Characterization of "Off-Target" Immune Modulation Induced by Live Attenuated Yellow Fever Vaccine.

BACKGROUND: Live attenuated vaccines alter immune functions and are associated with beneficial outcomes. We previously demonstrated that live attenuated yellow fever virus (YFV) vaccine (LA-YF-Vax) dampens T-cell receptor (TCR) signaling in vitro via an RNA-based mechanism. We examined study participants before and after LA-YF-Vax to assess TCR-mediated functions in vivo. METHODS: Serum samples and peripheral blood mononuclear cells (PBMCs) were obtained before and after LA-YF-Vax (with or without additional vaccines) or quadrivalent influenza vaccine. TCR-mediated activation was determined by interleukin 2 release or phosphorylation of the lymphocyte-specific Src kinase. TCR-regulating phosphatase (protein tyrosine phosphatase receptor type E [PTPRE]) expression was also measured. RESULTS: Compared with prevaccination findings, LA-YF-Vax recipient PBMCs demonstrated transient reduction in interleukin 2 release after TCR stimulation and PTPRE levels, unlike in control participants who received quadrivalent influenza vaccine. YFV was detected in 8 of 14 participants after LA-YF-Vax. After incubation of healthy donor PBMCs in serum-derived extracellular vesicles prepared from LA-YF-Vax recipients, TCR signaling and PTPRE levels were reduced after vaccination, even in participants without detectable YFV RNA. CONCLUSIONS: LA-YF-Vax reduces TCR functions and PTPRE levels after vaccination. Extracellular vesicles from serum recapitulated this effect in healthy cells. This likely contributes to the reduced immunogenicity for heterologous vaccines after LA-YF-Vax administration. Identification of specific immune mechanisms related to vaccines should contribute to understanding of the "off-target," beneficial effects of live vaccines.

Evaluation of Latex Agglutination Test for Rapid Identification of Staphylococcus aureus Isolated from Pyogenic Wound Infections at a Tertiary Care Hospital.

Background Staphylococcus aureus infections are increasingly reported worldwide. It is a major clinical problem and imposes significant morbidity and mortality due to widespread emergence of multidrug resistant pathogens like methicillin resistant Staphylococcus aureus. Thus, rapid and reliable identification of Staphylococcus aureus is essential for timely and effective management of patient. Objective The performance of Latex agglutination test (Staphaurex Plus) was compared to conventional method tube coagulase test which is gold standard too for the identification of Staphylococcus aureus. Method This study was conducted at B.P. Koirala Institute of Health Sciences. Following standard microbiological methods, isolation and identification was done in the Department of Microbiology. MRSA detection was performed following Clinical and Laboratory Standard Institute. All the isolates of Staphylococci were subjected for Latex agglutination test and was performed according to manufacturer's instructions using Staphaurex Plus kit. Result Out of 377 (methicillin sensitive Staphylococcus aureus - 142; methicillin resistant Staphylococcus aureus - 233; Coagulase Negative Staphylococci -2) isolates of Staphylococci, Latex agglutination test was found to be positive in 138 (97.1%) of methicillin sensitive Staphylococcus aureus (MSSA) and 220 (94.4%) of methicillin resistant Staphylococcus aureus (MRSA). Overall sensitivity, specificity, positive predictive value and negative predictive value of Latex agglutination test was found to be 95.46%, 100%, 100%, 10.52% respectively. Conclusion In conclusion, Latex agglutination test is a rapid and reliable test for the identification of Staphylococcus aureus.

Prevalence of Microalbuminuria in Patients of Essential Hypertension and its Correlation with Left Ventricular Hypertrophy and Carotid Artery Intima­media Thickness.

