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BACKGROUND: Hepatitis C virus (HCV) genotyping is important for treatment and epidemiological purposes. The objective of this study was to evaluate the performance of AmpliSens(®) HCV-1/2/3-FRT kit in comparison to sequencing method for genotyping. METHODS: A total of 17 samples collected from December 2009 to January 2011 were analyzed. Reverse transcriptase polymerase chain reaction (PCR) was performed, followed by sequencing technique. Results were analyzed based on sequence information in GenBank. A second genotyping method (AmpliSens(®) HCV-1/2/3-FRT) was done, which differentiates HCV genotypes by means of real-time hybridization-fluorescence detection. RESULTS: From 17 samples, four were untypeable by AmpliSens(®) HCV-1/2/3-FRT. Eleven of 13 (84.6%) results showed concordant genotypes. A specimen that was determined as genotype 3a by sequencing was genotype 1 by the AmpliSens(®) HCV-1/2/3-FRT. Another specimen that was genotype 1 by sequencing was identified as genotype 3 by AmpliSens(®) HCV-1/2/3-FRT. CONCLUSION: HCV genotyping with AmpliSens(®) HCV-1/2/3-FRT using real-time PCR method provides a much simpler and more feasible workflow with shorter time compared to sequencing method. There was good concordance compared to sequencing method. However, more evaluation studies would be required to show statistical significance, and to troubleshoot discordant results. AmpliSens(®) HCV-1/2/3-FRT does differentiate between genotype but not until subtype level.
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Genotyping Techniques , Hepacivirus/genetics , Cross-Sectional Studies , Genotype , Sequence Analysis, RNAABSTRACT
Introduction: The COVID-19 pandemic is a major global public health crisis. More than 2 years into the pandemic, effective therapeutic options remain limited due to rapid viral evolution. Stemming from the emergence of multiple variants, several monoclonal antibodies are no longer suitable for clinical use. This scoping review aimed to summarize the preclinical and clinical evidence for bebtelovimab in treating newly emerging SARS-CoV-2 variants. Methods: We systematically searched five electronic databases (PubMed, CENTRAL, Embase, Global Health, and PsycINFO) from date of inception to September 30, 2022, for studies reporting on the effect of bebtelovimab in SARS-CoV-2 infection, using a combination of search terms around -bebtelovimabâ, -LY-CoV1404â, -LY3853113â, and -coronavirus infectionâ. All citations were screened independently by two researchers. Data were extracted and thematically analyzed based on study design by adhering to the stipulated scoping review approaches. Results: Thirty-nine studies were included, thirty-four non-clinical studies were narratively synthesized, and five clinical studies were meta-analyzed. The non-clinical studies revealed bebtelovimab not only potently neutralized wide-type SARS-CoV-2 and existing variants of concern such as B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma), and B.1.617.2 (Delta), but also retained appreciable activity against Omicron lineages, including BA.2.75, BA.4, BA.4.6, and BA.5. Unlike other monoclonal antibodies, bebtelovimab was able to bind to epitope of the SARS-CoV-2 S protein by exploiting loop mobility or by minimizing side-chain interactions. Pooled analysis from clinical studies depicted that the rates of hospitalization, ICU admission, and death were similar between bebtelovimab and other COVID-19 therapies. Bebtelovimab was associated with a low incidence of treatment-emergent adverse events. Conclusion: Preclinical evidence suggests bebtelovimab be a potential treatment for COVID-19 amidst viral evolution. Bebtelovimab has comparable efficacy to other COVID-19 therapies without evident safety concerns.
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COVID-19 , SARS-CoV-2 , Humans , Pandemics , Antibodies, Monoclonal/adverse effects , Antibodies, Neutralizing/therapeutic useABSTRACT
Background: Bacterial vaginosis (BV) is a common infection in women of reproductive age group because of vaginal dysbiosis. The impact of BV during pregnancy is still not well defined. The objective of this study is to assess the maternal-fetal outcome in women with BV. Materials and Methods: A prospective cohort study over one-year duration was conducted from December, 2014 until December, 2015, involving 237 women who presented with abnormal vaginal discharge, preterm labour or preterm prelabour rupture of membrane between 22- and 34-weeks period of gestation. Vaginal swabs were sent for culture and sensitivity, BV® Blue testing and PCR for Gardnerella vaginalis (GV). Results: BV was diagnosed in 24/237 (10.1%) cases. The median gestational age was 31.6 weeks. GV was isolated from 16 out of 24 (66.7%) in the BV positive group. There was a significantly higher preterm birth rate, below 34 weeks (22.7% vs. 6.2%, p = 0.019) in women with BV. There was no statistically significant difference in maternal outcome such as clinical chorioamnionitis or endometritis. However, placental pathology revealed more than half (55.6%) of women with BV had histologic chorioamnionitis. Neonatal morbidity was significantly higher with exposure to BV, with a lower median birth weight, higher rate of neonatal intensive care unit admission (41.7% vs. 19.0%, p = 0.010), increased intubation for respiratory support (29.2% vs. 7.6%, p = 0.004) and respiratory distress syndrome (33.3% vs. 9.0%, p = 0.002). Conclusion: More research is needed to formulate guidelines for prevention, early detection and treatment of BV during pregnancy to reduce intrauterine inflammation and the associated adverse fetal outcomes.
