Anticataract effects of S. cumini and A. marmelos on goat lenses in an experimental diabetic cataract model.

BACKGROUND: Cataractogenesis in diabetes mellitus is mainly due to generation of free radicals causing oxidative stress. Antioxidants are known to delay cataractogenesis. Indigenous plants are potential promising sources of antioxidants. OBJECTIVES: The present study was done in goat lenses for exploring local antioxidant and anticataract potential of Syzygium cumini (Jamun) and Aegle marmelos (Bael) and comparing their activities. MATERIAL AND METHODS: "Lens organ culture technique" was employed using "tissue culture medium 199" (TC 199). Lenses were divided into four groups of 30 each. Group 1 was "Normal Control". In remaining 3 groups, experimental diabetic cataract was produced using dextrose (110 mM). Group 2: "Toxic Control" (untreated experimental diabetic cataract lenses). Group 3: S. cumini seed extract (0.25%) treated lenses. Group 4: A. marmelos leaf extract (0.25%) treated lenses. Biochemical parameters measured in lens homogenates included total soluble lens proteins (index of cataractogenesis), malondialdehyde (index of lipid peroxidation), and superoxide dismutase, glutathione reductase, and glutathione peroxidase (indices of antioxidant enzyme activity). Lens morphology was compared in all groups. RESULTS: S. cumini and A. marmelos showed significantly increased activity of all three antioxidant enzymes, preserved total soluble proteins and decreased malondialdehyde (MDA). Lens morphology was well preserved with these extracts. S. cumini aqueous seed extract scored better over A. marmelos. CONCLUSION: In goat lenses with dextrose-induced experimental diabetic cataract, S. cumini and A. marmelos showed antioxidant and anticataract properties and preservation of lens morphology (p < 0.0001 to 0.05). S. cumini showed better anticataract activity than A. marmelos.

Sensitivity and specificity of rapid HIV testing of pregnant women in India.

OBJECTIVE: Efforts to prevent HIV transmission from mother to infants in settings like India may benefit from the availability of reliable methods for rapid and simple HIV screening. Data from India on the reliability of rapid HIV test kits are limited and there are no data on the use of rapid HIV tests for screening of pregnant women. METHODS: Pregnant women attending an antenatal clinic and delivery room in Pune agreed to participate in an evaluation of five rapid HIV tests, including (a) a saliva brush test (Oraquick HIV-1/2, Orasure Technologies Inc.), (b) a rapid plasma test (Oraquick HIV-1/2) and (c) three rapid finger prick tests (Oraquick HIV-1/2; HIV-1/2 Determine, Abbott; NEVA HIV-1/2 Cadila). Results of the rapid tests were compared with three commercial plasma enzyme immunoassay (EIA) tests (Innotest HIV AB EIA, Lab systems/ELISCAN HIV AB EIA, UBI HIV Ab EIA). RESULTS: Between September 2000 and October 1, 2001, 1258 pregnant women were screened for HIV using these rapid tests. Forty-four (3.49%) of the specimens were HIV-antibody-positive by at least two plasma EIA tests. All of the rapid HIV tests demonstrated excellent specificity (96-100%). The sensitivity of the rapid tests ranged from 75-94%. The combined sensitivity and specificity of a two-step algorithm for rapid HIV testing was excellent for a number of combinations of the five rapid finger stick tests. CONCLUSION: In this relatively low HIV prevalence population of pregnant women in India, the sensitivity of the rapid HIV tests varied, when compared to a dual EIA algorithm. In general, the specificity of all the rapid tests was excellent, with very few false positive HIV tests. Based upon these data, two different rapid HIV tests for screening pregnant women in India would be highly sensitive, with excellent specificity to reliably prevent inappropriate use of antiretroviral therapy for prevention of vertical HIV transmission.

Effect of ketoacids on H(2)O(2) induced cataract.

