The spleen is the graveyard of CD4+ cells in patients with immunological failure of visceral leishmaniasis and AIDS.

BACKGROUND: Visceral leishmaniasis (VL), or kala-azar, is a common comorbidity in patients with AIDS in endemic areas. Many patients continue to experiences relapses of VL despite virological control, but with immunological failure. These patients remain chronically symptomatic with hypersplenism, for example with anemia, leukopenia, and thrombocytopenia, and are at risk of severe co-infection due to low CD4+ count. Therefore, in this study, splenectomized patients with VL and HIV infection were investigated to understand why the CD4+ count fails to recover in these patients, evaluating the importance of spleen mass for hypersplenism and immunological failure. METHODS: From a retrospective open cohort of 13 patients who had previously undergone splenectomy as salvage therapy for relapsing VL, 11 patients with HIV infection were investigated. This study compared the patients' complete blood cell count (CBC) and CD4+ and CD8+ cell counts before and after splenectomy with respect to spleen weight. RESULTS: CBC was substantially improved after splenectomy, indicating hypersplenism. However, to the best of our knowledge, this is the first study to show that spleen mass is strongly and negatively correlated with CD4+ cell count (ρ = -0.71, P = 0.015). CONCLUSIONS: This finding was unexpected, as the spleen is the most extensive lymphoid tissue and T-lymphocyte source. After reviewing the literature and reasoning, we hypothesized that the immunological failure was secondary to CD4+ loss initially by apoptosis in the spleen induced by productive HIV infection and, subsequently, by pyroptosis sustained by parasitic infection in spleen macrophages.

Association between clinical outcomes, peripheral blood and cytomorphologic features of bone marrow in visceral leishmaniasis.

INTRODUCTION: An intracellular parasite of mononuclear phagocytes, mainly distributed in the bone marrow and the spleen, causes visceral leishmaniasis. Complete blood count (CBC) reveals the poorly understood pathogenesis of anemia, leukopenia and thrombocytopenia. Our study aimed to compare the CBC with bone marrow cytomorphological features and their association with clinical outcomes to clarify this relevant issue. METHODS: The CBC and bone marrow of 118 patients were described by two hematologists and compared to check their association with each other and mortality. RESULTS: Peripheral cytopenias were common findings, particularly anemia, as seen in almost all patients. No relationship was found between values of hemoglobin, neutrophils and platelet count with fatal outcomes. The bone marrow was normocellular in 61.9% of the cases. Dysplasia figures were frequent and 49.1% of the samples had dysgranulopoiesis. Additionally, erythroid hyperplasia was found in 72% of the patients with severe anemia. Patients with reduced bone marrow cellularity, erythroid hypercellularity and dyserythropoiesis seem to have a riskier disease. CONCLUSION: The study results suggest that the bone marrow of patients with visceral leishmaniasis manifests a reactional pattern to the inflammatory event, thereby modulating cytokines and other colony growth factors. This compensatory response may be dysplastic and ineffective and generate peripheral cytopenias of varying intensity. Further studies are needed to clarify the signaling pathways involved, which may be used as therapeutic tools in the future.

Relationship Between Zinc, Selenium, and Magnesium Status and Markers of Metabolically Healthy and Unhealthy Obesity Phenotypes.

Our objective was to investigate the relationship between zinc, selenium, and magnesium status and markers of metabolically healthy and unhealthy obesity phenotypes. This was a cross-sectional study with 140 women: metabolically healthy obese women (n = 35), metabolically unhealthy obese women (n = 28), and normal-weight women (n = 77). We have calculated the body mass index, waist-hip ratio, waist-height ratio and some adiposity indices. Additionally, we evaluated endocrine-metabolic parameters and estimated the dietary intake of energy, macronutrients, zinc, selenium, and magnesium. The mineral concentrations in plasma, erythrocytes, and urine were assessed. In obese patients, there was a significant decrease in dietary zinc, selenium, and magnesium intake per kilogram of body weight, as well as lower mineral concentrations in both plasma and erythrocytes. Additionally, these patients exhibited higher urinary mineral levels compared to the control group, regardless of whether they had healthy or unhealthy phenotypes. We observed a significant correlation between deficiencies in zinc, selenium, and magnesium and obesity-associated metabolic disorders, including dyslipidemias and redox status disturbances. This study highlights a connection between deficiencies in zinc, selenium, and magnesium and metabolic disorders linked to obesity, including dyslipidemias, alterations in redox status, and thyroid hormonal dysfunction.

High seroprevalence of Leishmania infantum is linked to immune activation in people with HIV: a two-stage cross-sectional study in Bahia, Brazil.

