Monitoring the Long-Term Effectiveness of Miltefosine in Indian Post-Kala-Azar Dermal Leishmaniasis.

Post-kala-azar dermal leishmaniasis (PKDL), the dermal sequel to visceral leishmaniasis (VL), is characterized by hypopigmented macules (macular) and/or papules and nodules (polymorphic). Post-kala-azar dermal leishmaniasis plays a significant role in disease transmission, emphasizing the need for monitoring chemotherapeutic effectiveness. Accordingly, this study aimed to quantify the parasite burden in PKDL patients after treatment with miltefosine by a quantitative polymerase chain reaction (qPCR). A Leishmania kinetoplastid gene-targeted qPCR was undertaken using DNA from skin biopsy specimens of patients with PKDL at three time points, i.e., at disease presentation (week 0, n = 157, group 1), upon completion of treatment (week 12, n = 39, group 2), and at any time point 6 months after completion of treatment (week ≥36, n = 54, group 3). A cycle threshold (Ct) <30 was considered the cutoff for positivity, and load was quantified as the number of parasites/µg genomic DNA (gDNA); cure was considered when samples had a Ct >30. The parasite load at disease presentation (group 1) was 10,769 (1,339-80,441)/µg gDNA (median [interquartile range]). In groups 2 and 3, qPCR results were negative in 35/39 cases (89.7%) and 48/54 cases (88.8%), respectively. In the 10/93 (10.8%) qPCR-positive cases, the parasite burdens in groups 2 and 3 were 2,420 (1,205-5,661)/µg gDNA and 22,195 (5,524-100,106)/µg gDNA, respectively. Serial monitoring was undertaken in 45 randomly selected cases that had completed treatment; all cases in groups 2 or 3 had a Ct >30, indicating cure. Overall, qPCR confirmed an 89.2% cure (as 83/93 cases showed parasite clearance), and the persistent qPCR positivity was attributed to nonadherence to treatment or unresponsiveness to miltefosine and remains to be investigated.

Portable smartphone-based molecular test for rapid detection of Leishmania spp.

PURPOSE: Leishmaniasis, caused by the parasite of the genus Leishmania, is a neglected tropical disease which is endemic in more than 60 countries. In South-East Asia, Brazil, and East Africa, it mainly occurs as kala-azar (visceral leishmaniasis, VL), and subsequently as post kala-azar dermal leishmaniasis (PKDL) in a smaller portion of cases. As stated per WHO roadmap, accessibility to accurate diagnostic methods is an essential step to achieve elimination. This study aimed to test the accuracy of a portable minoo device, a small battery-driven, multi-use fluorimeter operating with isothermal technology for molecular diagnosis of VL and PKDL. METHODS: Fluorescence data measured by the device within 20 min are reported back to the mobile application (or app) via Bluetooth and onward via the internet to a backend. This allows anonymous analysis and storage of the test data. The test result is immediately returned to the app displaying it to the user. RESULTS: The limit of detection was 11.2 genome copies (95% CI) as determined by screening a tenfold dilution range of whole Leishmania donovani genomes using isothermal recombinase polymerase amplification (RPA). Pathogens considered for differential diagnosis were tested and no cross-reactivity was observed. For its diagnostic performance, DNA extracted from 170 VL and PKDL cases, comprising peripheral blood samples (VL, n = 96) and skin biopsies (PKDL, n = 74) from India (n = 108) and Bangladesh (n = 62), was screened. Clinical sensitivity and specificity were 88% and 91%, respectively. CONCLUSION: Minoo devices can offer a convenient, cheaper alternative to other molecular diagnostics. Its easy handling makes it ideal for use in low-resource settings to identify parasite burden.

Molecular monitoring of treatment efficacy in human visceral leishmaniasis.

BACKGROUND: Focused efforts of the visceral leishmaniasis elimination program have led to a drastic decline in cases, and the present challenge is disease monitoring, which this study aimed to assess. METHODS: A Leishmania kinetoplastid-targeted qPCR quantified parasite load at disease presentation, and following treatment completion (n=49); an additional 80 cases were monitored after completion of treatment. RESULTS: The parasite load at disease presentation was 13 461.00 (2560.00-37764.00)/µg gDNA, which upon completion of treatment reduced in 47 of 49 cases to 1(1-1)/µg gDNA, p<0.0001. In 80 cases that presented >2 months post-treatment, their parasite burden similarly decreased to 1(1-1)/µg gDNA except in 6 of 80 cases, which were qPCR positive. CONCLUSION: In 129 cases of visceral leishmaniasis, qPCR by quantification of parasite burden proved effective for monitoring treatment.