Background Urinary albumin excretion has been associated to cardiovascular events and increased mortality in hypertensive patients. There is limited information among Nepalese patients about the implications of microalbuminuria (MA) in the setting of hypertension and potential cardiovascular morbidity. Objective To investigate the prevalence of microalbuminuria in patients with essential hypertension and its connection with left ventricular hypertrophy (LVH) and carotid artery intima-media thickness (cIMT). Method The study involved 80 hypertension individuals in total. All patients in the study had basic biochemical tests, routine urine evaluations, echocardiography, and carotid artery intima-media thickness measurements performed, and the data were analyzed. Result The prevalence of microalbuminuria was present in 37.5% cases of essential hypertension. The mean left ventricular mass index (LVMI) was significantly higher in patients with increased microalbuminuria as compared to patient with normal microalbuminuria. In addition, a significant positive correlation between microalbuminuria and left ventricular hypertrophy was also observed. Furthermore, mean carotid artery intima-media thickness was found to be higher in patients with microalbuminuria (p < 0.001), with 76.7% of the patients with microalbuminuria having elevated mean carotid artery intima-media thickness. The carotid artery intima-media thickness had a positive correlation with both microalbuminuria and left ventricular hypertrophy. Conclusion Microalbuminuria assessment in hypertensive patients is an important test for the evaluation of target organ damage. This study shows that microalbuminuria is common in hypertension patients, particularly those with left ventricular hypertrophy. Microalbuminuria was found to be associated with left ventricular hypertrophy and carotid artery intima-media thickness.

Detection of bla NDM-1 Encoding Imepenemase among the Imipenem-Resistant Gram-Negative Bacilli Isolated from Various Clinical Samples at a Tertiary Care Hospital of Eastern Nepal: A Descriptive Cross-Sectional Study.

BACKGROUND: Carbapenem resistance among Gram-negative isolates caused by the production of the metallo-ß-lactamase (MBL) enzyme is being increasingly reported worldwide. One of the newly emerged metallo-ß-lactamases is New Delhi metallo-ß-lactamase. Data regarding its occurrence in hospital setting and percentage prevalence among different Gram-negative bacterial isolates are lacking in our part. This study has been undertaken for determining the presence of the bla NDM-1 gene among the clinical isolates of imipenem-resistant Gram-negative bacteria in a tertiary care center in Dharan, Nepal. METHODS: A total of 75 imipenem-resistant Gram-negative isolates were studied. These were screened for metallo-ß-lactamase (MBL) production by phenotypic assays such as double-disc synergy test (DDST) and combined disc diffusion test (CDDT). PCR was performed for the molecular detection of gene NDM-1. Ten-disc method was performed to detect the presence of ESBL, AmpC, carbapenamase, and K1 ß-lactamase production. RESULTS: Using the molecular method, bla NDM-1 was detected in 36% of the isolates. Phenotypically, double-disc synergy test (DDST) and combined disc diffusion test (CDST) detected MBL production in 38.7% and 37.3% of the isolates, respectively. Ten-disc method detected ESBL in 26.6% of the isolates, but none of the isolates was found to be AmpC, carbapenamase, and K1 ß-lactamase producers. CONCLUSION: A high percentage of the NDM-1 producer was noted among imipenem-resistant GNB. Apart from performing only antimicrobial sensitivity test, phenotypic and molecular screening should be employed to find out the actual number of metallo-ß-lactamase producers and the existence of the resistance gene.

Understanding the effects of pasture type and stocking rate on the hydrology of the Southern Great Plains.

