Discovery of 4 exonic and 1 intergenic novel susceptibility loci for leprosy.

Seven new risk coding variants have been identified through an exome-wide association study (EWAS), which studied the contributions of protein-coding variants to leprosy susceptibility. But some potential susceptibility loci were not studied in the previous EWAS study because of the project consideration. Seventeen unstudied potential susceptibility loci of the previous EWAS were validated in 3169 cases and 9814 controls in this study. Four disease-associated exonic loci were identified: rs671 in ALDH2 (P = 2.0 × 10-20 , odds ratio [OR] = 1.35), rs13259978 in SLC7A2 (P = 1.74 × 10-8 , OR = 1.28), rs925368 in GIT2 (P = 9.18 × 10-17 , OR = 1.44), and rs75680863 in TCN2 (P = 8.37 × 10-21 , OR = 0.74). Potentially implicating ZFP36L1 as a new susceptibility gene, 1 intergenic single nucleotide polymorphism (SNP), rs1465788 (P = 7.81 × 10-6 , OR = 0.88), was also suggested to be associated with leprosy. A luciferase reporter assay showed that the rs1465788 risk allele notably decreased the transcription activity of the flanking sequence. These findings suggest the possible involvement of lipid metabolism, NF-κB homeostasis and macrophage antimicrobial pathways in leprosy pathogenesis.

Langerin (CD207)-positive cells in leprosy: Possible implications for pathogenesis of the disease with special emphasis on dermal immunoreactivity.

Leprosy is a disease caused by Mycobacterium leprae, which is characterized by two distinct poles, the tuberculoid pole and the lepromatous pole, depending on the immune response to the bacillus. Langerin-positive cells are dendritic cells that appear to play an essential role in the development of the disease. These cells are specialized in the processing and presentation of antigens, exerting an important function in the activation of the immune system. To evaluate the expression of langerin-positive cells (CD207+) in skin lesion fragments of patients with a diagnosis of M. leprae infection and to associate the expression of these cells with the polar forms of the disease. Langerin-positive cells were detected in larger numbers in lesions of patients with the tuberculoid form compared to those with the lepromatous form. The presence of a larger number of these cells in patients with the tuberculoid form suggests an important participation of langerin-positive cells, capturing antigens and favoring an effective immune response to infection with M. leprae.

Can different stages of leprosy treatment influence the profile of oral health? Oral status in leprosy.

BACKGROUND: The aim of study was to evaluate the oral health status, salivary flow and halitosis among individuals diagnosed with leprosy as compared with healthy subjects. MATERIAL AND METHODS: A sample of 160 individuals was allocated into four groups, as follows: (G1) individuals with complete leprosy treatment; (G2) individuals diagnosed with leprosy and under multi-drug therapy; (G3) individuals diagnosed with leprosy not yet under treatment; and (G4) healthy individuals. Then individuals were submitted to periodontal clinical examination (visible plaque index, bleeding index, depth of probing and clinical attachment level); DMFT index (decayed-missing-filled teeth index); evaluation of salivary flow and halitosis using a halimeter equipment (Interscan Corp, Chatsworth, CA, USA). RESULTS: The data were analyzed using Kruskal-Wallis and chi-square tests. The mean DMFT was found to be higher than 6.6, which is considered very high, with no significant difference between groups (P>0.05). As for salivary flow, 76.2% of the subjects presented normal flow rates, while 10% and 13.7% showed low and very low salivary flow rates, respectively, with hyposalivation being mostly observed in Groups 1 and 2. The highest prevalence of noticeable odor was found in healthy individuals (G4), and the most prevalent periodontal diagnosis was gingivitis (63.1%) in Group 3 (individuals with leprosy not yet under multi-drug therapy) followed by periodontitis (25%) in Group 1 (individuals who had completed leprosy treatment). CONCLUSIONS: It was observed that individuals with a history of leprosy present poor oral health similar to that of systemically healthy individuals.

Ultrasound elastography assessment of the median nerve in leprosy patients.

