Oxidative stress and anti-oxidant status in leprosy patients.

Severe oxidative stress has been reported in leprosy patients because of malnutrition and poor immunity. The purpose of this study was to investigate the serum lipid peroxidation products, serum LDH and important free radical scavenging enzymes, i.e. superoxide dismutase (SOD), and catalase and anti-oxidant glutathione levels and total anti-oxidant status, in different types of leprosy patients. The subjects for this study were normal human volunteers (NHVs, n=14), paucibacillary leprosy patients (PB, n=18), untreated MB patients (MB1, n=18), MB patients under treatment (MB2, n=19), and MB patients released from treatment (RFT) (MB3, n=28). The levels of lipid peroxidation product, malondialdehyde (MDA), and LDH increased significantly (p<0.001) in MB (MB1, MB2, MB3) patients, and both gradually decreased with clinical improvement following MDT. The levels of SOD, catalase and glutathione, and the total anti-oxidant status decreased significantly in MB (MB1, MB2, MB3) patients (p<0.001), in comparison with NHVs. They gradually increased with clinical improvement with MDT. There was no significant variation of these parameters in PB leprosy patients in comparison with healthy volunteers. High free radical activity and low anti-oxidant levels observed in MB (MB1, MB2, MB3) leprosy patients indicate that there is an oxidative stress in MB cases, irrespective of the treatment status and suggest a suitable anti-oxidant therapy to prevent possible tissue injury.

Assessment of antioxidant activity of some antileprotic drugs.

DPPH (alpha,alpha-diphenyl-beta-picryl hydrazyl) (CAS 217-591-8), a stable free radical can be used to determine the antioxidant activity (AOA) of some drugs. In the present study the DPPH method was used for the first time to test AOA of dapsone (CAS 80-08-0), clofazimine (CAS 2030-63-9) and rifampicin (CAS 13282-42-1) in vitro and deproteinated blood method. Ascorbic acid (CAS 50-81-7) was used as a control in the study, which showed concentration-dependent antioxidant activity. Rifampicin showed a per se effect but it showed concentration dependent decrease in the DPPH absorbance. Ascorbic acid, dapsone and rifampicin showed DPPH scavenging activity both in vitro and deproteinated blood method. Clofazimine did not have any influence on DPPH. This method may be extended to different drugs for testing their AOA in biological fluids.

Pro- and anti-oxidant effects of some antileprotic drugs in vitro and their influence on super oxide dismutase activity.

The effect of ofloxacin (CAS 82419-36-1), dapsone (CAS 80-08-0), rifampicin (CAS 13282-46-1) and clofazimine (CAS 2030-63-9) on the generation of superoxide anions was studied in vitro. The drugs were incubated with a superoxide generating system (photochemical reaction between riboflavin and dianisidine). The change in optical density was measured. The optical density was increased with dapsone and ofloxacin and decreased in presence of clofazimine and rifampicin. This result indicates that ofloxacin and dapsone have an antioxidant property, whereas clofazimine and rifampicin behave as pro-oxidants.

Characterization of leprosy based on the nasal lipid profile.

While extracting the M. leprae from the nasal flushings of leprosy patients it was found that these organisms were trapped in the waxy layer, between the aqueous and the chloroform layers. Thin layer chromotography (TLC) analysis of this layer, using chloroform-methanol-water system, revealed different spots when sprayed with acid alcohol and heated at 160 degrees C. The TLC profile of lipids of lepromatous and borderline (MB according to the WHO terminology) leprosy patients was distinctly different from that of tuberculoid leprosy patients and normal human volunteers. A simple, economical and fast procedure to characterize patients belonging to different spectra has been developed.

Characterization of clofazimine metabolites in humans by HPLC-electrospray mass spectrometry.

Clofazimine (CAS 2030-63-9) is an important drug used in the treatment of leprosy. Its important metabolites are investigated by thin layer chromatography (TLC), HPLC (diode array) and HPLC-electrospray mass spectrometry. The resulting analytical data, extraction, isolation and characterization methods are presented. Their applicability is described for human urine analysis.

Disposition of rifampicin following intranasal and oral administration.

