Association of acute toxic encephalopathy with litchi consumption in an outbreak in Muzaffarpur, India, 2014: a case-control study

Background Outbreaks of unexplained illness frequently remain under-investigated. In India, outbreaks of an acute neurological illness with high mortality among children occur annually in Muzaffarpur, the country’s largest litchi cultivation region. In 2014, we aimed to investigate the cause and risk factors for this illness. Methods In this hospital-based surveillance and nested age-matched case-control study, we did laboratory investigations to assess potential infectious and non-infectious causes of this acute neurological illness. Cases were children aged 15 years or younger who were admitted to two hospitals in Muzaffarpur with new-onset seizures or altered sensorium. Age-matched controls were residents of Muzaffarpur who were admitted to the same two hospitals for a non-neurologic illness within seven days of the date of admission of the case. Clinical specimens (blood, cerebrospinal fluid, and urine) and environmental specimens (litchis) were tested for evidence of infectious pathogens, pesticides, toxic metals, and other non-infectious causes, including presence of hypoglycin A or methylenecyclopropylglycine (MCPG), naturally-occurring fruit-based toxins that cause hypoglycaemia and metabolic derangement. Matched and unmatched (controlling for age) bivariate analyses were done and risk factors for illness were expressed as matched odds ratios and odds ratios (unmatched analyses). Findings Between May 26, and July 17, 2014, 390 patients meeting the case definition were admitted to the two referral hospitals in Muzaffarpur, of whom 122 (31%) died. On admission, 204 (62%) of 327 had blood glucose concentration of 70 mg/dL or less. 104 cases were compared with 104 age-matched hospital controls. Litchi consumption (matched odds ratio [mOR] 9·6 [95% CI 3·6 – 24]) and absence of an evening meal (2·2 [1·2–4·3]) in the 24 h preceding illness onset were associated with illness. The absence of an evening meal significantly modified the effect of eating litchis on illness (odds ratio [OR] 7·8 [95% CI 3·3–18·8], without evening meal; OR 3·6 [1·1–11·1] with an evening meal). Tests for infectious agents and pesticides were negative. Metabolites of hypoglycin A, MCPG, or both were detected in 48 [66%] of 73 urine specimens from case-patients and none from 15 controls; 72 (90%) of 80 case-patient specimens had abnormal plasma acylcarnitine profiles, consistent with severe disruption of fatty acid metabolism. In 36 litchi arils tested from Muzaffarpur, hypoglycin A concentrations ranged from 12·4 μg/g to 152·0 μg/g and MCPG ranged from 44·9 μg/g to 220·0 μg/g. Interpretation Our investigation suggests an outbreak of acute encephalopathy in Muzaffarpur associated with both hypoglycin A and MCPG toxicity. To prevent illness and reduce mortality in the region, we recommended minimising litchi consumption, ensuring receipt of an evening meal and implementing rapid glucose correction for suspected illness. A comprehensive investigative approach in Muzaffarpur led to timely public health recommendations, underscoring the importance of using systematic methods in other unexplained illness outbreaks

Immune Checkpoint Targets for Host-Directed Therapy to Prevent and Treat Leishmaniasis

Leishmaniasis encompasses a group of diseases caused by protozoan parasites belonging to the genus Leishmania. These diseases range from life threatening visceral forms to self-healing cutaneous lesions, and each disease manifestations can progress to complications involving dissemination of parasites to skin or mucosal tissue. A feature of leishmaniasis is the key role host immune responses play in disease outcome. T cells are critical for controlling parasite growth. However, they can also contribute to disease onset and progression. For example, potent regulatory T cell responses can develop that suppress antiparasitic immunity. Alternatively, hyperactivated CD4+ or CD8+ T cells can be generated that cause damage to host tissues. There is no licensed human vaccine and drug treatment options are often limited and problematic. Hence, there is an urgent need for new strategies to improve the efficacy of current vaccine candidates and/or enhance both antiparasitic drug effectiveness and subsequent immunity in treated individuals. Here, we describe our current understanding about host immune responses contributing to disease protection and progression in the various forms of leishmaniasis. We also discuss how this knowledge may be used to develop new strategies for host-directed immune therapy to prevent or treat leishmaniasis. Given the major advances made in immune therapy in the cancer and autoimmune fields in recent years, there are significant opportunities to ride on the back of these successes in the infectious disease domain. Conversely, the rapid progress in our understanding about host immune responses during leishmaniasis is also providing opportunities to develop novel immunotherapy strategies that could have broad applications in diseases characterized by inflammation or immune dysfunction

Altered IL-7 signaling in CD4+ T cells from patients with visceral leishmaniasis.

