Stimulation with Ebola VLPs Containing and Lacking RIG-I

Ebola virus (EBOV), a filovirus family member, is a highly pathogenic virus that causes Ebola hemorrhagic fever (EHF) resulting in documented mortality rates in humans as high as 50%. Currently, the basic EBOV virus-like particle (VLP) vaccine contains the Ebola virus (EBOV) matrix VP40 and attachment glycoprotein (GP). VLPs are morphologically and biochemically similar to parental virus, yet because they lack a genome and cannot replicate, are safe enough to be used as vaccines. We hypothesize that addition of a constitutionally active retinoic acid-inducible gene 1 (RIG-I) would enhance the ability of the vaccine to induce interferon-dependent immune functions yielding an improved vaccine. Expression of EBOV VP40 in 293T cells induces the spontaneous production of VLPs into the media supernatant and if expressed with EBOV GP, will produce VLPs studded with the attachment GP. Recombinant chimeric constitutively active (ca)RIG-I-VP40 matrix and a nonfunctional mutant L58A (mu)RIG-I-VP40 matrix genes were constructed to produce VLPs containing constitutively active and nonfunctional RIG-I. Supernatant from 293Ts transfected with caRIG-I-VP40, muRIG-I-VP40 or VP40 along with GP expression plasmids were tested for the presence of VLPs. Western blotting of purified VLPs confirmed the presence of RIG-I in caRIG-I-VP40 and muRIG-I-VP40, but not VP40 containing VLPs. Monocyte-like and PMA-differentiated macrophage-like THP-1 Dual cells were treated with nothing, VP40+GP, caRIG-I-VP40+GP, muRIG-I-VP40+GP VLPs as well as LPS and a Vaccinia virus (VACV-70) positive controls and tested for induction of interferon (IFN) signaling. CaRIG-I containing, but not muRIG-I containing VLPs induced interferon signaling from both macrophages and monocytes. The addition of RIG-I to EBOV VLPs revealed an increase of interferon in human peripheral blood monocytes

Accuracy of bedside index for severity in acute pancreatitis ‘BISAP’ score in predicting outcome of acute pancreatitis

Introduction: Early identification of severe acute pancreatitis is of paramount importance in the management and for improving outcomes. Bedside index for severity in acute pancreatitis (BISAP) is a simple and accurate score for stratification in acute pancreatitis. This study was conducted to find out the accuracy of BISAP score in predicting outcomes of acute pancreatitis in local population. Method: We prospectively analyzed 96 patients with acute pancreatitis from February 2019 to December 2019. Revised Atlanta classification was used to stratify mild, moderately severe and severe pancreatitis. BISAP score was calculated within 24 hours of admission. Accuracy was measured by area under receiver operating curve (AUC). Result: Out of 96 patients, alcohol related acute pancreatitis accounted for 74.7%. There were 63.2% of mild AP, 37.3% of moderately severe AP, 9.4% of severe AP and 15.8 % of pancreatic necrosis. The AUC for moderately severe AP, severe AP and pancreatic necrosis were 0.77 (CI 0.68-0.87), 0.95 (CI 0.90-0.99) and 0.87 (CI 0.79-0.96) respectively. The statistically significant BISAP cut off for diagnosing sever AP was≥3, and ≥2 for moderately sever AP and pancreatic necrosis. There was positive correlation between revised Atlanta severity of acute pancreatitis and length of hospital stay (r=0.41). Mortality was 3.3 % which was seen in BISAP score 3 or above. Conclusion: BISAP is a simple predictive model in identifying patient at a risk of developing different severity of pancreatitis and its outcome in our population

A Global Collaboration to Develop and Pilot Test a Mobile Application to Improve Cancer Pain Management in Nepal

INTRODUCTION: Quality palliative care, which prioritizes comfort and symptom control, can reduce global suffering from non-communicable diseases, such as cancer. To address this need, the Nepalese Association of Palliative Care (NAPCare) created pain management guidelines (PMG) to support healthcare providers in assessing and treating serious pain. The NAPCare PMG are grounded in World Health Organization best practices but adapted for the cultural and resource context of Nepal. Wider adoption of the NAPCare PMG has been limited due to distribution of the guidelines as paper booklets. METHODS: Building on a long-standing partnership between clinicians and researchers in the US and Nepal, the NAPCare PMG mobile application (“app”) was collaboratively designed. Healthcare providers in Nepal were recruited to pilot test the app using patient case studies. Then, participants completed a Qualtrics survey to evaluate the app which included the System Usability Scale (SUS) and selected items from the Mobile App Rating Scale (MARS). Descriptive and summary statistics were calculated and compared across institutions and roles. Regression analyses to explore relationships (α = 0.05) between selected demographic variables and SUS and MARS scores were also conducted. RESULTS: Ninety eight healthcare providers (n = 98) pilot tested the NAPCare PMG app. Overall, across institutions and roles, the app received an SUS score of 76.0 (a score > 68 is considered above average) and a MARS score of 4.10 (on a scale of 1 = poor, 5 = excellent). 89.8% (n = 88) “agreed” or “strongly agreed” that the app will help them better manage cancer pain. Age, years of experience, and training in palliative care were significant in predicting SUS scores (p-values, 0.0124, 0.0371, and 0.0189, respectively); institution was significant in predicting MARS scores (p = 0.0030). CONCLUSION: The NAPCare PMG mobile app was well-received, and participants rated it highly on both the SUS and MARS. Regression analyses suggest end-user variables important to consider in designing and evaluating mobile apps in lower resourced settings. Our app design and pilot testing process illustrate the benefits of cross global collaborations to build research capacity and generate knowledge within the local context