INTRODUCTION: We sought to compare median nerve elasticity between leprosy patients (LPs) and healthy volunteers (HVs) using ultrasound elastography (UE). METHODS: Two radiologists independently measured the strain ratio of the median nerve/flexor digitorum superficialis muscle (MN/FDSM) of 18 LP and 18 HV using real-time freehand UE. Statistical analysis included intra-class correlation coefficients (ICC) and Mann-Whitney test. RESULTS: The MN/FDSM strain ratios of the LP and HV were 2.66 ± 1.30 and 3.52 ± 0.93, respectively (P < 0.05). We observed a significantly lower MN/FDSM strain ratio in LP with reactions (types 1 and 2 cutaneous reactions associated with or without neuritis) (2.30 ± 0.91) compared with LP without reactions (3.60 ± 1.70). We found no significant differences between HV and LP without reactions. The intra- and inter-observer ICCs were 0.50 (95% confidence interval [CI], 0.11-0.72) and 0.34 (95% CI, 0.28-0.52), respectively. CONCLUSIONS: MN/FDSM strain ratios were significantly lower in LP with reactions. UE may be useful for nerve elasticity evaluation in leprosy. Muscle Nerve 56: 393-398, 2017.

International open trial of uniform multidrug therapy regimen for leprosy patients: Findings & implications for national leprosy programmes.

BACKGROUND & OBJECTIVES: Uniform therapy for all leprosy patients will simplify leprosy treatment. In this context, we evaluated six-month multidrug therapy (MDT) currently recommended for multibacillary (MB) patients as uniform MDT (U-MDT) in a single-arm open trial under programme conditions. Primary objective was to determine efficacy to prevent five-year cumulative five per cent relapse. Secondary objectives were to assess acceptability, safety and compliance. METHODS: Newly detected, treatment-naive leprosy patients were enrolled in India (six sites) and P. R. China (two sites). Primary outcome was clinically confirmed relapse of occurrence of one or more new skin patches consistent with leprosy, without evidence of reactions post-treatment. Event rates per 100 person years as well as five-year cumulative risk of relapse, were calculated. RESULTS: A total of 2091 paucibacillary (PB) and 1298 MB leprosy patients were recruited from the 3437 patients screened. Among PB, two relapsed (rate=0.023; risk=0.11%), eight had suspected adverse drug reactions (ADRs) (rate=0.79) and rate of new lesions due toreactions was 0.24 (n=23). Rates of neuritis, type 1 and type 2 reactions were 0.39 (n=37), 0.54 (n=51) and 0.03 (n=3), respectively. Among MB, four relapsed (rate=0.07; risk=0.37%) and 16 had suspected ADR (rate=2.64). Rate of new lesions due to reactions among MB was 1.34 (n=76) and rates of neuritis, type 1 and type 2 reactions were 1.37 (n=78), 2.01 (n=114) and 0.49 (n=28), respectively. Compliance to U-MDT was 99 per cent. Skin pigmentation due to clofazimine was of short duration and acceptable. INTERPRETATION & CONCLUSIONS: We observed low relapse, minimal ADR and other adverse clinical events. Clofazimine-related pigmentation was acceptable. Evidence supports introduction of U-MDT in national leprosy programmes. [CTRI No: 2012/ 05/ 002696].

Gene set signature of reversal reaction type I in leprosy patients.

Leprosy reversal reactions type 1 (T1R) are acute immune episodes that affect a subset of leprosy patients and remain a major cause of nerve damage. Little is known about the relative importance of innate versus environmental factors in the pathogenesis of T1R. In a retrospective design, we evaluated innate differences in response to Mycobacterium leprae between healthy individuals and former leprosy patients affected or free of T1R by analyzing the transcriptome response of whole blood to M. leprae sonicate. Validation of results was conducted in a subsequent prospective study. We observed the differential expression of 581 genes upon exposure of whole blood to M. leprae sonicate in the retrospective study. We defined a 44 T1R gene set signature of differentially regulated genes. The majority of the T1R set genes were represented by three functional groups: i) pro-inflammatory regulators; ii) arachidonic acid metabolism mediators; and iii) regulators of anti-inflammation. The validity of the T1R gene set signature was replicated in the prospective arm of the study. The T1R genetic signature encompasses genes encoding pro- and anti-inflammatory mediators of innate immunity. This suggests an innate defect in the regulation of the inflammatory response to M. leprae antigens. The identified T1R gene set represents a critical first step towards a genetic profile of leprosy patients who are at increased risk of T1R and concomitant nerve damage.

