Search

Early diagnosis of nerve damage in leprosy is quite important for preventing the deformities. With an aim to find a good marker we have investigated Krox-20 as early peripheral nerve damage indicator in leprosy. Ambulatory patients of Kediri Leprosy Hospital, Malang, Indonesia have been studied. Degree of disability was measured based on WHO's criteria. Immunohistochemistry was.used to study the expression of Krox-20 on Schwann cells on nerve twigs in skin biopsies from leprosy cases. Receiver Operating Characteristic (ROC) curve was used to determine cut off value. Of all 79 leprosy patients studied, 36 patiens had degree of disability 0, and 43 patients had degree of disability 1. Analysis of ROC curve shown that cut off value for Krox- 20, was 8 and the cases in whom degree of disability was 0 (zero) had significantly higher mean values than those with degree of disability 1 (mean: 12.56 vs 4.24 (p<0.05)). Krox-20 appears to be a potentially good biomarker to identify early peripheral nerve damage in leprosy.

The aim of the study is to assess the role of saliva as a diagnostic tool for measurement of total antioxidant capacity in children with leprosy and children born to leprosy parent. One hundred fifty children in the age group of 4-15 years were split into three equal groups: children with leprosy (CL) and children born to leprotic parents (CLP) and healthy children. Vitamin C level was measured in saliva of children spectrophotometrically at 695nm by Phosphomolybdenum method. Data were determined with student's unpaired t test and one way ANOVA. The result of the study showed that children with leprosy exhibited significantly decreased salivary total antioxidant capacity as compared to healthy controls. Antioxidant Vitamin C was higher in the Paucibacillary leprosy (PB) than those of Multibacillary type (MB) (P < 0.001). As age advanced, there was a gradual increase in total antioxidant capacity in both the control and study groups and the results were highly significant statistically. Saliva is an easy medium.

Intracellular localization of antileprosy drugs dapsone (DDS) and rifampicin (RFP) was carried out on skin and nerve lesions obtained from multidrug treated, multi (BL-LL)- and pauci (BT-TT) bacillary cases of leprosy using immunocytochemical techniques. Intracellular localization of the above drugs especially in macrophages and Schwann cells was aimed by using rabbit raised anti DDS and RFP polyclonal antibody in an indirect peroxidase assay. Our study records both intra and extracellular staining with anti DDS and RFP antibodies in the skin as well as nerve lesions of MB and PB cases treated with MDT. All the nerves under investigation had moderate to severe pathology and hence free diffusion of the drug could be attributed to the broken barrier. Basal lamina around the Schwann cell did not seem to form a barrier. It was also noted that the drug (metabolite) persisted over a long period of time).

Six male bacteriologically highly positive patients of lepromatous leprosy with ENL reaction not adequately controlled by conventional antireaction drugs were put on thalidomide 400 mg per day in four divided doses. Reaction was controlled between 13th to 18th day of therapy. There was no change in the bacteriological status. Liver functions, renal functions and hemogram were normal before therapy and remained unaltered at the end of treatment. Apart from fatigue, drowsiness and occassional constipation, thalidomide had no adverse effect. Control of ENL reaction by thalidomide in these patients is probably due to its immunosuppressive effect, more likely by its stablising action on lysosomes.

Twenty one patients of leprosy were studied for gastrointestinal symptoms and submitted to detailed intestinal absorption tests and jejunal biopsy before and after the institution of Clofazimine therapy. Fifteen patients were administered 100 mg orally daily for 3 months, while six patients with ENL received 300 mg of the drug for 6 weeks initially and then 100 mg daily. Mild diarrhoea and abdominal symptoms occurred in four patients, fecal fat excretion increased in one patient but Schilling's and d-Xylose tests did not alter. No significant changes were produced by Clofazimine therapy in jejunal mucosa. Clofazimine crystals were seen in the lamina properia of one patient, the overlying mucosa was normal. No correlation was found between the abnormality in mucosal pattern, crystal deposition, absorption parameters or symptomatology and doses of drug taken.

The study of hydration power of stratum corneum of lesions show highly significant poor water-uptake at low temperatures (p less than 0.001), a defect not recorded after removal of water soluble fractions. Secondly, the lesions show significant poor water-diffusive power (p less than 0.001). The findings suggest a qualitative alteration in the stratum corneum of leprosy patients, probably in its water-soluble protein fraction.

Nine patients of leprosy, 5 BL and 4 LL who developed jaundice during the course of disease were investigated. Two LL patients developed jaundice during ENL reaction. There was slight hepatomegaly in 5 patients and moderate splenomegaly in 3 only. There was significant alterations in liver enzymes and serum bilirubin in all patients. The abnormalities of the enzymes levels persisted for abnormally long periods even when the serum bilirubin had come down and the patients had become asymptomatic. Blood for HBsAg and anti-HAV IgM was negative in all the patients except one in whom HBsAg was positive. Drugs could not be implicated as the cause of jaundice, all patients maintained recovery even after restarting antileprosy drugs. The possibility of non A, non B viruses producing hepatitis during the course of disease is brought out. Course of prolonged jaundice in leprosy is compared with other diseases which could result in a similar situation.

