RESULTS: In this study, ten clinical isolates were obtained from corneal scrapings. Rns genotype and intra-genotypic variation at the DF3 region of the isolates were identified. Results revealed that all clinical isolates belonged to the T4 genotype, with T4/6 (4 isolates), T4/2 (3 isolates), T4/16 (2 isolates) and one new genotype T4 sequence (T4/36), being determined. The axenic clinical isolates were cytopathogenic to rabbit corneal fibroblasts. MBP and AhLBP mRNA expression are directly correlated to Acanthamoeba cytopathic effect.
CONCLUSIONS: All ten Malaysian clinical isolates were identified as genotype T4 which is predominantly associated with AK. Measuring the mRNA expression of Acanthamoeba virulent markers could be useful in the understanding of the pathogenesis of Acanthamoeba keratitis.
MATERIALS AND METHODS: Murine MC3T3-E1 preosteoblastic cells were cultured in the different concentrations of AnTT (0.001-1 µg/mL) up to 24 days. Expression of osteoblastic differentiation markers was measured by qPCR (osterix [OSX], collagen 1 alpha 1 [COL1α1], alkaline phosphatase [ALP], and osteocalcin [OCN]) and by fluorometric assay for ALP activity. Detection of collagen and mineralized nodules was done via Direct Red staining and Alizarin Red staining, respectively.
RESULTS: The results showed that osteoblastic differentiation-related genes, such as OSX, COL1α1, ALP, and OCN, were significantly increased in the AnTT-treated groups compared to the vehicle group in a time-dependent manner (P<0.05). Type 1 collagen level was increased from day 3 to day 15 in the AnTT-treated groups, while ALP activity was increased from day 9 to day 21 in the AnTT-treated groups (P<0.05). Enhanced mineralization was observed in the AnTT-treated groups via increasing Alizarin Red staining from day 3 to day 21 (P<0.05).
CONCLUSION: Our results suggest that AnTT enhances the osteogenic activity by promoting the bone formation-related genes and proteins in a temporal and sequential manner.
METHODOLOGY: A retrospective cross-sectional study was employed to identify patients with positive AR bacteria between March 2019 and March 2022. The bacterial isolates and patients' data were identified from laboratory and medical records departments retrospectively. Binary logistic regression analysis was performed to identify the factors associated with AR and deaths. Multinominal logistic regression was applied to confirm the factors associated with AR classification.
RESULTS: AR Gram-negative bacteria decreased during and after the pandemic. However, S. aureus showed a negligible increase in resistance rate after pandemic, while E. faecium, recorded a higher-than-average resistance rate during the pandemic. The prevalence of pan drug resistance (PDR) during the pandemic (85.7%) was higher than before (0%) and after (14.3%), p = 0.001. The length of stay and time were significant predictors for AR classification. The odds of multi drug resistance (MDR) development to PDR during the pandemic were 6 times higher than before and after (OR = 6.133, CI =, p = 0.020). Age, nationality, COVID-19 infection, smoking, liver disease, and type and number of bacteria were associated with death of patients with positive AR.
CONCLUSIONS: Further studies are recommended to explore the prevalence of PDR and to justify the increased rates of E. faecium AR during the COVID-19 pandemic.