Cataract is one of the leading causes of visual disability often leading to blindness in the elderly population. One of the causes is oxidation of proteins present in lens, by hydrogen peroxide (H(2)O(2)). In the present study 100 goat lenses were analyzed to determine the protective efficacy of ketoacids, against the oxidative insult by H(2)O(2). The ketoacids used were (pyruvate, alpha ketoglutarate and oxaloacetate), that are constantly produced endogenously. The lenses were incubated as control and experimental groups in TC-199 media for 72 hrs. H(2)O(2) concentration of 10mM was used to induce cataract. The biochemical parameters measured were levels of malondialdehyde (MDA), a lipid peroxidation product and activity of glutathione peroxidase (G-Px), an enzymatic antioxidant. The results showed a significant increase in the levels of MDA and significant decrease in the activity of G-Px in the cataractous lenses as compared to control. After addition of ketoacids (pyruvate (10mM), alpha ketoglutarate (20mM) and oxaloacetate (20mM)) separately, the levels of MDA decreased significantly and the activity of G-Px increased significantly. The results suggest that the ketoacids can be very promising antioxidants for the treatment of cataract. They may also be useful in treating other disabilities related to acute and chronic oxidative stress.

Elimination of superoxide dismutase interference in fructosamine assay.

OBJECTIVE: Superoxide dismutase (SOD) (EC 1.15.1.1) is reported to decrease the reduction of nitroblue tetrazolium (NBT) in the fructosamine assay. The study was undertaken to find a method to eliminate this interference. DESIGN AND METHODS: We studied the NBT reduction in the presence and absence of added bovine erythrocyte SOD during fructosamine assay. Formation of reduced NBT decreased with the increasing concentration of SOD. Various inhibitors of SOD were experimented with for effectively eliminating this interference. RESULTS: Cyanide eliminates the interference due to SOD, but is unsuitable because in it's presence glucose becomes reducing under the conditions of fructosamine assay. SOD inhibitors such as EDTA and Azide did not eliminate the effect of SOD. Guanidine. HCl gives opalescence in the reaction mixture. Addition of 2M HCl to the serum and incubation at 37 degrees C for 10 min eliminated the effect of added SOD (70 kU/L). The correlation between deltaA10-20 min of serum treated with HCI in presence and absence of added SOD is y = 0.9011x + 0.01055 with r = 0.9789 and S.E. (y) 0.007686. CONCLUSION: SOD does not interfere in globin bound fructosamine assay as acid-acetone treatment in preparation of heme free globin inhibits SOD. Pretreatment of serum with HCl can satisfactorily eliminate the interference due to SOD in fructosamine assay. The acid treatment could be used to inhibit SOD in various other reactions that are followed with NBT reduction.

Efficacy of stimuliv, an indigenous compound formulation, against hepatotoxicity of antitubercular drugs--a double blind study.

Anti-tubercular drugs are known to cause hepatotoxicity, which may lead to noncompliance to drug therapy. Stimuliv, an indigenous compound formulation, is reported to be useful in liver disorders. Efficacy of prophylactic administration of stimuliv against anti-tubercular drugs-induced hepatotoxicity was studied in this double blind randomized clinical trial. One hundred and forty-five newly diagnosed patients of tuberculosis were included in the study. Out of these, sixty three patients were treated with stimuliv (2 tablets thrice daily), sixty received the placebo, while twenty-two dropped out of the study. The patients were assessed clinically and biochemically at two-week intervals over a period of two months. In stimuliv-treated group, the incidence and severity of hepatotoxicity was significantly less (p < 0.05) as compared to placebo-treated group. In addition, patients treated with stimuliv had better appetite and weight gain. Stimuliv treatment may be recommended in newly diagnosed adult patients of tuberculosis.

Efficacy of potassium and magnesium in essential hypertension: a double-blind, placebo controlled, crossover study.

