Association of viable Mycobacterium leprae with Type 1 reaction in leprosy.

UNLABELLED: The working hypothesis is that, viable Mycobacterium leprae (M. leprae) play a crucial role in the precipitation of Type 1 reaction (T1R) in leprosy. MATERIAL AND METHODS: A total of 165 new multibacillary patients were studied. To demonstrate presence of viable M. leprae in reactional lesion (T1R+), three tests were used concurrently viz. growth in the mouse foot pad (MFP), immunohistochemical detection of M. leprae secretory protein Ag85, and 16s rRNA--using in situ RT-PCR. Mirror biopsies and non reactional lesions served as controls (T1R-). FINDINGS: A significantly higher proportion of lesion biopsy homogenates obtained at onset, from T1R(+) cases have shown unequivocal growth in MFP, proving the presence of viable bacteria, as compared to T1R(-) (P < 0.005). In contrast, few Mirror biopsies were positive in both T1R(+) and T1R(-). With respect to Ag85, while the overall positivity was higher in T1R(+) (74%), however the intensity of staining (Grade 2+) was disproportionately higher in T1R(+) BT-BB lesions 11/20 (55%). In the rebiopsies obtained during a repeat episode of T1R, Ag 85 as well as 16s rRNA, positivity (62% & 100%) was higher in T1R(+). It is inferred therefore 'viable' bacteria are an essential component in T1R and difference in the quality of bacilli, not the quantity or the ratio of dead to viable play a role in the precipitation of T1R. In conclusion, the findings show that 'metabolically active' M. leprae is a component/prerequisite and the secretory protein Ag 85, might be the trigger for precipitation of T1R.

Type 1 reaction masquerading clinically as ENL: A Case Report.

Attention is drawn to a Type 1 reaction masquerading clinically as ENL. Histology showed no evidence of ENL but suggested heightened T-cell activity (CMI), a characteristic feature of Type 1 reaction. We present a case of a 29 year old man diagnosed as lepromatous leprosy with recurrent Type 2 reaction treated with thalidomide for 2 years. The patient was referred to our institute from a teaching hospital. Skin biopsies were carried out during two separate eruptive episodes 2 months apart. Histopathology showed heightened T-cell activity, but no evidence of ENL.

BCG immunotherapy as an adjunct to chemotherapy in BL-lL patients--its effect on clinical regression, reaction severity, nerve function, lepromin conversion, bacterial/antigen clearance and 'persister' M. leprae.

BACKGROUND AND OBJECTIVE: Multidrug therapy in leprosy has failed to eliminate the problem of persister bacilli. Clearance of bacterial antigens is extremely slow which could predispose to continued nerve damage even after release from treatment. In the present study the immunomodulatory efficacy of BCG vaccine administered post-MDT in BL-LL leprosy patients was investigated in depth with a view to determining if augmenting chemotherapy with immunotherapy would help in faster clearance of M. leprae/antigens, bring down the level of persisters and minimise the occurrence/severity of reaction and nerve damage. METHODS: This is a placebo-controlled study in treated BL-LL patients. The patients are matched with respect to age, sex, bacteriological index and history of reaction, stratified and allocated to the two groups. One group (Gr A) received two doses of BCG-MOSCOW (3-33 x 10(5) cells) and the other (Gr B) normal saline (0.85%), injected intra-dermally at 3 month intervals. The Primary outcomes assessed at the end of 6 months were bacterial/antigen clearance, lepromin conversion, granuloma clearance and the occurrence of persisters. The secondary outcomes were clinical regression, occurrence and severity of reaction and changes in nerve functions. MATERIAL: A total of 107 BL-LL patients comprised of 49 in Gr A and 58 in Gr B; of which 36 and 42 respectively completed the study as per protocol, and are included in the final analysis. FINDINGS: The study findings show that both the primary and the secondary out comes were comparable in the two groups. Two doses of BCG administered post-MDT (Gr A) did not significantly alter the level of persisters or help in hastening the bacterial/antigen clearance, clinical regression of lesions and granuloma clearance. Lepromin conversion rates were also comparable. While the frequency of lepra reaction/neuritis following the intervention was comparable, the severity of reactions was significantly higher in Gr A. On the positive side neural functions assessed by nerve conduction studies showed that deterioration of motor nerve conduction was significantly lower in the BCG arm. Since all patients developing moderate to severe reactions, immediately received a course of corticosteroids, it is possible that timely use of it might have helped.