IFN-λ3 is induced by Leishmania donovani and can inhibit parasite growth in cell line models but not in the mouse model, while it shows a significant association with leishmaniasis in humans.

The intracellular protozoan parasite Leishmania donovani causes debilitating human diseases that involve visceral and dermal manifestations. Type 3 interferons (IFNs), also referred to as lambda IFNs (IFNL, IFN-L, or IFN-λ), are known to play protective roles against intracellular pathogens at the epithelial surfaces. Herein, we show that L. donovani induces IFN-λ3 in human as well as mouse cell line-derived macrophages. Interestingly, IFN-λ3 treatment significantly decreased parasite load in infected cells, mainly by increasing reactive oxygen species production. Microscopic examination showed that IFN-λ3 inhibited uptake but not replication, while the phagocytic ability of the cells was not affected. This was confirmed by experiments that showed that IFN-λ3 could decrease parasite load only when added to the medium at earlier time points, either during or soon after parasite uptake, but had no effect on parasite load when added at 24 h post-infection, suggesting that an early event during parasite uptake was targeted. Furthermore, the parasites could overcome the inhibitory effect of IFN-λ3, which was added at earlier time points, within 2-3 days post-infection. BALB/c mice treated with IFN-λ3 before infection led to a significant increase in expression of IL-4 and ARG1 post-infection in the spleen and liver, respectively, and to different pathological changes, especially in the liver, but not to changes in parasite load. Treatment with IFN-λ3 during infection did not decrease the parasite load in the spleen either. However, IFN-λ3 was significantly increased in the sera of visceral leishmaniasis patients, and the IFNL genetic variant rs12979860 was significantly associated with susceptibility to leishmaniasis.

Inequalities in adequate maternal healthcare opportunities: evidence from Bangladesh Demographic and Health Survey 2017-2018.

OBJECTIVES: This study investigated the inequalities in access to maternal healthcare services in Bangladesh. DESIGN AND SETTING: This study used cross-sectional data from the nationally representative Bangladesh Demographic and Health Survey conducted in 2017-2018. The survey encompassed diverse regions and households across Bangladesh. The study used the Human Opportunity Index (HOI) and Shapley's decomposition technique to measure the inequality in access to maternal healthcare opportunities. PARTICIPANTS: This study included 20 127 women aged 15-49 years. Among them, 5012 women had live births in the preceding 3 years of the survey, forming the study sample. PRIMARY AND SECONDARY OUTCOME MEASURES: This study has no secondary outcome variable. The primary dependent variable is 'adequate maternal healthcare', a dichotomous variable. RESULTS: Household wealth status contributed the highest to inequality in accessing adequate maternal healthcare services (41.4%) such as receiving at least four antenatal care (ANC) visits (39.7%), access to proper ANC (50.7% and 44.0%) and health facility birth (43.4%). Maternal educational status contributes the second highest inequality among all factors in accessing adequate maternal healthcare (29.5%). Adequate maternal healthcare presented the lowest coverage rate and opportunity index among all (approximately 24% with HOI=17.2). CONCLUSIONS: We found that attained adequate maternal healthcare had the lowest coverage and widest dissimilarity, while wealth index, education and place of residence are the major factors that contribute to inequalities in accessibility to maternal healthcare services in Bangladesh. These findings underscore a need for pro-poor interventions to narrow the economic inequalities between the poor and rich in terms of accessibility to maternal healthcare services. The results indicate the need for the Bangladeshi government and its health department to strengthen their commitment to improving female education. Investments should be made in initiatives that facilitate the proximity of maternal healthcare services to women in rural areas.

Evaluation of Recombinase Polymerase Amplification assay for monitoring parasite load in patients with kala-azar and post kala-azar dermal leishmaniasis.