Grassland is one of the major biomes in the United States (US) and the world. In the US, the majority of grasslands are concentrated in the Great Plains and has undergone through significant interventions or management changes over the last few decades. A key economy-driven intervention in the Southern Great Plains (SGP) include the introduction of new forage species and conversion of native grassland to introduced pasture to increase productivity and its nutritive value for improved cattle production. Since water is one of the fundamental resources needed to sustain grassland productivity, it is important to understand how such pasture conversion and prevailing cattle grazing practices affect water balance and biomass production in a given pasture system. In this study, the Nutrient Tracking Tool (NTT) with its core APEX (Agricultural Policy Environmental eXtender) model was used to assess the hydrological impacts of the pasture introduction, i.e., native pasture (little bluestem, Schizachyrium halapense) vs. introduced pasture (old world bluestem, Bothriochloa caucasica), and the stocking rate in the SGP. Monthly evapotranspiration (ET) and biomass estimates from NTT compared well with observed data at two USDA-ARS experimental pastures (native and introduced) near El Reno, Oklahoma, for the years 2015 and 2016. Simulated long-term average annual hydrologic fluxes (i.e., ET, runoff, and groundwater recharge) from the introduced pasture were slightly lower than the observed data but not significantly different than those from the native pasture under the current management conditions. NTT predicted higher water yield (runoff and recharge) and significantly lower ET for the introduced pasture than the native pasture. Results suggest that grazing has the potential to alter the hydrological balance in the SGP. For example, the increase in stocking rate within the carrying capacity of the farm decreases ET and increases runoff and groundwater recharge for both pastures. Comparison of estimated biomass production between native and introduced pastures indicated that introduced pastures are more efficient in using the available water and thus produce a higher forage biomass per unit of water in the SGP. This study highlighted the potential significance of considering hydrological and other biophysical impacts of new forage introduction and stocking rate changes for the sustainable management of grazing and pasture systems in the SGP.

Chlamydia trachomatis in women with pelvic inflammatory disease (PID): report from a tertiary center in eastern Nepal.

Chlamydia trachomatis is an important agent of pelvic inflammatory disease (PID) globally. Laboratory diagnosis, which is vital for early and appropriate treatment, remains a challenge in resource-limited settings. Our study was undertaken to detect C. trachomatis in women with clinical features of PID. Three endocervical swabs, each obtained from 100 women clinically diagnosed with PID, were subjected to C. trachomatis antigen detection, microscopy and bacteriological culture. Logistic regression was used to assess the risk factors associated with PID. C. trachomatis antigen was present in 6%. The use of hormonal contraception, previous history of PID and a smoking habit were found to have statistically significant association in those who tested positive. Adjunctive use of rapid Chlamydia antigen test with a routinely practiced syndromic approach is beneficial for timely and appropriate antimicrobial therapy in women with PID.

Adipose tissue uncoupling protein 1 levels and function are increased in a mouse model of developmental obesity induced by maternal exposure to high-fat diet.

With brown adipose tissue (BAT) becoming a possible therapeutic target to counteract obesity, the prenatal environment could represent a critical window to modify BAT function and browning of white AT. We investigated if levels of uncoupling protein 1 (UCP1) and UCP1-mediated thermogenesis are altered in offspring exposed to prenatal obesity. Female CD-1 mice were fed a high-fat (HF) or standard-fat (SF) diet for 3 months before breeding. After weaning, all pups were placed on SF. UCP1 mRNA and protein levels were quantified using quantitative real-time PCR and Western blot analysis, respectively, in brown (BAT), subcutaneous (SAT) and visceral (VAT) adipose tissues at 6 months of age. Total and UCP1-dependent mitochondrial respiration were determined by high-resolution respirometry. A Student's t-test and Mann-Whitney test were used (significance: P<0.05). UCP1 mRNA levels were not different between the HF and SF offspring. UCP1 protein levels, total mitochondrial respiration and UCP1-dependent respiration were significantly higher in BAT from HF males (P=0.02, P=0.04, P=0.005, respectively) and females (P=0.01, P=0.04, P=0.02, respectively). In SAT, the UCP1 protein was significantly lower in HF females (P=0.03), and the UCP1-dependent thermogenesis was significantly lower from HF males (P=0.04). In VAT, UCP1 protein levels and UCP1-dependent respiration were significantly lower only in HF females (P=0.03, P=0.04, respectively). There were no differences in total respiration in SAT and VAT. Prenatal exposure to maternal obesity leads to significant increases in UCP1 levels and function in BAT in offspring with little impact on UCP1 levels and function in SAT and VAT.

Lack of correlation between the promastigote back-transformation assay and miltefosine treatment outcome.