Evaluation of Phenological Traits of Pear Varieties in Warm Temperate Region of Nepal

Phenological study of a fruit tree is important for timely and proper orchard management. This study was carried out in orchard located at Warm Temperate Horticulture Centre (WTHC), Kathmandu, Nepal from January to August, 2020 with the objective to identify the different timing of phenological stages in 18 different varieties of pear. Data on days from swollen bud to bud burst, swollen bud to green cluster, swollen bud to white bud, swollen bud to bloom, swollen bud to petal fall, swollen bud to fruit set, fruit length, and fruit diameter were recorded. The phenological and fruit traits were found highly significant. The shortest flowering period (days from swollen bud to petal fall) was observed in varieties Sinko and Yakumo (13.33 days) and the longest period was found in Pharping local (Green) (35.00 days) followed by Pharping local (Brown) (34.00 days). The longest fruit (58.80 mm) was recorded in Anjou followed by Bartlett (52.90 mm) and Yakumo (49.80 mm). Similarly, the highest fruit diameter (57.60 mm) was recorded in Anjou followed by Yakumo (55.80 mm) and Atago (51.60 mm). On the basis of total variability, pear varieties were grouped into five distinct clusters.  Cluster 2 showed that Anjou had the highest fruit length and fruit diameter. Cluster 5 illustrated that the longest flowering period was observed in the Pharping local (Green) followed by Pharping local (Brown). Based on phenological traits Sinko and Yakumo and based on fruits traits Anjou were found potential varieties for cultivation under warm temperate region of Nepal

Stimulation with Ebola VLPs Containing and Lacking RIG-I

Ebola virus (EBOV), a filovirus family member, is a highly pathogenic virus that causes Ebola hemorrhagic fever (EHF) resulting in documented mortality rates in humans as high as 50%. Currently, the basic EBOV virus-like particle (VLP) vaccine contains the Ebola virus (EBOV) matrix VP40 and attachment glycoprotein (GP). VLPs are morphologically and biochemically similar to parental virus, yet because they lack a genome and cannot replicate, are safe enough to be used as vaccines. We hypothesize that addition of a constitutionally active retinoic acid-inducible gene 1 (RIG-I) would enhance the ability of the vaccine to induce interferon-dependent immune functions yielding an improved vaccine. Expression of EBOV VP40 in 293T cells induces the spontaneous production of VLPs into the media supernatant and if expressed with EBOV GP, will produce VLPs studded with the attachment GP. Recombinant chimeric constitutively active (ca)RIG-I-VP40 matrix and a nonfunctional mutant L58A (mu)RIG-I-VP40 matrix genes were constructed to produce VLPs containing constitutively active and nonfunctional RIG-I. Supernatant from 293Ts transfected with caRIG-I-VP40, muRIG-I-VP40 or VP40 along with GP expression plasmids were tested for the presence of VLPs. Western blotting of purified VLPs confirmed the presence of RIG-I in caRIG-I-VP40 and muRIG-I-VP40, but not VP40 containing VLPs. Monocyte-like and PMA-differentiated macrophage-like THP-1 Dual cells were treated with nothing, VP40+GP, caRIG-I-VP40+GP, muRIG-I-VP40+GP VLPs as well as LPS and a Vaccinia virus (VACV-70) positive controls and tested for induction of interferon (IFN) signaling. CaRIG-I containing, but not muRIG-I containing VLPs induced interferon signaling from both macrophages and monocytes. These results lead us to conclude that supplemented CaRIG-I would be ideal for robust induction of interferon-dependent immune functions, which may improve vaccine efficacy

Association between Body Mass Index and Gastroesophageal Reflux Symptoms in Nepalese Adult Population. A Single Centered Hospital Based Study

