Detection of Biofilm Production and Antibiotic Susceptibility Pattern among Clinically Isolated Staphylococcus aureus.

Aim: The increasing antibiotic resistance and the ability to form biofilms in medical devices have become the leading cause of severe infections associated with Staphylococcus aureus (S. aureus). Since the bacteria living in biofilms can exhibit 10- to 1,000-fold increase in antibiotic resistance and implicate chronic infectious diseases, the detection of S. aureus ability to form biofilms is of great importance for managing, minimizing, and effectively treating infections caused by it. This study aimed to compare the tube and tissue culture methods to detect biofilm production and antibiotic susceptibility in MRSA and MSSA. Materials and Methods: The S. aureus isolates were identified by the examination of the colony morphology, Gram staining, and various biochemical tests. Antimicrobial susceptibility testing of all isolates was performed by the modified Kirby-Bauer disc diffusion method as recommended by CLSI guidelines. MRSA screening was performed phenotypically using a cefoxitin disc (30 µg). Isolates were tested for inducible resistance using the D-test, and two phenotypic methods detected biofilm formation. Results: Among 982 nonrepeated clinical specimens, S. aureus was isolated from 103 (10.48%). Among 103 clinical isolates of S. aureus, 54 (52.42%) isolates were MRSA, and 49 (47.57%) were MSSA. Among 54 MRSA isolates, the inducible MLSB phenotype was observed in 23/54 (42.59%) with a positive D-test. By TCP method, 26 (48.1%) MRSA isolates were strong biofilm producers, whereas, among all MSSA isolates, only 6 (12.2%) were strong biofilm producers. Conclusion: MRSA showed strong biofilm production in comparison with MSSA. The TCP method is a recommended reliable method to detect the biofilm among S. aureus isolates, and the TM method could be useful for the screening of biofilm production in S. aureus in the routine clinical laboratory.

Giant endometrial polyp in a post-menopausal woman: a rare case report from Nepal.

Introduction: Endometrial polyps (EPs) result from the overgrowth of endometrial glands and stroma. Giant endometrial polyps, defined as those exceeding 4 cm, are rare, and their association with phytoestrogen (PE) intake is infrequently reported. Case presentation: The authors present a case of a giant endometrial polyp in a 59-year-old post-menopausal woman from Nepal. The patient presented with lower abdominal pain and a history of vaginal spotting. She was not under any drugs or medications, including hormones, but had a regular intake of PE-rich foods. Imaging revealed a giant endometrial polyp and a uterine fibroid. Total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH-BSO) were performed and histopathology examination confirmed the diagnosis of endometrial polyp and fibroid. Discussion: In our case, the patient's increased age and PE-rich diet were identified as potential risk factors for the giant endometrial polyp. Giant endometrial polyps are rare, with limited cases reported to date, often associated with tamoxifen or raloxifene use. Phytoestrogens can exhibit oestrogenic effects, contributing to endometrial polyps. This case emphasizes the importance of further research to explain the relationship between phytoestrogen intake and giant endometrial polyps. Conclusion: Giant endometrial polyps are uncommon, and their association with phytoestrogen intake remains underexplored. Clinicians should consider dietary factors in history while evaluating endometrial polyps, and further research is necessary to explore the potential role of phytoestrogens in the development of giant endometrial polyps.

Acute oral poisoning due to pretilachlor herbicide - a rare case report from Nepal.

Introduction: Acute oral intoxication of pretilachlor, a synthetic chloroacetanilide herbicide, can present similar clinical manifestations of organophosphorus toxicity in humans. Case presentation: A 15-year-old male was admitted after suicidal ingestion of pretilachlor poison, with decreased consciousness and blood-mixed vomiting. Discussion: Pretilachlor is a colorless and odorless liquid that can cause neurotoxicity and carcinogenicity due to its prolonged exposure. The effects of acute oral exposure are mild and may differ from chronic exposure. Individuals exposed to chloroacetanilides may not show symptoms or experience vomiting and neurological issues. Clinical manifestations such as vomiting, excessive lacrimation, bowel and bladder incontinence, bradycardia, and hypotension can be observed in both organophosphate poisoning and pretilachlor poisoning, making accurate diagnosis challenging, particularly in resource-limited settings like ours. There is no specific antidote for pretilachlor poisoning. Treatment focuses on symptomatic care and monitoring the patient's hemodynamics as per standard protocol. Conclusion: This case underscores the need for prompt stabilization, vigilant monitoring, and supportive care to ensure timely recovery in pretilachlor poisoning cases despite similarities with organophosphate poisoning. It emphasizes the importance of educating and raising awareness among physicians about potential mimickers like organophosphates.

