Clofazimine-induced cutaneous hyperpigmentation as a source of stigma in the treatment of leprosy: A cross-sectional study.

OBJECTIVES: Cutaneous hyperpigmentation is one of the main adverse effects encountered in patients undergoing leprosy treatment with multidrug therapy (WHO-MDT). This adverse effect has been described as intolerable and capable of contributing to social stigma. The objectives of this study were to quantify the variation in skin colour induced by clofazimine during and after treatment and to assess the related stigma. METHODS: This observational cross-sectional study objectively measured skin colour in 51 patients by reading the individual typology angle (ITA°) with a spectrophotometer, followed by the application of the Stigma Scale of the Explanatory Model Interview Catalogue (EMIC). RESULTS: Skin hyperpigmentation was observed in 100% of the individuals. They showed more negative ITA° values in lesion areas than non-lesion areas, particularly in sun-exposed regions. Clofazimine-induced cutaneous hyperpigmentation was not homogeneous and seemed to follow the lesion locations. The mean EMIC score was 18.8 points. CONCLUSION: All patients presented skin hyperpigmentation caused by clofazimine, detectable through spectrophotometry. Hyperpigmentation strongly impacted the social domain, indicating the intersectionality of disease and skin colour stigma, contributing to the social isolation of these patients. Health authorities should consider the negative impact of clofazimine on treatment adherence.

MicroRNA biomarkers in leprosy: insights from the Northern Brazilian Amazon population and their implications in disease immune-physiopathology.

Leprosy, or Hansen's Disease, is a chronic infectious disease caused by Mycobacterium leprae that affects millions of people worldwide. Despite persistent efforts to combat it leprosy remains a significant public health concern particularly in developing countries. The underlying pathophysiology of the disease is not yet fully understood hindering the development of effective treatment strategies. However, recent studies have shed light on the potential role of microRNAs (miRNAs), small non-coding RNA molecules that can regulate gene expression, as promising biomarkers in various disease, including leprosy. This study aimed to validate a set of nine circulating miRNAs to propose new biomarkers for early diagnosis of the disease. Hsa-miR-16-5p, hsa-miR-106b-5p, hsa-miR-1291, hsa-miR-144-5p, and hsa-miR-20a-5p showed significant differential expression between non-leprosy group (non-LP) and leprosy group (LP), accurately discriminating between them (AUC > 0.75). In addition, our study revealed gender-based differences in miRNA expression in LP. Notably, hsa-miR-1291 showed higher expression in male LP, suggesting its potential as a male-specific biomarker. Similarly, hsa-miR-16-5p and hsa-miR-20a-5p displayed elevated expression in female LP, indicating their potential as female-specific biomarkers. Additionally, several studied miRNAs are involved in the dysregulation of apoptosis, autophagy, mitophagy, cell cycle, and immune system in leprosy. In conclusion, the validation of miRNA expression highlights several miRNAs as potential biomarkers for early diagnosis and provides new insights into the pathogenesis of the disease.

Specialized active leprosy search strategies in an endemic area of the Brazilian Amazon identifies a hypermutated Mycobacterium leprae strain causing primary drug resistance.

Introduction: Leprosy, an infectious disease caused by Mycobacterium leprae, remains a public health concern in endemic countries, particularly in Brazil. In this study, we conducted an active surveillance campaign in the hyperendemic city of Castanhal in the northeastern part of the state of Pará using clinical signs and symptoms combined with serological and molecular tools to diagnose new cases and to identify drug resistance of circulating M. leprae strains and their distribution in the community. Methods: During an active surveillance of one week, we enrolled 318 individuals using three different strategies to enroll subjects for this study: (i) an active survey of previously treated cases from 2006 to 2016 found in the Brazil National Notifiable Disease Information System database (n = 23) and their healthy household contacts (HHC) (n = 57); (ii) an active survey of school children (SC) from two primary public schools in low-income neighborhoods (n = 178), followed by visits to the houses of these newly diagnosed SC (n = 7) to examine their HHC (n = 34) where we diagnosed additional new cases (n = 6); (iii) and those people who spontaneously presented themselves to our team or the local health center with clinical signs and/or symptoms of leprosy (n = 6) with subsequent follow-up of their HHC when the case was confirmed (n = 20) where we diagnosed two additional cases (n = 2). Individuals received a dermato-neurological examination, 5 ml of peripheral blood was collected to assess the anti-PGL-I titer by ELISA and intradermal earlobe skin scrapings were taken from HHC and cases for amplification of the M. leprae RLEP region by qPCR. Results: Anti-PGL-I positivity was highest in the new leprosy case group (52%) followed by the treated group (40.9%), HHC (40%) and lowest in SC (24.6%). RLEP qPCR from SSS was performed on 124 individuals, 22 in treated cases, 24 in newly diagnosed leprosy cases, and 78 in HHC. We detected 29.0% (36/124) positivity overall in this sample set. The positivity in treated cases was 31.8% (7/22), while in newly diagnosed leprosy cases the number of positives were higher, 45.8% (11/23) and lower in HHC at 23.7% (18/76). Whole genome sequencing of M. leprae from biopsies of three infected individuals from one extended family revealed a hypermutated M. leprae strain in an unusual case of primary drug resistance while the other two strains were drug sensitive. Discussion: This study represents the extent of leprosy in an active surveillance campaign during a single week in the city of Castanhal, a city that we have previously surveyed several times during the past ten years. Our results indicate the continuing high transmission of leprosy that includes fairly high rates of new cases detected in children indicating recent spread by multiple foci of infection in the community. An unusual case of a hypermutated M. leprae strain in a case of primary drug resistance was discovered. It also revealed a high hidden prevalence of overt disease and subclinical infection that remains a challenge for correct clinical diagnosis by signs and symptoms that may be aided using adjunct laboratory tests, such as RLEP qPCR and anti-PGL-I serology.

