Toxic effects of alcohol intake on prostate of rats

BACKGROUND. The present report was carried out to determine whether alcohol intake could induce prostate lesions.METHODS. We tested male rats for 300 days. Animals were divided into three groups: controls received only tap water as liquid diet; the chronic alcohol intake group received only ethanol solution in semivoluntary research; and the withdrawal group received the same treatment as chronic alcohol intake until 240 days, after which they reverted to drinking water.RESULTS. Chronic alcohol intake increased lipoperoxide concentrations and acid phosphatase activities. Cu-Zn superoxide dismutase (SOD) was decreased at 60 days, but approached controls values at 300 days following treatment. The serum increased alkaline phosphatase, and alanine transaminase activities reflected the chronic toxic effect of ethanol.CONCLUSIONS. Since SOD activity was unable to scavenge superoxide radical and lipoperoxide formation, we can conclude that superoxide is an important intermediate in prostate damage of chronic alcohol intake. (C) 1997 Wiley-Liss, Inc.

REACTIVE OXYGEN GENERATION BY AZOMETHINE-H - A NEW ANTIMALARIAL DRUG

Superoxide radical (O-2(-)) is a free radical that may be involved in various toxic processes. Cu-Zn superoxide dismutase catalyzes the dismutation of the superoxide free radical and protects cells from oxidative damage. A rat bioassay validated for the identification of the toxic effects of azomethine H revealed increased serum activities of amylase, alanine transaminase, and alkaline phosphatase. The lipoperoxide and bilirubin concentrations were also increased in animals that received azomethine H (1 g/kg) from ascorbic or hydrochloric acid solutions. Azomethine H increased Cu-Zn superoxide dismutase activity. This elevation of Cu-Zn superoxide dismutase activity was highest on the 7th day and was at levels comparable with those of control rats from day 60 onwards. Superoxide is an important intermediate in the action and toxicity of azomethine H.

Alendronate therapy in cyclosporine-induced alveolar bone loss in rats

Background and Objective: Cyclosporine A is an immunosuppressive drug that is widely used in organ transplant patients as well as to treat a number of autoimmune conditions. Bone loss is reported as a significant side-effect of cyclosporine A use because this can result in serious morbidity of the patients. As we have shown that cyclosporine A-associated bone loss can also affect the alveolar bone, the purpose of this study was to evaluate the effect of the concomitant administration of alendronate on alveolar bone loss in a rat model.Material and Methods: Forty Wistar rats (10 per group) were given cyclosporine A (10 mg/kg, daily), alendronate (0.3 mg/kg, weekly), or both cyclosporine A and alendronate, for 60 d. The control group received daily injections of sterile saline. The expression of proteins associated with bone turnover, including osteocalcin, alkaline phosphatase and tartrate-resistant acid phosphatase (TRAP), and also the calcium levels, were evaluated in the serum. Analysis of the bone volume, alveolar bone surface, the number of osteoblasts per bone surface and the number of osteoclasts per bone surface around the lower first molars was also performed.Results: the results indicate that cyclosporine A treatment was associated with bone resorption, represented by a decrease in the bone volume, alveolar bone surface and the number of osteoblasts per bone surface and by an increase in the number of osteoclasts per bone surface and TRAP-5b. These effects were effectively counteracted by concomitant alendronate administration.Conclusion: It is concluded that concomitant administration of alendronate can prevent cyclosporine A-associated alveolar bone loss.

Leukocyte and thrombocyte reference values for channel catfish (Ictalurus punctatus Raf), with an assessment of morphologic, cytochemical, and ultrastructural features

Background: Hematology tests are useful to evaluate physiologic disturbances in fish and can provide important information for the diagnosis and prognosis of disease. Objectives: the primary purpose of this study was to define reference intervals for thrombocytes and leukocytes in healthy channel catfish (Ictalurus punctactus). In addition, the morphologic, cytochernical, and ultrastructural features of blood cells were assessed. Methods: Blood samples (0.5 mL were collected into EDTA from 40 clinically healthy catfish on a commercial fish farm in Jaboticabal, Brazil. Thrombocyte, total WBC, and differential WBC counts were determined and reference intervals were calculated as the 25-95th percentiles of data. Thrombocyte and leukocyte morphology was assessed in blood smears stained with May Griinwald-Giemsa-Wright and ultrastructurally by transmission electron microscopy. Cytochemical staining patterns were described using periodic acid-Schiff (PAS), peroxidase, nonspecific esterase, alkaline phosphatase, and toluidine blue. Results: Reference intervals were as follows: thrombocytes 58,802-99,569/mu L; total WBCs 27,460-41,523/mu L; lymphocytes 5380-11,581/mu L; monocytes 2949-7459/mu L; neutrophils 12,529-22,748/mu L, and basophils 736-2003/mu L. Neutrophils were positive for peroxidase and PAS; monocytes were positive for nonspecific esterase; and basophils were positive with toluidine blue. Conclusion: the morphologic and staining features of neutrophils and monocytes of channel catfish are similar to those of mammals, and the presence of basophils in this species was verified. These reference intervals and morphologic findings provide a foundation for future investigations on the functions and alterations of blood cells in channel catfish.