Normal threshold values for a monofilament sensory test in sural and radial cutaneous nerves in Indian and Nepali volunteers.

The monofilament test (MFT) is a reliable method to assess sensory nerve function in leprosy and other neuropathies. Assessment of the radial cutaneous and sural nerves, in addition to nerves usually tested, can help improve diagnosis and monitoring of nerve function impairment (NFI). To enable the detection of impairments in leprosy patients, it is essential to know the monofilament threshold of these two nerves in normal subjects. The radial cutaneous, sural, ulnar, median and posterior tibial nerves of 245 volunteers were tested. All nerves were tested at three sites on both left and right sides. Normal monofilament thresholds were calculated per test-site and per nerve. We assessed 490 radial cutaneous and 482 sural nerves. The normal monofilament was 2 g (Filament Index Number (FIN) 4.31) for the radial cutaneous and 4 g (FIN 4.56) for the sural nerve, although heavy manual laborers demonstrated a threshold of 10 g (FIN 5.07) for the sural nerve. For median and ulnar nerves, the 200 mg (FIN 3.61) filament was confirmed as normal while the 4 g (FIN 4.56) filament was normal for the posterior tibial. Age and occupation have an effect on the mean touch sensitivity but do not affect the normal threshold for the radial cutaneous and sural nerves. The normal thresholds for the radial cutaneous and sural nerves are determined as the 2 g (FIN 4.31) and the 4 g (FIN 4.56) filaments, respectively. The addition of the radial cutaneous and sural nerve to sensory nerve assessment may improve the diagnosis of patients with impaired sensory nerve function.

Reliability of clinical nerve function assessment in peripheral neuropathies.

INTRODUCTION: Sensory and/or motor nerve function impairment as a consequence of neuropathy is often assessed using electroneurophysiological tests. However, in low-resource countries where the required equipment is rarely available, manual muscle strength testing (MMST) and monofilament testing (MFT) offer very reliable alternatives. In six leprosy programmes in four Asian countries, a multi-centre randomised clinical trial (RCT) was carried out to assess the effect of corticosteroids on neuropathy in leprosy-affected people. The sensory and motor nerve function was tested using MMST and MFT, including new test sites for the sural and radial cutaneous nerves (MFT) and the posterior tibial and common peroneal nerves (MMST). The reliability studies of the MMST and MFT tests of the TENLEP (Treatment of Early Neuropathy in LEProsy) trials are presented here. METHODS: Two assessors in each centre independently used the MFT and MMST in 30 leprosy-affected people. RESULTS: Reliability is good to very good for MFT in nearly all nerves. MMST also shows good to very good agreement, with a few exceptions. CONCLUSION: Our study confirms that MMST and MFT can be performed reliably, and that the new tests also have acceptable reliability.

Evaluation of light-touch sensation in the buccal mucosa of leprosy patients.

OBJECTIVE: The aim of this study is to evaluate the light-touch sensation of the oral mucosa in leprosy patients. MATERIALS AND METHODS: A cross-sectional study included 228 adults, 133 being leprosy patients and 95 normal controls. To assess light-touch sensation, the five-filament standard Semmes-Weinstein kit in eight regions of the oral mucosa was used. Chi-square test was used to verify the differences in responses between the studied groups. RESULTS: Normal sensation was predominant in both groups, and diminished sensation was found also in the nonleprosy group. Normal controls showed diminished light-touch sensation in at least one point, which may be attributed to the definition of sensation normality. The chi-square test revealed no significant difference between the two groups (p = 0.43171). Severe diminished sensation was detected only in the leprosy group. CONCLUSION: It is concluded that altered light-touch sensation in the oral cavity may not be a common feature in leprosy. CLINICAL RELEVANCE: Decreased or lost sensation in skin lesions is a paramount in leprosy diagnosis; however, clinicians must be aware that this seems not to be true in the oral mucosa in leprosy cases for diagnosis purpose.

A Study of Liver Function Tests in leprosy.