OBJECTIVES: Widespread antimony resistance in the Indian subcontinent has enforced a therapy shift in visceral leishmaniasis treatment primarily towards miltefosine and secondarily also towards paromomycin. In vitro selection of miltefosine resistance in Leishmania donovani turned out to be quite challenging. Although no increase in IC50 was detected in the standard intracellular amastigote susceptibility assay, promastigote back-transformation remained positive at high miltefosine concentrations, suggesting a more 'resistant' phenotype. This observation was explored in a large set of Nepalese clinical isolates from miltefosine cure and relapse patients to assess its predictive value for patient treatment outcome. METHODS: The predictive value of the promastigote back-transformation for treatment outcome of a set of Nepalese L. donovani field isolates (n = 17) derived from miltefosine cure and relapse patients was compared with the standard susceptibility assays on promastigotes and intracellular amastigotes. RESULTS: In-depth phenotypic analysis of the clinical isolates revealed no correlation between the different susceptibility assays, nor any clear link to the actual treatment outcome. In addition, the clinical isolates proved to be phenotypically heterogeneous, as reflected by the large variation in drug susceptibility among the established clones. CONCLUSIONS: This in vitro laboratory study shows that miltefosine treatment outcome is not necessarily exclusively linked with the susceptibility profile of pre-treatment isolates, as determined in standard susceptibility assays. The true nature of miltefosine treatment failures still remains ill defined.

Prevalence of depression and utilization of health care in single and multiple morbidity: a population-based cohort study.

BACKGROUND: This study aimed to determine whether depression in patients with long-term conditions is associated with the number of morbidities or the type of co-morbidity. Method A cohort study of 299 912 participants aged 30-100 years. The prevalence of depression, rates of health-care utilization and costs were evaluated in relation to diagnoses of diabetes mellitus (DM), coronary heart disease (CHD), stroke and colorectal cancer. RESULTS: The age-standardized prevalence of depression was 7% in men and 14% in women with no morbidity. The frequency of depression increased in single morbidities including DM (men 13%, women 22%), CHD (men 15%, women 24%), stroke (men 14%, women 26%) or colorectal cancer (men 10%, women 21%). Participants with concurrent diabetes, CHD and stroke had a very high prevalence of depression (men 23%, women 49%). The relative rate of depression for one morbidity was 1.63 [95% confidence interval (CI) 1.59-1.66], two morbidities 1.96 (95% CI 1.89-2.03) and three morbidities 2.35 (95% CI 2.03-2.59). Compared to those with no morbidity, depression was associated with higher rates of health-care utilization and increased costs at any level of morbidity. In women aged 55 to 64 years without morbidity, the mean annual health-care cost was £513 without depression and £1074 with depression; when three morbidities were present, the cost was £1495 without depression and £2878 with depression. CONCLUSIONS: Depression prevalence and health-care costs are more strongly associated with the number of morbidities than the nature of the co-morbid diagnosis.

In vitro activity of cefoperazone-sulbactam combination against gram negative bacilli.

Cefoperazone is a â-lactam antimicrobial and Sulbactam is an irreversible â-lactamase inhibitor. The objective of this study was to know the susceptibility pattern of gram negative bacilli (GNB) towards cefoperazone-sulbactum. All GNB isolated from different clinical samples during the period of May, 2010 to Aug, 2010 were tested for susceptibility to cefoperazone-sulbactum, meropenem, ceftazidime, cefotaxime, ceftriaxone, chloromphenicol, cotrimoxazole, ampicillin, amikacin, nalidixic acid, ciprofloxacin, carbenicillin and piperacillin using standard Kirby-Bauer disc diffusion antimicrobial susceptibility testing method. The susceptibilities were recorded according to CLSI guidelines. A total of 406 GNB were isolated (urine: 66.7%, pus: 19.2%, and blood: 7.9%). Escherichia coli (54.4%) was most frequently isolated organisms followed by Acinetobacter species (17.7%), Klebsiella pneumoniae (9.1%) and Pseudomonas species (6.1%). Overall, 11.8% of isolates showed resistance to cefoperazone-sulbactam. Frequencies of isolates showing resistance to meropenem and amikacin were 14.7% and 26.25% respectively. Only 3.9% of Escherichia coli isolates showed resistance to cefoperazone-sulbactam. For other organisms, their lowest frequency ranging from 0-20%, exhibited resistance to meropenem. In Pseudomonas spp, in-vitro activity of amikacin was also better as only 11.1% isolates showed resistance to it. This study demonstrated the in-vitro synergistic effect of cefoperazonerazone-sulbactam and meropenem having good activity against GNB compared to the activity of other commonly tested antimicrobials. Cefoperazone-sulbactam can be recommended for the clinical practice against GNB exhibiting resistant to other antimicrobials as it is cheaper alternative to meropenem. Our results also focused on the continuous surveillance of the trends and features of resistance of common antimicrobials.

