Purpose: This study aimed to evaluate and optimize microbial DNA extraction methods from urine, a non-invasive sample source, to enhance DNA quality, purity, and reliability for urinary microbiome research and biomarker discovery in bladder cancer. Materials and Methods: A total of 302 individuals (258 with genitourinary cancers and 44 with benign urologic diseases) participated in this study. Urine samples were collected via sterile catheterization, resulting in 445 vials for microbial analysis. DNA extraction was performed using three protocols: the standard protocol (SP), water dilution protocol (WDP), and chelation-assisted protocol (CAP). DNA quality (concentration, purity, and contamination levels) was assessed using NanoDrop spectrophotometry.Microbial analysis was conducted on 138 samples (108 cancerous and 30 benign) using 16S rRNA sequencing. Prior to sequencing on the Illumina MiSeq platform, Victor 3 fluorometry was used for validation. Results: WDP outperformed other methods, achieving significantly higher 260/280 and 260/230 ratios, indicating superior DNA purity and reduced contamination, while maintaining reliable DNA yields. CAP was excluded due to poor performance across all metrics. Microbial abundance was significantly higher in WDP-extracted samples (p<0.0001), whereas SP demonstrated higher alpha diversity indices (p<0.01), likely due to improved detection of low-abundance taxa. Beta diversity analysis showed no significant compositional differences between SP and WDP (p=1.0), supporting the reliability of WDP for microbiome research. Conclusions WDP is a highly effective and reliable method for microbial DNA extraction from urine, ensuring high-quality and reproducible results. Future research should address sample variability and crystal precipitation to further refine microbiome-based diagnostics and therapeutics.

INTRODUCTION: We aimed to understand the awareness and attitudes of elderly Southeast Asians towards telehealth services during the coronavirus disease 2019 (COVID-19) pandemic in this study. METHODS: In this qualitative study, 78 individuals from Singapore (51.3% female, mean age 73.0 ± 7.6 years) were interviewed via telephone between 13 May 2020 and 9 June 2020 during Singapore's first COVID-19 'circuit breaker'. Participants were asked to describe their understanding of telehealth, their experience of and willingness to utilise these services, and the barriers and facilitators underlying their decision. Transcripts were analysed using thematic analysis, guided by the United Theory of Acceptance Use of Technology framework. RESULTS: Of the 78 participants, 24 (30.8%) were able to describe the range of telehealth services available and 15 (19.2%) had previously utilised these services. Conversely, 14 (17.9%) participants thought that telehealth comprised solely home medication delivery and 50 (51.3%) participants did not know about telehealth. Despite the advantages offered by telehealth services, participants preferred in-person consultations due to a perceived lack of human interaction and accuracy of diagnoses, poor digital literacy and a lack of access to telehealth-capable devices. CONCLUSION Our results showed poor overall awareness of the range of telehealth services available among elderly Asian individuals, with many harbouring erroneous views regarding their use. These data suggest that public health education campaigns are needed to improve awareness of and correct negative perceptions towards telehealth services in elderly Asians.
Humans ; COVID-19/epidemiology* ; Female ; Telemedicine ; Aged ; Male ; Singapore/epidemiology* ; Qualitative Research ; Health Knowledge, Attitudes, Practice ; SARS-CoV-2 ; Aged, 80 and over ; Middle Aged ; Pandemics ; Awareness ; Asian People ; Southeast Asian People

Purpose: T1 high grade (T1HG) bladder cancer (BC) is a type of non-muscle invasive BC (NMIBC) that is recognized as an aggressive subtype with a heightened propensity for progression. Current risk stratification methods for NMIBC rely on clinicopathological indicators; however, these approaches do not adequately capture the aggressive nature of T1HG BC. Thus, new, more accurate biomarkers for T1HG risk stratification are needed. Here, we enrolled three different patient cohorts and investigated expression of collagen type VI alpha 1 (COL6A1), a key component of the extracellular matrix, at different stages and grades of BC, with a specific focus on T1HG BC. Materials and Methods: Samples from 298 BC patients were subjected to RNA sequencing and real-time polymerase chain reaction. Results: We found that T1HG BC and muscle invasive BC (MIBC) exhibited comparable expression of COL6A1, which was significantly higher than that by other NMIBC subtypes. In particular, T1HG patients who later progressed to MIBC had considerably higher expression of COL6A1 than Ta, T1 low grade patients, and patients that did not progress, highlighting the aggressive nature and higher risk of progression associated with T1HG BC. Moreover, Cox and Kaplan–Meier survival analyses revealed a significant association between elevated expression of COL6A1 and poor progression-free survival of T1HG BC patients (multivariate Cox hazard ratio, 16.812; 95% confidence interval, 3.283–86.095; p=0.001 and p=0.0002 [log-rank test]). Conclusions These findings suggest that COL6A1 may be a promising biomarker for risk stratification of T1HG BC, offering valuable insight into disease prognosis and guidance of personalized treatment decisions.

