Mir Misbahuddin

Efficacy of Spirulina Extract Plus Zinc in Patients of Chronic Arsenic Poisoning: A Randomized Placebo-Controlled Study

Background. Millions of people in Bangladesh, India, Taiwan, and Chile are consuming high concentration of arsenic through drinking water, and thousands of them have already developed chronic arsenic poisoning. There is no specific treatment. Some authors suggest the use of vitamins and minerals for more than 6 months. The present placebo-controlled double-blind study was conducted to evaluate effectiveness of spirulina extract plus zinc in the treatment of chronic arsenic poisoning. Methods. Forty-one patients of chronic arsenic poisoning were randomly treated orally by either placebo (17 patients) or spirulina extract (250 mg) plus zinc (2 mg) (24 patients) twice daily for 16 weeks. Each patient was supplied with arsenic-safe drinking water by installing a locally made water filter at household level. Effectiveness of spirulina extract plus zinc was evaluated by comparing changes in skin manifestations (clinical scores), arsenic contents in urine and hair, between the placebo- and spirulina extract plus zinc-treated groups. Results. The concentrations of total arsenic in water (without filtration) of placebo- and spirulina extract plus zinc-treated groups were 150.1 ±18.3 and 161.7 ± 23.9 μg/l, respectively. Intake of these high concentrations of arsenic lead to increased excretion of arsenic in urine (72.1 ± 14.5 μg/l in placebo-treated group and 78.4 ± 19.1 μg/l in spirulina plus zinc-treated group). After 2 weeks of using filtered water, there were significant reduction of both arsenic intake through water and urinary arsenic excretion (8.3 ± 3.6 μg/l and 18.4 ± 7.3 μg/l in placebo group; 9.7 ± 5.4 μg/l and 21.6 ± 5.8 μg/l) in spirulina extract plus zinc-treated group. There was a sharp increase in urinary excretion of arsenic (138 ± 43.6 μg/l) at 4 weeks following spirulina plus zinc administration and the effect was continued for another 2 weeks. Spirulina extract plus zinc removed 47.1% arsenic from scalp hair. Spirulina extract had no major adverse effect that required physicians attention. The clinical scores (median) for melanosis before and after treatment with placebo was not statistically significant (p > 0.05), whereas in spirulina extract plus zinc-treated group it was statistically significant (p < 0.01). In cases of keratosis, the median clinical scores before and after treatment was not statistically significant (p > 0.05) in placebo-treated group. In spirulina extract plus zinc-treated group, the clinical scores for keratosis before and after treatment was statistically significant (p < 0.05). Conclusions: Results show that spirulina extract (250 mg) plus zinc (2 mg) twice daily for 16 weeks may be useful for the treatment of chronic arsenic poisoning with melanosis and keratosis.

Muscarinic receptor-mediated increase in cytoplasmic free Ca2+ in isolated bovine adrenal medullary cells: effects of TMB-8 and phorbol ester TPA

The change in cytoplasmic free calcium, [Ca2+]iin isolated bovine adrenal medullary cells during stimulation by acetylcholine (ACh) in Ca2+‐free incubation medium was measured using the fluorescent Ca2+ indicator quin2. ACh (1–100 μM) caused an increase in [Ca2+]i by mobilization of Ca2+ from the intracellular pool. Nicotine (10 μM) did not increase [Ca2+]iin the absence of extracellular Ca2+. Pretreatment of the cells with atropine (10 μM) completely inhibited ACh‐induced increase in [Ca2+]i, whereas pretreatment with hexamethonium (100 μM) did not. The intracellular Ca2+ antagonist 8‐(N,N‐diethylamino)octyl‐3,4,5trimethoxybenzoate (TMB‐8), inhibited ACh‐induced increase in [Ca2+]i. The activator of protein kinase C 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA), but not its ‘inactive’ analog 4α‐phorbol‐12,13‐didecanoate (PDD), also inhibited ACh‐induced increase in [Ca2+]i. These findings suggest that in bovine adrenal medullary cells, stimulation of muscarinic ACh receptor causes an increase in [Ca2+]i by mobilizing Ca2+ from the intracellular pool and that protein kinase C is involved in ‘termination’ or ‘down regulation’ of this response.

Water hyacinth removes arsenic from arsenic-contaminated drinking water.

Abstract Water hyacinth (Eichhornia crassipes) removes arsenic from arsenic-contaminated drinking water. This effect depends on several factors, such as the amount of water hyacinth, amount of arsenic present in the water, duration of exposure, and presence of sunlight and air. On the basis of the present study, the authors suggest that water hyacinth is useful for making arsenic-contaminated drinking water totally arsenic free. Water hyacinth provides a natural means of removing arsenic from drinking water at the household level without monetary cost.