Leprosy is a chronic progressive granulomatous disease caused by Mycobacterium leprae. Hepatic involvement is seen in early stages of the disease. Administration of the hepatotoxic drugs like Rifampicin and Dapsone may further deteriorate the liver function. The present study was undertaken to evaluate hepatic status by studying the various liver function tests in leprosy patients and compared to healthy controls. Thirty untreated leprosy patients (18 Multibacillary, 12 Paucibacillary) with duration of illness varying from one month to three years were selected as cases. Twenty healthy age and sex matched persons were taken as controls. Hepatic functional status was evaluated by estimation of serum total bilirubin, alanine transaminase, aspartate transaminase, alkaline phosphatase, total protein, albumin and globulin. Independent sample 't' test was used to compare the data. P < 0.05 was considered as significant. Except for serum total protein, there was a statistically significant difference between the mean values of all the parameters in cases when compared to controls. The present study revealed minimal derangement in hepatic function in leprosy patients. Therefore monitoring of liver function tests is very important to assess the functional status of the liver before administration of therapy in leprosy.

Disabilities in leprosy--The new concepts.

The concept of disabilities has undergone changes in recent years and disability is no longer a mere physical dysfunction. It includes activity limitations, stigma, discrimination, and social participation restrictions. In addition to the presence of an illness or impairment, the understanding of disability now explores the relationship between disease/illness/impairment, the persons functioning within daily activities/social roles, and the social, cultural, and physical environments that enable or limit an individual's ability to participate fully in his or her community and daily lives. International Classification of Functioning Disability and Health (ICF) has recognized several dimensions of disability viz., body structure and function (and impairment thereof), activity (and activity restrictions) and participation (and participation restrictions). It also recognizes the role of physical and social environmental factors in affecting disability outcomes and has shifted the focus from the cause of disability to its effect, thereby emphasizing the role of the environment (physical, cultural, social, political) rather than focusing on disability as a 'medical' or 'biological' dysfunction. There is not much information available about these relationships in leprosy related disabilities. Studies are required in different patient groups having different socio-cultural background to develop a better understanding of these issues. Accordingly the need for services can be worked out for rehabilitation of the patients released from the treatment and "Cure".

The International Classification of Functioning, Disability and Health to map leprosy-related disability in rural and remote areas in Indonesia.

The International Classification of Function, Disability, and Health (ICF-WHO, 2001) recognizes several dimensions of disability, such as body structure and function (and impairment thereof), activity (and activity restrictions) and participation (and participation restriction) and their interactions with contextual factor (personal and environmental). In this study, we map and analyse the relationship between the components of ICF in leprosy patients from two rural areas in Indonesia: Lewoleba (East Nusa Tenggara) and Likupang (North Minahasa). This study was part of a community outreach program by the KATAMATAKU team from Universitas Indonesia. The body structure was graded using the WHO hand and feet disability grade and the number of enlarged nerves, while the body function was measured by the Jebsen Taylor Hand Function Test (JTT) and Timed-up and Go (TUG). Activity limitation and participation restriction were measured using the Screening Activity Limitation Safety Awareness (SALSA) Scale and Participation Scale (P-scale), respectively. There were 177 leprosy patients from the two regions and 150 patients with complete data were included in the analysis. We found 82% (95% CI: 75.08%-87.32%) of subjects with multibacillary leprosy, 10.67% (95% CI: 6.67%-16.62%) of subjects with grade 2 WHO hand disability, and 9.33% (95% CI: 5.64%-15.06%) of subjects with grade 2 WHO foot disability. Assessment using the SALSA Scale showed 29.33% of subjects with limitation activity and 11.33% with participation restriction. Age was shown to have positive correlations with SALSA, JTT, and TUG. Inter-dimensional analysis showed that the SALSA scale had significant positive correlations with the number of nerve enlargements, P-scale, JTT, and TUG. SALSA scores of grade 2 WHO hand and foot disability were also significantly higher than grades 1 and 0. The participation scale also had a positive correlation with JTT but not TUG. Hand disability seemed to affect societal participation while foot did not. We used the ICF to describe and analyse dimensions of leprosy-related disability in Indonesia.

