Decreased Level of Vitamin D is Associated with Rheumatoid Arthritis Patients from Western Region of Nepal

Background: Rheumatoid Arthritis (RA) is a progressive, chronic type of autoimmune disease and the role of vitamin D in the pathogenesis of RA is under investigation. Objective: The objective of this study was to determine the vitamin D deficiency in patients with RA as compared to healthy controls and to assess the relationship between serum vitamin D and anti-cyclic citrullinated peptide (anti-CCP) antibody levels in patients with newly diagnosed rheumatoid arthritis. Methods: The study was conducted between January 2017 to February 2018 at Fishtail Hospital and Research centre. A total of 63 patients with early RA diagnosed and a control group of 56 healthy participants, not on vitamin D supplements were recruited from Department of Internal Medicine. Five ml of blood samples were drawn from cubital veins. Blood glucose, creatinine, uric acid, calcium, RA factor, 25-hydroxy vitamin D, anti-CCP antibody, and erythrocyte sedimentation rates were measured in a centralized laboratory of Fishtail Hospital and Research centre. Results: The level of serum 25-hydroxy vitamin D were significantly lower in RA group (20.03 ±9.97 ng/mL) in comparison to the control group (24.46 ±8.45 ng/mL) (p<0.003). Our result indicates the prevalence of vitamin D deficiency was more in RA group compared with control group (47.61% vs. 33.16%, p <0.002). The level of Anti-CCP is significantly high in RA group than control group. In RA patients serum 25-hydroxy vitamin D levels were negatively correlated to anti-CCP antibody levels (rs = 0.72, p <0.001), and erythrocyte sedimentation rate (rs = 3.95, p <0.005). Conclusion: In RA patient vitamin D deficiency is quite common and serum 25-hydroxy vitamin D level was negatively correlated to anti-CCP antibody level. Our results suggest that vitamin D level is a motivation factor rather than a consequence of RA activity

Epidemiology and Clinical Outcome of Snakebite in Western Nepal: A Retrospective Study

Introduction: Snakebite is an environmental hazard associated with significant morbidity and mortality. It is an important medical emergency and cause of significant numbers of hospital admissions in many parts of the Asian region. In this study, we assess the epidemiology and clinical outcome of snake bite. Methods: This was a retrospective study of all patients with snake bites admitted to the Department of Internal Medicine, Manipal Teaching Hospital, Pokhara, kaski, Nepal. A total numbers of 265 snake bite cases in the period of 2013 to 2016 were enrolled in this study. Snake bite cases by person, place and time along with month of snake bite and time of bite, were analyzed. We also identified the types of snake and site of the bite. Sign and symptoms were clinically observed and the management of the snake bite cases was clinically done. Prothrombin time (PT) test along with INR value was performed by Medical Technologist at the Department of Laboratory, Manipal Teaching Hospital. Data was entered in to the Microsoft excel and analyzed by SPPS version 21.0. Percentages were applied to find the results. Results: Total numbers of snake bite cases were 265. More than half, 60.4% of the snake bite cases were females. Regarding the age group, nearly half, 47.9% were in the age group of 20 - 40 years and 9.8% cases were in the age group of 60 years and above. In this study, 50.6% bite cases were held at the day time and most of the bites were reported/ observed in the limb, 53.6% in lower limb, and 43.4% in the upper limb. Very few bites were in the head, neck and trunk. Our result shows 49.1% were green snake and 30.9% snake were unidentified. When we observed the sign and symptoms, 153 (57.7%) cases showed local swelling, 83 (31.3%) showed fang mark. Hematological manifestation were 144 (54.3%) cases and complication observed in 145 (54.7%) cases. Snake bite cases were managed after PT/INR test, INR. Antibiotic were prescribed in 154 cases and in 135 (50.9%) cases blood was transfused. There were no fetal cases noticed among hospital admitted snake bite cases. Conclusion: There is gross disparity in the management and outcomes of snake bite in different hospitals. Snake bite cases should manage in tertiary care hospital as early as possible

Fetal Outcome in Pregnancies Complicated with Polyhydraminos: Study Done in Pokhara, Nepal

Introduction: Amniotic fluid plays an important role in the development of fetus. Any abnormality in the production amniotic fluid may have adverse effect on the fetus and the mother. Polyhydraminos is one of the common complications occurring during pregnancy and complicates around 0.2 - 2% of pregnancies. Methods: It is a prospective study conducted in Manipal Teaching Hospital, Pokhara, Nepal from January 2013 to December 2015. All the pregnant ladies irrespective of gestational age with amniotic fluid index (AFI) 25 cm or more were enrolled for the study. According to the AFI, polyhydraminos was classified as mild (25 – 30 cm), moderate (30.1 - 35 cm) and severe (>35 cm). Fetal outcome, mode of delivery, presence of congenital anomalies, NICU admission and maternal glucose intolerance were recorded. Results: Out of 8232 deliveries, 24 were diagnosed and admitted with the diagnosis of polyhydraminos. Mild polyhydraminos, 50% (n=12) occurred after 37 weeks of gestation and 12.5% (n=3) had severe polyhydraminos. All pregnant ladies 50% (n=12), beyond 37 weeks gestation had cesarean section, whereas 25% (n=6) had vaginal deliveries. 33.3% (n=8) had preterm labor, 12.5% (n=3) had premature rupture of membrane, 25% (n=6) had congenital anomalies, one IUFD, one case of Rh isoimmunisation and one case of twin pregnancy. NICU admission needed in 20.5% (n=5). Pregnant ladies with impaired glucose intolerance were 8.3% (n=2). Conclusions: Polyhydraminos is associated with increased incidence of cesarean section, preterm labor, fetal malformation and NICU admission