Five Cases of Idiopathic Sciatic Mononeuropathy: Clinical, Electrophysiological, Radiological, and Histological Features.

Background: Common etiologies of sciatic mononeuropathy are compressive, infiltrative, traumatic, or diabetic. However, in a proportion of patients, the etiology remains elusive despite extensive serological, electrophysiological, radiological, and histological investigations. Methods: Patients with unexplained sciatic mononeuropathy were studied with regard to their clinical, radiological, pathological, and treatment aspects. Results: We could identify five cases of sciatic mononeuropathy wherein the etiology remained unknown even after a comprehensive evaluation. The compressive, metabolic, hematological, and immune causes were ruled out with necessary investigations. The clinical, electrophysiological, radiological, and histological features of these patients are discussed. Conclusion: The etiology of sciatic mononeuropathy can remain obscure in certain instances in spite of the comprehensive workup. The role of investigations and the exclusion process of various diagnostic entities are discussed.

One year follow up of a cohort of suspected leprosy cases: findings from a leprosy 'Selective Special Drive' in Gadchiroli district, Maharashtra, India.

OBJECTIVES: The study involves a follow-up visit in 2010, to hyper-endemic Gadchiroli district of Maharashtra, India, to evaluate the current status of those suspected in 2009 of having skin/nerve lesions suggestive of leprosy, and to study the interactions between such people and the State leprosy programme. DESIGN: The study cohort comprised of those confirmed with leprosy (n = 151 and 157/233 absentee 'suspects' who were not examined by the study team in 2009 in 14 of 45 Primary Health Centres (PHCs). At follow-up, the treatment status of the confirmed cases was checked from PHC registers and cross-checked by direct questioning of patients and their views were sought on PHC leprosy services. The 157 absentee 'suspects' were queried about the reasons for their absence. RESULTS: Thirty nine 'absentee suspects' were found to have leprosy. A notable feature of the follow-up visit was that 114 people in the communities, other than those listed as 'suspects' by Community Health Workers (CHWs), voluntarily sought out the team for their opinion on hypopigmented/anaesthetic lesions, which resulted in a further 39 new cases being brought to light. (Total new cases = 78). The follow-up revealed discrepancies (100% vs. 75%) between PHC records and testimonies of the registered patients about regularity of treatment; irregularity of MDT supply was cited by some for dropping out of treatment. Other reasons proffered for irregularity were lepra reaction, fear of stigma, ignorance about leprosy and preference for faith healers. Medical Officers of PHCs were not trained in the management of lepra reactions; that task, along with disability care being entrusted to a paramedical worker of an NGO during periodic visits. CONCLUSIONS: There are remediable lacunae in the recording and dispensing of MDT by the State apparatus, as well as a need for refresher training in leprosy diagnosis for PHC staff, and in lepra reaction management for medical officers. The large number (78) of new cases detected in the follow-up, in part of Gadchiroli district strongly suggests more to-be-discovered cases in the communities.

Perceptions, health seeking behaviour and access to diagnosis and treatment initiation among previously undetected leprosy cases in rural Maharashtra, India.

OBJECTIVES: To study sociodemographic profiles, perceptions about leprosy and health seeking patterns among adult leprosy patients and parents of children with leprosy detected through a prevalence survey conducted earlier, in rural areas of Panvel tehsil in Maharashtra. METHODS: The study was cross-sectional and used mixed (qualitative and quantitative) methods. Of the 97 confirmed rural leprosy cases who had been detected through the initial prevalence survey, 58 newly detected adult leprosy cases and parents of 22 children detected with leprosy were interviewed with a semistructured interview schedule between May 2008 and March 2009. FINDINGS: The study revealed that most of the leprosy patients belonged to the poor socioeconomic strata. Nearly 58% of the adult patients reported that they had been detected through the survey within 3 months of noticing their symptom(s) for the first time. Despite having been diagnosed and receiving treatment, only 48% of adult cases knew their condition as leprosy, reflecting their poor knowledge of the disease and lack of communication between providers and patients. The symptom 'patch on the skin' seems to have percolated in the community. Despite approaching the private or public sector for help in the first instance, many patients and children remained undiagnosed and untreated for leprosy. CONCLUSION: Active surveys for leprosy case detection should substitute the self-reporting approach until IEC measures are sufficiently effective to achieve a significant impact on transmission. Nevertheless both approaches will need the presence of staff with active diagnostic skills and optimal drug availability at PHCs.

