OBJECTIVES: To establish a simple, efficient, and optimized method to produce a G6PDViangchan variant and characterize the phenotypes of recombinant human wild-type G6PD and G6PDViangchan.
METHODS: G6PD was amplified by polymerase chain reaction (PCR) from a human cDNA plasmid, and the gene for G6PDViangchan was amplified by initiating a mutation at location 871 (G>A) through site-directed mutagenesis. Protein expression and western blotting were conducted after successful cloning. The enzymatic activity of both proteins was assessed spectrophotometrically after purification.
RESULTS: Both amplicons were successfully cloned into a pET26b(+) expression vector and transformed into Escherichia coli BL21 (DE3) cells for overexpression as C-terminally histidine-tagged recombinant proteins. Western blotting confirmed that both proteins were successfully produced at similar levels. The enzymes were purified by immobilized metal (Co) affinity chromatography. Postpurification assay of enzyme activity revealed about 2-fold differences in the levels of specific activity between the wild-type G6PD (155.88 U/mg) and G6PDViangchan (81.85 U/mg), which is consistent with earlier reports. Analysis in silico showed that the coding change in G6PDViangchan has a substantial effect on protein folding structure.
CONCLUSIONS: We successfully cloned, expressed, and purified both wild-type G6PD and G6PDViangchan proteins. Such a protocol may be useful for creating a model system to study G6PD deficiency disease.
METHODS: This is a single-center, cross-sectional study using in-patient data maintained by the Case-Mix Unit of a teaching hospital in Malaysia from 2016 to 2017. The study included all patients with International Classification of Disease (ICD) code 164 (stroke, not specified as hemorrhage or infarct). The significance of association was determined using nonparametric tests in the form of the Mann-Whitney U test and the Kruskal-Wallis test.
RESULTS: A total of 162 stroke patients from 2016 to 2017 from Case-Mix database were included in the study. The age ranged from 31 to 97 years old. The minimum and maximum LOS for patients with stroke ranged from 1 to 17 days. The severity of illness was found to be significantly associated with longer LOS (p < 0.001); however, age, sex, and presence of co-morbidities did not show any significant association.
CONCLUSION: Despite its limitations, this study is an essential first step to examine the characteristics of patients with stroke and to determine the factors associated with LOS.
Lay summary: Although male infertility affects 1:15 men, there is no obvious reason in the vast majority of cases. Reactive oxidative species (ROS) are highly active molecules containing oxygen and are natural byproducts of normal metabolism. However, high concentrations of ROS have been shown to damage sperm, which negatively impacts a couple's ability to conceive. Carnitines are natural antioxidants found in the body that counterbalance the damaging effects of ROS. We conducted a comprehensive review of published studies to assess whether carnitine supplements are safe and effective in improving sperm quality and pregnancy rates. Our analysis shows that carnitines improve sperm swimming and production of normal-shaped sperm cells but do not affect sperm count or pregnancy rates, although there are only a few studies and scientific evidence is limited. Whilst it is possible that carnitines may benefit male infertility, more evidence is required regarding chances of pregnancy after carnitine therapy.
METHODS: PubMed, Scopus, and Web of Science databases were searched using the keywords "EBV or Epstein Barr virus and Oral cancer or Oral squamous cell carcinoma" for published case-control studies in the English language upto August 2019.
RESULTS: The search yielded 985 articles out of which 966 articles were excluded by screening their titles and abstracts as they were irrelevant or duplicates. Based on the full-text assessment of the remaining 19 articles, only 7 satisfied the inclusion criteria and were included in the qualitative analysis, out of which only 4 were compatible to be included in the meta-analysis. The diagnostic modalities used included immunohistochemistry, in situ hybridization and polymerase chain reaction. The diagnostic targets included latent membrane protein (LMP)-1, EBV determined nuclear antigen-1, EBV-encoded small non-polyadenylated RNA-2. The meta-analysis showed that there is an association between the EBV and OSCC.
CONCLUSIONS: Determining the association of EBV with OSCC is highly tedious due to the contrasting data obtained from individuals' studies which in turn is due to the wide variations in the sensitivity and specificity of the diagnostic modalities used and diagnostic targets selected. Although the meta-analysis revealed an association between EBV and OSCC, the number and the quality of the studies included in the meta-analysis are limited, thus the association requires further validation for any conclusive inference.
Methods: We performed a systematic review and meta-analysis to compare the effects of PMRT to no PMRT for elderly patients with intermediate-risk breast cancer. We searched PubMed for eligible studies from Jan 2008 to Dec 2018. We assessed the methodological quality of the included studies using the ROBINS-I tool and performed the meta-analysis with random effects model. The primary outcome of interest was overall survival (OS); secondary outcomes were breast cancer specific survival (BCSS), loco-regional (LRR) and distant disease recurrence (DDR).
Results: We found 2 retrospective cohort studies with 743 patients directly comparing PMRT to no PMRT. Both studies were judged to have serious risk of bias in their methodological quality. The pooled results suggest that PMRT was associated with a 20% relative reduction in the hazard in death, ranging from 41% relative reduction, a substantial negative association to 10% relative increase, a small positive association (HR 0.80, 95% CI: 0.59-1.1, P=0.62, I2=0%). PMRT was also associated with a 17% relative reduction in the hazard for breast cancer related death, ranging from 52% relative reduction, a substantial negative association to 41% relative increase, a substantial positive association (HR 0.83, 95% CI: 0.48-1.41, P=0.48, I2=0%). One study did not observe any significant differences in LRR and DDR between the two groups.
Conclusions: The survival benefits from PMRT in unselected elderly patients with intermediate risk breast cancer is unclear. Further research to better select elderly patients who may benefit from PMRT is warranted. Patients with a multiple pathological risk factors suggestive of high risk of loco-regional recurrence post-mastectomy should consider PMRT.