Genetic fingerprinting and profile analysis of virulence genes in XDR clinical isolates of Klebsiella Pneumoniae.

OBJECTIVE: Klebsiella pneumoniae, a leading cause of hospital- and community-acquired infections, requires the acquisition of drug resistance and virulence for survival in evading the host's immune responses. The investigation looks at the molecular diversity of 30 clinical isolates using rep-PCR DNA fingerprinting and the association between drug resistance and virulence genes. MATERIALS AND METHODS: Isolates were cultured using basic bacteriological techniques, while antimicrobial susceptibility test and ID confirmation were made by Vitek 2 Compact Automated System (BioMerieux, Marcy L'Etoile, France). DNA used for PCR reactions was extracted with Qiagen DNA extraction kit according to the manufacturer's guidelines, while molecular genotyping by REP-PCR was carried out according to previously described methods. Some virulence genes and capsular serotypes, ybtS, mrkD, entB, rmpA, K2, Kfu, alls, iutA, and magA, were amplified by multiplex PCR. RESULTS: The majority (60%) of the isolates were MDR, others were XDR (37%) and susceptible strains (3%). Additionally, resistance was high (73%) for meropenem and lower (40%) for imipenem, while 97% were susceptible to ertapenem and azithromycin. DNA fingerprinting by rep-PCR showed polymorphic loci sizes that ranged from 100-2000 bp-, and phylogenetic analysis placed the isolates in seven clades with 40-96%. There was a 75% phylogenetic relatedness to antimicrobial resistance, but no specific pattern of relatedness was seen between virulence genes for the K. pneumoniae strain. Regulator mucoid phenotype A (rmpA) gene was not amplified in any of the isolates. CONCLUSIONS: This study further confirms the presence of both MDR and XDR K. pneumonia strains. Isolates exhibited genetic diversity in phylogenetic patterns and carriage of virulence with relatedness by susceptibility to antimicrobials. Therefore, circulation could pose a risk to public health.

Subchronic administration of mitoxantrone and the influence of enzyme inhibitors on its induced cardiotoxicity in mice: role of NRF-2/CYP2E1.

OBJECTIVE: Mitoxantrone (MTX)- induced cardiotoxicity is a clinical concern that is limiting its use. The aim of this paper, therefore, was to investigate the subchronic administration of MTX plus nonspecific/specific inhibitors of CYP450/2E1, to assess the extent of oxidative-induced injury by measuring levels of oxidative cardiac and injury biomarkers in mice and to evaluate the effects of CYP2E1 on caspase 3 activity and nuclear factor erythroid 2-related factor-2 (NRF-2). MATERIALS AND METHODS: Mice (n = 32) were divided into four treatment groups of eight: control, MTX, MTX + 4-methlypyrazole (4MP) and MTX + disulfiram (Disf). After 6 weeks of treatments, blood and heart samples were collected. RESULTS: Liquid chromatography-mass spectrometry (LCMS) analysis of MTX-treated plasma samples revealed several metabolites with different retention times. Cardiac antioxidant enzymes and creatine kinase (CK) levels were not significantly different among the groups. However, cardiac troponin and caspase 3 activity were significantly raised, with increased CYP2E1 expressions and reduced NRF-2 expression. Tissue damage was observed in all the treatment groups, including MTX, leading to the conclusion that MTX-induced cardiotoxicity was mediated by CYP2E1 activity, which initiated caspase 3 production, and decreased NRF-2 expression. CONCLUSIONS: Therefore, agents that inhibit CPY2E1 expression might attenuate MTX-induced cardiotoxicity by increasing NRF-2 expression.

Toxicological evaluation of therapeutic and supra-therapeutic doses of Cellgevity® on reproductive function and biochemical indices in Wistar rats.