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Aim: Sanitation and cleanliness are essential factors in reducing the spread of pathogens and preventing healthcare-associated infections. Disinfectants are associated with better hygiene outcomes to reduce pathogen transmission risk and minimize risks to healthcare workers (HCWs) and patients. Methods: A literature search was undertaken using the electronic databases Scopus, Web of Science, Ovid and Google Scholar. The inclusion criteria for this study are observational and original research studies dating from the five-year period 2017-2021. Other inclusion criteria are full text, English language, qualitative or quantitative studies relevant to the research question. The exclusion criteria are animal studies, systematic reviews, conference proceedings, abstracts, projection modelling studies, in-vivo or in-vitro studies, and books. Results: Five study nations included the United States of America (USA), the United Kingdom (UK), China, India and South Korea, together with Malaysia. These nations have existing policies, regulations and guidelines regarding the use of disinfectants. HCWs should be aware of the national laws and guidelines that govern the purchase, distribution and use of disinfectants. They should also understand the different roles of the agencies involved, so the context for the guidance provided is clear. Coordination and collaboration across various stakeholders are required for creating solid policies. Conclusion: Product research and innovation are indispensable, as appropriate personal protective equipment and safety measures for HCWs and patients have top priority in every nation. Hence, clear guidelines for handling disinfectants, in addition to health education about scientific-evidence-based disinfectants, are required.
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Introduction. Non-contact condensing lenses (NCLL) are a requirement in ophthalmology examinations. To date, there have been no studies on the types of bacteria found on handheld lenses used to examine patients in an ophthalmology clinic.Hypothesis. The BD BACTEC Peds Plus broth (BACTEC-PP) culture method can isolate more organisms as compared to conventional culture plates (CCP) from NCLL.Aim. To evaluate the organism spectrum cultured from NCCL for fundus examination and to compare the results between BACTEC-PP and CCP. The isolation results were then related to the participant's knowledge and hand hygiene practices (HHP).Methodology. This is a comparative cross-sectional study involving consenting trainee ophthalmologists from a single centre, whose preferred NCCL was swabbed from January to December 2019. The respondents completed the adapted World Health Organization Hand Hygiene Knowledge and Perception Questionnaire, and their HHP were observed by Infective Control Unit nurses. Positive bacterial growth using both methods, in addition to hand hygiene practices, were compared.Results. All samples had positive yields by at least one method. BACTEC-PP had a higher yield of 47 (90.4%) isolates compared to CCP with 37 (71.2%) isolates, P=0.041. CoNS sp (38.9â%) was the most common isolate with both methods, followed by Bacillus sp (25.3â%). At the same time, three fungi were detected with CCP only (3.2â%). There was a significant correlation in bacterial isolation with years of training, with fewer isolates among seniors in both BACTEC-PP (P=0.049) and CCP (P=0.034). There were no significant correlations between HHP and positive yields from either culture method.Conclusions. NCCL used by trainee ophthalmologists are typically contaminated by at least one bacteria, with CoNS sp being the most commonly isolated. More positive cultures occurred in lenses from junior trainees. Contamination was not correlated with knowledge or HHP. BACTEC-PP has significantly higher yields than CCP for bacterial isolation from NCCL, but did not isolate any fungus.