From periphery to center, untold story of pure neuritic leprosy: an electrophysiological study.

BACKGROUND: Pure neuritic leprosy (PNL) is uncommon form of leprosy involving peripheral nerves. Some isolated case reports have shown imaging changes in the central nervous system (CNS) and also impairment in visual evoked potential (VEP), somatosensory evoked potential (SSEP) and brain stem auditory-evoked potentials (BAEPs) parameters in PNL, but there is lack of large study. This prospective observational study evaluates impairment in these central conduction studies among PNL patients. METHODS: We screened patients with leprosy presenting with features of neuropathy and/or thickened nerves. Patients with bacilli-positive nerve biopsies were included in the study and subjected to routine tests along with nerve conduction study (NCS), VEP, tibial SSEP and BAEPs. Parameters of these studies were analyzed based on data from previous studies. RESULTS: Of 76 patients screened for PNL 49 had positive findings in biopsy. Most of patients were male and mean age group was 46.35 ± 15.35 years. Mononeuritis multiplex was most common NCS pattern in 46.93% (23/49) patients. We found abnormal VEP in 13 out of 35 patients (37.14%). Similarly abnormal SSEP and BAEPs among 42.85% and 40% patients respectively. DISCUSSION: This study shows that in PNL significant number of patients have subclinical CNS involvement. Exact pathophysiology of CNS involvement is not known till now but study of VEP, SSEP and BAEPs parameter may help in early diagnosis of PNL.

Isolated median neuropathy as the first symptom of leprosy.

INTRODUCTION: Focal peripheral neuropathy of the median nerve is mainly caused by a traumatic event or pressure, but it may also be produced by systemic illnesses. Among the latter, leprosy is a rare cause. METHODS: Six cases of isolated median neuropathy as the first sign of leprosy were selected from patients with an exclusively neurological complaint as the initial symptom. The patients, evaluated at the National Leprosy Reference Center in Rio de Janeiro, Brazil, followed routine and specialized procedures. RESULTS: Three of the patients had pure neural leprosy, and 3 had skin lesions. Clinical median nerve function impairment was confirmed by neurophysiological testing and histopathology. Both mononeuritis and mononeuritis multiplex were observed. CONCLUSIONS: This case series demonstrates an additional form of presentation of leprosy, which, if not diagnosed and treated in time, may lead to permanent disability.

The prevention of leprosy related disability as an integral component of the government health delivery programme in Indonesia: perspectives on implementation.

This paper presents a record of three interviews with groups of Ministry of Health personnel and consultants that took place in Jakarta, Indonesia in May 2012. Those contributing to the first interview were provincial and district supervisors with responsibility for leprosy. Those contributing to the second interview were consultants, three of whom were seconded to the Ministry of Health and one was a WHO consultant. A third interview was conducted with the Head and a technical staff member of the Sub Directorate of Leprosy and Yaws Control Programme, Ministry of Health, Indonesia. Leprosy control in Indonesia had been targeted for further enquiry after it became apparent, through an earlier survey of national programme managers and consultants, that the programme had been relatively successful in integrating POD into the government health delivery programme. The perspectives of significant representatives and actors in the national programme were recorded through the interviews undertaken in Jakarta. Limitations This report does not purport to be a study of integration of leprosy services in Indonesia. The perspectives of representatives and significant actors are offered here to enhance understanding of factors that contributed to POD becoming a routine component of general health care in Indonesia. It is also declared here that no independent verification of statements was undertaken and that the effectiveness of measures taken to integrate leprosy related POD has not been independently evaluated.

Disability adjusted working life years (DAWLYs) of leprosy affected persons in India.