OBJECTIVE: To evaluate the antihypertensive activity of potassium given alone or in combination with magnesium in patients with mild hypertension. DESIGN: A double blind, randomised, placebo controlled, crossover trial of 32 weeks' duration. SETTINGS: Cardiology outpatient department, Sassoon General Hospitals, Pune, India. PATIENTS: 37 Adults with mild hypertension (diastolic blood pressure less than 110 mm Hg). INTERVENTION: Patients received either placebo or potassium 60 mmol/day alone or in combination with magnesium 20 mmol/day in a crossover design. No other drug treatment was allowed. MEASUREMENTS: Blood pressure and heart rate assessed at weekly intervals and biochemical parameters at monthly intervals. RESULTS: Potassium alone or in combination with magnesium produced a significant reduction in systolic and diastolic blood pressures (p less than 0.001) and a significant reduction in serum cholesterol concentration (p less than 0.05); other biochemical variables did not change. Magnesium did not have an additional effect. Urinary potassium excretion increased significantly in the groups who received potassium alone or in combination with magnesium. The drug was well tolerated and compliance was satisfactory. CONCLUSION: Potassium 60 mmol/day lowers arterial blood pressure in patients with mild hypertension. Giving magnesium as well has no added advantage.

Effect of inhibition of lens membrane Na+-K+ ATPase by ouabain pre-treatment--an in vitro study.

The precise factors responsible for the cataract formation are not known. The role of Na+-K+-ATPase in maintaining ionic concentration of lens and lens membrane permeability and its involvement in the formation of cataracts has been of recent interest. Thus the present study was undertaken to study the effect of pre-treatment with Ouabain, a known Na-K-ATPase inhibitor, on the intra-lenticular ionic concentration and the rule of lens membrane permeability in cataractogenesis. Fresh goat lenses were used for the experimental work. Isolated lens culture technique was used. The electrolytes were estimated before and 24 hours after Ouabain pre-treatment. The electrolyte pattern showed significant changes after pre-treatment with Ouabain. Lens sodium ion concentration increased significantly with a concommitant decrease in potassium ion concentration. Intra-lenticular chloride concentration also showed a significant increase as compared to control. Calcium and magnesium ion concentrations also showed slight increase after the inhibition of Na+-K+-ATPase system by Ouabain.

Correlation between serum choline esterase and serum albumin in leprosy.

The clinical material included 255 cases of leprosy consisting of Tuberculoid leprosy (74), Lepromatous leprosy (116), Lepromatous leprosy with lepra reaction. Liver biopsy could be performed on 50 cases of Lepromatous leprosy. Specific granulomatous changes and parenchymal cell damage were the significant findings. Serum choline esterase and serum albumin are synthesized in liver. Serum Choline esterase levels in the present study decreased abruptly with exacerbation of the disease but the serum albumin levels declined gradually and slowly. Possible hypothesis to explain the correlation and uneven fall in activity is discussed at the cellular level.

L.D.H. isoenzymes sub-unit ratio in leprosy and the effect of clofazimine treatment on L.D.H. isoenzymes sub-unit.

The clinical material for our studies of serum total LDH activity and LDH isoenzymes in leprosy included 255 patients consisting of Tuberculoid (74), Lepromatous leprosy (116), and Lepromatous leprosy with lepra reaction (65). 20 patients with suspected DDS resistance and repeated attacks of lepra reactions were selected for Clofazimine studies. All leprosy patients exhibited higher total LDH activity as compared to normals. M/H ratio was significantly increased in patients of Lepromatous leprosy and correlated closely with the clinical severity and advancement of disease. Tuberculoid leprosy patients showed values close to normals. Hence M/H ratio could demarcate two polar types of leprosy i.e. Tuberculoid and Lepromatous leprosy. Clofazimine treatment over a period of one year in patients with suspected DDS resistance and repeated attacks of lepra reaction decreased total LDH activity and M/H ratio considerably. Fall in M/H ratio during Clofazimine treatment could be attributed to the clearance of 'M' subunits by the drug due to removal of blockade of R.E.S. system produced by lepra bacilli.

Thalidomide in lepra reaction (ENL) in lepromatous leprosy patients.

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.

Once monthly rifampicin (1200 mg) plus daily dapsone (100 mg) and clofazimine (100 mg) in the initial treatment of lepromatous leprosy.

