Co-administration of adjuvants along with Moringa oleifera attenuates beryllium-induced oxidative stress and histopathological alterations in rats.

CONTEXT: Moringa oleifera Lam. (Moringaceae) is a rich source of antioxidants. All parts of the plant are medicinally important and have been used as traditional medicine for a variety of human ailments in India. OBJECTIVE: Therapeutic efficacy of adjuvants with M. oleifera (MO) root extract was investigated against beryllium-induced oxidative stress. MATERIALS AND METHODS: Hydroalcoholic (50% v/v) root extract of M. oleifera (150 mg/kg, p.o.) alone and combinations of M. oleifera with either piperine (2.5 mg/kg, p.o.) or curcumin (5.0 mg/kg, p.o.) daily for 1 week were administered in experimental rats against beryllium toxicity (1.0 mg/kg, i.p. daily for 5 weeks). Oxidative stress parameters including blood sugar, G-6-Pase in liver, and DNA damage were analyzed. Histopathological changes in liver and kidney were also observed. RESULTS: Beryllium enhanced lipid peroxidation (LPO), depleted reduced glutathione (GSH) and antioxidant enzymes activities, decreased blood sugar and G-6-Pase activity, and did not damage DNA. Histologically, liver was observed with structural loss and disintegration of hepatocytes, heavy vacuolation in hepatocytes, and kidney was observed with constriction of glomeruli and hypertrophy in epithelial cells of uriniferous tubules. Therapy of M. oleifera with piperine was effective; however, combination of M. oleifera with curcumin showed better therapeutic effect by reduction of LPO, elevated GSH level, maintained antioxidant enzymes activities, restored blood sugar, and G-6-Pase activity in liver together with almost normal histoarchitecture of liver and kidney. DISCUSSION AND CONCLUSION: Curcumin enhanced therapeutic efficacy of M. oleifera root extract and showed better antioxidant potential against beryllium toxicity.

Case report: heavy metal burden presenting as Bartter syndrome.

CONTEXT: Maternal transfer of heavy metals during fetal development or lactation possibly contributed to the clinical manifestations of Bartter syndrome and developmental delay in the offspring. CASE PRESENTATION: An 11-month-old child diagnosed with Bartter syndrome and failure to thrive was treated concurrently for elevated metal burden while he was undergoing standard medical interventions. Treatment with body-weight doses of meso-2,3-dimercaptosuccinic acid (DMSA) reduced the body burden of lead, beryllium, copper, mercury, and cadmium at the three- and sixth-month follow-up tests. During the course of the six-month treatment, the patient gained 2.4 kg (5.2 lb) and grew approximately 9.5 cm (3.75 in). His weight shifted from significantly below the 5th percentile in weight to within the 5th percentile, and from below the 5th to within the 10th percentile for length. DISCUSSION: The child's acquisition of lead, beryllium, and copper correspond to his mother's history of stained glass assembly and occurred during fetal development or lactation, since there were no other identifiable sources that could have contributed to the heavy metal burden. Tests for known genetic mutations leading to Bartter syndrome were all negative. RELEVANCE TO CLINICAL PRACTICE: This case report highlights the potential benefit of DMSA for treatment of heavy metal body burden in infants who present with Bartter syndrome.

Use of proliferation tests to evaluate the effects of complexing agents on beryllium toxicity.