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Bacteria , Fungi , Bacteriological Techniques/methods , Cross-Sectional Studies , Culture Media , Hospitals , HumansABSTRACT
INTRODUCTION: Gardnerella vaginalis (GV)-associated bacterial vaginosis is recognised for its detrimental effects on pregnancy resulting in poor obstetric and neonatal outcomes. There is limited knowledge of the effects on placental histomorphology following GV infection in pregnancy. We investigated the effects of GV infection on the placenta, particularly with regards to the syncytiotrophoblasts and vascular development, and related these to neonatal outcomes. METHODS: A prospective cohort study involving GV-positive pregnant women presented with abnormal vaginal discharge, with gestational age-matched healthy pregnant women controls. Placental sampling was performed upon delivery and examined histologically. Vascular endothelial growth factor-A (VEGF-A) and hypoxia-inducible factor-1α (HIF-1α) mRNA and protein expression were analysed by real-time PCR and immunohistochemistry respectively. The standard measures in neonatal outcomes were recorded. RESULTS: Placentas from GV-positive mothers were found to have significant histological evidence of maternal and/or fetal inflammatory response compared with the controls (17/28: 60.7% vs 2/20: 10%) (p = 0.0011). There was an increase in the percentage of syncytial nuclear aggregates (SNAs) per villus (47.4 ± 11.09%) in placentas from GV-positive mothers (p < 0.0001). VEGF-A was significantly increased in specifically, the villous endothelial cells of placentas with GV infection, but no difference in the immunoexpression of HIF-1α in these cells between groups. However, these were not associated with adverse neonatal outcomes. DISCUSSION: Increased placental VEGF-A expression associated with increased SNAs in pregnant women with GV infection of the genital tract may be an intrauterine response towards placental vascular remodeling, that may also serve as a protective role in moderating birth outcomes.
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Vaginosis, Bacterial , Vascular Endothelial Growth Factor A , Endothelial Cells/metabolism , Female , Gardnerella vaginalis/metabolism , Humans , Infant, Newborn , Placenta/metabolism , Placentation , Pregnancy , Prospective Studies , Vaginosis, Bacterial/metabolism , Vaginosis, Bacterial/pathology , Vascular Endothelial Growth Factor A/metabolismABSTRACT
The looming severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a long-lasting pandemic of coronavirus disease 2019 (COVID-19) around the globe with substantial morbidity and mortality. N-acetylcysteine, being a nutraceutical precursor of an important antioxidant glutathione, can perform several biological functions in mammals and microbes. It has consequently garnered a growing interest as a potential adjunctive therapy for coronavirus disease. Here, we review evidence concerning the effects of N-acetylcysteine in respiratory viral infections based on currently available in vitro, in vivo, and human clinical investigations. The repurposing of a known drug such as N-acetylcysteine may significantly hasten the deployment of a novel approach for COVID-19. Since the drug candidate has already been translated into the clinic for several decades, its established pharmacological properties and safety and side-effect profiles expedite preclinical and clinical assessment for the treatment of COVID-19. In vitro data have depicted that N-acetylcysteine increases antioxidant capacity, interferes with virus replication, and suppresses expression of pro-inflammatory cytokines in cells infected with influenza viruses or respiratory syncytial virus. Furthermore, findings from in vivo studies have displayed that, by virtue of immune modulation and anti-inflammatory mechanism, N-acetylcysteine reduces the mortality rate in influenza-infected mice animal models. The promising in vitro and in vivo results have prompted the initiation of human subject research for the treatment of COVID-19, including severe pneumonia and acute respiratory distress syndrome. Albeit some evidence of benefits has been observed in clinical outcomes of patients, precision nanoparticle design of N-acetylcysteine may allow for greater therapeutic efficacy.
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OBJECTIVE: Pregnant women with bacterial vaginosis due to Gardnerella vaginalis (GV) infection presents with a wide-ranging disease symptomatology. We speculate this may be due to interaction that varies between host immune response and the pathogen. We studied the oxidative burst in polymorphonuclear leukocytes (PMNL)s from maternal blood (MB) and cord blood (CB) upon phagocytosis of GV and compared against E. coli and Group B Streptococcus (GBS). RESULTS: The PHAGOBURST™ assay detects fluorescence from oxidized dihydrorhodamine during oxidative burst. The average percentage of PMNL showing oxidative burst was almost two-fold greater with GBS (99.5%) and E. coli (98.2%) than GV (56.9%) (p < 0.01) in MB, but a similar proportion of PMNL with burst activity was seen in CB (84.7%). The mean fluorescence intensity (MFI) of oxidative burst in MB PMNL with GV was lower compared to E. coli but comparable to GBS. The MFI of CB PMNL (1580 ± 245.8) was significantly higher than MB PMNL (1198 ± 262.1) with GV, p = 0.031. The live-cell imaging showed neutrophil oxidative burst upon phagocytosis of GV produces hypochlorous acid (HOCl). Overall, the HOCL-mediated microbicidal activity against GV is more variable and less robust than E. coli and GBS, especially in maternal than CB PMNL.
