Protocol for isolation of signet ring cells from human gastric mucosa.

More than 90% of individuals with germline pathogenic CDH1 variants will harbor occult, microscopic foci of signet ring cell carcinomas capable of progressing to advanced diffuse-type gastric cancer. Here, we present a protocol for high viability suspension of signet ring cells from human gastric tissue. We describe the steps for gastric mucosa isolation and tissue dissociation. We then detail procedures for embedding cells into HistoGel for immunohistochemistry staining and additional applications such as flow cytometry and single-cell sequencing.

Study of diagnostic utility of Xpert MTB/Rif test on pleural fluid in the evaluation of patients presenting with Pleural Tuberculosis in Nepal.

Introduction: Existing tests for the diagnosis of pleural tuberculosis (TB) have major limitations in terms of accuracy, time to diagnosis and drug resistance testing. A test which can diagnose pleural TB and detect resistance, like Xpert MTB/Rif, would be optimal for rapid diagnosis and treatment. Methods: A prospective observational study was done in a tertiary care hospital in Eastern Nepal. Fifty-one patients with clinic-radiologic suspicion of pleural TB were included. The results of pleural fluid Xpert MTB/Rif were compared with two Composite Reference Standards. Composite Reference Standard-1 consisted of positive pleural fluid smear, positive culture, positive histology of pleural biopsy, and positive sputum results. Composite Reference Standard-2 included those with Composite Reference Standard-1 and those with high ADA values (>40 U/l) with response to anti-tubercular treatment at 8 weeks of follow-up. Results: Thirty-six patients were diagnosed as Pleural TB. Nine fulfilled Composite Reference Standard-1. Pleural fluid Xpert MTB/Rif was positive in five cases with Composite Reference Standard-1 and nine cases with Composite Reference Standard-2. The sensitivity, specificity, positive predictive value and negative predictive value with reference to Composite Reference Standard-1 were 55.56%, 88.10%, 50%, and 90.24%, respectively. Using Composite Reference Standard-2 as reference, sensitivity, specificity, positive predictive value and negative predictive value were 25%, 93.33%, 90%, and 34.15%, respectively. Two cases were diagnosed Xpert Rif resistant on pleural fluid. Conclusion: Due to low sensitivity, the Xpert MTB/Rif test cannot be recommended as initial test of diagnosis in a high prevalence setting. At the same time its clinical utility lies in testing of patients suspected to have drug-resistant pleural tuberculosis.

Success of Using Iodopovidone as a Sclerosing Agent for Chemical Pleurodesis in a Tertiary Care Center: A Descriptive Cross-Sectional Study.

INTRODUCTION: Pleurodesis is a procedure to achieve symphysis between the two layers of pleura aimed at preventing the accumulation of either air or fluid in the pleural space. In Nepal, intrapleural instillation of the chemical sclerosing agent is more commonly done as  thoracoscopy facility is not easily available. However, iodopovidone is rarely used for this purpose in Nepal. The study aims to find the prevalence of success using iodopovidone as the chemical sclerosing agent. METHODS: The study included cases undergoing pleurodesis over a two-year period. The clinicodemographic data, diagnosis, treatment effect and treatment response were analyzed. The treatment response was graded as Treatment Success (Complete Response or Partial Response) and Treatment Failure. RESULTS: Pleurodesis was done in a total of 54 cases. Of those, 39 cases were Secondary Spontaneous Pneumothorax, 11 were Malignant Pleural Effusion, 3 were Primary Spontaneous Pneumothorax, and 1 was a case of Hepatic Hydrothorax. Among Secondary Spontaneous Pneumothorax, Pleurodesis was successful in 37 (95%) out of 39 cases, with 35 (90%) having a Complete Response and 2 (5%) having a Partial Response while 2 (5%) had Treatment failure. Among Malignant Pleural Effusion, treatment success was achieved in 6 (55%) out of 11, whereas 5 (45%) failed the treatment. The commonest complication was burning sensation, and the commonest pain scale was "distressing." CONCLUSIONS: This study highlights the safety and ease of use of iodopovidone as an agent for chemical pleurodesis. It confirms the high rate of success of pleurodesis in cases of pneumothorax as found in other studies. In contrast, the success rate is understandably lower in cases of Malignant Pleural effusion.

Bronchoscopic visualization of a cavity in entirety: an unusual finding.

It is unusual to be able to visualize an entire cavity with such clarity.

Study of microbiological and antibiotic sensitivity pattern of ventilator associated pneumonia (VAP) in ICU of a tertiary care hospital in Nepal.