BackgroundCD4+ T cells play a central role in control of L. donovani infection, through IFN-γ production required for activation of macrophages and killing of intracellular parasites. Impaired control of parasites can in part be explained by hampered CD4+ T cells effector functions in visceral leishmaniasis (VL) patients. In a recent studies that defined transcriptional signatures for CD4+ T cells from active VL patients, we found that expression of the IL-7 receptor alpha chain (IL-7Rα; CD127) was downregulated, compared to CD4+ T cells from endemic controls (ECs). Since IL-7 signaling is critical for the survival and homeostatic maintenance of CD4+ T cells, we investigated this signaling pathway in VL patients, relative to ECs.MethodsCD4+ T cells were enriched from peripheral blood collected from VL patients and EC subjects and expression of IL7 and IL7RA mRNA was measured by real time qPCR. IL-7 signaling potential and surface expression of CD127 and CD132 on CD4+ T cell was analyzed by multicolor flow cytometry. Plasma levels of soluble IL-7 and sIL-7Rα were measured by ELISA.ResultTranscriptional profiling data sets generated previously from our group showed lower IL7RA mRNA expression in VL CD4+ T cells as compared to EC. A significant reduction was, however not seen when assessing IL7RA mRNA by RT-qPCR. Yet, the levels of soluble IL-7Rα (sIL-7Rα) were reduced in plasma of VL patients compared to ECs. Furthermore, the levels of soluble IL-7 were higher in plasma from VL patients compared to ECs. Interestingly, expression of the IL-7Rα protein was higher on VL patient CD4+ T cells as compared to EC, with activated CD38+ CD4+ T cells showing higher surface expression of IL-7Rα compared to CD38- CD4+ T cells in VL patients. CD4+ T cells from VL patients had higher signaling potential baseline and after stimulation with recombinant human IL-7 (rhIL-7) compared to EC, as measured by phosphorylation of STAT5 (pSTAT5). Interestingly, it was the CD38 negative cells that had the highest level of pSTAT5 in VL patient CD4+ T cells after IL-7 stimulation. Thus, despite unaltered or potentially lowered IL7RA mRNA expression by CD4+ T cells from VL patients, the surface expression of the IL-7Rα was higher compared to EC and increased pSTAT5 was seen following exposure to rhIL-7. Accordingly, IL-7 signaling appears to be functional and even enhanced in VL CD4+ T cells and cannot explain the impaired effector function of VL CD4+ T cells. The enhanced plasma IL-7 may serve as part of homeostatic feedback mechanism regulating IL7RA expression in CD4+ T cells

Human papillomavirus infection & anal cytological abnormalities in HIV-positive men in eastern India

Abstract Background Oncogenic Human papillomavirus (HPV) infections are closely associated with anal cancer which is high among human immunodeficiency virus (HIV) infected males. There are no data regarding anal HPV infection and cytological abnormalities in HIV positive males receiving free therapy in the national program. Thus, this cross-sectional study was performed to assess the prevalence and risk factors of anal HPV infection and cytological abnormalities in HIV positive males. Methods We screened 126 HIV-positive male patients attending the antiretroviral treatment center (ART) between 2014 and 2015 with anal papanicolaou smear cytology and HPV-DNA testing. HPV-DNA was detected by using polymerase chain reaction (PCR) method with two consensus primer sets E6 and MY09/11 and further analyzed for the presence of various HPV genotype by Sanger sequencing. Risk factors associated with anal cytological abnormalities and HPV infection was analyzed by using univariate and multivariate logistic regression models. Results Out of 126, 52 were on antiretroviral therapy. 91% were married to female partners but during the study 48 (38%) gave positive history of anal intercourse with other men. Anal cytology was done in 95 patients, out of which 60 (63.15%) had cytological abnormalities. LSIL (low-grade squamous intraepithelial lesions) was present in 27 (45%), ASCUS (atypical squamous cells of undetermined significance) in 31 (52%) and ASC-H (atypical squamous cells cannot exclude a high-grade squamous intraepithelial lesion) in 2 (3.33%). In multivariate analysis, the risk factors for cytological abnormality were presence of history of anal intercourse (OR, 6.1; 95% CI, 2.0–18.7) and WHO stage III & IV (OR, 2.7; 95% CI, 1.1–7.5). HPV-DNA was detected in 33/119 (27.73%) patients. The most prevalent HPV type in the study was HPV-16 (10.08%), other HPV types detected were 18,31,35,17,66,72,52,68 and 107 (17.65%). Conclusions High prevalence of anal cytological abnormalities in our study suggests that regular anal Pap smear screening should be done in HIV positive males in the ART center

Type I Interferons Suppress Anti-parasiticImmunity and Can Be Targeted to Improve Treatment of Visceral Leishmaniasis

Type I interferons (IFNs) play critical roles in anti-viral and anti-tumor immunity. However, they also suppress protective immune responses in some infectious diseases.This work was made possible through NIH Tropical Medicine Research Centers (TMRC) grant U19 AI074321 and Queensland State Government funding. The research was supported by grants and fellowships from the National Health and Medical Research Council of Australia (NHMRC; 1037304, 1058685, 1132975, and 1154265) as well as Australian post-graduate awards through Griffith University Institute of Glycomics and School of Natural Sciences (to P.B. and S.S.N., respectively), junior research fellowships from the Indian Council of Medical Research (to S.S.S. and S.B.C.), and an INSPIRE Faculty fellowship (LBSM-109/IF-14 to R.K.) provided by the Indian government Department of Science and Technology (DST)