Development and evaluation of different PCR-based typing methods for discrimination of Leishmania donovani isolates from Nepal.

INTRODUCTION: Leishmania donovani, the causative agent of visceral leishmaniasis in the Indian subcontinent, has been reported to be genetically homogeneous. In order to support ongoing initiatives to eliminate the disease, highly discriminative tools are required for documenting the parasite population and dynamics. METHODS: Thirty-four clinical isolates of L. donovani from Nepal were analysed on the basis of size and restriction endonuclease polymorphisms of PCR amplicons from kinetoplast minicircle DNA, 5 nuclear microsatellites, and nuclear loci encoding glycoprotein 63, cysteine proteinase B, and hydrophilic acylated surface protein B. We present and validate a procedure allowing standardized analysis of kDNA fingerprint patterns. RESULTS: Our results show that parasites are best discriminated on the basis of kinetoplast minicircle DNA (14 genotypes) and 1 microsatellite defining 7 genotypes, while the remaining markers discriminated 2 groups or were monomorphic. Combination of all nuclear markers revealed 8 genotypes, while extension with kDNA data yielded 18 genotypes. CONCLUSION: We present tools that allow discrimination of closely related L. donovani strains circulating in the Terai region of Nepal. These can be used to study the micro-epidemiology of parasite populations, determine the geographical origin of infections, distinguish relapses from re-infection, and monitor the spread of particular variants.

Iron deficiency in a male with multiple myeloma.

A 58-year-old man with long standing unexplained iron deficiency anaemia presented with severe hypercalcaemia and was subsequently diagnosed with multiple myeloma. Anaemia in multiple myeloma is usually normochromic and normocytic.1 To our knowledge, there are no reported cases in the English literature of true iron deficiency associated with multiple myeloma. Our case highlights the importance of pursuing the aetiology of iron deficiency, especially in a middle aged male. It also reinforces the fact that severe hypercalcaemia usually has a malignant aetiology.

Amphiphilic triblock copolymer poly(p-dioxanone-co-L-lactide)-block-poly(ethylene glycol), enhancement of gene expression and inhibition of lung metastasis by aerosol delivery.

We describe the development of an aerosol system for topical gene delivery to the lungs of C57BL/6 mice. This system is based on the combination of the commercial cationic lipid Lipofectin with a novel amphiphilic triblock copolymer, poly(p-dioxanone-co-L-lactide)-block-poly(ethylene glycol) (PPDO/PLLA-b-PEG, and abbreviated in the text as polymeric micelles). After optimizing conditions for DNA delivery to the lungs of mice using the combination of polymeric micelles with Lipofectin and LacZ DNA, we used the Lipofectin/polymeric micelle system to deliver the tumor suppressor gene PTEN to the lungs of C57BL/6 mice bearing the B16-F10 melanoma. Lipofectin/PTEN/polymeric micelles significantly improved gene expression of PTEN in the lungs of mice with no evidence of cell toxicity or acute inflammation. Importantly, lung metastasis, as measured by lung weight, was significantly reduced (P<0.001), as were total tumor foci in the lungs (P<0.001) and size of individual tumor nodules in animals treated with Lipofectin/PTEN/polymeric micelles compared with control animals. Survival time was also extended. These results suggest that the Lipofectin/polymeric micelle system is appropriate for enhancing gene delivery in vivo and that it can be applied as a non-invasive gene therapy for lung cancer.