Primary omental ectopic pregnancy: A rare case report.

Primary abdominal pregnancy is the rarest form of ectopic pregnancy in which the fertilized ovum is directly implanted into peritoneal cavity. This condition poses significantly high risk of maternal morbidity and mortality. Here, we present a case of 5 weeks of primary omental pregnancy managed by laparotomy.

Association of Hematological and Biochemical Parameters with Serological Markers of Acute Dengue Infection during the 2022 Dengue Outbreak in Nepal.

Background: Nepal faced a major dengue outbreak in 2022. The majority of hospitals and laboratories had limited resources for dengue confirmation and had to rely on rapid dengue diagnostic tests. The purpose of the study is to find the predictive hematological and biochemical parameters in each serological phase of dengue infection (NS1 and IgM) that may assist in dengue diagnosis, severity assessment, and patient management via the use of rapid serological tests. Method: A laboratory-based cross-sectional study was conducted among dengue patients. Rapid antigen (NS1) and serological test (IgM/IgG) was performed to diagnose positive dengue cases. Furthermore, hematological and biochemical investigations were carried out and compared between NS1 and/or IgM-positive participants. A logistic regression analysis was used to identify the validity of the hematological and biochemical characteristics for dengue diagnosis as well as patient management. Receiver-operating characteristic (ROC) curve analysis was used to define the best cut-off, sensitivity, and specificity. Result: Multiple logistic regression showed thrombocytopenia (ORA = 1.000; p = 0.006), leukopenia (ORA = 0.999; p < 0.001), glucose level (ORA = 1.028; p = 0.029), aspartate aminotransferase (ORA = 1.131; p = 0.001), and monocytosis (ORA = 2.332; p = 0.020) as significant parameters in the NS1-only positive group. Similarly, thrombocytopenia (ORA = 1.000; p = 0.001), glucose level (ORA = 1.037; p = 0.004), and aspartate aminotransferase (ORA = 1.141; p < 0.001) were significant in IgM-only positive patients. Moreover, thrombocytopenia (ORA = 1.000; p < 0.001), leukopenia (ORA = 0.999; p < 0.001), glucose (ORA = 1.031; p = 0.017), aspartate aminotransferase (ORA = 1.136; p < 0.001), and lymphopenia (ORA = 0.520; p = 0.067) were independent predictors in both NS1 + IgM positive groups. Platelets consistently demonstrated a higher area under the curve with increased sensitivity and specificity throughout all models, while aspartate aminotransferase (AUC = 0.811) and glucose (AUC = 0.712) demonstrated better results when single IgM positivity was observed. The total leukocyte count performed better when both NS1 + IgM were positive (AUC = 0.814). Conclusion: Hence, thrombocytopenia, elevated AST, high glucose level, leukopenia with monocytosis, and leukopenia with lymphopenia may predict dengue diagnosis and its severity during an active infection. Therefore, these laboratory parameters can be used to complement less sensitive rapid tests, improve dengue diagnosis, and help with proper patient management.

Anti-SARS-CoV-2 Antibody Screening in Healthcare Workers and Its Correlation with Clinical Presentation in Tertiary Care Hospital, Kathmandu, Nepal, from November 2020 to January 2021.