BACKGROUND & OBJECTIVES: Disability-adjusted life years (DALYs) have been accepted as a useful method to estimate the burden of disease, and can be adapted to determine the number of productive years lost due to the disability. DALY has been reported for many studies but not for leprosy. Hence this study was carried out in three States of India. In view of the fact that in this study, productive working years are used, the term is modified as DAWLY. METHODS: A representative random sample of 150 leprosy affected persons, 50 from each States of Uttar Pradesh, West Bengal and Chhattisgarh, was chosen, and data were collected on detailed work-life history, occupation, time when leprosy was discovered, reported and treatment started, break of job/loss of income due to leprosy. The loss of wages and durations were used to compute the life-years lost due to leprosy, and summarized over the average total duration of 42 years of productive work-life from 18 to 60 years. The percentage losses were determined and differences tested for statistical significance. RESULTS: The overall mean (± SE) disability adjusted working life years was 28.6 (±0.67), a reduction of 13.4 yr from the ideal productive working life period of 42 yr. The youngest patients with disability had a reduction of 41.4 per cent, as compared to the oldest patients. There was a significant increase in loss based on year for those whose disability started earlier (P=0.0024). INTERPRETATION & CONCLUSIONS: On an average, 30 per cent of the leprosy affected person's work life is lost due to disability.

New sonographic measures of peripheral nerves: a tool for the diagnosis of peripheral nerve involvement in leprosy.

To evaluate ultrasonographic (US) cross-sectional areas (CSAs) of peripheral nerves, indexes of the differences between CSAs at the same point (∆CSAs) and between tunnel (T) and pre-tunnel (PT) ulnar CSAs (∆TPTs) in leprosy patients (LPs) and healthy volunteers (HVs). Seventy-seven LPs and 49 HVs underwent bilateral US at PT and T ulnar points, as well as along the median (M) and common fibular (CF) nerves, to calculate the CSAs, ∆CSAs and ∆TPTs. The CSA values in HVs were lower than those in LPs (p < 0.0001) at the PT (5.67/9.78 mm2) and T (6.50/10.94 mm2) points, as well as at the M (5.85/8.48 mm2) and CF (8.17/14.14 mm2) nerves. The optimum CSA- receiver operating characteristic (ROC) points and sensitivities/specificities were, respectively, 6.85 mm2 and 68-85% for the PT point, 7.35 mm2 and 71-78% for the T point, 6.75 mm2 and 62-75% for the M nerve and 9.55 mm2 and 81-72% for the CF nerve. The ∆CSAs of the LPs were greater than those of the HVs at the PT point (4.02/0.85; p = 0.007), T point (3.71/0.98; p = 0.0005) and CF nerve (2.93/1.14; p = 0.015), with no difference found for the M nerve (1.41/0.95; p = 0.17). The optimum ∆CSA-ROC points, sensitivities, specificities and p-values were, respectively, 1.35, 49%, 80% and 0.003 at the PT point, 1.55, 55-85% and 0.0006 at the T point, 0.70, 58-50% and 0.73 for the M nerve and 1.25, 54-67% and 0.022 for the CF nerve. The ∆TPT in the LPs was greater than that in the HVs (4.43/1.44; p <0.0001). The optimum ∆TPT-ROC point was 2.65 (90% sensitivity/41% specificity, p < 0.0001). The ROC analysis of CSAs showed the highest specificity and sensitivity at the PT point and CF nerve, respectively. The PT and T ∆CSAs had high specificities (> 80%) and ∆TPT had the highest specificity (> 90%). New sonographic peripheral nerve measurements (∆CSAs and ∆TPT) provide an important methodological improvement in the detection of leprosy neuropathy.

Genetic and immunological evidence implicates interleukin 6 as a susceptibility gene for leprosy type 2 reaction.

In leprosy, type 1 reaction (T1R) and type 2 reaction (T2R) are major causes of nerve injury and permanent disabilities. A previous study on plasma levels of 27 cytokines in patients with T1R or T2R and controls with nonreactional leprosy identified the gene for interleukin 6 (IL-6) as a candidate for genetic analysis. Two nested case-control studies were built from a cohort of 409 patients with leprosy from central Brazil, monitored for T1R and T2R. There was evidence for association between T2R and IL-6 tag single-nucleotide polymorphisms rs2069832 (P = .002), rs2069840 (P = .03), and rs2069845 (P = .04), with information on the entire IL-6 locus, as well as functional IL-6 variant rs1800795 (P = .005). Moreover, IL-6 plasma levels in patients with T2R correlated with IL-6 genotypes (P = .04). No association was found between IL-6 variants and T1R. Identifying genetic predictive factors for leprosy reactions may have a major impact on preventive strategies.