Occupational exposure to beryllium may cause chronic beryllium disease (CBD), a granulomatous interstitial pneumonitis caused by a cell-mediated immune response with delayed hypersensitivity initiated by an electrostatic interaction with the MHC class II human leukocyte antigen (HLA). Increased research efforts focus on the development of a CBD treatment by chelation therapy. This work presents an in vitro evaluation of the beneficial effects of beryllium chelation with different organic substrates. We have used a standard beryllium lymphocyte proliferation test (BeLPT) adapted for mouse splenocytes. Three complexing agents, 4,5-dihydroxy-1,3-benzenedisulfonic acid (tiron), nitrilotripropionic acid (NTP) and nitrilotriacetic acid (NTA), were tested using different protocols of the splenocyte proliferation test (SPT). We studied their corrective effect (beryllium pre-exposed splenocytes), their protective effect (ligand pre-exposed splenocytes) and their combined effects at fixed Be:L ratio of 1:2, at fixed Be concentration and at fixed L concentration. We also studied the effect of tiron in preventing splenocyte sensitization to beryllium. All three complexing agents showed a corrective effect and proved efficient in the combined effects, except NTA in the fixed Be:L ratio. Only NTP and tiron showed a significant protection at lower beryllium concentrations, while NTA was not significant. Splenocytes pre-exposed to chelated beryllium did not show sensitization while splenocytes pre-exposed to beryllium were sensitized. We observed a strong correlation between the efficiency of the complexing agent and its affinity towards beryllium. Both tiron and NTP showed a similar affinity towards the beryllium ion that is 10(7) higher than that of NTA.

Studies on the genotoxic effect of beryllium chloride and the possible protective role of selenium/vitamins A, C and E.

The genotoxic potential of beryllium chloride (BeCl2) was evaluated in vivo in mice using different endpoints. Chromosomal aberrations in bone marrow cells and in spermatocytes as well as sperm abnormalities were determined in the tested mice. The protective role of an orally administered drug consisting of selenium and vitamins A, C and E (selenium-ACE) was also studied. For analysis of chromosomal aberrations, both single and repeated oral treatments for a period of 3 weeks were performed. The doses used were 93.75, 187.50, 375, and 750 mg BeCl2/kg bw, which corresponds to 1/16, 1/8, 1/4, and 1/2 of the experimental LD50. BeCl2 induced a statistically significant increase in the percentage of chromosomal aberrations in both somatic and germ cells, with a dose- and time-response. The percentage of induced chromosomal aberrations was significantly reduced in all BeCl2-treated groups after oral administration of selenium-ACE. Beryllium chloride also induced a significant increase in the percentage of abnormal sperm. This percentage reached values of 9.62 +/- 0.32 and 5.56 +/- 0.31 in mice treated with the highest test dose of BeCl2 and with BeCl2+selenium-ACE, respectively, compared with 1.96 +/- 0.14 for the control. In conclusion, the results demonstrate the genotoxic effect of beryllium chloride and confirm the protective role of selenium-ACE against the genotoxicity of beryllium chloride.

Influence of alpha-tocopherol, propolis and piperine on therapeutic potential of tiferron against beryllium induced toxic manifestations.

The therapeutic potential of the chelator tiferron (sodium-4,5-dihydroxy-1,3-benzene disulphonate; 300 mg kg(-1), i.p.) and adjuvants, i.e. alpha-tocopherol (25 mg kg(-1), p.o.), propolis (a honey-bee hive product; 200 mg kg(-1), p.o.) and piperine (10 mg kg(-1), p.o.) were evaluated individually and in combination against beryllium induced biochemical alterations and oxidative stress consequences. Female albino rats were exposed to beryllium nitrate (1 mg kg(-1), i.p.) daily for 28 days followed by treatment with the above mentioned therapeutic agents for 5 consecutive days. Administration of beryllium altered blood biochemical variables with significant depletion in hemoglobin, blood sugar, total serum protein, albumin and significant enhancement in the release of serum transaminases. A significantly increased lipid peroxidation and a decreased level of glutathione after beryllium exposure indicated oxidative stress in the liver and kidney. Beryllium exposure decreased total protein and glycogen contents, whereas triglycerides and cholesterol increased significantly in liver and kidney. Individual administration of all the four compounds showed significant therapeutic potential in reverse of some of the biochemical parameters mentioned above. Furthermore, the combination of tiferron with alpha-tocopherol, propolis or piperine, respectively, could reverse all the variables significantly more towards the control. None of the test compounds showed any significant change in choleretic activity (bile flow and bile solids), indicating that these compounds had no adverse effects at these dose levels. It was concluded that all the combinations of tiferron and adjuvants played a beneficial role in reducing beryllium induced systemic toxicity at relatively lower doses and the combination of tiferron and propolis showed a more pronounced therapeutic potential.