Visceral leishmaniasis is an opportunistic disease in HIV-1 infected individuals, unrecognized as a determining factor for AIDS diagnosis. The growing geographical overlap of HIV-1 and Leishmania infections is an emerging challenge worldwide, as co-infection increases morbidity and mortality for both infections. Here, we determined the prevalence of people living with HIV (PWH) with a previous or ongoing infection by Leishmania infantum and investigated the virological and immunological factors associated with co-infection. We adopted a two-stage cross-sectional cohort (CSC) design (CSC-I, n = 5,346 and CSC-II, n = 317) of treatment-naïve HIV-1-infected individuals in Bahia, Brazil. In CSC-I, samples collected between 1998 and 2013 were used for serological screening for leishmaniasis by an in-house Enzyme-Linked Immunosorbent Assay (ELISA) with SLA (Soluble Leishmania infantum Antigen), resulting in a prevalence of previous or ongoing infection of 16.27%. Next, 317 PWH were prospectively recruited from July 2014 to December 2015 with the collection of sociodemographic and clinical data. Serological validation by two different immunoassays confirmed a prevalence of 15.46 and 8.20% by anti-SLA, and anti-HSP70 serology, respectively, whereas 4.73% were double-positive (DP). Stratification of these 317 individuals in DP and double-negative (DN) revealed a significant reduction of CD4+ counts and CD4+/CD8+ ratios and a tendency of increased viral load in the DP group, as compared to DN. No statistical differences in HIV-1 subtype distribution were observed between the two groups. However, we found a significant increase of CXCL10 (p = 0.0076) and a tendency of increased CXCL9 (p = 0.061) in individuals with DP serology, demonstrating intensified immune activation in this group. These findings were corroborated at the transcriptome level in independent Leishmania- and HIV-1-infected cohorts (Swiss HIV Cohort and Piaui Northeast Brazil Cohort), indicating that CXCL10 transcripts are shared by the IFN-dominated immune activation gene signatures of both pathogens and positively correlated to viral load in untreated PWH. This study demonstrated a high prevalence of PWH with L. infantum seropositivity in Bahia, Brazil, linked to IFN-mediated immune activation and a significant decrease in CD4+ levels. Our results highlight the urgent need to increase awareness and define public health strategies for the management and prevention of HIV-1 and L. infantum co-infection.

Recombinant protein KR95 as an alternative for serological diagnosis of human visceral leishmaniasis in the Americas.

In the Americas, visceral leishmaniasis (VL) is caused by the protozoan Leishmania infantum, leading to death if not promptly diagnosed and treated. In Brazil, the disease reaches all regions, and in 2020, 1,933 VL cases were reported with 9.5% lethality. Thus, an accurate diagnosis is essential to provide the appropriate treatment. Serological VL diagnosis is based mainly on immunochromatographic tests, but their performance may vary by location, and evaluation of diagnostic alternatives is necessary. In this study, we aimed to evaluate the performance of ELISA with the scantily studied recombinant antigens, K18 and KR95, comparing their performance with the already known rK28 and rK39. Sera from parasitologically confirmed symptomatic VL patients (n = 90) and healthy endemic controls (n = 90) were submitted to ELISA with rK18 and rKR95. Sensitivity (95% CI) was, respectively, 83.3% (74.2-89.7) and 95.6% (88.8-98.6), and specificity (95% CI) was 93.3% (85.9-97.2) and 97.8% (91.8-99.9). For validation of ELISA with the recombinant antigens, we included samples from 122 VL patients and 83 healthy controls collected in three regions in Brazil (Northeast, Southeast, and Midwest). When comparing the results obtained with the VL patients' samples, significantly lower sensitivity was obtained by rK18-ELISA (88.5%, 95% CI: 81.5-93.2) compared with rK28-ELISA (95.9%, 95% CI: 90.5-98.5), but the sensitivity was similar comparing rKR95-ELISA (95.1%, 95% CI: 89.5-98.0), rK28-ELISA (95.9%, 95% CI: 90.5-98.5), and rK39-ELISA (94.3%, 95% CI: 88.4-97.4). Analyzing the specificity, it was lowest with rK18-ELISA (62.7%, 95% CI: 51.9-72.3) with 83 healthy control samples. Conversely, higher and similar specificity was obtained by rKR95-ELISA (96.4%, 95% CI: 89.5-99.2), rK28-ELISA (95.2%, 95% CI: 87.9-98.5), and rK39-ELISA (95.2%, 95% CI: 87.9-98.5). There was no difference in sensitivity and specificity across localities. Cross-reactivity assessment, performed with sera of patients diagnosed with inflammatory disorders and other infectious diseases, was 34.2% with rK18-ELISA and 3.1% with rKR95-ELISA. Based on these data, we suggest using recombinant antigen KR95 in serological assays for VL diagnosis.

Scientometric review of research on Neglected Tropical Diseases: a 31-year perspective from the Journal of the Brazilian Society of Tropical Medicine.

BACKGROUND: To analyze the temporal evolution of research on Neglected Tropical Diseases (NTDs) published by the Journal of the Brazilian Society of Tropical Medicine (JBSTM). METHODS: We performed an analysis of the scientific production in JBSTM on NTDs using an advanced search, which included authors' descriptors, title, and abstract, and by combining specific terms for each NTDs from 1991 to 2021. Data related to authors, countries of origin, institutions, and descriptors, were evaluated and analyzed over time. Bibliographic networks were constructed using VOSviewer 1.6.16. RESULTS: The JBSTM published 4,268 scientific papers during this period. Of these 1,849 (43.3%) were related to NTDs. The number of publications on NTDs increased by approximately 2.4-fold, from 352 (total 724) during 1991-2000 to 841 (total 2,128) during 2011-2021, despite the proportional reduction (48.6% versus 39.5%). The most common singular NTDs subject of publications included Chagas disease (31.4%; 581/1,849), leishmaniasis (25.5%, 411/1,849), dengue (9.4%, 174/1,849), schistosomiasis (9.0%; 166/1,849), and leprosy (6.5%, 120/1,849), with authorship mostly from Brazil's South and Southeast regions. CONCLUSIONS: Despite the proportional reduction in publications, JBSTM remains an important vehicle for disseminating research on NTDs during this period. There is a need to strengthen the research and subsequent publications on specific NTDs. Institutions working and publishing on NTDs in the country were concentrated in the South and Southeast regions, requiring additional investments in institutions in other regions of the country.

Relationship Between Markers of Chronic Inflammation and Copper Nutritional Status in Obese Women.