Stimulation by vasoactive intestinal polypeptide of catecholamine synthesis in isolated bovine adrenal chromaffin cells: Possible involvement of protein kinase C

In isolated bovine adrenal medullary cells, vasoactive intestinal polypeptide (VIP) stimulated 14C-catecholamine synthesis from 14C-tyrosine, but not from 14C-DOPA. This stimulatory effect of VIP on 14C-catecholamine synthesis was not dependent upon extracellular Ca2+. VIP did not affect the intracellular cyclic AMP (cAMP) level. The stimulatory effect of VIP on 14C-catecholamine synthesis was additive with that of carbamylcholine, which was dependent upon extracellular Ca2+, but not with that of phorbol ester 12-O-tetradecanoyl phorbol 13-acetate (TPA), an activator of protein kinase C. Moreover, 1-(isoquinolinyl-sulfonyl)-2-methylpiperazine (H-7), an inhibitor of protein kinase C, inhibited not only TPA-stimulated, but also VIP-stimulated 14C-catecholamine synthesis from 14C-tyrosine. These results suggested that VIP stimulated catecholamine synthesis by activation of tyrosine hydroxylase and that protein kinase C was involved in this stimulatory mechanism.

Stimulatory effect of vasoactive intestinal polypeptide on catecholamine secretion from isolated guinea pig adrenal chromaffin cells

The stimulatory effect of vasoactive intestinal polypeptide (VIP) on catecholamine (CA) secretion from isolated guinea pig adrenal chromaffin cell was studied. VIP (1-10 microM) induced dose-dependent CA secretion, which was slow and continued for at least 30 min. This VIP-induced CA secretion was dependent on the presence of Ca2+ in the medium, but no significant increase in Ca2+ uptake by the cells was observed during their stimulation with VIP. Studies on the intracellular free Ca2+ level ([Ca2+]i) using fura-2 showed that acetylcholine and muscarine induced a marked increase in the [Ca2+]i, but that VIP induced only a slight increase. Thus VIP may induce CA secretion by increasing the sensitivity of the secretion of CA to Ca2+.

Muscarinic stimulation of guinea pig adrenal chromaffin cells stimulates catecholamine secretion without significant increase in Ca2+ uptake

In isolated guinea pig adrenal chromaffin cells, not only nicotine, but also muscarine stimulated catecholamine (CA) secretion, the stimulation by muscarine being the greater. The secretions of CA by muscarine and nicotine were both dependent on the presence of Ca2+ in the medium, but only the latter was associated with a rapid increase in 45Ca2+ uptake. Experiments with the fluorescent Ca2+ indicator quin 2, showed that muscarine caused an increase in cytoplasmic free Ca2+ concentration [( Ca2+]i). Moreover, the intracellular Ca2+ antagonist 8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate (TMB-8) inhibited both CA secretion and increase in [Ca2+]i induced by muscarine. These results indicate that in isolated guinea pig adrenal chromaffin cells, nicotine stimulated CA secretion by increasing Ca2+ uptake by the cells, whereas muscarine stimulated CA secretion by mobilizing Ca2+ from the intracellular pool.

Accumulation of arsenic in tissues of iron-deficient rats.

Toxic concentration of arsenic (400 microg/kg body weight/day) was administered orally for 8 weeks to both iron-supplemented and iron-deficient rats. The results showed greater amount of arsenic in both the duodenum and liver of iron-deficient rats, compared to iron-supplemented ones. Duodenum containing high concentration of arsenic in iron-deficient rats showed significant reduction of ferrireductase (the enzyme responsible for the conversion of ferric iron to ferrous one) activity. Our results suggest that anemic rats are more prone to develop arsenic poisoning following chronic ingestion of high content of arsenic.

Stimulation by vasoactive intestinal polypeptide of muscarinic receptor-mediated catecholamine secretion from isolated guinea pig adrenal medullary cells

The effects of vasoactive intestinal polypeptide (VIP) on catecholamine (CA) secretion by isolated guinea pig adrenal medullary cells were studied. VIP (1 microM) alone induced only a slight secretion of CA, but it stimulated ACh-induced CA secretion. At concentrations of 0.01-1 microM, it stimulated muscarine-induced CA secretion, but not nicotine-induced CA secretion. It did not affect high K+ or Ca2+ ionophore-induced CA secretion. The stimulatory effect of VIP on muscarine-induced CA secretion was observed at muscarine concentrations of 2 200 microM and was detectable after 2 min incubation.