Amelioration of beryllium induced alterations in hepatorenal biochemistry and ultramorphology by co-administration of tiferron and adjuvants.

Influence of adjuvants i.e., alpha-tocopherol (25 mg/kg, p.o.) and piperine (10 mg/kg, p.o.) on therapeutic potential of chelator tiferron (300 mg/kg, i.p.) was evaluated to encounter toxicogenic events of beryllium exposure. Albino rats were exposed to beryllium nitrate (1 mg/kg, i.p.) daily for 28 days followed by treatment of aforesaid therapeutic agents for 5 consecutive days. Results were considered to be significant at p < or =0.01 and p < or =0.05. Exposure to beryllium increased its concentration in liver, kidney and serum causing significant alterations in the activity of CYP-450 2E1 system, microsomal lipid peroxidation and protein; alkaline phosphtase, lactate dehydrogenase, gamma-glutamyl transpeptidase, bilirubin, creatinine and urea in serum; activity of acid phosphatase, alkaline phosphatase, adenosine triphosphatase, glucose-6-phosphatase and succinic dehydrogenase in liver and kidney. Beryllium exposure also induced severe alterations in histopathology and ultramorphology of liver and kidney proving its toxic consequences at cellular level. Tiferron along with adjuvants dramatically reversed alterations of all variables more towards control rather than individual treatment. Study concluded that tiferron in combination with alpha-tocopherol and piperine respectively was beneficial in diluting beryllium induced systemic toxicity; however, combination of tiferron and piperine presented more pronounced therapeutic potential.

Analysis of time-dependent recovery from beryllium toxicity following chelation therapy and antioxidant supplementation.

Efforts have been made to minimize the toxic effect caused by beryllium. Adult cyclic rats of Sprague Dawley strain were administered a bolus dose of 50mg/kg beryllium nitrate intramuscularly. The chelation therapy with glutathione (GSH), dimercapto propane sulfonic acid (DMPS)+ selenium (Se) and D-Penicillamine (DPA) + Se was given for 3 days followed by a rest of 1,3 and 7 days respectively. The results revealed a significant fall in the blood sugar level, serum alkaline phosphatase activity, serum proteins. A significant rise in the transaminases i.e. aspartate aminotranferase and alanine aminotranferase pattern is indicative of leakage of enzymes from liver resulting in alterations in the cell permeability. A rise in the hepatic lipid peroxidation activity is a direct indication of oxidative damage resulting in free radical generation. Results of the distribution studies by atomic absorption spectrophotometry reveal an increased concentration of beryllium in liver and kidney followed by lung and uterus. The relative ability of 3 chelating agents to act as antagonists for acute beryllium poisoning have been examined in liver, kidney, lungs and uterus. The appreciable change in the beryllium concentration in various organs is duration-dependent during the entire period being highly significant after 7 days rest. From the biochemical assays, and distribution studies it can be assumed that DPA+Se was the most effective therapeutic agent followed by DMPS+Se and GSH. Thus it can be concluded that DPA+Se is a better therapeutic agent as compared to DMPS+Se and GSH.

Role of chelating agents and antioxidants in beryllium induced toxicity.

The present study was conducted to evaluate the therapeutic effectiveness of chelating agents [glutathione, 2,3 dimercapto propane sulfonic acid (DMPS) and D-penicillamine (DPA)] in combination with antioxidant (sodium selenite) in beryllium induced toxicity in female rats. A bolus dose of 50mg/kg-beryllium nitrate was administered singly followed by chelation therapy with GSH, DMPS + Se and DPA + Se at various durations of 1,3 and 7 days respectively. Results revealed a significant fall in the glycogen content, whereas, a marginal fall in the protein was also observed. The enzymatic activity of alkaline phosphatase and adenosine triphosphatase was depleted; on the contrary, there was a significant rise in the acid phosphatase and glucose-6-phosphatase pattern. A rise in the hepatic lipid peroxidation activity is a direct indication of oxidative damage resulting in free radical generation. The distribution of the metal by atomic absorption spectrophotometry revealed an increased concentration of beryllium in liver and kidney, followed by lung and uterus. The relative ability of three chelating agents to act as antagonists, for acute beryllium poisoning, have been examined in liver, kidney, lungs and uterus. The appreciable change in the beryllium concentration in various organs is duration dependent during the entire period being highly significant at 7 days regimen. Biochemical and distribution studies reveal that DPA + Se was the most effective therapeutic agent followed by DMPS + Se and GSH.