The half time of disappearance of dapsone and monoacetyl dapsone and the acetylator phenotype of the leprosy patients who harboured dapsone sensitive and dapsone resistant M. leprae was assessed in 27 subjects. Sixteen patients were rapid acetylators, five were slow and six were intermediate acetylators. The mean T 1 1/2 lives of dapsone (30.26 +/- 11.0) and monoacetyl dapsone (31.11 +/- 12.0) were also studied in the above patients. The percentage of different acetylators in both resistant and sensitive groups were similar showing no correlation between the emergence of drug resistance and the phenotype of the patient. The mean time of disappearance of DDS and MAD in the different acetylators did not show significant difference. The ratios of MAD/DDS in an individual at 3, 6 or 24 hours after the dose were similar. The mean T 1 1/2 lives of DDS and MAD in resistant and sensitive patients also showed no difference. Neither T 1 1/2 lives of DDS or MAD nor the acetylator phenotype seem to influence the emergence of dapsone resistance.

Croystat sections of dermal lesions from 33 untreated patients with leprosy were studied by indirect immunofluorescence using monoclonal antibodies to human fibronectin. Macrophages in borderline (BL) and lepromatous (LL) leprosy showed intense staining with antifibronectin antibodies. In the tuberculoid lesions cells of the mononuclear phagocyte series in an epithelioid cell granuloma stained for fibronectin. The lymphocytes surrounding these granulomas also showed the presence of fibronectin. These results suggest that the granuloma of leprosy consists of macrophages expressing fibronectin and the lymphocytes in the lesion also appear to express this protein. In six borderline cases, the sub epidermal collagen band showed intense staining with antifibronectin antibodies. In addition, the distribution of Ia-like antigens and fibronectin was studied on the plastic adherent cells obtained from peripheral blood of 11 untreated patients with leprosy. Results indicate that higher percentage of adherent cells from lepromatous patients express fibronectin in comparison to adherent cells from tuberculoid patients or controls. However, no difference was observed in the expression of the Ia-like antigens by the adherent cells, from these patients.

Dapsone (DDS) in urine of 250 leprosy patients collected on surprise visits were screened by simple paper spot, tile tests and sensitive Enzyme linked immunosorbent assay (ELISA) and Haemagglutination inhibition (HI) tests. The urinary DDS concentration as well as DDS/C ratios were also studied. Simultaneously, 50 microliter of blood was collected from each of these patients and its dapsone content was estimated by HPLC. Urine samples with means of 25 to 30 micrograms/ml DDS and 55-64 micrograms/mg DDS/C ratios were found to give positive tests by any of the above screening procedures, while their mean blood DDS concentration was found to be 0.91 microgram/ml. The corresponding values for those specimens giving negative tests were 3.8 to 5.7 micrograms DDS per ml and 9 to 13 micrograms/mg DDS/C ratio. The blood DDS concentration in this group was ranging from 0.16 to 0.18 micrograms/ml. The findings are discussed in relation to their metabolic significance and their application in a leprosy control programme.

In order to determine the suitability of the armadillos as a model of human leprosy for chemotherapeutic studies, especially in evaluating newer anti-leprosy drugs, uninfected Armadillos were used to study the metabolic disposition of DDS. Serum DDS levels ranged from 500 ng/ml at 3 hours to 13 ng/ml at 96 hours after intravenous administration of DDS (1 mg/kg). In an ad libitum feeding trial of DDS it was found that the level of serum DDS varied according to the dose of DDS, and even at a dose of 0.0001%, the animals maintained MIC of DDS against M. leprae. Finally, it was demonstrated that armadillos acetylate DDS to MADDS and 7-9% of DDS is acetylated by armadillos.

The existence, distribution and behaviour of degradation products of M. leprae in leprosy lesions were investigated in tissue specimens fixed in neutral formalin and embedded in paraffin. Cytopathologic findings using tissue imprints were unsatisfactory. Sections were stained with hematoxylin-eosin, acid-fast stains, silver methenamine and by an immunochemical (PAP) technique using serial paraffin sections. A comparison in respect of the distribution of the bacilli within the macrophages showed considerable differences between the superficial and deep granulomas. This corresponds roughly with the central, intermediary and peripheral locations. In a small granuloma seen in BL lesions, there were two zones: central and peripheral. In a large LL granuloma, three zones were seen, central, intermediary and peripheral zones. It is suggested that the degradation of disintegrated particles of bacilli might be due to the lysosymal activity of macrophages. The phagocytized bacilli are slowly degraded with long incubation periods, but the undigested debris remains inside the phagosomes. The chemical complexity of cytoplasm, cell wall and lipid fractions of M. leprae, and it is such that the lipid fractions of M. leprae mask some other antigenic components, which may be responsible for the cellular response and lysosymal production. According to our findings we believe that chemotherapy kills M. leprae but degraded products are not removed. These components are chemically complex and digested with difficulty. Lysosymal enzymes could be inhibited from productions by the bacterial debris or the lipid fractions could serve as a mask to delay lysosymal production in the cell. These aspects need further study.

The quantity of clofazimine absorbed from the gastrointestinal tract when administered to lepromatous leprosy patients at varying single doses of 600 mg., 400 mg., 300 mg., and 100 mg. has been worked out by determining the amount of clofazimine present in total faecal excreta. Except in 100 mg. dose where the percentage absorption was 62.5 +/- 17 in all other case the values were around 45%. The efficacy of daily administration of 100 mg. clofazimine is discussed in this first article.