Clinical, bacteriological and histopathological study of 62 referral relapse cases between Jan 2004 and Dec 2009 at the Foundation for Medical Research, Mumbai.

UNLABELLED: Sixty two patients with relapsed leprosy seen between Jan 2004 and Dec 2009 were studied using clinical, bacteriological and histopathological parameters. The findings thus obtained were correlated to parameters such as trend and source of referral, clinical characteristics at diagnosis, treatment received, other events during or after RFT and duration between cessation of treatment and relapse. FINDINGS: Referrals per year have doubled since 2006. Most patients were referred by NGOs (58%), followed by Govt. hospitals (16%) and then by GPs (25%); 76% had received one of the WHO - MDT regimens including 16 treated with 24 months or more MB - MDT, 23 with 12 months MB - MDT and eight with 6 months PB - MDT. Of the remaining 14 cases, four had received DDS mono-therapy, seven had single dose of Rifampicin, Ofloxacin and Minocycline (ROM) and four Rifampicin and Ofloxacin (RO) daily for 28 days. The average incubation time of relapse, defined as duration between cessation of treatment and relapse was (SD) + 6-4 years. 59% of patients had positive slit skin smears on relapse. Relapse for the second time occurred in six BL cases including five from group 2 and one RO treated patient and 11/23 cases from group 2 conferred to BT-BB leprosy. Clinical features at diagnosis and on relapse were comparable in 47% of cases. CONCLUSION: All leprosy patients, regardless of their type and MDT regime, carry 'risk of relapse'. A shorter treatment duration reduces the incubation time to relapse. In group 2 (treated with 12 months MB-MDT regime) 11/23 were BT-BB cases and 5/23 (21%) were relapse for the second time, which further supports our earlier documented findings and maybe the efficacy of WHO-MDT regime is poor in a small subset of patients.

Neuropathies of leprosy.

Neuropathies form an integral part of the symptomatology of leprosy. Neuropathies of leprosy take various forms and shapes. At one end is the cutaneous nerve involvement adjacent to the anaesthetic skin patch and the other is of symmetrical pansensory neuropathy and the devastating sensory ataxia of leprous ganglionits. Lepra reactions add to the spectrum. Hosts immunological status largely decides the clinical manifestations seen in nerves and skin. A wide array of diagnostic techniques like ultrasonography, magnetic resonance neurography, serological markers, molecular tests, skin biopsy and in selected cases, the nerve biopsy with special stains and electron microscopy are obtainable to help the clinical diagnosis. The unsuspecting clinician, lack of community awareness and limited availability of diagnostic tests are important adverse factors in the total outcome. Multi drug therapy is efficacious and corticosteroids reduce the impact of nerve damage in leprosy. The efficacy, dose and duration of corticosteroid therapy are presently inexact and other immune suppressants like azathioprine are being evaluated. Chronic disabilities and residual deficits require attention of multiple specialties. In the coming time, focus on prevention could lead to favourable results. This review will discuss the classification systems, common and uncommon clinical features, diagnostic armamentarium and therapeutic and preventive aspects of neuropathies of leprosy.

The effect of corticosteroids usage on bacterial killing, clearance and nerve damage in leprosy; part 3--Study of two comparable groups of 100 multibacillary (MB) patients each, treated with MDT + steroids vs. MDT alone, assessed at 6 months post-release from 12 months MDT.