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Gardnerella vaginalis , Neutrophils , Escherichia coli , Female , Fetal Blood , Humans , Phagocytosis , Pregnancy , Respiratory BurstABSTRACT
PURPOSE: The objectives of this study were to determine the prevalence of microbial contamination of multi-user preserved ophthalmic drops (POD) in Ophthalmology Outpatient Clinic (OOC), to compare the rate of contamination between the dropper tip and the residual contents in the bottle, and to identify the contaminating organisms. METHODS: This was an observational cross-sectional study using a convenience sampling method conducted in the OOC of Universiti Kebangsaan Malaysia Medical Center, Malaysia. The samples of POD bottles were divided into groups obtained after 14 days (T14) and after 30 days (T30) of use. The contamination rate at the dropper tip and in the residual contents was determined and the contaminating organisms were identified. RESULTS: A total of 140 of 149 extended-use POD bottles were included. The prevalence of contamination was 30%. There was a statistically significant difference in the rate of contamination between samples T14 and T30 (19% and 11%, respectively; p=0.046). Proparacaine and tropicamide showed higher contamination rates in the T14 samples (p=0.027 and p=0.497, respectively) than in the T30 samples. The site of contamination was higher at the dropper tip than in the residual contents (p>0.05). Coagulase-negative Staphylococcus species were the most frequently identified contaminants (89%). CONCLUSION: The dropper tip was more contaminated than the residual contents, and coagulase-negative Staphylococcus species, which are common commensal flora of the ocular conjunctiva and skin, were the most frequently identified organisms.
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Background: Gardnerella vaginalis (GV) is most frequently associated with bacterial vaginosis and is the second most common etiology causing intrauterine infection after Ureaplasma urealyticum. Intrauterine GV infection adversely affects pregnancy outcomes, resulting in preterm birth, fetal growth restriction, and neonatal pneumonia. The knowledge of how GV exerts its effects is limited. We developed an in vivo animal model to study its effects on fetal development. Materials and Methods: A survival mini-laparotomy was conducted on New Zealand rabbits on gestational day 21 (28 weeks of human pregnancy). In each dam, fetuses in the right uterine horn received intra-amniotic 0.5 × 102 colony-forming units of GV injections each, while their littermate controls in the left horn received sterile saline injections. A second laparotomy was performed seven days later. Assessment of the fetal pups, histopathology of the placenta and histomorphometric examination of the fetal lung tissues was done. Results: Three dams with a combined total of 12 fetuses were exposed to intra-amniotic GV, and 9 fetuses were unexposed. The weights of fetuses, placenta, and fetal lung were significantly lower in the GV group than the saline-inoculated control group [mean gross weight, GV (19.8 ± 3.8 g) vs. control (27.9 ± 1.7 g), p < 0.001; mean placenta weight, GV (5.5 ± 1.0 g) vs. control (6.5 ± 0.7 g), p = 0.027; mean fetal lung weight, GV (0.59 ± 0.11 g) vs. control (0.91 ± 0.08 g), p = 0.002. There was a two-fold increase in the multinucleated syncytiotrophoblasts in the placenta of the GV group than their littermate controls (82.9 ± 14.9 vs. 41.6 ± 13.4, p < 0.001). The mean alveolar septae of GV fetuses was significantly thicker than the control (14.8 ± 2.8 µm vs. 12.4 ± 3.8 µm, p = 0.007). Correspondingly, the proliferative index in the interalveolar septum was 1.8-fold higher in the GV group than controls (24.9 ± 6.6% vs. 14.2 ± 2.9%, p = 0.011). The number of alveoli and alveolar surface area did not vary between groups. Discussion: Low-dose intra-amniotic GV injection induces fetal growth restriction, increased placental multinucleated syncytiotrophoblasts and fetal lung re-modeling characterized by alveolar septal hypertrophy with cellular proliferative changes. Conclusion: This intra-amniotic model could be utilized in future studies to elucidate the acute and chronic effects of GV intrauterine infections.
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Bacterial culture and biochemical testing (CBtest) have been the cornerstone of pathogen identification in the diagnostic microbiology laboratory. With the advent of Sanger sequencing and later, next-generation sequencing, 16S rRNA next-generation sequencing (16SNGS) has been proposed to be a plausible platform for this purpose. Nevertheless, usage of the 16SNGS platform has both advantages and limitations. In addition, transition from the traditional methods of CBtest to 16SNGS requires procurement of costly equipment, timely and sustainable maintenance of these platforms, specific facility infrastructure and technical expertise. All these factors pose a challenge for middle-income countries, more so for countries in the lower middle-income range. In this review, we describe the basis for CBtest and 16SNGS, and discuss the limitations, challenges, advantages and future potential of using 16SNGS for bacterial pathogen identification in diagnostic microbiology laboratories of middle-income countries.