Interleukin 2 is an Upstream Regulator of CD4+ T Cells from Visceral Leishmaniasis Patients with Therapeutic Potential

Control of visceral leishmaniasis (VL) caused by Leishmania donovani requires interferon-γproduction by CD4+ T cells. In VL patients, antiparasitic CD4+ T-cell responses are ineffective for unknown reasons. In this study, we measured the expression of genes associated with various immune functions in these cells from VL patients and compared them to CD4+ T cells from the same patients after drug treatment and from endemic controls. We found reduced GATA3, RORC, and FOXP3 gene expression in CD4+ T cells of VL patients, associated with reduced Th2, Th17, and FOXP3+CD4+ T regulatory cell frequencies in VL patient blood. Interleukin 2 (IL-2) was an important upstream regulator of CD4+ T cells from VL patients, and functional studies demonstrated the therapeutic potential of IL-2 for improving antiparasitic immunity. Together, these results provide new insights into the characteristics of CD4+ T cells from VL patients that can be used to improve antiparasitic immune responses.</p

An outline of the development of the Czech sport financing after 1989

Title: An outline of the development of the Czech sport financing after 1989 Objectives: The main aim of this thesis is to analyse the development of the czech sport financing from the selected resources after 1989 till the present situation and it's prospect included in The concept sports funding 2016 - 2025 SPORT 2025 Methods: The basic method used in this thesis is the method of document analysis. Fundamental documents include legislative documents, especially The Lottery Act and its updating and The Act on Promotion of Sports. Aditional resources represent analytic and comparative studies prepared by ČOV, ČUS and MŠMT conception. Another method is statistic analysis of quantitative data, in form of termporal lines, which are processed into basic charts and diagrams. Results: In recent 25 years have been significant changes in sports funding established. The turning point is considered the year 2011, when czech sport lost two stable financial resources. Primarily the company Sazka, a.s. went bankrupt, secondarily the amendment of the lottery act was accepted and has discontinued offtakes for public utility objects. Since then an optimal sports funding system is sought. State sports support shows a long-term downward trend and belongs to the lowest rank in the EU comparison. efforts to improve..

The effects of different immune and drug therapy combinations.

<p><i>L</i>. <i>donovani</i>-infected mice (low-dose challenge with 5 x 10⁶ parasites) were treated either with control Ab, anti-GITR mAb, anti-IL-10R mAb or both mAbs with (black bars) or without (open bars) a sub-optimal dose of sodium stibogluconate (Sb^v). A group receiving a full dose of Sb^v was also included. Infected mice were treated with anti-GITR mAb on day 14 p.i., and anti-IL-10R mAb on days 14, 19 and 24 p.i., with or without drug, as indicated. Rat IgG was used as a control. Liver parasite burdens were measured at day 28 p.i. (A). Spleen cells were also isolated at this time and cultured with parasite antigen for 24 hours before measuring levels of IFNγ (B), TNF(C) and IL-10 (D) in culture supernatants. Data are represented as the mean +/- SEM, and statistical differences of p < 0.05 (*), p < 0.01 (**) and p < 0.001 (***) are indicated (n = 6 mice per group, one independent experiment from 1–3 independent experiments).</p

Increased frequency of GITR-positive CD4⁺ T cells in VL patients.

<p><b>A.</b> The relative expression of GITR mRNA in PBMC of VL patients was measured by qPCR before treatment (Pre-drug; n = 7) and 28 days after the commencement of treatment (Post-drug; n = 7),as well as in healthy endemic control (EC; n = 5) samples. <b>B</b>. PBMC’s from VL patients before drug treatment (VL; n = 7) and healthy endemic controls (EC; n = 5) were gated on CD3ε⁺ CD4⁺ T cells and the frequency of GITR-positive CD4⁺ T cells was measured by FACS. Box and whisker plots show the box extending from the 25^th to 75^th percentiles with the line in the middle of the box representing the median and whiskers going down to the smallest value and up to the largest. Statistical differences of p < 0.05 (*) and p < 0.01 (**) are indicated.</p

Parasite inoculum determines combination treatment outcome.

<p>Parasite burdens at day 28 p.i., were measured in the livers and spleens of mice, as indicated, infected with a high (<b>A</b>) or low (<b>B</b>) dose of parasite inoculum. Infected mice were treated with mAb alone or a combination of anti-GITR mAb on day 14 p.i., and anti-IL-10R mAb on days 14, 19 and 24 p.i.. Rat IgG was used as a control. Data are represented as the mean +/- SEM, and statistical differences of p < 0.05 (*), p <0.01 (**) and p < 0.001 (***) are indicated (n = 15 mice per group from 3 independent experiments).</p