OBJECTIVE: To investigate effects of therapeutic usage of corticosteroids on M. leprae killing and clearance, on clearance of granuloma and on nerve damage in multibacillary (MB) leprosy patients. DESIGN: From a cohort of 400 untreated MB patients, a comparable group of 100 each receiving MDT + steroids (group A) vs MDT alone (group B) were assessed at 18 months as compared to month zero with respect to clinical and granuloma regression, M. leprae killing and clearance, and nerve functions. Analysis was performed using SPSS version 10.0. The significance of association was tested using Chi square and Fisher's exact tests. RESULTS: Regression of lesions assessed clinically and by histopathology was seen in 52% and 53% patients in group A and 46% and 63% in B respectively (P not significant). Clearance of bacteria assessed by bacteriological index (BI) in slit skin smears (SSS) and extent and intensity of antigen using anti-BCG staining were also comparable in the two groups. Multiplication of M. leprae in the mouse foot pad (MFP) indicating the presence of viable bacilli was seen in 14% and 16% of SSS positive BL-LLs patients in groups A and B respectively (P not significant). The occurrence of viable M. leprae was higher among patients with repeat reaction (19%) than single (11%). Using clinical tests (nerve palpation, monofilament and voluntary muscle testing), the proportion of sensory and motor nerves showing improvement or deterioration were similar in the two groups. However using nerve conduction studies, the overall proportion of nerves showing deterioration (22%) was significantly higher than improvement (9%) (P < 0.001). CONCLUSIONS: Treatment with MDT + corticosteroids does not adversely affect the clearance of granuloma, M. leprae and/or its antigens and M. leprae killing. However the continued presence of viable bacteria in > 14% of BL-LLs patients indicate that 12 months of MDT may be insufficient for complete bacterial killing. In both groups nerve conduction studies indicated that deterioration of nerves was high suggesting, MDT + corticosteroids was not very efficacious in the prevention or reversal of nerve damage. A better immuno-modulatory drug or a modified corticosteroid regime is needed.

Optimizing Investigations for Evaluation of Enlargements of the Roots, Plexuses and Nerves: A Study of 133 Patients.

BACKGROUND AND AIMS: A wide variety of neurological diseases result in clinical and/or radiological enlargement of nerves, roots and plexuses. With the advancement in techniques and use of magnetic resonance neurography (MRN), aided by electrophysiology, proximal segments of the lower motor neuron (LMN) can be well studied. The relative merits of investigative modalities have not been well defined and comprehensive information on this subject is sparse. METHODS: This retrospective study included data from January 2010 to June 2018. Patients having clinical and/or radiological enlargements of lower motor neuron were included. Clinical and laboratory work up, electrophysiology, MRN and biopsy studies were documented and analyzed. RESULTS: 133 patients fulfilled the inclusion criteria. The diagnostic categories were of leprosy (32%), immune neuropathies (27.8%), nerve infiltrations (8.2%), inherited neuropathies (9%), diabetic radiculopathies (9%) and others (12.7%). MRN was essential to diagnosis in 24.8% and supportive in 31.5% patients. Electrophysiology was essential in diagnosis in 70.6%, biopsy in 45.8% and genetic studies in 6.4% patients. CONCLUSION: The manuscript presents a large cohort of diseases causing enlargement of LMN with clinical and investigative aspects of 7 patients of the most unusual condition of chronic immune sensorimotor polyradiculopathy (CISMP) and details of 7 other patients with chronic mononeuropathies at non-entrapment sites. A table of comparative utility and an algorithm depicting the optimization of investigations has been presented.

Hypophosphorylation of NF-H and NF-M subunits of neurofilaments and the associated decrease in KSPXK kinase activity in the sciatic nerves of swiss white mice inoculated in the foot pad with mycobacterium leprae.

OBJECTIVE: To study the phosphorylation state of neurofilament (NF) proteins and activity of KSPXK kinase in the sciatic nerves of Swiss white (S/W) mice inoculated in the hind foot pads with M. leprae. DESIGN: Test group includes S/W mice inoculated in the foot pads with freshly harvested human derived (viable) M. leprae. Control groups were constituted by (1) Age matched un-inoculated mice, (2) Mice similarly inoculated with M. smegmatis and (3) heat killed M. leprae. Phosphorylation state of NF was studied using Western blot analysis and phosphor-specific NF antibody (SMI 31; Sternberger Monoclonals, Inc.). The KSPXK kinase activity was assayed by using KSPXK fusion protein in a radiometric method using gamma(22)P ATP. RESULTS: Several fold increase in M. leprae numbers was seen in viable M. leprae group while M. smegmatis failed to show any fold increase in the foot pads of S/W mice. Western immunoblot analysis of cytoskeletal preparation from sciatic nerves of uninoculated mice and mice inoculated with M. smegmatis showed immunoreactivity to SMI 31 antibody and protein bands corresponding to both NF-H and NF-M at all the time points from 4-20 months post inoculation. In case of viable M. leprae; SMI 31 reactive protein bands were seen at 4 months but not at any of the later intervals, i.e., between 6-20 months. With heat killed M. leprae transient loss of immunoreactivity to SMI 31 was seen. Decrease in KSPXK kinase activity was recorded in sets inoculated with viable and heat killed M. leprae, and corroborated with loss of immunoreactivity seen in WBs reacted with SMI 31 antibody. CONCLUSIONS: Alterations in the sciatic nerve NF cytoskeleton was seen following inoculation in the hind foot pad with both viable and heat killed M. leprae. The hypophosphorylation of NF observed in this study corroborates with the earlier observations in human leprous nerves.