Adipose tissue dysfunction causes the development of metabolic complications, such as low-grade chronic inflammation, which may to alter copper homeostasis in obese individuals. Thus, the objective of this study is to analyze the relationship between markers of chronic inflammation and copper nutritional status in obese women. Cross-sectional study involved women aged 20-50 years, divided into two groups: case (BMI > 35 kg/m2) and control (18.5 > BMI > 24.9 kg/m2). Plasma and erythrocyte copper concentrations were determined by inductively coupled plasma optical emission spectrometry (ICP-OES) method. Activity of superoxide dismutase (SOD) enzyme in the erythrocytes was determined with an automatic biochemical analyzer. Serum concentrations of interleukin (IL)-6, IL-8, IL-12, IL-10, and IL-1ß and tumor necrosis factor-alpha (TNF-α) were determined by using flow cytometer. Serum IL-6 concentrations were 105% higher in the case group compared to eutrophic women. Plasma copper concentrations were 20.5% higher, and erythrocyte copper concentrations were 23.5% lower in patients with obesity. In addition, erythrocyte SOD activity was 20% lower in obese participants when compared to eutrophic women. Our study identified significant negative correlation between the cytokines TNF-α and IL-10 and the SOD activity in the case group, suggesting a possible influence of chronic inflammation on copper distribution in obese individuals.

Scientometric review of research on Neglected Tropical Diseases: a 31-year perspective from the Journal of the Brazilian Society of Tropical Medicine

ABSTRACT Background: To analyze the temporal evolution of research on Neglected Tropical Diseases (NTDs) published by the Journal of the Brazilian Society of Tropical Medicine (JBSTM). Methods: We performed an analysis of the scientific production in JBSTM on NTDs using an advanced search, which included authors' descriptors, title, and abstract, and by combining specific terms for each NTDs from 1991 to 2021. Data related to authors, countries of origin, institutions, and descriptors, were evaluated and analyzed over time. Bibliographic networks were constructed using VOSviewer 1.6.16. Results: The JBSTM published 4,268 scientific papers during this period. Of these 1,849 (43.3%) were related to NTDs. The number of publications on NTDs increased by approximately 2.4-fold, from 352 (total 724) during 1991-2000 to 841 (total 2,128) during 2011-2021, despite the proportional reduction (48.6% versus 39.5%). The most common singular NTDs subject of publications included Chagas disease (31.4%; 581/1,849), leishmaniasis (25.5%, 411/1,849), dengue (9.4%, 174/1,849), schistosomiasis (9.0%; 166/1,849), and leprosy (6.5%, 120/1,849), with authorship mostly from Brazil's South and Southeast regions. Conclusions: Despite the proportional reduction in publications, JBSTM remains an important vehicle for disseminating research on NTDs during this period. There is a need to strengthen the research and subsequent publications on specific NTDs. Institutions working and publishing on NTDs in the country were concentrated in the South and Southeast regions, requiring additional investments in institutions in other regions of the country.

Evidence map of diagnosis, treatment, prognosis, prevention, and control in visceral leishmaniasis.

Objective: To develop an evidence map on visceral leishmaniasis prevention, control, diagnosis, treatment, and prognosis. Methods: Systematic reviews on visceral leishmaniasis were searched using MEDLINE/PubMed and Virtual Health Library. After selection, each included systematic review was assessed, characterized, and categorized by intervention type and by outcomes, according to the methodology offered by the PAHO/WHO Latin American and Caribbean Center on Health Sciences Information (BIREME). The methodological quality was assessed using the AMSTAR2 tool to determine the confidence level of the evidence obtained. Results: Among the prevention and control interventions, insecticide spraying, bednets, dog collars, and dog culling were the most assessed, emphasizing that insecticidal dog collars can reduce visceral leishmaniasis incidence in dogs. Regarding diagnosis, polymerase chain reaction (PCR), rK39 immunochromatographic test (rK39 ICT), and direct agglutination test (DAT) presented high sensitivity and specificity. As for treatment, pentavalent antimonials and amphotericin B were the most analyzed drugs and showed therapeutic success; however, serious adverse events can occur due to their use. The prognostic factors identified were anemia, edema, bleeding, jaundice, age, and HIV coinfection. Conclusions: The evidence map developed shows rK39 ICT and DAT as promising diagnostic alternatives and reinforces the efficacy of liposomal amphotericin B and pentavalent antimonials. Insecticide-impregnated dog collars appear as a promising measure for the control of visceral leishmaniasis, but there is also a need for future studies and reviews with higher methodological quality, especially on prevention and control interventions.

Development of Immunological Assays Based on Leishmania donovani Antigen for Diagnosis of Canine Visceral Leishmaniasis and Their Multicenter Evaluation in Brazil and Italy.

Canine visceral leishmaniasis (CVL) due to Leishmania infantum infection is a zoonotic disease prevalent in the areas of South America and the Mediterranean. Infected dogs as reservoirs can contribute to disease transmission and can be a scourge to public health. Therefore, early diagnosis of infected dogs may play a pivotal role in circumscribing disease progression. Invasive tissue aspiration and insufficient serological methods impair a single assay for prompt CVL diagnosis. In the present study, we aimed to evaluate the potential of Leishmania donovani isolated membrane protein, LAg, for the diagnosis of CVL through immunological assays. Initially, enzyme-linked immunosorbent assay was done with Brazilian dog sera to evaluate the performance of LAg in diagnosing CVL and found sensitivity and specificity of 92.50% and 95%, respectively. The study further confirmed the diagnostic efficacy of LAg in a dipstick format. The dipstick test of canine sera from three centers in Brazil and one center in Italy collectively showed sensitivity values in the range of 53.33% to 100% in recognizing symptomatic dogs and specificity values between 75% and 100% to rule out healthy dogs. Moreover, a rapid immunochromatographic test was developed and optimized using LAg. This test was able to identify 94.73% of CVL of Brazilian origin with specificity of 97.29%. The current results highlight the reactive potential of the L. donovani antigen, LAg, for L. infantum CVL diagnosis and support our previous findings, which suggest the utility of LAg for the diagnosis of both L. donovani and L. infantum human VL in a variety of endemic regions. LAg as a diagnostic candidate may be employed to identify comprehensive CVL cases in epidemiological areas.

