0.01) suppressed by pre-treatment of the rats with indomethacin (30 mg/kg, i.p.). The extract significantly reduced gastric lesion formation in pylorus ligated rats, but this was not associated with an increase in gastric mucus production or with a reduction in acid content, volume of gastric secretion or pepsin activity of the gastric juice.

In vivo antitumor activity of 4-amino 4-methyl 2-pentyne 1-al, an inhibitor of aldehyde dehydrogenase.

4-amino-4-methyl-2-pentyne-1-al (AMPAL), a new irreversible inhibitor of aldehyde dehydrogenase (ALDH) has been assayed for its in vitro and in vivo antitumor activity. In vitro, AMPAL inhibits the proliferation and the ALDH activity of L1210 and RBL5 cell lines. In vivo, AMPAL significantly increases the mean survival time of mice i.p. grafted with leukemia (L1210, P815, MBL2, EL4, RBL5 cell lines) or carcinoma cells (Krebs cell line), without haematopoetic toxicity. No carcinostatic effect was observed against the P388 leukemia and the 3LL Lewis lung carcinoma. A possible relationship between the ALDH isoenzyme activity of the tumor and its sensitivity to AMPAL is discussed in the light of previous reports concerning the role of aldehydes in cell growth control.