Sensitivity and specificity of nerve palpation, monofilament testing and voluntary muscle testing in detecting peripheral nerve abnormality, using nerve conduction studies as gold standard; a study in 357 patients.

OBJECTIVE: To determine sensitivity and specificity of clinical tools viz. nerve palpation (NP), monofilament (MF), and voluntary muscle testing (VMT), for assessing peripheral nerve function impairment (NFI) in leprosy, using nerve conduction studies (NCS) as gold standard. STUDY POPULATION AND METHODS: 357 untreated multibacillary (MB) leprosy patients were assessed using above tests. The nerves assessed were left and right ulnar, median, radial cutaneous, sural, common peroneal and posterior tibial. The concordance between the clinical and NCS tests was done for each nerve. The sensitivity and specificity of clinical tests for detecting nerve impairment was determined, using NCS as gold standard. Analysis was performed using SPSS version 10.0. RESULTS: The sensitivity of NP ranged between 71% to 88% for all nerves, except the median (43%) and sural (59%) nerves. Specificity was > 60% for all, but low for ulnar (34%) and common peroneal (40%) nerves. The specificity of MF testing was > 80% and of VMT assessment was >90% for all nerves. The sensitivity of MF testing ranged between 35-44%, while of VMT assessment was very low i.e. 4-5%, the maximum was for the ulnar nerve (25%). Detection sensitivity of MF testing and VMT assessment improved two fold when combined with NP and was closely comparable to NCS test findings. CONCLUSIONS: Both MF testing and VMT assessment showed good specificity, but moderate to low sensitivity. NP was less specific but more sensitive than MF testing and VMT assessment. Combining NP with MF testing and VMT assessment gives a two fold improvement in the sensitivity for assessing nerve damage and could therefore serve as the most useful clinical tools for diagnosis of leprosy and detecting nerve damage at field level.

Detection of previously undetected leprosy cases in a defined rural and urban area of Maharashtra, Western India.

INTRODUCTION: Leprosy has been a major public health problem in India for centuries. In India, between 2001 and 2005, the prevalence of leprosy was reduced by 80%. This sharp decline in the prevalence of leprosy alongside the cessation of active surveillance for detection of leprosy cases has raised a sense of alarm in the scientific community. MATERIALS AND METHODS: This is a total population survey aiming to estimate the prevalence of undetected active cases of leprosy in the community in defined rural (Panvel Taluka, Raigad District) and urban (M-East Ward, Mumbai) areas by health workers from Kushth Nivaran Samiti (Panvel) and Lok Seva Sangam (Mumbai). Those provisionally diagnosed with leprosy were subjected to an independent verification using clinical, bacteriological and histopathological investigations at the Foundation for Medical Research. FINDINGS: A population of 196,694 and 600,247 was covered in defined rural and urban areas respectively. In the rural area on examining 178,646 individuals, 120 provisionally diagnosed leprosy cases were detected, of which 65 were paucibacillary (PB) and 55 were multibacillary (MB) based on the WHO operational classification used by health workers at field level. In the urban area, of the 512,434 individuals who were examined, 134 provisionally diagnosed leprosy cases were detected with 92 PB and 42 MB cases. Among the clinically confirmed cases, 35.6% (32/90) and 34.9% (36/109) in rural and urban areas respectively were children. CONCLUSIONS: There are large numbers of undetected leprosy cases in the community with a high proportion of MB patients and children among them. This indicates active transmission pointing to the need for a paradigm shift in leprosy care services and control programme.

Clinical characteristics of neuropathic pain in leprosy and associated somatosensory profiles: a deep phenotyping study in India.