Acute appendicitis represents one of the most common causes of acute abdomen that may warrant emergency surgery. Necrotizing fasciitis complicated by acute appendicitis is considered a rare complication with life-threatening implications. A 65-year-old man presented with abdominal pain, where a scan, revealed a perforated appendicitis complicated by an anterior abdominal wall collection. He underwent percutaneous drainage, but subsequently developed extensive necrotizing fasciitis requiring extensive debridement and reconstruction. The rapid progression of necrotizing fasciitis calls for early recognition and prompt intervention. The key management principles employed were broad spectrum antibiotics and aggressive surgical debridement. This case demonstrated the use of vacuum-assisted closure dressing and multidisciplinary care in wound healing and coverage. To ensure early diagnosis and intervention for acute appendicitis complicated by necrotizing fasciitis, a high degree of clinical suspicion and awareness of this complication is required.

Objectives: Vibrio parahaemolyticus is a major foodborne pathogen in aquatic animals and a threat to human health worldwide. This study investigated the prevalence, antimicrobial resistance, antimicrobial resistance genes (ARGs), and biofilm formation of V. parahaemolyticus strains isolated from fish mariculture environments in Cat Ba Island, Vietnam. Methods: In total, 150 rearing water samples were collected from 10 fish mariculture farms in winter and summer. A polymerase chain reaction assay was used to identify V. parahaemolyticus, its virulence factors, and ARGs. The antimicrobial resistance patterns and biofilm formation ability of V. parahaemolyticus strains were investigated using the disk diffusion test and a microtiter plate-based crystal violet method, respectively. Results: Thirty-seven V. parahaemolyticus isolates were recovered from 150 samples. The frequencies of the tdh and trh genes among V. parahaemolyticus isolates were 8.1% and 21.6%, respectively. More than 90% of isolates were susceptible to ceftazidime, cefotaxime, and chloramphenicol, but over 72% were resistant to ampicillin, tetracycline, and erythromycin. Furthermore, 67.57% of isolates exhibited multidrug resistance. The presence of ARGs related to gentamicin (aac(3)-IV), tetracycline (tetA) and ciprofloxacin (qnrA) in V. parahaemolyticus isolates was identified. Conversely, no ARGs related to ampicillin or erythromycin resistance were detected. Biofilm formation capacity was detected in significantly more multidrug-resistant isolates (64.9%) than non-multidrug-resistant isolates (18.9%). Conclusion Mariculture environments are a potential source of antibiotic-resistant V.parahaemolyticus and a hotspot for virulence genes and ARGs diffusing to aquatic environments. Thus, the prevention of antibiotic-resistant foodborne vibriosis in aquatic animals and humans requires continuous monitoring.

INTRODUCTION: We investigated the knowledge, attitudes and practice (KAP) towards coronavirus disease 2019 (COVID-19) and its related preventive measures in Singaporeans aged ≥60 years. METHODS: This was a population-based, cross-sectional, mixed-methods study (13 May 2020-9 June 2020) of participants aged ≥ 60 years. Self-reported KAP about ten symptoms and six government-endorsed preventive measures related to COVID-19 were evaluated. Multivariable regression models were used to identify sociodemographic and health-related factors associated with KAP in our sample. Associations between knowledge/attitude scores and practice categories were determined using logistic regression. Seventy-eight participants were interviewed qualitatively about the practice of additional preventive measures and data were analysed thematically. RESULTS: Mean awareness score of COVID-19 symptoms was 7.2/10. The most known symptom was fever (93.0%) and the least known was diarrhoea (33.5%). Most participants knew all six preventive measures (90.4%), perceived them as effective (78.7%) and practised 'wear a mask' (97.2%). Indians, Malays and participants living in smaller housing had poorer mean scores for knowledge of COVID-19 symptoms. Older participants had poorer attitudes towards preventive measures. Compared to Chinese, Indians had lower odds of practising three out of six recommendations. A one-point increase in score for knowledge and attitudes regarding preventive measures resulted in higher odds of always practising three of six and two of six measures, respectively. Qualitative interviews revealed use of other preventive measures, for example, maintaining a healthy lifestyle. CONCLUSIONS Elderly Singaporeans displayed high levels of KAP about COVID-19 and its related preventive measures, with a positive association between levels of knowledge/attitude and practice. However, important ethnic and socioeconomic disparities were evident, indicating that key vulnerabilities remain, which require immediate attention.
Humans ; Aged ; COVID-19/epidemiology* ; SARS-CoV-2 ; Health Knowledge, Attitudes, Practice ; Cross-Sectional Studies ; Singapore/epidemiology* ; Surveys and Questionnaires