Secretion of arsenic, cholesterol, vitamin E, and zinc from the site of arsenical melanosis and leucomelanosis in skin

Background. Melanosis and leucomelanosis with or without keratosis are the earliest symptoms of arsenicosis. Uneven distribution of arsenical melanosis and leucomelanosis in skin led us to investigate the possibility of preferential secretion of arsenic and three constituents of sweat; cholesterol, vitamin E, and zinc. Methods. Twenty-four-hour skin secretion was collected from skin lesions and unaffected sites of 20 patients. Skin secretions were collected from 20 people exposed to arsenic-contaminated drinking water and 20 healthy, unexposed individuals. Results. The secretion of arsenic from the skin of healthy controls (mean ± SE; unit: μg/in.2 of skin/24 h; chest: 0.6 ± 0.2; back: 0.3 ± 0.1; abdomen: 0.5 ± 0.2) was increased several folds in arsenic-exposed controls (chest: 8.4 ± 1.8; back: 8.3 ± 1.9; abdomen: 6.7 ± 1.8) and patients (chest: 9.2 ± 1.3; back: 7.8 ± 1.3; abdomen: 5.2 ± 1.0). There was no difference in the skin lesions and unaffected sites in patients. However, the secretion of cholesterol was significantly lower in the chest of patients (190 ± 36) and healthy controls (686 ± 100) (p < 0.001). Secretions of vitamin E were low in healthy controls (chest: 8.5 ± 3.1; back: 5.2 ± 1.7; and abdomen: 8.7 ± 2.4), higher in arsenic-exposed controls (chest: 30.2 ± 8.1; back: 16.3 ± 8.9; and abdomen: 24.8 ± 9.3), and highest in patients [chest: 91.4 ± 14.9 (p < 0.0001 vs. control); back: 72.4 ± 13.2 (p < 0.001 vs. control); and abdomen: 46.8 ± 12.9]. Chronic intake of arsenic led to several folds higher secretion of zinc both in patients and in arsenic-exposed controls. One molecule of arsenic appears to be co-secreted with two molecules of zinc. Conclusion. Arsenic skin lesions showed no alteration in secretion of arsenic, although the secretion of cholesterol, vitamin E, and zinc was changed. Potential implications are discussed.

Factors Involved in the Development of Chronic Arsenic Poisoning in Bangladesh

gladesh are exposed to toxic levels (> 10 μg/l) of arsenic in their drinking water.1,2 Of these, approximately 30,000 individuals suffer from chronic arsenic poisoning. When all members of a family drink arseniccontaminated water from the same source, it is surprising that none, or only 1 or 2, may exhibit the signs and symptoms of chronic poisoning. The role of confounding factors such as age, sex, cigarette smoking, malnutrition, and impaired liver or kidney function in this outcome remains unclear. Design: To determine why some individuals suffered from chronic arsenic poisoning when others similarly exposed did not, we compared groups of family members with and without chronic arsenic poisoning, and a control group. Participants: Twelve patients who presented with signs of chronic arsenic poisoning were compared—in terms of age, sex, smoking habits, nutritional status, and liver and kidney function—with 23 of their family members who drank contaminated water from the same source but had no clinical manifestations of poisoning. Control data were from 15 individuals who consumed water from another source with an arsenic level ≤ 10 μg/l. Setting: Our study was conducted in a village about 35 km from the city of Dhaka, in an area where most of the residents obtained water from shallow tubewells that contained toxic concentrations of arsenic (mean concentration: 587.7 μg/l; highest concentration measured: 1,588.3 μg/l). Protocol: Diagnosis of chronic arsenic poisoning was based on (a) hyperpigmentation or “raindrop” pigmentation of the skin, (b) hyperkeratosis, and (c) a high concentration of arsenic in the urine. Blood samples were collected from patients and controls for measurement of hemoglobin concentration, glutathione (GSH) level in red blood cells, serum alanine transaminase, and creatinine level. Patients, family members, and controls were requested to provide 24-hr urine samples, in containers we provided, to estimate the total arsenic/day excreted. Urine collection was repeated for 3 consecutive days to obtain an average. We also calculated subjects’ body mass index (BMI) values, and assessed smoking habits, alcohol intake, daily water intake, and level of nutrition, by means of a prepared questionnaire. Arsenic concentrations in drinking water, and in subjects’ urine, were estimated by spectrophotometry. Results: Among the patients diagnosed with chronic arsenic poisoning, 11 (92%) were male and 1 (8%) was female. Family members who drank from the same source comprised 11 (47.8%) males and 12 (52.2%) females. All the controls were male. The mean ages of the 3 groups were similar: 39.3 yr, 31.4 yr, and 34.2 yr, respectively. Among the patients with chronic arsenic poisoning, 4 (33.3%) had a history of cigarette smoking. Similarly, 5 (21.7%) family members were smokers. There were 4 smokers (26.7%) among the control individuals. All the smokers were males who smoked more than 20 cigarettes/day. None of the subjects had a history of alcohol consumption. The BMI of those with chronic arsenic poisoning was 19.6, vs. 19.0 for their family members. This difference was not statistically significant. The mean BMI for controls was 23.2. The mean hemoglobin concentrations were 13.4 gm/dl, 13.1 gm/dl, and 14.7 gm/dl in patients, family members, and controls, respectively. Hemoglobin levels correlated well with other hematological indices and peripheral blood films. The average daily intake of water by the patients with Factors Involved in the Development of Chronic Arsenic