Point-of-care ultrasound of peripheral nerves in the diagnosis of Hansen's disease neuropathy.

Introduction: Hansen's disease (HD) is the most common cause of treatable peripheral neuropathy in the world that may or may not involve skin manifestations, and physical examination based on simplified neurologic evaluation is a subjective and inaccurate procedure. High-resolution ultrasound (HRUS) can be used to evaluate peripheral nerves and is a validated technique of good reproducibility, permitting a detailed and precise examination. Objectives: We proposed to establish objective criteria for absolute values of the measurement of the CSA of peripheral nerves and their indices of the ΔCSA and ΔTpT in the diagnosis of Hansen's disease neuropathy as compared with healthy voluntaries. Materials and methods: In municipalities from different regions of Brazil, we randomly selected 234 volunteer Brazilian patients diagnosed with leprosy to be submitted to peripheral nerve echography and compared with 49 healthy Brazilian volunteers. Results: Hansen Disease assessed by high resolution ultrasound is a primarily neural disease that leads to multiple hypertrophic mononeuropathy characterized by CSA values exceeding normal limits (Med CT = 10.2 mm2; UT = 9.8 mm2; UPT = 9.3 mm2; CFFH = 18.3 mm2; T = 9.6 mm2), and the pattern of asymmetry (ΔCSA>2.5 mm2 with RR 13) and focality (ΔTPT > 2.5 mm2 with RR 6.4) of this thickening has higher sensitivity (76,1%) and specificity (87,8 %) for its early diagnosis that laboratory tests. Analyzing each subject, the percentage of thickened nerves detected among the total number of nerves assessed was higher among patients with HD than among healthy individuals (p < 0.0001). Individuals with two or more thickened nerves were at 24.1 times higher relative risk (95% CI: 6.74-88.98) of HD.

Silent peripheral neuropathy determined by high-resolution ultrasound among contacts of patients with Hansen's disease.

Introduction: Hansen's disease (HD) primarily infects peripheral nerves, with patients without HD being free of peripheral nerve damage. Household contacts (HHCs) of patients with HD are at a 5-10 times higher risk of HD than the general population. Neural thickening is one of the three cardinal signs that define a case of HD according to WHO guidelines, exclusively considering palpation examination that is subjective and may not detect the condition in the earliest cases even when performed by well-trained professionals. High-resolution ultrasound (HRUS) can evaluate most peripheral nerves, a validated technique with good reproducibility allowing detailed and accurate examination. Objective: This study aimed to use the peripheral nerve HRUS test according to the HD protocol as a diagnostic method for neuropathy comparing HHCs with healthy volunteers (HVs) and patients with HD. Methods: In municipalities from 14 different areas of Brazil we selected at random 83 HHC of MB-patients to be submitted to peripheral nerve ultrasound and compared to 49 HVs and 176 HD-patients. Results: Household contacts assessed by HRUS showed higher median and mean absolute peripheral nerve cross-sectional area (CSA) values and greater asymmetries (ΔCSA) compared to HVs at the same points. Median and mean absolute peripheral nerve CSA values were higher in patients with HD compared to HCCs at almost all points, while ΔCSA values were equal at all points. Mean ± SD focality (ΔTpT) values for HHCs and patients with HD, respectively, were 2.7 ± 2.2/2.6 ± 2.2 for the median nerve, 2.9 ± 2.7/3.3 ± 2.9 for the common fibular nerve (p > 0.05), and 1.3 ± 1.3/2.2 ± 3.9 for the ulnar nerve (p < 0.0001). Discussion: Considering HRUS findings for HHCs, asymmetric multiple mononeuropathy signs (thickening or asymmetry) in at least 20% of the nerves evaluated could already indicates evidence of HD neuropathy. Thus, if more nerve points are assessed in HHCs (14 instead of 10), the contacts become more like patients with HD according to nerve thickening determined by HRUS, which should be a cutting-edge tool for an early diagnosis of leprosy cases.