INTRODUCTION: Ventilator-associated pneumonia (VAP) is the most frequent intensive care unit (ICU) acquired infection among patients receiving mechanical ventilation. Accurate clinical and microbiologic diagnosis of VAP is essential not only for selection of appropriate antimicrobials but also to prevent their misuse. As the organisms and their sensitivity pattern may differ in every ICU, the knowledge of the resident flora and their behaviour should be known for successful treatment. METHODS: The study was conducted to evaluate the organisms responsible for VAP and their Antibiotic Sensitivity Pattern for the study setting. A prospective, open, epidemiological clinical study was performed in a tertiary care hospital in Nepal. 100 patients admitted to ICU and Mechanically Ventilated were evaluated about VAP. Clinical Pulmonary Infection Score (CPIS) was used to diagnose VAP. RESULTS: Among 60 patients ventilated for more than 48 hours, 25 (41.6%) developed VAP. The VAP was caused predominantly by Klebsiella pneumonia in 34.5% of cases, followed by Acinetobacter calcoaceticus baumanni in 27.6%, Acinetobacter wolffi and Pseudomonas aeruginosa in 13.8% each and Escheresia coli in 10.3%. The most sensitive antibiotics were Colistin, followed by Polymyxin B and Amikacin with sensitivity rates of 67%, 60% and 58%, respectively. CONCLUSION: Based on these results, an empiric approach to antibiotic treatment can be made tailored to the specific settings. Given the magnitude of drug resistance and its implicated financial and societal burden, there is an urgent need for broad implementation of Antibiotic Stewardship programs across all health care settings.

Prevalence of Iron Deficiency Anemia in Biochemically Defined Moderate to Severely Anemic patients in a Tertiary Care Centre.

INTRODUCTION: Anemia due to iron deficiency and chronic diseases is common occurrence in developing country like Nepal, the latter seen in patients with various inflammatory, autoimmune, and malignant disorders . The Intensive method of marrow iron examination, which this study has employed, provides clinically useful iron status classification in cases of functional iron deficiency. The aim of the study is to find out the prevalence of iron deficiency anemia in biochemically defined moderate to severe anemic patients in tertiary care center. METHODS: A descriptive cross-sectional study was done in 43 patients who underwent bone marrow aspiration for evaluation of any cause and had moderate to severe anemia at the same time over a period of one year from Nov 2015 to 2016. Ethical clearance was obtained from Institutional Review Committee. The bone marrow iron stores were assessed by"intensive method" apart from the routinely used Gale's method. Data was collected and entry were done in Statistical Package for Social Sciences version 24. Point estimate at 95% Confidence Interval was calculated along with frequency and proportion for binary data. RESULTS: The intensive grading system demonstrated normal marrow iron store in 13 (30.2%), depleted iron stores in 3 (7%), functional iron deficiency in 14 (32.6%), and combined deficiency in 13 (30.2%) patients. Mean log ferritin concentration was lower in patients with depleted iron stores (2.2µg/l) than in those with normal (2.7µg/l), and functional iron deficiency (2.4µg/l). The mean log ferritin in combined deficiency was lower than the mean log ferritin concentration in iron store deficiency (1.9µg/l). CONCLUSIONS: The prevalence of functional iron deficiency anemia was greatest when the intensive method for assessment of bone marrow iron was used, thus differentiating four different iron status categories, including functional iron deficiency, from actual iron store deficiency, avoiding unnecessary iron supplementation in the former group.

Incidence and Outcome of Ventilator Associated Pneumonia in ICU of a Tertiary Care Hospital in Nepal.

INTRODUCTION: Ventilator associated pneumonia is an important intensive care unit acquired infection in mechanically ventilated patients. Early and correct diagnosis of Ventilator associated pneumonia is difficult but is an urgent challenge for an optimal antibiotic treatment. METHODS: A prospective observational study was conducted in Intensive Care Unit of a tertiary care hospital in Nepal. Consecutive patients were considered during the study period, who met the criteria were included for the study. Clinical Pulmonary Infection Score was used to diagnose Ventilator associated pneumonia. RESULTS: Among 60 patients ventilated for more than 48 hours, 25 (41.6%) developed ventilator associated pneumonia. The incidence was 25 VAPs per 100 ventilated patients or 26 VAPs per 1000 ventilator days during the period of study. Days on ventilator and duration in ICU were higher in the VAP group. There was a trend towards increasing mortality in the VAP group (P value=0.06). CONCLUSIONS: There exists a high rate of VAP in our Intensive Care Unit. Targeted strategies aimed at reducing Ventilator associated pneumonia should be implemented to improve patient outcome and reduce length of Intensive Care Unit stay and costs.