Evidence map of diagnosis, treatment, prognosis, prevention, and control in visceral leishmaniasis

[ABSTRACT]. Objective. To develop an evidence map on visceral leishmaniasis prevention, control, diagnosis, treatment, and prognosis. Methods. Systematic reviews on visceral leishmaniasis were searched using MEDLINE/PubMed and Virtual Health Library. After selection, each included systematic review was assessed, characterized, and catego- rized by intervention type and by outcomes, according to the methodology offered by the PAHO/WHO Latin American and Caribbean Center on Health Sciences Information (BIREME). The methodological quality was assessed using the AMSTAR2 tool to determine the confidence level of the evidence obtained. Results. Among the prevention and control interventions, insecticide spraying, bednets, dog collars, and dog culling were the most assessed, emphasizing that insecticidal dog collars can reduce visceral leishmaniasis incidence in dogs. Regarding diagnosis, polymerase chain reaction (PCR), rK39 immunochromatographic test (rK39 ICT), and direct agglutination test (DAT) presented high sensitivity and specificity. As for treatment, pentavalent antimonials and amphotericin B were the most analyzed drugs and showed therapeutic success; however, serious adverse events can occur due to their use. The prognostic factors identified were anemia, edema, bleeding, jaundice, age, and HIV coinfection. Conclusions. The evidence map developed shows rK39 ICT and DAT as promising diagnostic alternatives and reinforces the efficacy of liposomal amphotericin B and pentavalent antimonials. Insecticide-impreg- nated dog collars appear as a promising measure for the control of visceral leishmaniasis, but there is also a need for future studies and reviews with higher methodological quality, especially on prevention and control interventions.

[RESUMEN]. Objetivo. Elaborar un mapa de evidencia sobre la prevención, el control, el diagnóstico, el tratamiento y el pronóstico de la leishmaniasis visceral. Métodos. Se realizaron búsquedas de revisiones sistemáticas sobre la leishmaniasis visceral en MEDLINE/ PubMed y la Biblioteca Virtual en Salud. Tras la selección, cada revisión sistemática incluida fue sometida a evaluación, caracterización y categorización según tipo de intervención y resultados, de acuerdo con la metodología ofrecida por el Centro Latinoamericano y del Caribe de Información en Ciencias de la Salud de la OPS/OMS (BIREME). La calidad metodológica se evaluó con la herramienta AMSTAR2 para determinar el nivel de confianza de la información obtenida. Resultados. Entre las intervenciones de prevención y control, las más evaluadas fueron la fumigación con insecticidas, los mosquiteros, los collares para perros y el sacrificio de perros y se hizo hincapié en que los collares insecticidas para perros pueden reducir la incidencia de leishmaniasis visceral en perros. En cuanto al diagnóstico, la reacción en cadena de la polimerasa (PCR, por su sigla en inglés), la prueba inmunocro- matográfica rK39 (rK39 ICT) y la prueba de aglutinación directa (DAT, por su sigla en inglés) mostraron alta sensibilidad y especificidad. Con respecto al tratamiento, los fármacos más analizados que arrojaron éxito terapéutico fueron los antimoniales pentavalentes y la anfotericina B; sin embargo, su uso puede provocar efectos adversos graves. Los factores pronósticos que se identificaron fueron anemia, edema, sangrado, ictericia, edad y coinfección por el VIH. Conclusiones. El mapa de evidencia elaborado presenta la prueba inmunocromatográfica rK39 y la prueba de aglutinación directa como alternativas diagnósticas prometedoras, y consolida la eficacia de la anfoteri- cina B liposomal y los antimoniales pentavalentes. Los collares de perro impregnados de insecticida parecen ser una medida prometedora para el control de la leishmaniasis visceral, si bien también son necesarios estudios y revisiones adicionales de mayor calidad metodológica, especialmente sobre intervenciones de prevención y control.

[RESUMO]. Objetivo. Desenvolver um mapa de evidências de prevenção, controle, diagnóstico, tratamento e prognóstico da leishmaniose visceral. Métodos. Foram realizadas buscas por revisões sistemáticas sobre leishmaniose visceral no MEDLINE/ PubMed e na Biblioteca Virtual em Saúde. Após a seleção, cada revisão sistemática incluída foi avaliada, caracterizada e classificada por tipo de intervenção e por desfechos, de acordo com a metodologia oferecida pelo Centro Latino-Americano e do Caribe de Informação em Ciências da Saúde (BIREME) da OPAS/OMS. A qualidade metodológica foi avaliada utilizando a ferramenta AMSTAR2 para determinar o nível de confiança das evidências obtidas. Resultados. Entre as intervenções de prevenção e controle, pulverização com inseticida, mosquiteiros, colei- ras para cães e abate de cães foram as mais frequentemente avaliadas, com destaque para as coleiras inseticidas na redução da incidência de leishmaniose visceral em cães. Quanto ao diagnóstico, a reação em cadeia da polimerase (PCR), o teste imunocromatográfico rK39 (rK39 ICT) e o teste de aglutinação direta (DAT, sigla em inglês para direct agglutination test) apresentaram alta sensibilidade e especificidade. Em relação ao tratamento, os antimoniais pentavalentes e a anfotericina B foram os medicamentos mais analisa- dos e demonstraram sucesso terapêutico, embora seu uso possa resultar em eventos adversos graves. Os fatores prognósticos identificados foram anemia, edema, sangramento, icterícia, idade e coinfecção com HIV. Conclusões. O mapa de evidências desenvolvido mostra o rK39 ICT e o DAT como alternativas promisso- ras para o diagnóstico e reforça a eficácia da anfotericina B lipossomal e dos antimoniais pentavalentes. As coleiras impregnadas com inseticida aparecem como medida promissora para o controle da leishmaniose vis- ceral, mas estudos e revisões futuras com mais qualidade metodológica, especialmente sobre intervenções de prevenção e controle, são necessários.