[Combined use of serologic markers and spatial analysis for epidemiological surveillance of leprosyUso conjunto de los marcadores serológicos y del análisis espacial en la vigilancia epidemiológica de la lepra]. / Uso combinado de marcadores sorológicos e análise espacial na vigilância epidemiológica da hanseníase.

OBJECTIVE: To evaluate the combined use of serologic markers and spatial analysis to increase the sensitivity of leprosy epidemiological surveillance. METHOD: This cross-sectional study was performed with neighbors of leprosy cases and neighbors and family members of schoolchildren with a positive anti-phenolic glycolipid I (PGL-I) test in Diamantina, state of Minas Gerais, Brazil. Neighbors were those living within a 100-m radius of schoolchildren or leprosy cases. Data collection included a semi-structured interview, dermatologic examination, and rapid ML Flow test. All residential addresses were georeferenced. Multivariate regression and spatial analysis were performed with anti-PGL-I seropositivity as the dependent variable. RESULTS: The study included 1 491 individuals: 1 009 (67.7%) family members and neighbors of schoolchildren with positive serology test and 482 (32.3%) neighbors of leprosy cases. Of the total, 421 (28.2%) were positive for anti-PGL-I. Seropositivity was higher among family members and neighbors of seropositive schoolers (P < 0.001), among people with family income of 1 minimum wage (P < 0.001), among the youngest participants (P < 0.001), and among those living in homes with one to five rooms (P = 0.007). The seropositivity rate was higher in the geographic area corresponding to seropositive schoolchildren (P < 0.001), that is, the spot with the highest number of cases was different from the spot with the highest rate of seropositivity. CONCLUSIONS: The combined use of serologic markers and spatial analysis allowed us to easily identify operational weaknesses of services and a possible occult leprosy endemism in the municipality's urban census tracts. Tracing of social contact and neighbors, active search, educational campaigns, school surveys, and territorial analyses facilitate the early diagnosis of leprosy.

OBJETIVO: Evaluar el uso conjunto de los marcadores serológicos y del análisis espacial para ampliar la sensibilidad de la vigilancia epidemiológica de la lepra. MÉTODO: Este estudio transversal se realizó con vecinos de personas con casos de lepra y con familiares y vecinos de escolares con resultados positivos en las pruebas serológicas del antígeno glicolípido fenólico I (PGL-I) en Diamantina (Minas Gerais, Brasil). Se definieron como vecinos las personas residentes en un radio de hasta 100 metros de distancia de los escolares y de las personas con casos de lepra. Para la recopilación de datos se realizaron entrevistas semiestructuradas, exámenes dermatoneurológicos y pruebas serológicas rápidas ML Flow. Todas las direcciones se ubicaron por georreferencia. Se realizó una regresión multivariante y un análisis espacial, y se empleó la reactividad serológica anti-PGL-I como variable dependiente. RESULTADOS: Se estudiaron 1 491 personas: 1 009 (67,7%) familiares y vecinos de escolares con resultados positivos en las pruebas serológicas y 482 (32,3%) vecinos de las personas con casos de lepra. Del total, 421 (28,2%) presentaron seropositividad anti-PGL-I. La posibilidad de presentar seropositividad fue mayor en los familiares y vecinos de los escolares seropositivos (P < 0,001), en las personas con ingresos familiares de un salario mínimo (P < 0,001), en las personas más jóvenes (P < 0,001) y en los residentes en domicilios con una a cinco habitaciones (P = 0,007). La tasa de seropositividad fue mayor en la zona geográfica correspondiente a los escolares seropositivos (P < 0,001), por lo que hubo divergencia entre el foco de mayor concentración de casos y el de mayor seropositividad. CONCLUSIONES: El uso conjunto de los marcadores serológicos y del análisis espacial permitió determinar las fragilidades operativas de los servicios y una posible endemia oculta de lepra en los sectores urbanos del municipio incluidos en el censo correspondiente. Las actividades de rastreo de contactos sociales y cercanos, la búsqueda activa de casos, las campañas educativas, las encuestas escolares y el análisis del territorio facilitan el diagnóstico temprano de la lepra.