BACKGROUND: The potential reservoirs of visceral leishmaniasis (VL) in South Asia include asymptomatic and relapsed cases of VL, along with patients with post kala-azar dermal leishmaniasis (PKDL). Accordingly, accurate estimation of their parasite load is pivotal for ensuring disease elimination, presently targeted for 2023. Serological tests cannot accurately detect relapses and/or monitor treatment effectiveness, and therefore, parasite antigen/nucleic acid based detection assays remain the only viable option. An excellent option is the quantitative polymerase chain reaction (qPCR) but the high cost, technical expertise and time involved precludes its wider acceptability. Accordingly, the recombinase polymerase amplification (RPA) assay operated in a mobile suitcase laboratory has emerged not simply as a diagnostic tool for leishmaniasis but also to monitor the disease burden. METHODOLOGY/PRINCIPAL FINDINGS: Using total genomic DNA isolated from peripheral blood of confirmed VL cases (n = 40) and lesional biopsies of PKDL cases (n = 64), the kinetoplast-DNA based qPCR and RPA assay was performed and parasite load expressed as Cycle threshold (Ct) and Time threshold (Tt) respectively. Using qPCR as the gold standard, the diagnostic specificity and sensitivity of RPA in naïve cases of VL and PKDL was reiterated. To assess the prognostic potential of the RPA, samples were analyzed immediately at the end of treatment or ≥6 months following completion of treatment. In cases of VL, the RPA assay in terms of cure and detection of a relapse case showed 100% concordance with qPCR. In PKDL following completion of treatment, the overall detection concordance between RPA and qPCR was 92.7% (38/41). At the end of treatment for PKDL, 7 cases remained qPCR positive, whereas RPA was positive in only 4/7 cases, perhaps attributable to their low parasite load. CONCLUSIONS/SIGNIFICANCE: This study endorsed the potential of RPA to evolve as a field applicable, molecular tool for monitoring parasite load, possibly at a point of care level and is worthy of consideration in resource limited settings.

Socioeconomic correlates of Adequate Maternal Care in Bangladesh: Analysis of the Bangladesh Demographic and Health Survey 2017-18.

Introduction: In recent times, Bangladesh has made significant improvements in various health outcomes, but not so much in maternal death. The current flat trend in reducing maternal mortality in Bangladesh has been mainly due to the lower coverage of maternal health care. To improve the coverage, it is essential to find biosocial factors related to adequate maternal health care. Therefore, this study is aimed at finding out the socioeconomic correlates of adequate maternal health care in Bangladesh. Methods: The study used data from the Bangladeshi demographic and health survey 2017-18. The total unweighted sample of 4012 women who reported pregnancy before three years of the survey was analyzed. A composite binary indicator of adequate maternal care has been constructed using the variables-access to maternal care service, four antenatal care visits, at least one visit with qualified providers, and institutional delivery. A binary logistic regression model was employed to find out the socioeconomic correlate of adequate maternal care. Results: Only 24.4% percent of sample women received adequate maternal care. The result of the logistic regression model shows that urban, Khulna, Rajshahi, and Rangpur were associated with an increase in the odds of having adequate maternal care. High education and health care decisions taken by the partner or husband were also associated with an increased odd of adequate maternal care. Islam and lower wealth status were associated with a lower probability of adequate maternal care. Conclusions: Policymakers and health administration should pay attention to the variation in the utilization of maternal health care across residence, region, religion, education, and wealth status to ensure safe motherhood.

Performance Evaluation of Machine Learning Algorithm for Classification of Unintended Pregnancy among Married Women in Bangladesh.

Intended pregnancy is one of the significant indicators of women's well-being. Globally, 74 million women become pregnant every year without planning. Unintended pregnancies account for 28% of all pregnancies among married women in Bangladesh. This study aimed to investigate the performance of six different machine learning (ML) algorithms applied to predict unintended pregnancies among married women in Bangladesh. From BDHS 2017-18, only 1129 pregnant women aged 15-49 were eligible for this study. An independent χ 2 test had performed before we considered six popular ML algorithms, such as logistic regression (LR), random forest (RF), support vector machine (SVM), k-nearest neighbor (KNN), naïve Bayes (NB), and elastic net regression (ENR) to predict the unintended pregnancy. Accuracy, sensitivity, specificity, Cohen's Kappa statistic, and area under curve (AUC) value were used as model evaluation. The bivariate analysis result showed that women aged 30-49 years, poor, not educated, and living in male-headed households had a higher percentage of unintended pregnancy. We found various performance parameters for the classification of unintended pregnancy: LR accuracy = 79.29%, LR AUC = 72.12%; RF accuracy = 77.81%, RF AUC = 72.17%; SVM accuracy = 76.92%, SVM AUC = 70.90%; KNN accuracy = 77.22%, KNN AUC = 70.27%; NB accuracy = 78%, NB AUC = 73.06%; and ENR accuracy = 77.51%, ENR AUC = 74.67%. Based on the AUC value, we can conclude that of all the ML algorithms we investigated, the ENR algorithm provides the most accurate classification for predicting unwanted pregnancy among Bangladeshi women. Our findings contribute to a better understanding of how to categorize pregnancy intentions among Bangladeshi women. As a result, the government can initiate an effective campaign to raise contraception awareness.