Biomarkers of disease severity in patients with visceral leishmaniasis co-infected with HIV.

Visceral leishmaniasis (VL) is caused by the protozoan Leishmania spp, transmitted by sand fly bites. VL is one of the deadliest tropical infection diseases, yet the coinfection with HIV virus drastically increases relapses, treatment failure and mortality. The concomitant action of these two pathogens leads to high cellular activation independently of the progression to AIDS. In addition, microbial translocation and bacterial infections are thought to contribute worsening the clinical picture. Identifying biomarkers associated with disease severity is of interest for clinical management of patients with VL-HIV/AIDS. Thus, we analyzed in the sera several markers including interleukins (IL-1ß, IL-6, IL-8, and IL-17), interferon-γ (IFN- γ), tumor necrosis factor (TNF), lipopolysaccharide (LPS), soluble CD14 (sCD14), macrophage migration inhibitory factor (MIF) and intestinal fatty acid-binding protein (IFABP). These markers were compared with disease severity in 24 patients with VL/HIV presenting different clinical outcomes. Disease severity was defined by the probability of death calculated using a score set system derived by the Kala-Cal® software. Probability of death ranged from 3.7% to 97.9%, with median of 28.8%. Five patients died (20%). At the univariate analysis, disease severity was correlated with TNF, IFN-γ and sCD14. LPS was positively correlated with sCD14 specifically in patients with low CD4+ count (CD4+ T-cell <200 cells/mL). Most importantly, the multivariate analysis including LPS, CD4+count and sCD14 showed that sCD14 was the only independent predictor for disease severity and death. Altogether, our results indicated that sCD14 is a powerful marker of pathogenicity and death for patients with VL-HIV/AIDS.

C-reactive protein for the diagnosis and prognosis of visceral leishmaniasis caused by Leishmania infantum

Acute-phase reaction (APR) and its marker C-reactive protein (CRP) are heightened in some infectious diseases. APR may contribute to clinical manifestations of systemic inflammation such as hemorrhages, anemia and edema. These symptoms are shared by visceral leishmaniasis (VL), a disease caused by the protozoa Leishmania infantum and L. donovani. The former is zoonotic, hitting mostly children and the immunosuppressed, with higher mortality. As APR and CRP have not been studied in VL caused by L. infantum, we decided to investigate their role as diagnostic and prognostic tools in Brazil. We measured CRP in 136 patients before the treatment of VL and 128 who survived and returned 30 days later and compared it to the clinical presentation, HIV status, and disease severity. Sensitivity for the disease was 97.8% (95%CI: 93.7 - 99.5) and specificity was 85.9% (95%CI: 78.7 - 91.0) with the cut-off of 10mg/L. There was no association of CRP concentration with demographic, clinical and laboratory data. The correlation between pre-and post-treatment levels existed but was poor. Marginal association with the presence of parasites in the bone marrow and death was noticed. The role of APR in the pathogenesis of VL and disease severity remains to be explored. However, the study reveals the significant role of CRP for VL caused by L. infantum and should be routinely required for the diagnosis and follow-up.

Evidence map of diagnosis, treatment, prognosis, prevention, and control in visceral leishmaniasis / Mapa de evidencia sobre el diagnóstico, tratamiento, pronóstico, prevención y control de la leishmaniasis visceral / Mapa de evidências de diagnóstico, tratamento, prognóstico, prevenção e controle da leishmaniose visceral

ABSTRACT Objective. To develop an evidence map on visceral leishmaniasis prevention, control, diagnosis, treatment, and prognosis. Methods. Systematic reviews on visceral leishmaniasis were searched using MEDLINE/PubMed and Virtual Health Library. After selection, each included systematic review was assessed, characterized, and categorized by intervention type and by outcomes, according to the methodology offered by the PAHO/WHO Latin American and Caribbean Center on Health Sciences Information (BIREME). The methodological quality was assessed using the AMSTAR2 tool to determine the confidence level of the evidence obtained. Results. Among the prevention and control interventions, insecticide spraying, bednets, dog collars, and dog culling were the most assessed, emphasizing that insecticidal dog collars can reduce visceral leishmaniasis incidence in dogs. Regarding diagnosis, polymerase chain reaction (PCR), rK39 immunochromatographic test (rK39 ICT), and direct agglutination test (DAT) presented high sensitivity and specificity. As for treatment, pentavalent antimonials and amphotericin B were the most analyzed drugs and showed therapeutic success; however, serious adverse events can occur due to their use. The prognostic factors identified were anemia, edema, bleeding, jaundice, age, and HIV coinfection. Conclusions. The evidence map developed shows rK39 ICT and DAT as promising diagnostic alternatives and reinforces the efficacy of liposomal amphotericin B and pentavalent antimonials. Insecticide-impregnated dog collars appear as a promising measure for the control of visceral leishmaniasis, but there is also a need for future studies and reviews with higher methodological quality, especially on prevention and control interventions.