Uso combinado de marcadores sorológicos e análise espacial na vigilância epidemiológica da hanseníase

[RESUMO]. Objetivo. Avaliar o uso combinado de marcadores sorológicos e análise espacial para ampliar a sensibilidade da vigilância epidemiológica da hanseníase. Método. Este estudo transversal foi realizado com vizinhos de casos de hanseníase e familiares e vizinhos de escolares com sorologia positiva anti-glicolipídeo fenólico I (PGL-I) em Diamantina, Minas Gerais, Brasil. Definiram- se como vizinhos as pessoas que residiam em um raio de até 100 metros de escolares e de casos de hanseníase. Para a coleta de dados, foram realizados entrevista semiestruturada, exame dermatoneurológico e teste sorológico rápido ML Flow. Todos os endereços foram georreferenciados. Foram realizadas regressão multivariada e análise espacial, tendo a sororreatividade anti-PGL-I como variável dependente. Resultados. Foram estudadas 1 491 pessoas: 1 009 (67,7%) familiares e vizinhos dos escolares com sorologia positiva e 482 (32,3%) vizinhos dos casos de hanseníase. Do total, 421 (28,2%) apresentaram soropositividade anti-PGL-I. A chance de soropositividade foi maior entre familiares e vizinhos dos escolares soropositivos (P < 0,001), entre pessoas com renda familiar de 1 salário-mínimo (P < 0,001), entre os mais jovens (P < 0,001) e entre os que residiam em domicílios com um a cinco cômodos (P = 0,007). A taxa de soropositividade foi maior em área geográfica correspondente aos escolares soropositivos (P < 0,001), ou seja, houve divergência entre o foco de maior concentração de casos e o de maior soropositividade. Conclusões. O uso combinado de marcadores sorológicos e análise espacial possibilitou identificar fragilidades operacionais dos serviços e uma possível endemia oculta de hanseníase nos setores censitários urbanos do município. Atividades de rastreamento de contatos sociais e vizinhos, busca ativa, campanhas educativas, inquéritos escolares e análise do território facilitam o diagnóstico precoce da hanseníase.

ABSTRACT Objective. To evaluate the combined use of serologic markers and spatial analysis to increase the sensitivity of leprosy epidemiological surveillance. Method. This cross-sectional study was performed with neighbors of leprosy cases and neighbors and family members of schoolchildren with a positive anti-phenolic glycolipid I (PGL-I) test in Diamantina, state of Minas Gerais, Brazil. Neighbors were those living within a 100-m radius of schoolchildren or leprosy cases. Data collection included a semi-structured interview, dermatologic examination, and rapid ML Flow test. All residential addresses were georeferenced. Multivariate regression and spatial analysis were performed with anti-PGL-I seropositivity as the dependent variable. Results. The study included 1 491 individuals: 1 009 (67.7%) family members and neighbors of schoolchildren with positive serology test and 482 (32.3%) neighbors of leprosy cases. Of the total, 421 (28.2%) were positive for anti-PGL-I. Seropositivity was higher among family members and neighbors of seropositive schoolers (P < 0.001), among people with family income of 1 minimum wage (P < 0.001), among the youngest participants (P < 0.001), and among those living in homes with one to five rooms (P = 0.007). The seropositivity rate was higher in the geographic area corresponding to seropositive schoolchildren (P < 0.001), that is, the spot with the highest number of cases was different from the spot with the highest rate of seropositivity. Conclusions. The combined use of serologic markers and spatial analysis allowed us to easily identify operational weaknesses of services and a possible occult leprosy endemism in the municipality’s urban census tracts. Tracing of social contact and neighbors, active search, educational campaigns, school surveys, and territorial analyses facilitate the early diagnosis of leprosy.

[RESUMEN]. Objetivo. Evaluar el uso conjunto de los marcadores serológicos y del análisis espacial para ampliar la sensibilidad de la vigilancia epidemiológica de la lepra. Método. Este estudio transversal se realizó con vecinos de personas con casos de lepra y con familiares y vecinos de escolares con resultados positivos en las pruebas serológicas del antígeno glicolípido fenólico I (PGL-I) en Diamantina (Minas Gerais, Brasil). Se definieron como vecinos las personas residentes en un radio de hasta 100 metros de distancia de los escolares y de las personas con casos de lepra. Para la recopilación de datos se realizaron entrevistas semiestructuradas, exámenes dermatoneurológicos y pruebas serológicas rápidas ML Flow. Todas las direcciones se ubicaron por georreferencia. Se realizó una regresión multivariante y un análisis espacial, y se empleó la reactividad serológica anti-PGL-I como variable dependiente. Resultados. Se estudiaron 1 491 personas: 1 009 (67,7%) familiares y vecinos de escolares con resultados positivos en las pruebas serológicas y 482 (32,3%) vecinos de las personas con casos de lepra. Del total, 421 (28,2%) presentaron seropositividad anti-PGL-I. La posibilidad de presentar seropositividad fue mayor en los familiares y vecinos de los escolares seropositivos (P < 0,001), en las personas con ingresos familiares de un salario mínimo (P < 0,001), en las personas más jóvenes (P < 0,001) y en los residentes en domicilios con una a cinco habitaciones (P = 0,007). La tasa de seropositividad fue mayor en la zona geográfica correspondiente a los escolares seropositivos (P < 0,001), por lo que hubo divergencia entre el foco de mayor concentración de casos y el de mayor seropositividad. Conclusiones. El uso conjunto de los marcadores serológicos y del análisis espacial permitió determinar las fragilidades operativas de los servicios y una posible endemia oculta de lepra en los sectores urbanos del municipio incluidos en el censo correspondiente. Las actividades de rastreo de contactos sociales y cercanos, la búsqueda activa de casos, las campañas educativas, las encuestas escolares y el análisis del territorio facilitan el diagnóstico temprano de la lepra.