This study investigated the clinical characteristics and somatosensory profiles of patients suffering from leprosy in Mumbai, India. A cross-sectional deep profiling study was conducted in 86 patients with leprosy (with and without pain) using an extensive battery of phenotyping measures including structured clinical examination, psychological state (General Health Questionnaire [GHQ-12]), and a quality-of-life condition-specific instrument (Brief Pain Inventory-short form). Quantitative sensory testing was performed according to the protocol of the German Research Network on Neuropathic Pain (DFNS) to assess the somatosensory profiles in the ulnar nerve innervation territory of all participants (dorsum of the hand). Reference data from 50 healthy Indian subjects were within the range of published DFNS values. Somatosensory profiles in leprosy patients with clinically or electroneurographically diagnosed neuropathy (with and without pain) revealed a profile of sensory loss to thermal and tactile stimuli combined with preservation of vibration and deep pressure detection. Sensory gain phenomena were not generally observed in patients with leprosy. In the group of subclinical neuropathy, a high degree of impaired thermal sensation was found, which could be clinically deployed to enhance identification of leprosy neuropathy at an early stage. Quantitative sensory testing can effectively document leprosy-associated neuropathy but does not distinguish between patients with or without pain. Patients with leprosy and neuropathic pain reported a poor quality of life and less psychological well-being compared with the pain-free patients with leprosy neuropathy.

The effect of corticosteroid usage on the bacterial killing, clearance and nerve damage in leprosy: a prospective cohort study: part 1--study design and baseline findings of 400 untreated multibacillary patients.

OBJECTIVE: To investigate possible adverse effects of therapeutic usage of corticosteroids on the killing and clearance of M. leprae and the clearance of granuloma, in patients with multibacillary (MB) leprosy. DESIGN: A cohort of 400 untreated MB patients were sub-grouped into those to be treated with corticosteroids (prednisolone 40 mg daily tapered to 5 mg over 12 weeks) along with MB-MDT for reaction and/or neuritis or silent neuropathy (SN) of <6 months duration (group A), and those with no reaction and to be treated with MDT only (group B). Clinical, bacteriological, histopathological and neurological test findings at fixed time points were compared. Analysis was performed using SPSS version 10.0. The significance of association was tested using Chi-square test. In the current report, we describe the study design and baseline findings of 400 untreated MB patients, with special emphasis on differences between patients in groups A and B. RESULTS: At baseline, applying Ridley-Jopling classification, 39% patients were BT, 20% BB, 24% BL, 12% sub-polar LL and 5% pure neural (PN). Overall, 60% patients were slit skin smear (SSS) negative and 33% presented with disability either grades 1 or 2. Overall 140/400 (35%) patients presented with reaction and/or neuritis and 11/400 (3%) presented with SN of <6 months duration. Comparing groups A and B, the percentage of patients presenting with DG2 was significantly higher in group A (43%). By clinical tests, monofilaments (MF) and voluntary muscle testing (VMT), the percentage of patients and nerves showing functional impairment was also significantly higher in group A. However, in the more sensitive nerve conduction velocity (NCV) test, the percentage of patients that showed nerve abnormalities was closely comparable; 94% and 91% in groups A and B respectively while number of affected nerves was higher in group A. CONCLUSION: At baseline, as recorded by NCV, peripheral nerve function abnormality was observed in almost all the MB patients regardless of reaction; but among those presenting with reaction or neuritis, the nerve damage was more severe and extensive.

Effect of cryo preservation on Mycobacterium leprae growth in the footpads of non-immunosuppressed mice.

OBJECTIVE: To investigate using the mouse footpad system, whether the use of cryopreservants help in retaining the viability of Mycobacterium leprae samples stored at three different temperatures of 4 degrees, -20 degrees and -70 degrees C for 30 days. DESIGN: Biopsies from eight untreated lepromatous leprosy cases were homogenised and inoculated into footpads of normal Swiss White mice within 24 hours (control) and remaining homogenates in each case was divided and stored at 4 degrees C, -20 degrees C and -70 degrees C respectively for 1 month, using either 10% skimmed milk (SM) or Roswell Park Memorial Institute media + 10% glycerol (RPMI) (test). Homogenates adjusted to contain 1 x 10(4) M. leprae/footpad was inoculated into 10 mice per set. Harvestings were done at 6th, 7th, 8th and 12th months. Footpad counts showing > 1 x 10(5) M. leprae at 6th month or later were considered as positive yield. RESULTS: Control All the cases showed > 100 fold growth and 100% take. Viability at 4 degrees C: Only one case (SM) showed a 100 fold increase and 23% take. Viability at -20 degrees C: Two cases showed fold growth that was 40-60 fold less with takes of 63% (SM) and 71% (RPMI) respectively. Viability at -70 degrees C: Positivity was 45% but the fold increase was less as compared to control and takes were between 80-20%, except one RPMI where take was 100%. CONCLUSION: The viability assessed using the mouse footpad was best and consistent in the inoculas that were injected within 24 hours of harvest from the host tissue (control group). None of the storage temperatures used matched with the controls with respect to bacterial yield or % takes. Among the three storage temperatures, -70 degrees C appeared to be better with 45% of the samples showing growth. There was no significant difference noted between the two preservatives used.