Background &amp; Objectives: Gastroesophageal reflux disease (GERD) is a common disorder, and studies have reported inconsistent association between high BMI and its elevated risk of GERD symptoms. The aim of the present study was to estimate the strength of the association between body mass index and reflux symptoms in Nepalese adults.Materials &amp; Methods: This was a cross-sectional study conducted at National Academy of Medical Science, Bir Hospital, Kathmandu, Nepal. All patients of age 18 years or above who underwent endoscopy and diagnosed to have esophagitis were recruited for the analysis. Symptoms were identified by using a self-administered validated questionnaire regarding GERD that identify the onset for GERD symptoms and grade the frequency and severity of symptoms experienced over a period of one year. BMI data, the cut off points were based on the WHO classification of overweight and obesity. A BMI value ?25–30 represents overweight and BMI &gt;30 indicates obesity. A BMI value &lt;25 is considered as normal. Severity of erosive esophagitis was graded according to the Los Angeles classification.Results: The Among 127 enrolled esophagitis subjects, about 43% were under 20 BMI, 43% overweight and about 24% were obese. Obesity measured by BMI revealed no significant association among age, sex, smoker, alcoholics, hernia and severity grade of esophagitis (p&gt;0.05). Ages with various categories (18-40, &gt;40-60,&gt;60) were significantly associated with the GERD symptoms (P=0.021). We categorize BMI in three categories. A BMI value &lt; 25 (normal), ?25–30 (overweight), and BMI &gt;30 (obesity). Categorical analysis of BMI with GERD symptoms confirmed the absence of any tendency towards an association (p&gt;0.05).Conclusion: The tendency of reflux symptoms towards BMI is null, and weight reduction may not be the adequate justifiable for the symptoms therapy.JCMS Nepal. 2015; 11(2):23-26</p

Association between Body Mass Index and Gastroesophageal Reflux Symptoms in Nepalese Adult Population. A Single Centered Hospital Based Study

<p class="Default"><strong>Background &amp; Objectives: </strong>Gastroesophageal reflux disease (GERD) is a common disorder, and studies have reported inconsistent association between high BMI and its elevated risk of GERD symptoms. The aim of the present study was to estimate the strength of the association between body mass index and reflux symptoms in Nepalese adults.</p><p class="Default"><strong>Materials &amp; Methods: </strong>This was a cross-sectional study conducted at National Academy of Medical Science, Bir Hospital, Kathmandu, Nepal. All patients of age 18 years or above who underwent endoscopy and diagnosed to have esophagitis were recruited for the analysis. Symptoms were identified by using a self-administered validated questionnaire regarding GERD that identify the onset for GERD symptoms and grade the frequency and severity of symptoms experienced over a period of one year. BMI data, the cut off points were based on the WHO classification of overweight and obesity. A BMI value ?25–30 represents overweight and BMI &gt;30 indicates obesity. A BMI value &lt;25 is considered as normal. Severity of erosive esophagitis was graded according to the Los Angeles classification.</p><p class="Default"><strong>Results: </strong>The Among 127 enrolled esophagitis subjects, about 43% were under 20 BMI, 43% overweight and about 24% were obese. Obesity measured by BMI revealed no significant association among age, sex, smoker, alcoholics, hernia and severity grade of esophagitis (p&gt;0.05). Ages with various categories (18-40, &gt;40-60,&gt;60) were significantly associated with the GERD symptoms (P=0.021). We categorize BMI in three categories. A BMI value &lt; 25 (normal), ?25–30 (overweight), and BMI &gt;30 (obesity). Categorical analysis of BMI with GERD symptoms confirmed the absence of any tendency towards an association (p&gt;0.05).</p><p class="Default"><strong>Conclusion: </strong>The tendency of reflux symptoms towards BMI is null, and weight reduction may not be the adequate justifiable for the symptoms therapy.</p><p class="Default">JCMS Nepal. 2015; 11(2):23-26</p

Prevalence of Organic Colonic Lesions by Colonoscopy in Patients Fulfilling ROME IV Criteria of Irritable Bowel Syndrome

Introduction: Irritable bowel syndrome occurs as recurrent abdominal pain that is related to defecation and associated with change in frequency and/or form of stool. Few Patients with IBS may have organic lesions detectable at colonoscopy. Methods: A cross sectional study was carried out in 140 consecutive patients of IBS fulfilling the ROME IV criteria. The study was conducted in Gastroenterology unit, Department of Medicine, Bir hospital, Kathmandu from July 2016 to September 2017. All patients underwent full colonoscopy along with biopsy from sigmoid colon and any visibly abnormal areas. Results: The average age of patients was 37.5 years with 76 (52.8%) males. Forty-two (30%) had IBS-D, 36 (26%) had IBS-C, 31 (22%) had IBS-M and 31 (22%) had IBS-U. Dyspepsia was seen in 16 (11.4%) and fear of TB/Malignancy/IBD was seen in 27 (19.2%). Organic lesions were seen in 39 (27.85%) patients. Nonspecific colitis was seen in 10 (7.1%) followed by ileal erosions in 7 (5%), polyps in 8 (5.7%), hemorrhoids in 6 (4.2%) and diverticula in 3 (2.1%). Only one (0.71%) patient had microscopic colitis and one (0.71%) had malignant lesion seen at histopathological examination. Females with IBS-D had more organic findings than males (P=0.03, RR=4.13, 95% CI=1.21-15.71). Conclusions: The prevalence of organic lesions in patients with IBS fulfilling ROME IV criteria is 27%. Dyspepsia is the most common comorbidity and fear of TB/malignancy/IBD is the most common reason for seeking health care. Females with IBS-D have a higher risk of detecting organic lesions by colonoscopy and histopathology examination