The therapeutic effect of rifampicin 1200 mg once monthly and 100 mg clofazimine daily for the first six months of treatment was evaluated in 30 patients of bacteriologically positive lepromatous leprosy patients. Moderate to marked clinical improvement was seen in all the patients and a very rapid bacteriological regression as indicated by the decrease in bacteriological and morphological indices of the skin within one week. Seven patients become MI negative at one month and three months and 13 at the end of nine months. Two patients became MI and BI negative at the end of six months and six at the end of nine months. These observations clearly establish the high therapeutic efficacy and practicability of the three drug regimen. Once monthly rifampicin is highly effective and well tolerated, and has many advantages like low cost, better patient compliance and reliability of the treatment. Addition of clofazimine to rifampicin and dapsone prevents the emergence of E.N.L. reactions which were seen during treatment with once monthly rifampicin and daily dapsone. This regimen is thus ideal for initial, intensive treatment of lepromatous leprosy and may help in preventing the spread of the disease and development of dapsone resistance.

The effect of clofazimine on liver function tests in lepra reaction (ENL).

Twenty patients with suspected DDS resistance and repeated attacks of lepra reactions were selected for the study. Clofazimine was administered in different doses over a period of 12 months. Elevated levels of transaminases and Alkaline phosphatase prior therapy attained values to near normalcy. Progressive fall in serum Bilirubin and Proteins with normal A/G ratio at the end of therapy was also observed. Clofazimine by its anti-inflammatory and antibacterial action could inhibit the process of liver damage and happened to have minimal deleterious effect on liver by studying the liver function tests.

Effect of one year clofazimine therapy on autonomic functions in lepromatous leprosy with lepra (ENL) reaction.

Twenty patients of either sex of Lepromatous leprosy with frequent type II (ENL) reactions were selected for the study after exclusion of autonomic disorders. ANS functions were evaluated before and after Clofazimine therapy at an interval of two months for one year. Sweat function test, valsalva manoeuvre, Histamine triple response, cold pressor test and Homatropine instillation were carried out in twenty normal healthy volunteers (controls) and the (ENL) patients. Clofazimine was administered initially 100 mg three times daily for one month and then gradually reduced to a dose of 100 mg biweekly for the last three months. Before the clofazimine therapy, eighteen patients had abnormal sweat functions, five patients had abnormal valsalva score, five patients had abnormal cold pressor response and all the eighteen patients had abnormal histamine triple response test. Homatropine instillation test was normal in all the patients. ANS functions did not improve significantly with Clofazimine therapy, except in one patients with abnormal cold pressor response who showed slight improvement in pressor response which, however, did not reach the normal value. ANS function tests indicate complete involvement of local axon reflex and hyporeactivity of sympathetic system and cardiac regulatory mechanisms in these patients. Parasympathetic system seems to be normal as seen from Homatropine instillation test.

Clofazimine in lepra (ENL) reaction, one year clinical trial.

Twenty patients of lepromatous leprosy with lepra reaction and suspected dapsone resistance were treated with tapering doses of clofazimine. Clinical assessment was carried out every week. Bacteriological examination was carried out every six month. Fifteen patients became reaction free at the end of three months and severity and frequency of reactions was reduced in other patients. Nerve tenderness, arthralgia, nodular eruptions and all other signs and symptoms except anaesthesia showed complete recovery in fifteen patients and severity of the reactions was reduced in others. Gynaecomastia regressed in two out of three patients within nine months. In a majority of patients, the BI and MI was reduced at the end of 3 months, and further reduced after 6 months, and in one case both BI and MI became negative. Clinical and bacteriological improvement is attributed to the antibacterial effect of clofazimine while reduction in incidence of (ENL) reactions was attributed to anti-inflammatory effect of clofazimine. Regression of gynaecomastia may be due to either improvement of involvement of testes or liver or both. Apart from dyschromia clofazimine did not produce any severe side effects or toxicity.

Serum lipoproteins in lepromatous leprosy.

Serum total cholesterol and the lipoprotein fractions were studied in forty subjects of lepromatous leprosy and age matched controls. The study revealed a significant decrease in serum cholesterol in the disease group as compared to control group. The decrease in cholesterol was 28.76% An alteration in serum lipoprotein fractions was observed in disease group. The beta-lipoprotein fraction showed a significant decrease along with a rise in alpha-lipoprotein fraction. A positive correlation was also observed between total cholesterol and beta-lipoproteins. The significance of the above findings are discussed.