Active search strategies, clinicoimmunobiological determinants and training for implementation research confirm hidden endemic leprosy in inner São Paulo, Brazil.

BACKGROUND: This study evaluates implementation strategies for leprosy diagnosis based on responses to a Leprosy Suspicion Questionnaire (LSQ), and analyzes immunoepidemiological aspects and follow-up of individuals living in a presumptively nonendemic area in Brazil. METHODOLOGY/PRINCIPAL FINDINGS: Quasi-experimental study based on LSQ throughout Jardinópolis town by community health agents, theoretical-practical trainings for primary care teams, dermatoneurological examination, anti-PGL-I serology, RLEP-PCR, and spatial epidemiology. A Leprosy Group (LG, n = 64) and Non-Leprosy Group (NLG, n = 415) were established. Overall, 3,241 LSQs were distributed; 1,054 (32.5%) LSQ were positive for signs/symptoms (LSQ+). Among LSQ+ respondents, Q2-Tingling (pricking)? (11.8%); Q4-Spots on the skin? (11.7%); Q7-Pain in the nerves? (11.6%); Q1-Numbness in your hands and/or feet? (10.7%) and Q8-Swelling of hands and feet? (8.5%) were most frequently reported symptoms. We evaluated 479 (14.8%) individuals and diagnosed 64 new cases, a general new case detection rate (NCDR) of 13.4%; 60 were among 300 LSQ+ (NCDR-20%), while 4 were among 179 LSQ negative (NCDR-2.23%). In LG, Q7(65%), Q2(60%), Q1(45%), Q4(40%) and Q8(25%) were most frequent. All 2x2 crossings of these 5 questions showed a relative risk for leprosy ranging from 3 to 5.8 compared with NLG. All patients were multibacillary and presented hypochromatic macules with loss of sensation. LG anti-PGL-I titers were higher than NLG, while 8.9% were positive for RLEP-PCR. The leprosy cases and anti-PGL-I spatial mappings demonstrated the disease spread across the town. CONCLUSIONS/SIGNIFICANCE: Implementation actions, primarily LSQ administration focused on neurological symptoms, indicate hidden endemic leprosy in a nonendemic Brazilian state.

Latent leprosy infection identified by dual RLEP and anti-PGL-I positivity: Implications for new control strategies.

The number of new cases of leprosy reported worldwide has remained essentially unchanged for the last decade despite continued global use of free multidrug therapy (MDT) provided to any diagnosed leprosy patient. In order to more effectively interrupt the chain of transmission, new strategies will be required to detect those with latent disease who contribute to furthering transmission. To improve the ability to diagnose leprosy earlier in asymptomatic infected individuals, we examined the combined use of two well-known biomarkers of M. leprae infection, namely the presence of M. leprae DNA by PCR from earlobe slit skin smears (SSS) and positive antibody titers to the M. leprae-specific antigen, Phenolic Glycolipid I (anti-PGL-I) from leprosy patients and household contacts living in seven hyperendemic cities in the northern state of Pará, Brazilian Amazon. Combining both tests increased sensitivity, specificity and accuracy over either test alone. A total of 466 individuals were evaluated, including 87 newly diagnosed leprosy patients, 52 post-treated patients, 296 household contacts and 31 healthy endemic controls. The highest frequency of double positives (PGL-I+/RLEP+) were detected in the new case group (40/87, 46%) with lower numbers for treated (12/52, 23.1%), household contacts (46/296, 15.5%) and healthy endemic controls (0/31, 0%). The frequencies in these groups were reversed for double negatives (PGL-I-/RLEP-) for new cases (6/87, 6.9%), treated leprosy cases (15/52, 28.8%) and the highest in household contacts (108/296, 36.5%) and healthy endemic controls (24/31, 77.4%). The data strongly suggest that household contacts that are double positive have latent disease, are likely contributing to shedding and transmission of disease to their close contacts and are at the highest risk of progressing to clinical disease. Proposed strategies to reduce leprosy transmission in highly endemic areas may include chemoprophylactic treatment of this group of individuals to stop the spread of bacilli to eventually lower new case detection rates in these areas.

Leprosy piRnome: exploring new possibilities for an old disease.