BACKGROUND: The misconception about dietary supplements being safe has led many into the in-patient wards. Cellgevity® (CGV) is a Max International premiere antioxidant supplement formula used by a large population. This study evaluated the effects of therapeutic and supra-therapeutic doses of CGV on reproductive function and biochemical indices in Wistar rats. METHODS: Seventy-two Wistar rats weighing 130 ± 15.8 g were grouped into two categories (male or female) of six rats per group. Control group received distilled water (10 ml/kg). Others received therapeutic (14.3 mg/kg or 28.6 mg/kg) and supra-therapeutic CGV doses (1000, 2000 or 3000 mg/kg) body weight per oral respectively. RESULTS: After 60 days, supra-therapeutic doses of CGV reduced sperm motility (p < 0.05) by 31.8%, 31.3% and 34.5% respectively and increased (p < 0.05) abnormality in sperms by 200%, 241% and 141.3% respectively. CGV altered male (luteinizing, follicle stimulating hormones and testosterone) and female reproductive hormones (luteinizing, follicle stimulating hormones estrogen and progesterone) respectively. Therapeutic doses of CGV elevated reduced glutathione, superoxide dismutase, catalase and glutathione S-transferase, although, this was exceeded by supra-therapeutic doses and more in females than male rats. Supra-therapeutic dose (3000 mg/kg CGV) decreased body weight in both male and female rats by 50% (F(1.5, 30) = 1.2, p = 0.041) and 62.7% (F(2.1, 30) = 0.38, p = 0.038) respectively in treated rats. Supratherapeutic (3000 mg/kg) dose of CGV increased (p < 0.05) creatinine level by 99.1% while serum total protein was reduced (p < 0.05) by 60.1% (2000 mg/kg) and 57.2% (3000 mg/kg) respectively in male animals. In Female rats, supra-therapeutic doses of CGV elevated creatinine levels by 72.2% (1000 mg/kg), 60.2% (2000 mg/kg) and 124.8% (3000 mg/kg) respectively and 3000 mg/kg produces elevated serum low density lipoprotein by 34.6% in treated rats. Serum cholesterol, triglycerides, albumin, alkaline phosphatase were unaltered by CGV dosing. Histology shows seminiferous tubules with reduced spermatogenic cells. Also, female rat kidney revealed acute tubular necrosis at highest dose used in this study. CONCLUSION: Overall, these data suggest that pro-oxidant potential of the supra-therapeutic CGV doses is evident. Hence, it is necessary that its administration be done with caution using appropriate doses.

Morinda lucida reduces contractility of isolated uterine smooth muscle of pregnant and non-pregnant mice.

The present work investigated the effect of Morinda lucida (M. lucida) extract on isolated uterine smooth muscle of pregnant and non-pregnant mice. Pregnant and non-pregnant mice were pretreated with oral stilboesterol (0.1 mg/kg body weight) and killed by cervical dislocation. Thin strips of the uterus were cut and mounted in a 20 ml organ bath containing De Jalon solution bubbled with 95%O2-5% CO2 gas mixture. The strips were connected to a force transducer coupled to a Grass 7D Polygraph for the recording of isometric tension. Effects of graded concentrations of oxytocin (OXY; 10-5-10-2 mol/L), acetylcholine (ACh; 10-9-10-5 mol/L) and M. lucida extract (0.015-1.5 mg/ml) were recorded. Fresh uterine strips were then incubated with M. lucida extract for 5 mins and cumulative response to OXY was repeated. Another set of fresh strips was incubated in L-NAME for 15 mins and the cumulative responses to M.lucida extract were repeated. OXY resulted in increased contractile responses in both pregnant and non-pregnant uterine muscles. M. lucida resulted in relaxation of the uterine smooth muscle in both pregnant and non-pregnant mice at all doses. However, at 1.5mg/ml, M. lucida completely blocked spontaneous uterine contractions. Following incubation with L-NAME, M. lucida extract led to a slightly greater relaxation of the uterine strips. In conclusion, M. lucida reduced contractility of uterine smooth muscle in both pregnant and non-pregnant mice as well as blocking contractile responses to OXY and Ach in uterine smooth muscle of pregnant and non-pregnant mice. There was no significant alteration of M. lucida activity by L-NAME suggesting that the action of the compound on uterine muscle is not associated with impaired nitric oxide synthase.

Comparative efficacy of dihydroartemisinin, chloroquine and combination of dihydroartemisinin and chloroquine or mefloquine in mice infected with Plasmodium berghei

Comparative efficacy of dihydroartemisinin alone, chloroquine alone, combination of dihydroartemisinin plus mefloquine and combination of dihydroartemisinin plus chloroquine was evaluated in mice. Parasite clearance time was very short in mice treated with dihydroartemisinin alone mean ± SD PCT was (1.64- ± 0.50 days). This was followed by combination of dihydroartemisinin with mefloquine (2.73 ± 0.47), Then combination of dihydroartemisinin with chloroquine (2.84 ± 0.50). The mice that were treated with chloroquine alone had PCT of 4.0 ± 2.32. There was significant difference between the dihydroartemisinin group and the chloroquine group (P<0.0002). There was also significant difference between the dihydroartemisinin group and combination of dihydroartemisinin plus mefloquine and also combination of dihydroartemisinin plus chloroquine (P<0.005). The combination therapy was more effective than when chloroquine was administered alone