Beryllium-induced toxicity and its prevention by treatment with chelating agents.

The efficacy of Tiron and calcium disodium EDTA in the treatment of experimental beryllium intoxication was investigated in rats. Beryllium nitrate was administered intramuscularly (50 mg kg(-1)) once only, provoking duration-dependent changes. Maximum changes were recorded after a 7-day regimen. Considerable inhibition was recorded in protein and glycogen contents, as well as in the activity of alkaline phosphatase, glucose-6-phosphatase, acid phosphatase and lipid peroxidation. These parameters were restored considerably with chelation therapy, but comparatively Tiron offered better protection. These findings were further confirmed by atomic adsorption spectrophotometry. Tiron was found to be significantly more effective than CaNa(2)EDTA in reducing the beryllium concentration in the liver, kidney and lungs.

Particle clearance and histopathology in lungs of C3H/HeJ mice administered beryllium/copper alloy by intratracheal instillation.

Beryllium/copper (BeCu) alloys are commonly used in the electronics, automotive, consumer, defense, and aerospace industries. Some individuals exposed occupationally to BeCu alloys have developed chronic beryllium disease. However, little is known of the toxicity and fate of BeCu alloys in the respiratory tract. To begin to address this question, we investigated the pulmonary toxicity and clearance of BeCu alloy (2% Be; 98% Cu) in mice. Groups of 40 female C3H/HeJ mice were administered 12.5, 25, and 100 microg BeCu alloy or 2 and 8 microg Be metal by intratracheal instillation. Mice were sacrificed at 1 h and 1, 7, 14, and 28 days postinstillation. Left lungs were evaluated for histopathological change. Right lungs were analyzed for Be and Cu content. Twenty-five percent of the high-dose BeCu mice and 7.5% of the mid-dose BeCu mice died within 24 h of dosing. Acute pulmonary lesions included acute alveolitis and interstitial inflammation. Type II epithelial cell hyperplasia and centriacinar fibrosis were present by 7 days after dosing. Lesions persisted through 28 days after instillation. No lesions attributable to alloy exposure were present in liver or kidney. Be metal instillation caused no deaths and minimal pulmonary changes over the time studied, indicating that the pulmonary lesions were due to Cu rather than Be. Cu cleared the lung with a half-time of 0. 5-2 days. Be cleared with a half-time of several weeks or longer. Results of this study suggest that exposure to BeCu alloy is more acutely toxic to lung than Be metal. The results of tissue analyses also indicate that, while the Cu component of the alloy clears the lung rapidly, Be is retained and may accumulate upon repeated exposure.

[Comparative informative value of chromosome aberrations and sister chromatid exchanges in the evaluation of metals in the environment]. / Sravnitel'naia informativnost' khromosomnykh aberratsii i sestrinskikh khromatidnykh obmenov pri otsenke metallov v okruzhaiushchei srede.

Simultaneous recording of the frequency of chromosomal aberrations and sister chromatid exchanges supplements data on the nature of genetic disorders in the sanitary and toxicological assessment of metals polluting the environment. A obvious threshold of genetic effects of metals was not revealed in the context of chromosomal aberrations and sister chromatid exchanges. Methods for detecting the genotoxicity of metals await further search.

Protective action of a herbal preparation Liv-52 against beryllium toxicity in rats.

An Ayurvedic medicine, Liv-52 has been studied as a prophylactic against beryllium induced toxicity in rats. Administration of beryllium per se caused severe degenerative and necrotic changes in liver, kidney, lungs and spleen. Its administration reduced glycogen content, activity of alkaline phosphatase and adenosine triphosphatase in these organs. On the contrary, activities of acid phosphatase and glucose-6-phosphatase were increased significantly. In Liv-52 primed rats significant recoupment was observed in all the parameters.