Leprosy is transmitted by dissemination of M.leprae which are lodged in the nose of the patients suffering from multibacillary (MB) type of the disease. Rifampicin, a potent bactericidal antileprotic drug is given orally to the patients with a view to make the infective cases non-infective. Earlier work by us has shown that intranasal administration of rifampicin helps in reducing the M.leprae load in the nose much faster than after conventional oral administration. In the present study, rifampicin concentrations in plasma/urine/nasal wash of healthy volunteers following oral and intranasal administration were determined. Following intranasal administration, rifampicin was not detectable in plasma and high concentrations were measured in the nasal wash. Following oral administration, rifampicin was not detectable in the nasal wash indicating that enough antibiotic levels are not available for clearing M.leprae from nose.

Tissue distribution and deposition of clofazimine in rat following subchronic treatment with or without rifampicin.

Tissue distribution and deposition characteristics of clofazimine (CAS 2030-63-9), an antileprotic drug in rats have been investigated following controlled sub-chronic administration (p.o.) for a period of 1-2 months. The drug was administered alone at a dose of 20 mg/kg body weight and in combination with rifampicin (CAS 13292-46-1) (20 mg/kg p.o.). Various tissues (liver, lung, spleen, small intestine, brain, heart, kidney, skin, stomach and subcutaneous fat) were analyzed for clofazimine in all the treated groups. High levels (range 0.9-3.6 mg/g of wet tissue) were observed in tissues having reticuloendothelial components. In other tissues the levels were relatively lower (range 3-114 micrograms/g of wet tissue). Histopathological studies revealed that clofazimine is deposited in many tissues in the form of reddish-orange crystals. Concomitant treatment with rifampicin did not significantly alter tissue distribution or deposition profile of clofazimine nor did it influence the histopathology.

New approach to curb the transmission of leprosy.

The effect of local treatment of nose of lepromatous type of patients with different formulations of rifampicin nasal drops/sprays was investigated in a large number of patients. The preparations were either sprayed or instilled into the nostrils after flushing the nostrils with normal saline at 37 degrees C. It was observed that 10 mg/ml of rifampicin was effective in reducing the BI and MI to zero in nose in seven days in majority of the patients. No untoward effect was seen in any of the patients. It is suggested that nasal sprays/drops may be able to prevent the transmission of hanseniasis, as nose is recognised to be an important portal of exit of M. leprae. Further when rifampicin drops/sprays are used as soon as the diagnosis is made, the nasal deformity may be prevented. It is believed that local treatment along with systemic therapy would go a long way in controlling the transmission of hanseniasis.

New approach to curb the transmission of leprosy

O efeito do tratamento local do nariz de paciente do tipo lepromatoso com diferentes formulaçöes de rifampicina em gotas ou "sprays" nasais, foi investigado em um grande número de pacientes. As preparaçöes foram ou aspergidas ou instiladas dentro das narinas depois de sua limpeza por jato com soluçäo salina normal a 37-C. Observou-se que 10 mg/ml de rifampicina eram eficazes na reduçäo do BI e do MI a zero no nariz em sete dias na maioria dos pacientes. Näo foi visto efeito desagradável em qualquer dos paciente. Sugere-se que "sprays"/gotas nasais possam prevenir a transmissäo da hanseníase, uma vez que o nariz é reconhecido ser uma importante porta de saida de M. leprae. Além disso a deformidade nasal pde ser prevenida quando a rifampicina "sprays"/gotas é usada täo logo o diagnóstico é feito. Acredita-se que o tratamento local juntamente com a terapia sistêmica contribuiriam muito no controle da transmissäo da hanseníase

How much non-infectious are the "non-infectious" lepromatous leprosy patients?

Nose forms an important site at which the M. leprae in lepromatous leprosy (LL) patients lodge and multiply. Nose forms an important reservoir for M. leprae, from where they may be transmitted to healthy contacts. Inspite of realizing the above fact, nose does not normally receive due importance during the chemotherapy of leprosy. LL patients, after regular treatment with dapsone or rifampin for about 20 wks and 3 wks respectively are normally considered non-infectious. From the present investigation it is clear that local treatment of the ones with a bactericidal agent should perhaps be necessary during chemotherapy of LL patients to make them non-infectious and to control the transmission of the disease.

Jala Neti a yoga technique for nasal comfort and hygiene in leprosy patients

The technique 'Jala Neti' as described in yoga is a general hygienic procedure to cleanse the nostrils effectively with a normal saline. The above mentioned technique was tried in three leprosy centres namely Jammikunt, Huzurabad and Warangal in Andhra Pradesh, India and was found to be acceptable to patients of leprosy. The technique has been found to be superior to the conventional method recommended to cleanse the nostrils in leprosy patients.