BACKGROUND: Antibody titer and the life span of antibodies against SARS-CoV-2 have been found to be associated with the clinical presentation in individuals. The extent of exposure of healthcare workers and the general public to SARS-CoV-2 needs to be assessed to monitor the COVID-19 pandemic. Thus, this study is an attempt in assessing the anti-SARS-CoV-2 antibody in health care workers. METHODS: This laboratory-based cross-sectional study was performed in Manmohan Memorial Medical College and Teaching Hospital, Kathmandu from November 2020 to January 2021. A total of 185 HCWs were enrolled in this study. Their serum samples were screened for anti-SARS-CoV-2 antibodies, and a structured questionnaire was administered to collect further information. Anti-SARS-CoV-2 antibody screening was performed using lateral flow immunoassay. The data were analyzed using SPSS version 20. RESULTS: Among 185 HCWs that participated in the study, 41 (22.2%) tested positive for the anti-SARS-CoV-2 antibody. Of these 41 HCWs, 37 tested positive for IgG only and 4 of them tested positive for both IgM and IgG antibodies. The presence of the previous history of SARS-CoV-2 infection (p < 0.001), the presence of flu-like symptoms within the last 6 months (p < 0.001), and the presence of positive contact history (p=0.002) were statistically significant with the presence of the antibody among HCWs. CONCLUSION: Healthcare workers carry a high burden of SARS-CoV-2 infection and are at risk of acquiring infection from their workplace. Anti-SARS-CoV-2 antibody screening among healthcare workers is highly recommended in multiple healthcare settings as it can help in monitoring transmission dynamics and evaluation of infection control policies.

STAT3-mediated autophagy dependence identifies subtypes of breast cancer where autophagy inhibition can be efficacious.

Autophagy is a protein and organelle degradation pathway that is involved in diverse diseases, including cancer. Recent evidence suggests that autophagy is a cell survival mechanism in tumor cells and that its inhibition, especially in combination with other therapy, could be beneficial but it remains unclear if all cancer cells behave the same way when autophagy is inhibited. We inhibited autophagy in a panel of breast cancer cell lines and found that some of them are dependent on autophagy for survival even in nutrient rich conditions without any additional stress, whereas others need autophagy only when stressed. Survival under unstressed conditions is due to cell type-specific autophagy regulation of STAT3 activity and this phenotype is enriched in triple-negative cell lines. This autophagy-dependency affects response to therapy because autophagy inhibition reduced tumor growth in vivo in autophagy-dependent but not in autophagy-independent breast tumors, whereas combination treatment with autophagy inhibitors and other agent was preferentially synergistic in autophagy-dependent cells. These results imply that autophagy-dependence represents a tumor cell-specific characteristic where autophagy inhibition will be more effective. Moreover, our results suggest that autophagy inhibition might be a potential therapeutic strategy for triple-negative breast cancers, which currently lack an effective targeted treatment.

Chloroquine sensitizes breast cancer cells to chemotherapy independent of autophagy.

Chloroquine (CQ) is a 4-aminoquinoline drug used for the treatment of diverse diseases. It inhibits lysosomal acidification and therefore prevents autophagy by blocking autophagosome fusion and degradation. In cancer treatment, CQ is often used in combination with chemotherapeutic drugs and radiation because it has been shown to enhance the efficacy of tumor cell killing. Since CQ and its derivatives are the only inhibitors of autophagy that are available for use in the clinic, multiple ongoing clinical trials are currently using CQ or hydroxychloroquine (HCQ) for this purpose, either alone, or in combination with other anticancer drugs. Here we show that in the mouse breast cancer cell lines, 67NR and 4T1, autophagy is induced by the DNA damaging agent cisplatin or by drugs that selectively target autophagy regulation, the PtdIns3K inhibitor LY294002, and the mTOR inhibitor rapamycin. In combination with these drugs, CQ sensitized to these treatments, though this effect was more evident with LY294002 and rapamycin treatment. Surprisingly, however, in these experiments CQ sensitization occurred independent of autophagy inhibition, since sensitization was not mimicked by Atg12, Beclin 1 knockdown or bafilomycin treatment, and occurred even in the absence of Atg12. We therefore propose that although CQ might be helpful in combination with cancer therapeutic drugs, its sensitizing effects can occur independently of autophagy inhibition. Consequently, this possibility should be considered in the ongoing clinical trials where CQ or HCQ are used in the treatment of cancer, and caution is warranted when CQ treatment is used in cytotoxic assays in autophagy research.