Cytogenetic effect of 235U neutrons and d(50)+Be fast neutrons.

Induction of chromosome aberrations in G0 lymphocytes of peripheral human blood exposed to 235U and d(50)+Be neutron radiation was studied. Dose--effect relationships for different types of chromosome aberrations were analyzed. Linear dependence of the effect was established for the studied neutron radiation, except for the yield of dicentrics exposed to d(50)+Be neutrons. Accordingly to the yield of dicentrics, the relative biological efficiency (RBE) of 235U and d(50)+Be neutrons was 19.5 and 4.14, respectively.

Liv-52 protection against beryllium toxicity in female albino rats.

Toxic effects of beryllium salts on the reproductive organs of cyclic adult female albino rats have been studied. An attempt was made to overcome these effects using an Ayurvedic medicine Liv-52 (Himalaya Drug Co., Bombay). Liv-52-primed rats (1 mL/rat/day for 15 days) were exposed to beryllium nitrate intravenously and were sacrificed at different time intervals. At autopsy ovary, uterus, cervix, and vagina were processed for biochemical and histopathologic examination. Histoarchitecture of the ovary, uterus, cervix, and vagina revealed severe necrotic changes with beryllium nitrate treatment. Tissue glycogen content and the activity of alkaline phosphatase were inhibited significantly after beryllium treatment. Total and esterified cholesterol levels increased significantly in these organs when exposed to beryllium salts. However, a significant improvement was observed in the biochemical parameters and histoarchitecture of these organs when beryllium was injected into Liv-52-primed animals.

Effect of feeding Gymnema sylvestre leaves on blood glucose in beryllium nitrate treated rats.

The feeding of powdered leaves of Gymnema sylvestre in the diet of rats for 10 days prior and 15 days after i.v. beryllium nitrate significantly protected the animals from the full fall of blood glucose seen in rats receiving beryllium nitrate alone. The feeding of the leaves for 25 days to normal rats did not alter blood glucose significantly. The leaves may contain a principle that could be useful as a prophylactic against beryllium toxicity.

Trace metals and neoplasia.

Numerous trace metals induce cancerous growths in various animal species in vivo and cause mutagenic or chromosomal transformations in cells-cultured cells in vitro. The most potent is probably nickel. The present review indicates that arsenic, cadmium, chromium, nickel and probably beryllium are associated with malignant neoplasms in humans. Inhalation of these metals during processing at refineries has lead to a greater incidence of pulmonary carcinoma as well as other forms of cancer. There is an inverse relationship between the amount of selenium in the environment and the death rate from cancer in humans. Evidence is presented in this review indicating that mutagenic metal ions alter the fidelity of DNA synthesis. This has been demonstrated with purified DNA polymerases using both synthetic and natural DNA templates in vitro, and by mutagenic or carcinogenic effects in vivo. The need for further studies of the molecular effects of metal ions on DNA replication, RNA transcription and translation is indicated by these results.

Biochemical mechanisms and morphological selectivity in hepatotoxicity: studies in cultures of hepatic-parenchymal and non-parenchymal cells.

Primary cultures of rat-liver parenchymal and non-parenchymal cells have been used to study some of the factors influencing the selective injury that can be caused in vivo by the direct-acting hepatotoxins beryllium, cadmium, ricin and modeccin to either liver-parenchymal or non-parenchymal cells. The studies on beryllium and cadmium compounds show that it is necessary to consider the chemical species generated in the culture medium, since particulate or colloidal forms are taken up predominantly by non-parenchymal cells whereas soluble forms more readily enter parenchymal cells. The studies with the glycoproteins ricin and modeccin illustrate the importance in their selective cell toxicity of specific membrane-recognition processes present in liver cells, particularly uptake in non-parenchymal cells through interactions with terminal mannose oligosaccharides in the toxins.