RESUMEN Objetivo. Elaborar un mapa de evidencia sobre la prevención, el control, el diagnóstico, el tratamiento y el pronóstico de la leishmaniasis visceral. Métodos. Se realizaron búsquedas de revisiones sistemáticas sobre la leishmaniasis visceral en MEDLINE/PubMed y la Biblioteca Virtual en Salud. Tras la selección, cada revisión sistemática incluida fue sometida a evaluación, caracterización y categorización según tipo de intervención y resultados, de acuerdo con la metodología ofrecida por el Centro Latinoamericano y del Caribe de Información en Ciencias de la Salud de la OPS/OMS (BIREME). La calidad metodológica se evaluó con la herramienta AMSTAR2 para determinar el nivel de confianza de la información obtenida. Resultados. Entre las intervenciones de prevención y control, las más evaluadas fueron la fumigación con insecticidas, los mosquiteros, los collares para perros y el sacrificio de perros y se hizo hincapié en que los collares insecticidas para perros pueden reducir la incidencia de leishmaniasis visceral en perros. En cuanto al diagnóstico, la reacción en cadena de la polimerasa (PCR, por su sigla en inglés), la prueba inmunocromatográfica rK39 (rK39 ICT) y la prueba de aglutinación directa (DAT, por su sigla en inglés) mostraron alta sensibilidad y especificidad. Con respecto al tratamiento, los fármacos más analizados que arrojaron éxito terapéutico fueron los antimoniales pentavalentes y la anfotericina B; sin embargo, su uso puede provocar efectos adversos graves. Los factores pronósticos que se identificaron fueron anemia, edema, sangrado, ictericia, edad y coinfección por el VIH. Conclusiones. El mapa de evidencia elaborado presenta la prueba inmunocromatográfica rK39 y la prueba de aglutinación directa como alternativas diagnósticas prometedoras, y consolida la eficacia de la anfotericina B liposomal y los antimoniales pentavalentes. Los collares de perro impregnados de insecticida parecen ser una medida prometedora para el control de la leishmaniasis visceral, si bien también son necesarios estudios y revisiones adicionales de mayor calidad metodológica, especialmente sobre intervenciones de prevención y control.

RESUMO Objetivo. Desenvolver um mapa de evidências de prevenção, controle, diagnóstico, tratamento e prognóstico da leishmaniose visceral. Métodos. Foram realizadas buscas por revisões sistemáticas sobre leishmaniose visceral no MEDLINE/PubMed e na Biblioteca Virtual em Saúde. Após a seleção, cada revisão sistemática incluída foi avaliada, caracterizada e classificada por tipo de intervenção e por desfechos, de acordo com a metodologia oferecida pelo Centro Latino-Americano e do Caribe de Informação em Ciências da Saúde (BIREME) da OPAS/OMS. A qualidade metodológica foi avaliada utilizando a ferramenta AMSTAR2 para determinar o nível de confiança das evidências obtidas. Resultados. Entre as intervenções de prevenção e controle, pulverização com inseticida, mosquiteiros, coleiras para cães e abate de cães foram as mais frequentemente avaliadas, com destaque para as coleiras inseticidas na redução da incidência de leishmaniose visceral em cães. Quanto ao diagnóstico, a reação em cadeia da polimerase (PCR), o teste imunocromatográfico rK39 (rK39 ICT) e o teste de aglutinação direta (DAT, sigla em inglês para direct agglutination test) apresentaram alta sensibilidade e especificidade. Em relação ao tratamento, os antimoniais pentavalentes e a anfotericina B foram os medicamentos mais analisados e demonstraram sucesso terapêutico, embora seu uso possa resultar em eventos adversos graves. Os fatores prognósticos identificados foram anemia, edema, sangramento, icterícia, idade e coinfecção com HIV. Conclusões. O mapa de evidências desenvolvido mostra o rK39 ICT e o DAT como alternativas promissoras para o diagnóstico e reforça a eficácia da anfotericina B lipossomal e dos antimoniais pentavalentes. As coleiras impregnadas com inseticida aparecem como medida promissora para o controle da leishmaniose visceral, mas estudos e revisões futuras com mais qualidade metodológica, especialmente sobre intervenções de prevenção e controle, são necessários.

[Epidemiological profile and time trend in visceral leishmaniasis: Piauí, Brazil, 2008-2018]. / Perfil epidemiológico e tendência temporal da leishmaniose visceral: Piauí, Brasil, 2008 a 2018.

Visceral leishmaniasis (VL) or kala azar is one of the main neglected tropical diseases, of major importance due to the lethal nature of this zoonosis caused by protozoa of genus Leishmania. The study aimed to analyze the time trend in VL cases in the state of Piauí, Brazil, from 2008 to 2018, within the total period from 1971 to 2018. Data were obtained from the Brazilian Information System for Notificable Diseases (SINAN) and the Brazilian Mortality Information System (SIM). For burden of disease analysis, data were collected from the Institute for Health Metrics and Evaluation through the Global Burden of Diseases, Injuries, and Risk Factors (GBD Compare). Prais-Winsten regression analysis was used. A total of 2,374 cases were recorded with probable infection in the state of Piauí and 2,492 cases in residents of the state. The historical series analyzed, from 1971 to 2018, reaffirms the cyclical nature of the disease, based on the identification of epidemic peaks in 1983-1984, 1993-1994, 2003-2004, and 2013-2014. No significant alterations were identified in the trends in incidence, mortality, and case-fatality coefficients in the state. However, there was a significant increase in the incidence coefficient in the Chapada das Mangabeiras Development Territory, with an annual percent change of 17.5%, and a decrease in the Vale do Sambito Development Territory (-18.3%) and Vale do Rio Guaribas Development Territory (-8,1%). VL has a high burden of disease, measured by disability-adjusted life years, and is among the leading neglected tropical diseases in the state of Piauí. The study concludes by recommending the improvement of surveillance and control measures for this disease.