Leprosy, which is caused by the human pathogen Mycobacterium leprae, causes nerve damage, deformity and disability in over 200,000 people every year. Because of the long doubling time of M. leprae (13 days) and the delayed onset of detectable symptoms, which is estimated to be approximately 3-7 years after infection, there is always a large percentage of subclinically infected individuals in the population who will eventually develop the disease, mainly in endemic countries. piRNAs comprise the largest group of small noncoding RNAs found in humans, and they are distinct from microRNAs (miRNAs) and small interfering RNAs (siRNAs). piRNAs function in transposon silencing, epigenetic regulation, and germline development. The functional role of piRNAs and their associated PIWI proteins have started to emerge in the development of human cancers and viral infections, but their relevance to bacterial diseases has not been investigated. The present study reports the piRNome of human skin, revealing that all but one of the piRNAs examined are downregulated in leprosy skin lesions. Considering that one of the best characterized functions of piRNAs in humans is posttranscriptional mRNA silencing, their functions are similar to what we have described for miRNAs, including acting on apoptosis, M. leprae recognition and engulfment, Schwann cell (SC) demyelination, epithelial-mesenchymal transition (EMT), loss of sensation and neuropathic pain. In addition to new findings on leprosy physiopathology, the discovery of relevant piRNAs involved in disease processes in human skin may provide new clues for therapeutic targets, specifically to control nerve damage, a prominent feature of leprosy that has no currently available pharmaceutical treatment.

Prevalência e distribuição espacial da infecção pelo Mycobacterium leprae em município de média endemicidade / Prevalence and spatial distribution of Mycobacterium leprae infection in a medium endemicity municipality

Objetivo: analisar o perfil sorológico e a distribuição espacial da infecção e do adoecimento pelo Mycobacterium leprae. Métodos: estudo transversal, conduzido com casos de hanseníase (n=42), contatos intradomiciliares (n=81) e escolares (n=358). Realizados entrevista, exame dermatoneurológico, sorológico e georreferenciamento dos endereços. Resultados: dos casos soropositivos, todos residiam em zona rural, com mais de duas pessoas no domicílio, 50,0% apresentavam alguma alteração de força nos pés. Dos contatos soropositivos, 75,0% eram idosos e casados. Dentre os escolares, 75,0% eram do sexo feminino, 69,4% residiam com mais de quatro pessoas e 80,6% dormiam no quarto com alguém. Foi possível observar áreas hiperendêmicas de casos no munícipio estudado e relação espaço-temporal entre eles (p=0,010). Além disso, concentração de casos e escolares em setores de menor renda familiar per capita. Conclusão: a análise sorológica revelou que a condição de convivência se relacionou com a infecção pelo Mycobacterium leprae, e a espacial demonstrou cenário de endemia oculta. (AU)

miRNome Expression Analysis Reveals New Players on Leprosy Immune Physiopathology.

Leprosy remains as a public health problem and its physiopathology is still not fully understood. MicroRNAs (miRNA) are small RNA non-coding that can interfere with mRNA to regulate gene expression. A few studies using DNA chip microarrays have explored the expression of miRNA in leprosy patients using a predetermined set of genes as targets, providing interesting findings regarding the regulation of immune genes. However, using a predetermined set of genes restricted the possibility of finding new miRNAs that might be involved in different mechanisms of disease. Thus, we examined the miRNome of tuberculoid (TT) and lepromatous (LL) patients using both blood and lesional biopsies from classical leprosy patients (LP) who visited the Dr. Marcello Candia Reference Unit in Sanitary Dermatology in the State of Pará and compared them with healthy subjects. Using a set of tools to correlate significantly differentially expressed miRNAs with their gene targets, we identified possible interactions and networks of miRNAs that might be involved in leprosy immunophysiopathology. Using this approach, we showed that the leprosy miRNA profile in blood is distinct from that in lesional skin as well as that four main groups of genes are the targets of leprosy miRNA: (1) recognition and phagocytosis, with activation of immune effector cells, where the immunosuppressant profile of LL and immunoresponsive profile of TT are clearly affected by miRNA expression; (2) apoptosis, with supportive data for an antiapoptotic leprosy profile based on BCL2, MCL1, and CASP8 expression; (3) Schwann cells (SCs), demyelination and epithelial-mesenchymal transition (EMT), supporting a role for different developmental or differentiation gene families, such as Sox, Zeb, and Hox; and (4) loss of sensation and neuropathic pain, revealing that RHOA, ROCK1, SIGMAR1, and aquaporin-1 (AQP1) may be involved in the loss of sensation or leprosy pain, indicating possible new therapeutic targets. Additionally, AQP1 may also be involved in skin dryness and loss of elasticity, which are well known signs of leprosy but with unrecognized physiopathology. In sum, miRNA expression reveals new aspects of leprosy immunophysiopathology, especially on the regulation of the immune system, apoptosis, SC demyelination, EMT, and neuropathic pain.

Evidence of hidden leprosy in a supposedly low endemic area of Brazil.