Assessment of Adenosine Deaminase Level and the Utility of Polymerase Chain Reaction in Diagnosis of Tuberculous Pleural Effusion

Background: Extrapulmonary tuberculosis is one of the major causes of exudative pleural effusion. The paucibacillary nature of effusion suggests the need of markers and methodologies for accurate diagnosis and prognosis of tuberculosis as well as to differentiate it from other non-tubercular causes of pleural effusion. Objectives: This study was focused to evaluate the utility of polymerase chain reaction (PCR) in detection of tuberculosis antigen and to assess the level of Adenosine Deaminase (ADA) in tubercular pleural effusion (TPE) and contrast it with other causes. Methods: This is a cross-sectional study where 100 samples of pleural effusion suspected to be tuberculosis were analyzed by PCR for the detection of IS6110 segment of DNA. The level of ADA was then determined and compared in both PCR positive and negative samples. The cut-off value of ADA was >40 U/L for TPE. Student t test was applied to compare the means with statistical significance set at p<0.05. Results: Out of 100 samples analyzed, 45% were positive for TPE and remaining 55% were non-tuberculosis pleural effusion as detected by PCR. The level of ADA was above cut-off (>40 U/L) in 43% TPE samples whereas all the non-tuberculosis effusion had ADA <40 U/L (p<0.001). Conclusion: The ADA level was significantly higher in TPE than in non-tuberculosis cases, and PCR was able to detect suspected cases of tuberculosis effusion in almost half of the cases. This finding suggests the diagnostic utility of combined use of ADA and PCR in diagnosis of TPE.  J-GMC-N | Volume 11 | Issue 01 | January-June 2018, Page: 5-

A Multi-Center Assessment of Thyroid Function Test Precision in Chemiluminescence Immunoassay (CLIA) Systems

Background: Chemiluminiscence immunoassay (CLIA) is exclusively pragmatic technology for the analysis of biomarker for diagnosis of thyroid disorders. However, performance characteristics of different chemiluminescence immunoassay (CLIA) systems supplied by different manufacturers in diverse set up for thyroid function test (TFT) has not yet been studied well. Objective: Our aim is to evaluate laboratory results by assessment of the reproducibility and repeatability of TFTs in three different diagnostic set up to assure the quality of thyroid hormone assay using chemiluminescence immunoassay (CLIA) instruments: Advia Centaur CP (Siemens), Access 2 (Beckman Coulter) and Liaison (Diasorin). Materials and Methods: Among the adult male and female individuals visited for thyroid hormone assay, 51 normal individuals were selected for the study. Three aliquots of serum samples were distributed to assess the reproducibility of three different CLIA equipments operated in three diagnostic centers. Additional three aliquots of serum were analyzed weekly for TFT (fFT3, fFT4 and TSH) to check the repeatability of assay in ADVIA Centaur CP set up. Assay precision was determined by reproducibility and repeatability of test results. Results: Results of TFTs of serum samples obtained from three different interlaboratory assays using different CLIA systems have achieved good precision showing minimal variance (P>0.05) and acceptable reproducibility. Results are also precise with adequate repeatability showing minimal variance (P >0.05) obtained from the three different intra-laboratory assays in a single CLIA system using ADVIA Centaur CP by same team. Conclusion: Our study elucidates the thyroid hormone assay performance of CLIA systems in three centers, which has shown assay precision with good reproducibility and repeatability of thyroid hormone assay. Thus, the analysis of precision as an essential component of quality control is necessary to deliver precise diagnostic services

The transmission of Leishmania infantum chagasi by the bite of the Lutzomyia longipalpis to two different vertebrates

<p>Abstract</p> <p>Background</p> <p>Sandflies are vectors of <it>Leishmania</it>, the causative agent of leishmaniasis in mammalian hosts, including humans. The protozoan parasite is transmitted by the sandfly bite during salivation that occurs at the moment of blood feeding. The components of vector saliva include anticlotting and vasodilatory factors that facilitate blood flow and immunomodulatory factors that inhibit wound healing and quell the immune response. Not surprisingly, these factors also play important roles in the establishment of <it>Leishmania </it>infection. To date, the majority of knowledge that has been generated regarding the process of <it>Leishmania </it>infection, including <it>L. infantum chagasi </it>transmission has been gathered by using intradermal or subcutaneous inoculation of purified parasites.</p> <p>Findings</p> <p>This study presents the establishment of a transmission model of <it>Leishmania infantum chagasi </it>by the bite of <it>Lutzomyia longipalpis</it>, the vector of American visceral leishmaniasis. The parasites were successfully transmitted by infected sandfly bites to mice and hamsters, indicating that both animals are good experimental models. The <it>L. infantum chagasi </it>dose that was transmitted in each single bite ranged from 10 to 10, 000 parasites, but 75% of the sandflies transmitted less than 300 parasites.</p> <p>Conclusions</p> <p>The strategy for initiating infection by sandfly bite of experimental animals facilitates future investigations into the complex and dynamic mechanisms of visceral leishmaniasis. It is important to elucidate the transmission mechanism of vector bites. This model represents a useful tool to study <it>L. infantum chagasi </it>infection transmitted by the vector.</p