A leishmaniose visceral (LV) ou calazar é uma das principais doenças tropicais negligenciadas, de grande importância devido ao caráter letal desta zoonose causada por protozoários do gênero Leishmania. Objetivou-se analisar a tendência temporal dos casos de LV no Estado do Piauí, Brasil, de 2008 a 2018, inserindo-o dentro do período total de 1971 a 2018. Utilizaram-se dados do Sistema de Informação de Agravos de Notificação (SINAN) e do Sistema de Informações sobre Mortalidade (SIM). Para análise da carga da doença, foram coletados dados do Instituto de Métricas e Avaliação em Saúde através do estudo Carga Global de Doenças, Lesões e Fatores de Risco. Foi empregada a análise de regressão Prais-Winsten. Foram registrados 2.374 casos com provável local de infecção no Piauí e 2.492 casos em residentes do estado. A série histórica analisada, de 1971 a 2018, reafirma o caráter cíclico da doença, pela presença dos picos epidêmicos identificados em 1983 e 1984, 1993 e 1994, 2003 e 2004 e 2013 e 2014. Não foram identificadas alterações significativas na tendência dos coeficientes de incidência, mortalidade e letalidade no estado. No entanto, houve aumento significativo do coeficiente de incidência no Território de Desenvolvimento (TD) Chapada das Mangabeiras (variação percentual anual de 17,5%) e redução no TD Vale do Sambito (-18,3%) e TD Vale do Rio Guaribas (-8,1%). A LV tem a maior carga de doença medida em anos de vida ajustados por deficiência entre as doenças tropicais negligenciadas do estado. Recomenda-se o aprimoramento das medidas de vigilância e controle do agravo.

La leishmaniasis visceral (LV) o kala azar es una de las principales enfermedades tropicales desatendidas de gran importancia, debido al carácter letal de esta zoonosis causada por protozoos del género Leishmania. El objetivo fue analizar la tendencia temporal de los casos de LV en el estado de Piauí, Brasil, de 2008 a 2018, incluyéndola dentro del período total de 1971 a 2018. Se utilizaron datos del Sistema Brasileño de Información de Enfermedades de Notificación (SINAN) y del Sistema de Información sobre Mortalidad (SIM). Para el análisis de la carga de la enfermedad, se recogieron datos del Instituto de Métricas y Evaluación en Salud, a través del estudio Carga Global de Enfermedades, Lesiones y Factores de Riesgo (GBD Compare). Se empleó el análisis de regresión Prais-Winsten. Se registraron 2.374 casos con probable lugar de infección en Piauí y 2492 casos en residentes del estado. La serie histórica analizada, de 1971-2018, reafirma el carácter cíclico de la enfermedad por la presencia de los picos epidémicos, identificados en 1983 y 1984, 1993 y 1994, 2003 y 2004 y 2013 y 2014. No se identificaron alteraciones significativas en la tendencia de los coeficientes de incidencia, mortalidad y letalidad en el estado. No obstante, hubo un aumento significativo del coeficiente de incidencia en el Territorio de Desarrollo (TD) Chapada das Mangabeiras (variación del porcentaje anual de 17,5%) y reducción en el TD Vale do Sambito (-18,3%) y TD Vale do Rio Guaribas (-8,1%). La LV tiene una mayor carga de enfermedad medida en años de vida ajustados por deficiencia entre las enfermedades tropicales olvidadas del estado. Se recomienda el perfeccionamiento de las medidas de vigilancia y control de la enfermedad.

Splenic Transcriptional Responses in Severe Visceral Leishmaniasis: Impaired Leukocyte Chemotaxis and Cell Cycle Arrest.

Structural changes in the spleen have been reported in several infectious diseases. In visceral leishmaniasis (VL), a severe parasitic disease caused by Leishmania spp., the loss of white pulp accompanies a severe clinical presentation. Hamster model reproduces aspects of human VL progression. In the early stages, a transcriptomic signature of leukocyte recruitment was associated with white pulp hyperplasia. Subsequently, impaired leukocyte chemotaxis with loss of T lymphocytes in the periarteriolar lymphoid sheath occurred. This differential gene expression was subsequently corroborated by transcriptomic profiling of spleens in severe human VL. At the latest stage, spleen disorganization was associated with increasing clinical signs of VL. White pulp disruption was accompanied by decreased DLK1 expression. The expression of CXCL13, CCR5, CCL19, CCR6, CCR7 and LTA decreased, likely regulated by CDKN2A overexpression. Our findings enlighten a pathway implying cell cycle arrest and decreased gene expression involved in spleen organization.

Nutritional status and vitamin A and zinc levels in patients with kala-azar in Piauí, Brazil.