OBJECTIVES: Show that hidden endemic leprosy exists in a municipality of inner São Paulo state (Brazil) with active surveillance actions based on clinical and immunological evaluations. METHODS: The study sample was composed by people randomly selected by a dermatologist during medical care in the public emergency department and by active surveillance carried out during two days at a mobile clinic. All subjects received a dermato-neurological examination and blood sampling to determine anti-PGL-I antibody titers by enzyme-linked immunosorbent assay (ELISA). RESULTS: From July to December 2015, 24 new cases of leprosy were diagnosed; all were classified as multibacillary (MB) leprosy, one with severe Lucio's phenomenon. Seventeen (75%) were found with grade-1 or 2 disability at the moment of diagnosis. Anti-PGL-I titer was positive in 31/133 (23.3%) individuals, only 6/24 (25%) were positive in newly diagnosed leprosy cases. CONCLUSIONS: During the last ten years before this study, the average new case detection rate (NCDR) in this town was 2.62/100,000 population. After our work, the NCDR was raised to 42.8/100,000. These results indicate a very high number of hidden leprosy cases in this supposedly low endemic area of Brazil.

Evidence of hidden leprosy in a supposedly low endemic area of Brazil

OBJECTIVES Show that hidden endemic leprosy exists in a municipality of inner São Paulo state (Brazil) with active surveillance actions based on clinical and immunological evaluations. METHODS The study sample was composed by people randomly selected by a dermatologist during medical care in the public emergency department and by active surveillance carried out during two days at a mobile clinic. All subjects received a dermato-neurological examination and blood sampling to determine anti-PGL-I antibody titers by enzyme-linked immunosorbent assay (ELISA). RESULTS From July to December 2015, 24 new cases of leprosy were diagnosed; all were classified as multibacillary (MB) leprosy, one with severe Lucio's phenomenon. Seventeen (75%) were found with grade-1 or 2 disability at the moment of diagnosis. Anti-PGL-I titer was positive in 31/133 (23.3%) individuals, only 6/24 (25%) were positive in newly diagnosed leprosy cases. CONCLUSIONS During the last ten years before this study, the average new case detection rate (NCDR) in this town was 2.62/100,000 population. After our work, the NCDR was raised to 42.8/100,000. These results indicate a very high number of hidden leprosy cases in this supposedly low endemic area of Brazil.

Leprosy in Children.

PURPOSE OF REVIEW: This manuscript aims to review the cutting-edge developments regarding to the diagnosis, management, and prevention of leprosy in children. RECENT FINDINGS: Leprosy transmission still occurs continuously in some endemic areas in the world. Leprosy in children below 15 years old is a robust indicator of active source of infection in the community where they live. A special focus on children to reduce disabilities and reduce transmission is one of the core areas of interventions of the global leprosy strategy 2016-2020. Ongoing research is trying to develop better diagnostic tests and to advance chemoprophylaxis and immunoprophylaxis approaches. Early diagnosis in children can be hard because of the wide range of clinical aspects of the skin lesions and mainly due to the difficulty of performing the clinical peripheral nerve evaluation. We must maintain leprosy expertise and improve the health professionals training for leprosy diagnosis, since we still have a long journey to reach leprosy elimination.

Estratégia saúde da família e hanseníase: análise territorial / Family health strategy and leprosy: a territorial analysis

OBJETIVOS: Correlacionar a cobertura territorial da Estratégia Saúde da Família (ESF) com a distribuição espacial dos casos de hanseníase em um município hiperendêmico do Norte do País. MÉTODOS: Foram mapeados os territórios da ESF e os casos de Hanseníase do período de 2004 a 2010. Utilizou-se um Sistema de Informações Geográficas(SIG) para visualizar a distribuição espacial dos casos de hanseníase em relação à cobertura da ESF do município de Castanhal-PA. RESULTADOS: Foram desenvolvidos os shapes de cobertura da ESF (aproximadamente 76,38% da população é atendida pelo programa) e de distribuição dos casos de hanseníase (aproximadamente 81,62% destes casos estão dentro da área de atuação da ESF no município de estudo). CONCLUSÃO: Através desta técnica, foram identificadas as áreas descobertas pela ESF, bem como a distribuição de casos de Hanseníase.

OBJECTIVES: Correlate the territorial coverage of Family Health Strategy (FHS) with the spatial distribution of the leprosy cases in a hiperendemic municipality of North Region of the Country. METHODS: We mapped the FHS territories and the leprosy cases in the period from 2004 to 2010. We used a Geographic Information System (GIS) to visualize the spatial distribution of leprosy cases in relationship to the FHS coverage in Castanhal-PA. RESULTS: Were developed the shapes of FHS coverage (approximately 76.38% of the population is attended by the program) and distribution of leprosy cases (approximately 81.62% of these cases are within the FHS operating area in the municipality of study). CONCLUSION: Using this technique, the areas uncovered by the FHS as well as the distribution of Leprosy cases have been identified.