Mediterranean diet supplemented with nuts reduces waist circumference and shifts lipoprotein subfractions to a less atherogenic pattern in subjects at high cardiovascular risk

[Objective]: The PREDIMED trial showed that Mediterranean diets supplemented with either extra-virgin olive oil or nuts reduced incident cardiovascular events compared to a control diet. Consumption of both supplemental foods has been associated with reduced LDL-cholesterol, but it is unknown whether they can shift lipoprotein subfractions to a less atherogenic pattern. We investigated changes in adiposity and lipoprotein subfractions after consumption of the PREDIMED diets.[Methods]: In a PREDIMED sub-cohort (n = 169), lipoprotein subclasses (particle concentrations and size) were determined by nuclear magnetic resonance spectroscopy at baseline and after intervention for 1 year.[Results]: Participants allocated to the Mediterranean diet supplemented with nuts showed significant reductions from baseline of waist circumference (mean [95% CI]; −5 cm [−7; −3]) and concentrations of medium-small (−27 nmol/l [−46; −8]) and very small LDL (−111 nmol/l [−180; −42]); decreased LDL particle number (a nuclear magnetic resonance-specific measurement) (−98 nmol/l [−184; −11]); and an increase of large LDL concentrations (54 nmol/l [18; 90]), with a net increase (0.2 nmol/l [0.1; 0.4]) of LDL size. The Mediterranean diets with olive oil and nuts increased large HDL concentrations (0.6 μM [0.0; 1.1] and 1.0 μM [0.4; 1.5], respectively). Compared to the other two intervention groups, participants in the nut-enriched diet showed significantly reduced waist circumference (p ≤ 0.006, both) and increased LDL size (p < 0.05, both).[Conclusion]: Lipoprotein subfractions are shifted to a less atherogenic pattern by consumption of Mediterranean diets enriched with nuts. The results contribute mechanistic evidence for the reduction of cardiovascular events observed in the PREDIMED trial.This study was funded in part by ISCIII (Spanish Ministry of Economy) through grants RTIC G03/140, RTIC RD 06/0045, and FIS PS09/01292 and grant CNIC 06/2007 from Centro Nacional de Investigaciones Cardiovasculares, Madrid, Spain. AS-V was supported by post-doctoral contract FIS CD07/0083. MF was supported by a joint contract of ISCIII and the Health Department of the Catalan Government (Generalitat de Catalunya), CP 06/00100

Optimization of DNA Extraction from Individual Sand Flies for PCR Amplification

Numerous protocols have been published for extracting DNA from phlebotomines. Nevertheless, their small size is generally an issue in terms of yield, efficiency, and purity, for large-scale individual sand fly DNA extractions when using traditional methods. Even though this can be circumvented with commercial kits, these are generally cost-prohibitive for developing countries. We encountered these limitations when analyzing field-collected Lutzomyia spp. by polymerase chain reaction (PCR) and, for this reason, we evaluated various modifications on a previously published protocol, the most significant of which was a different lysis buffer that contained Ca2+ (buffer TESCa). This ion protects proteinase K against autolysis, increases its thermal stability, and could have a regulatory function for its substrate-binding site. Individual sand fly DNA extraction success was confirmed by amplification reactions using internal control primers that amplify a fragment of the cacophony gene. To the best of our knowledge, this is the first time a lysis buffer containing Ca2+ has been reported for the extraction of DNA from sand flies.Centro Regional de Estudios Genómico

Infection Parameters in the Sand Fly Vector That Predict Transmission of Leishmania major

To identify parameters of Leishmania infection within a population of infected sand flies that reliably predict subsequent transmission to the mammalian host, we sampled groups of infected flies and compared infection intensity and degree of metacyclogenesis with the frequency of transmission. The percentage of parasites within the midgut that were metacyclic promastigotes had the highest correlation with the frequency of transmission. Meta-analysis of multiple transmission experiments allowed us to establish a percent-metacyclic “cutoff” value that predicted transmission competence. Sand fly infections initiated with variable doses of parasites resulted in correspondingly altered percentages of metacyclic promastigotes, resulting in altered transmission frequency and disease severity. Lastly, alteration of sand fly oviposition status and environmental conditions at the time of transmission also influenced transmission frequency. These observations have implications for transmission of Leishmania by the sand fly vector in both the laboratory and in nature, including how the number of organisms acquired by the sand fly from an infection reservoir may influence the clinical outcome of infection following transmission by bite