INTRODUCTION: Malnutrition and kala-azar (or visceral leishmaniasis) are significant public health problems in different parts of the world. Immunity and susceptibility to infectious and parasitic diseases are directly linked to the host's nutritional state, but little is known about the interaction between nutrition and kala-azar. This study aimed to evaluate nutritional status with kala-azar and correlate these findings with the clinical and laboratory manifestations of the disease, and zinc and retinol levels. METHODS: This was a cross-sectional study of 139 patients with kala-azar. Nutritional status classification was performed according to international recommendations. Parametric or nonparametric tests were applied whenever indicated in a two-sided test with a 5% significance level. RESULTS: Weight loss and malnutrition were more frequent in adults. Body mass index-for-age, fat area of the arm, and upper arm muscle area were significantly associated with probability of death. The presence of human immunodeficiency virus, hepatomegaly, and splenomegaly was correlated with nutritional assessment. Blood leukocyte and lymphocyte, serum creatine, and vitamin A levels were significantly higher in adult men. Vitamin A levels were highly associated with the level of hemoglobin and C-reactive protein (CRP) in multivariate analysis. All patients had reduced plasma zinc levels, but this finding had no association with the outcome variables. CONCLUSIONS: Malnutrition was correlated with severe disease and was more prevalent in older people with kala-azar. Vitamin A deficiency was associated with hemoglobin and CRP. Zinc levels were reduced in patients with kala-azar.

Genetic structure of allopatric populations of Lutzomyia longipalpis sensu lato in Brazil.

Lutzomyia longipalpis sensu lato is a complex of phlebotomine sand fly species, which are widespread in the Neotropics. They have a great medico-veterinary importance due their role as vectors of Leishmania infantum, the causative agent of visceral leishmaniasis. Morphological variations of Lu. longipalpis s.l. males were reported in the late 1960s in Brazil. Male populations can present either one pair of spots on third abdominal tergites or two pairs on third and fourth ones, namely 1S and 2S phenotypes, respectively. Since then, there has been much interest on the taxonomic status of Lu. longipalpis s.l. Thereafter, several lines of evidence have been congruent in suggesting the existence of an uncertain number of cryptic species within Lu. longipalpis s.l. in Brazil. Herein, a 525 bp-fragment of the period gene was used for assessing the genetic structure and phylogenetic relationship of Lu. longipalpis s.l. populations in Brazil. We performed two set of analyses, first we originally sequenced three populations (Passira, Santarém and Teresina) of Lu. longipalpis s.l. and compared them. Thereafter, we performed a global analysis including in our dataset other three pairs of sympatric populations of Lu. longipalpis s.l. from three Brazilian localities available in GenBank. Fixed single nucleotide polymorphisms (SNPs) sharing, maximum likelihood inference, genetic structure and haplotype analyses revealed the presence of two genetic groups, one composed of Teresina population, and the other encompassing Passira and Santarém populations. The global analysis reflected the first of its kind, and two prominent groups were observed: the clade I comprising Teresina 1S, Bodocó 1S, Caririaçu 1S and Sobral 1S; and the clade II encompassing Passira 2S, Santarém 1S, Bodocó 2S, Caririaçu 2S and Sobral 2S. Genetic differentiation data suggested a limited gene flow between populations of the clade I versus clade II. Our results disclosed the presence of two prominent genetic groups, which could reasonably represent populations of Lu. longipalpis s.l. whose males produce the same courtship song.

Influence of sex hormones on the immune response to leishmaniasis.

The differences in morbidity and mortality patterns and life expectancy between the sexes are well established in different infectious and parasitic conditions, such as in leishmaniases, in which biological, genetic, sexual and hormonal variations can modulate the immune response indicating greater infectivity, prevalence and clinical severity in men. In this regard, in seeking the understanding of factors related to protection and susceptibility to infection, this review aimed to discuss the influence of sex hormones on the immune response to leishmaniases. In the literature, sex hormone variations promote differences in the innate, humoral and cell-mediated immune response, leading to greater susceptibility, mortality and complications in males. Epidemiological estimates confirm these results, showing a predominance of the disease, in its different clinical forms, in men and suggesting that sexual variations influence immunomodulatory mechanisms since the prevalence of cases comprises the post-puberty and adulthood period. In this perspective, the action of sex hormones has been investigated in different clinical models, highlighting the potential of testosterone in immunosuppression, given its association with greater susceptibility and poor control of parasite load and the induction of cell apoptosis and attenuation of pro-inflammatory signalling pathways. Therefore, hormonal variations influence the immune response among males and females against leishmaniases, in which androgens may present immunosuppressive potential, while steroids present immunomodulatory characteristics.

Development and Clinical Evaluation of Serum and Urine-Based Lateral Flow Tests for Diagnosis of Human Visceral Leishmaniasis.

Visceral leishmaniasis (VL), a fatal parasitic infection, is categorized as being neglected among tropical diseases. The use of conventional tissue aspiration for diagnosis is not possible in every setting. The immunochromatography-based lateral flow assay (LFA) has attracted attention for a long time due to its ability to give results within a few minutes, mainly in resource-poor settings. In the present study, we optimized and developed the LFA to detect anti-Leishmania antibodies for VL diagnosis. The performance of the developed test was evaluated with serum and urine samples of Indian VL patients and Brazilian sera. The new test exploits well-studied and highly-sensitive purified antigens, LAg isolated from Leishmania donovani promastigotes and protein G conjugated colloidal-gold as a signal reporter. The intensity of the bands depicting the antigen-antibody complex was optimized under different experimental conditions and quantitatively analyzed by the ImageJ software. For the diagnosis of human VL in India, LFA was found to be 96.49% sensitive and 95% specific with serum, and 95.12% sensitive and 96.36% specific with urine samples, respectively. The sensitivity and specificity of LFA were 88.57% and 94.73%, respectively, for the diagnosis of Brazilian VL using patients' sera infected with Leishmania infantum. LFA is rapid and simple to apply, suitable for field usage where results can be interpreted visually and particularly sensitive and specific in the diagnosis of human VL. Serum and urine LFA may improve diagnostic outcomes and could be an alternative for VL diagnosis in settings where tissue aspiration is difficult to perform.