Effectiveness of 32 versus 20 weeks of prednisolone in leprosy patients with recent nerve function impairment: A randomized controlled trial.

BACKGROUND: While prednisolone is commonly used to treat recent nerve function impairment (NFI) in leprosy patients, the optimal treatment duration has not yet been established. In this "Treatment of Early Neuropathy in Leprosy" (TENLEP) trial, we evaluated whether a 32-week prednisolone course is more effective than a 20-week course in restoring and improving nerve function. METHODS: In this multi-centre, triple-blind, randomized controlled trial, leprosy patients who had recently developed clinical NFI (<6 months) were allocated to a prednisolone treatment regimen of either 20 weeks or 32 weeks. Prednisolone was started at either 45 or 60 mg/day, depending on the patient's body weight, and was then tapered. Throughout follow up, NFI was assessed by voluntary muscle testing and monofilament testing. The primary outcome was the proportion of patients with improved or restored nerve function at week 78. As secondary outcomes, we analysed improvements between baseline and week 78 on the Reaction Severity Scale, the SALSA Scale and the Participation Scale. Serious Adverse Events and the need for additional prednisolone treatment were monitored and reported. RESULTS: We included 868 patients in the study, 429 in the 20-week arm and 439 in the 32-week arm. At 78 weeks, the proportion of patients with improved or restored nerve function did not differ significantly between the groups: 78.1% in the 20-week arm and 77.5% in the 32-week arm (p = 0.821). Nor were there any differences in secondary outcomes, except for a significant higher proportion of Serious Adverse Events in the longer treatment arm. CONCLUSION: In our study, a 20-week course of prednisolone was as effective as a 32-week course in improving and restoring recent clinical NFI in leprosy patients. Twenty weeks is therefore the preferred initial treatment duration for leprosy neuropathy, after which likely only a minority of patients require further individualized treatment.

Clinical, bacteriological, and histopathological characteristics of newly detected children with leprosy: a population based study in a defined rural and urban area of Maharashtra, Western India.

BACKGROUND: Leprosy has been a major public-health problem in many developing countries for centuries. According to the National Leprosy Elimination Programme report of March 2012, there were a total of about 0.13 million cases of leprosy in India, 9.7% of which were children. Numerous studies have investigated child leprosy amongst reported cases however, studies pertaining to proportion and characteristics of undetected childhood cases in the community are very few. AIM: To examine the clinical, bacteriological, and histopathological characteristics of newly detected child leprosy cases in the community. METHODS: The population survey conducted from June to September 2007 and the defined rural areas, which included five primary health centers of Panvel Taluka, in Raigad district and urban areas, which included M-east ward of the municipal corporation of greater Mumbai of western Maharashtra, India. RESULTS: House-to-house survey yielded 32 and 37 so far, undetected child cases of leprosy in the rural and urban region, and the prevalence rate was 10.5 and 1.5 per 10,000, respectively. The age of child leprosy cases detected, ranged from 3 to 14 years with a mean of 10.06 ± 3.35 years in the rural and 9.97 ± 3.12 years in the urban area. Most of the cases were paucibacillary (62%). A large proportion of children (49%) had single skin lesion (SSL). Of the 19 SSL cases examined histopathologically, 15 (99%) showed features of borderline tuberculoid, 1 (5%) borderline lepromatous and 3 (16%) had indeterminate type of leprosy. Tuberculoid leprosy was not seen in any, indicating less likelihood of self-healing. Overall, three cases had deformity (grade 1 = 1 and grade 2 = 2) and 31% of multibacillary cases were smear positive. CONCLUSION: The clinical, bacteriological, and histopathological characteristics of newly detected child cases in the community evidently indicate the grave nature of the problem of undetected child leprosy, recent active transmission, and highlight implications on individual patients and the community. KEY MESSAGE: Most of the cases were paucibacillary (62%). A large proportion of children (49%) had SSL and (55%) had it on the face followed by arms and leg (27%) and trunk (17%). The mean duration of symptoms exceeded one year which can be attributed to poor knowledge of leprosy or barriers in access to health care or its utilization.