Background: One of the most important elements for improving the primary care quality is to meet the growing needs of patients and to increase patient satisfaction. In-depth studies on patients’ expectation is vital for healthcare facilities to achieve a more reliable and valid healthcare outcome. This study aims: to explore people’s expectations towards primary care services and related factors at Family Medicine Clinic, Hue University of Medicine and Pharmacy. Methods: A mixed method study on 308 patients visiting the Family Physician Clinic, Hue University of Medicine and Pharmacy. Results: The most important aspects were: “Doctor provides correct diagnoses and cure it.”, “Doctor provides careful examination”, “Doctor takes full history taking”, “Doctor clearly explains diagnosis and treatment plan”, “List of medication covered by health insurance is enough and qualified”, “Doctor/healthcare worker ensures patient confidentiality” and “Healthcare workers are friendly”. There were differences in the expectations of prioritizing primary care services in different target groups. Younger patients groups placed more expectations on facility factors than older patient groups; Patients in different occupational groups differed in their expectations of healthcare workers and the quality of medical care.Conclusion: Factors related to health care workers and quality of medical care were the most prioritizing expectations at the Family Medicine Clinic. It is necessarry to improve primary care services to meet the expectations of different target groups. 

As part of our continued study on the chemical constituents of Willughbeia edulis stems, a new dimeric lignan named edulignan (1) was isolated from its EtOAc-soluble extract. Based on NMR spectroscopic interpretation, the planar structure of 1 has been suggested to have two 2-substituted 4-chromanone subunits with different stereochemical configurations. In addition, the MS/MS analysis of the products obtained by acidcatalyzed hydrolysis of 1 was supportive of its structure. Unfornatually, the new compound 1 did not show α-glucosidase inhibitory activity with an IC 50 value > 250 μM.

INTRODUCTION: Hospital-based resuscitation interventions, such as therapeutic temperature management (TTM), emergency percutaneous coronary intervention (PCI) and extracorporeal membrane oxygenation (ECMO) can improve outcomes in out-of-hospital cardiac arrest (OHCA). We investigated post-resuscitation interventions and hospital characteristics on OHCA outcomes across public hospitals in Singapore over a 9-year period. METHODS: This was a prospective cohort study of all OHCA cases that presented to 6 hospitals in Singapore from 2010 to 2018. Data were extracted from the Pan-Asian Resuscitation Outcomes Study Clinical Research Network (PAROS CRN) registry. We excluded patients younger than 18 years or were dead on arrival at the emergency department. The outcomes were 30-day survival post-arrest, survival to admission, and neurological outcome. RESULTS: The study analysed 17,735 cases. There was an increasing rate of provision of TTM, emergency PCI and ECMO (P<0.001) in hospitals, and a positive trend of survival outcomes (P<0.001). Relative to hospital F, hospitals B and C had lower provision rates of TTM (≤5.2%). ECMO rate was consistently <1% in all hospitals except hospital F. Hospitals A, B, C, E had <6.5% rates of provision of emergency PCI. Relative to hospital F, OHCA cases from hospitals A, B and C had lower odds of 30-day survival (adjusted odds ratio [aOR]<1; P<0.05 for hospitals A-C) and lower odds of good neurological outcomes (aOR<1; P<0.05 for hospitals A-C). OHCA cases from academic hospitals had higher odds ratio (OR) of 30-day survival (OR 1.3, 95% CI 1.1-1.5) than cases from hospitals without an academic status. CONCLUSION Post-resuscitation interventions for OHCA increased across all hospitals in Singapore from 2010 to 2018, correlating with survival rates. The academic status of hospitals was associated with improved survival.
Hospitals, Public ; Humans ; Out-of-Hospital Cardiac Arrest/therapy* ; Percutaneous Coronary Intervention ; Prospective Studies ; Singapore/epidemiology*

Purpose: Fear of cancer recurrence (FCR) is a common psychological issue in breast cancer (BC) survivors during early survivorship but whether the same is true among long-term survivors has yet to be empirically evaluated. This study investigated FCR level, its associated factors, and impact on quality of life (QoL) in long-term BC survivors. Materials and Methods: Participants included women diagnosed with BC between 2004 and 2010 at two tertiary hospitals. Survey was conducted in 2020. The study measured FCR with the Fear of Cancer Recurrence Inventory and other patient-reported outcomes, including depression and cancer-related QoL. Logistic regression was used to identify factors associated with FCR, and structural equation modeling was conducted to explore the impact of FCR on other outcomes. Results: Of 333 participants, the mean age at diagnosis was 45.5, and 46% experienced FCR. Age at diagnosis ≤ 45 (adjusted odds ratio [aOR], 2.64; 95% confidence interval [CI], 1.51 to 4.60), shorter time since diagnosis (aOR, 1.75, 95% CI, 1.08 to 2.89), and having a history of recurrence (aOR, 2.56; 95% CI, 1.16 to 5.65) was associated with more FCR. FCR was significantly associated with an increased risk of depression (β=0.471, p < 0.001) and negatively impacted emotional functioning (β=–0.531, p < 0.001). In addition, a higher FCR level may impair overall health-related QoL in long-term BC survivors (β=–0.108, p=0.021). Conclusion Ten years after diagnosis, long-term BC survivors still experienced a high level of FCR. Further, the negative impact of FCR on QoL and increased depression risk require an FCR screening and appropriate interventions to enhance long-term BC survivors' QoL.