Patterns of organophosphorous poisoning attending a teaching hospital.

The objective of this was to study the pattern of patients of organophosphorous (OP) poisoning attending Kathmandu Medical College Teaching Hospital (KMCTH). This is a retrospective study of OP poisoning from hospital records were analyzed meticulously and data extracted. Data collected were: patient's demographics, motive for poisoning, type of OP poison, arrival time, time since ingestion, patient's vitals, predisposing factors, serum cholinesterase levels, treatment given, duration of hospital stay and mortality. A total of 47 patients of OP poisoning attended KMCTH from Aug 2003 to July 2005. 22(46.8%) cases were male and 25(53.2%) were female. The maximum number of patients were between the age of 20-40 (33-70.2%). Married patients outnumbered the unmarried (35/74.5% vs 12/25.5%). The most common motive for poisoning was suicidal, 41 cases (87.2%). Metacid (methyl-parathion) was the most commonly used OP compounds in 32 (68%) patients. Interpersonal marital relationship seemed to be the commonest predisposing factor, 23 cases (48.9%). The commonest time of presentation was between 6 pm-12 midnight. 7(57.4%) cases presented within 2 hours of ingestion of the poison. Serum cholinesterase level measured after full atropinisation was >50% of normal level in 17 cases (36.2%). 10 cases (21.3%) required respiratory support. Intermediate syndrome (IMS) was observed in 3 cases. Overall mortality occurred in 3 cases (6.4%). This study highlights the problem of OP poisoning in our region. Establishment of strict policies against the sale and availability of insecticides and pesticides which are freely available in the market is an effective way to control OP poisoning.

GeO2 fibers: preparation, morphology and photoluminescence property.

Nanomicron to submicron fibers of GeO(2) have been prepared using poly(vinyl acetate) and germanium dioxide sol by electrospinning followed by high temperature calcination. The morphology of the fibers have been studied by scanning electron microscopy, transmission electron microscopy, and atomic force microscopy. X-ray diffraction indicates that the fibers are single crystal with hexagonal alpha-phase quartz-like structure. At room temperature, the fibers show photoluminescence under excitation at 325 nm. The fibers may have potential applications in one-dimensional optoelectronic nanodevices.

Traditional herbal medicines used to treat wounds and injuries in Nepal.

In rural Nepalese societies, due to the lack of modern health services and facilities, folk herbal preparations are still the dominant method of therapy for common ailments. These remedies are fairly well accepted, easily available, bear a minimal cost and are generally the only available resource. Information on the curative properties of 42 plant species from 40 genera and 23 families that are used to treat wounds and injuries, has been documented. These herbal remedies are based on ancestral knowledge and personal experience. Their successful use indicates that they are alive and functioning in the rural localities. Although the techniques of preparation of drugs or dosage forms employed by traditional healers have generally been observed to be poor and in most cases do not comply with the requirements of modern pharmaceutical practices, there are trends towards the development and modernization of practice including the standardization of specific dosage, storage of drugs, use of specific types of containers and their labelling. A critical evaluation of the phytochemistry and pharmacology of the alleged curative plants and the traditional pharmaceutical practices employed has been strongly recommended. In rural Nepalese societies, the number and distribution of traditional herbal practitioners and faith healers greatly exceed all other health workers. Their influence should be applied to the task of motivating the rural populace to adopt authentic herbal remedies and other health-related hygienic behaviour.

Traditional phytotherapy among the Sherpas of Helambu, central Nepal.

The traditional phytotherapy of the Sherpas of the Helambu region, Central Nepal, has been investigated. Sixty-seven widely accepted prescriptions involving 49 plant species (one species of fungi, two species of ferns, one species of gymnosperms and forty-five species of angiosperms) are presented along with details on uses. Ethnobotanically, the Helambu Sherpas appear to be one of the least acculturated groups of people. The vast number of prescriptions available to treat ailments, from minor cuts and wounds to bone fractures, gynaecological complaints and tuberculosis, suggest ample opportunities for their critical scientific study.