Spatial epidemiology and serologic cohorts increase the early detection of leprosy.

BACKGROUND: Leprosy remains an important public health problem in some specific high-burden pockets areas, including the Brazilian Amazon region, where it is hyperendemic among children. METHODS: We selected two elementary public schools located in areas most at risk (cluster of leprosy or hyperendemic census tract) to clinically evaluate their students. We also followed anti-PGL-I seropositive and seronegative individuals and households for 2 years to compare the incidence of leprosy in both groups. RESULTS: Leprosy was detected in 11 (8.2 %) of 134 school children in high risk areas. The difference in the prevalence was statistically significant (p < .05) compared to our previous findings in randomly selected schools (63/1592; 3.9 %). The 2-year follow-up results showed that 22.3 and 9.4 % of seropositive and seronegative individuals, respectively, developed leprosy (p = .027). The odds of developing overt disease in seropositive people were 2.7 times that of negative people (p < .01), indicating that a follow-up of 10 seropositives has a >90 % probability to detect at least one new case in 2 years. The odds of clinical leprosy were also higher in "positive houses" compared to "negative houses" (p < .05), indicating that a follow-up of ten people living in households with at least one seropositive dweller have a 85 % probability to detect at least one new case in 2 years. CONCLUSIONS: Targeted screening involving school-based surveillance planned using results obtained by spatial analysis and targeted household and individual continuous surveillance based on serologic data should be applied to increase the early detection of new leprosy cases.

Spatial analysis spotlighting early childhood leprosy transmission in a hyperendemic municipality of the Brazilian Amazon region.

BACKGROUND: More than 200,000 new cases of leprosy were reported by 105 countries in 2011. The disease is a public health problem in Brazil, particularly within high-burden pockets in the Amazon region where leprosy is hyperendemic among children. METHODOLOGY: We applied geographic information systems and spatial analysis to determine the spatio-temporal pattern of leprosy cases in a hyperendemic municipality of the Brazilian Amazon region (Castanhal). Moreover, we performed active surveillance to collect clinical, epidemiological and serological data of the household contacts of people affected by leprosy and school children in the general population. The occurrence of subclinical infection and overt disease among the evaluated individuals was correlated with the spatio-temporal pattern of leprosy. PRINCIPAL FINDINGS: The pattern of leprosy cases showed significant spatio-temporal heterogeneity (p<0.01). Considering 499 mapped cases, we found spatial clusters of high and low detection rates and spatial autocorrelation of individual cases at fine spatio-temporal scales. The relative risk of contracting leprosy in one specific cluster with a high detection rate is almost four times the risk in the areas of low detection rate (RR = 3.86; 95% CI = 2.26-6.59; p<0.0001). Eight new cases were detected among 302 evaluated household contacts: two living in areas of clusters of high detection rate and six in hyperendemic census tracts. Of 188 examined students, 134 (71.3%) lived in hyperendemic areas, 120 (63.8%) were dwelling less than 100 meters of at least one reported leprosy case, 125 (66.5%) showed immunological evidence (positive anti-PGL-I IgM titer) of subclinical infection, and 9 (4.8%) were diagnosed with leprosy (8 within 200 meters of a case living in the same area). CONCLUSIONS/SIGNIFICANCE: Spatial analysis provided a better understanding of the high rate of early childhood leprosy transmission in this region. These findings can be applied to guide leprosy control programs to target intervention to high risk areas.

High rates of undiagnosed leprosy and subclinical infection amongst school children in the Amazon Region

Leprosy in children is correlated with community-level factors, including the recent presence of disease and active foci of transmission in the community. We performed clinical and serological examinations of 1,592 randomly selected school children (SC) in a cross-sectional study of eight hyperendemic municipalities in the Brazilian Amazon Region. Sixty-three (4%) SC, with a mean age of 13.3 years (standard deviation = 2.6), were diagnosed with leprosy and 777 (48.8%) were seropositive for anti-phenolic glycolipid-I (PGL-I). Additionally, we evaluated 256 house-hold contacts (HHCs) of the students diagnosed with leprosy; 24 (9.4%) HHC were also diagnosed with leprosy and 107 (41.8%) were seropositive. The seroprevalence of anti-PGL-I was significantly higher amongst girls, students from urban areas and students from public schools (p < 0.0001). Forty-five (71.4%) new cases detected amongst SC were classified as paucibacillary and 59 (93.6%) patients did not demonstrate any degree of physical disability at diagnosis. The results of this study suggest that there is a high rate of undiagnosed leprosy and subclinical infection amongst children in the Amazon Region. The advantages of school surveys in hyperendemic areas include identifying leprosy patients at an early stage when they show no physical disabilities, preventing the spread of the infection in the community and breaking the chain of transmission.