Isoflavone-supplemented soy yoghurt associated with resistive physical exercise increase bone mineral density of ovariectomized rats

Objective: To determine the effects produced by the ingestion of a fermented soy product (soy yoghurt), supplemented with isoflavones and associated with a resistive exercise program, on the bone metabolism of mature ovariectomized (Ovx) and sham-ovariectomized (sham-Ovx) rats.Methods: A total of 56 rats were used. They were divided into 2 sedentary control groups, the Ovx control group (C-Ovx) and the sham-Ovx control group (C-Sovx), each with 7 sedentary animals, and 2 treated groups, Ovx and sham-Ovx, with 21 animals each. These two treated groups were subdivided into three subgroups of seven animals each, which received the following treatments: consuming the soy yoghurt + sedentary, only subjected to resistive exercise, and consuming the soy yoghurt + resistive exercise. Both the program of resistive exercise and the consumption of soy yoghurt (at 3 mL/(kg body weight day)) continued for 12 weeks. The soy yoghurt was supplemented with isoflavones at 50 mg/100 g of product. The animals were sacrificed and their right-side femurs and tibias removed and assessed for bone mineral density (BMD). The alkaline phosphatase activity (AP) was determined in the blood serum.Results: There was a significant increase in both femur and tibia BMD values and in serum alkaline phosphatase activity in all the treated subgroups, compared with the control groups (p < 0.05).Conclusion: the ingestion of the soy yoghurt supplemented with isoflavones was capable of preventing a loss of bone mass in Ovx rats and of increasing bone mass in sham rats, whilst the resistive exercise program was effective in augmenting the bone mass in sham and Ovx rats. (c) 2007 Elsevier B.V.. All rights reserved.

Constituintes sangüíneos de bovinos recém-nascidos das raças Nelore (Bos indicus) e Holandesa (Bos taurus) e de bubalinos (Bubalus bubalis) da raça Murrah

Newborn Holstein-Friesian and Nelore cattle and Murrah buffalo blood samples were collected for hemograma and serum constituents testing. Results indicated that: 1. buffalo red blood cell (RBC) counts were lower than RBC counts for cattle during the first 30 days after birth; 2. buffalo leukocytes, neutrophil and lymphocyte counts increased in older animals; 3. The highest fibrinogen levels occurred in older calves; 4. buffalo glucose levels were lower than Nelore cattle and decreased in the older calves; 5. buffalo iron, urea, and aspartate aminotransferase levels were higher than in cattle with the highest concentration at the time of birth; 6. cattle bilirubin and alanine aminotranferase levels were higher than buffalo with the highest value at the time of birth; 7. alkaline phosphatase and gammaglutamyltransferase activities were lower in the older calves; 8. buffalo magnesium levels were higher than cattle with lower concentrations at the time of birth..

Constituintes sangüíneos de vacas das raças Nelore (Bos indicus) e Holandesa (Bos taurus) e de bubalinas (Bubalus bubalis) da raça Murrah durante a gestação, no dia do parto e no puerpério

Blood constituents of Nelore, Hosltein and buffalo cows were determined during pregnancy, at the time of calving and at post-partum. It was verified that. 1. The pregnancy and post-partum periods had no influence in the erythrocyte values, but at calving the red blood cell counts and hemoglobin levels of Holstein and buffalo cows were lower than for Nelore cattle; 2. leukocyte counts were similar among groups; 3. total protein levels of Nelore cows were lower than Holstein and buffalo. The albumin levels were rite lowest at rite time of calving compared to late pregnancy; 4. glicose levels were lower ill buffalo cows during pregnancy as compared with Holstein cows. The glicemia of Nelore cows was lower as compared to Holstein cow's irt late pregnancy; 5. urea and creatinine levels were higher in buffalo cows than cattle. The urea and creatinine levels were greater in buffalo cows with maximum values at the post-partum and at the parturition, respectively; 6. bilirrubin levels were higher in bovine than buffalo cows; 7. aspartate aminotransferase activity was greater in buffalo cows and increased at the time of calving; 8. alkaline phosphatase activity was increased during pregnancy and decreased after the time of calving; 9. gammnglutamyltransferase activity,vas the highest for buffalo cows after calving; IO. calcium levels were the highest for Holstein cows at the post-partum and the phosphorus levels were higher in buffalo cows, which had the highest magnesium levels at the parturation.

In vitro biocompatibility of a novel membrane of the composite poly(vinylidene-trifluoroethylene)/barium titanate

This study was aimed at investigating the in vitro biocompatibility of a novel membrane of the composite poly(vinylidene-trifluoroethylene)/barium titanate (P(VDF-TrFE)/BT). Osteoblastic cells were obtained from human alveolar bone fragments and cultured under standard osteogenic condition until subconfluence. First passaged cells were cultured on P(VDF-TrFE)/BT and expanded polytetrafluoroethylene (e-PTFE - control) membranes in 24-well plates. Cell adhesion and spreading were evaluated at 30 min, and 4 and 24 h. For proliferation assay, cells were cultured for 1, 7, and 10 days. Cell viability was detected by trypan blue at 7 and 10 days. Total protein content and alkaline phosphatase (ALP) activity were measured at 7, 14, and 21 days. Cultures were stained with Alizarin red at 21 days, for detection of mineralized matrix. Data were compared by ANOVA and Student t test. Cell attachment (p = 0.001), cell number (p = 0.001), and ALP activity (p = 0.0001) were greater on P(VDF-TrFE)/BT. Additionally, doubling time was greater on P(VDF-TrFE)/BT (p = 0.03), indicating a decreased proliferation rate. Bone-like nodule formation took place only on P(VDF-TrFE)/BT. The present results showed that both membranes are biocompatible. However, P(VDF-TrFE)/BT presented a better in vitro biocompatibility and allowed bone-like nodule formation. Therefore, P(VDF-TrFE)/BT could be an alternative membrane to be used in guided tissue regeneration. (c) 2006 Wiley Periodicals, Inc.

THE ESSENTIAL ROLE OF ZINC IN GROWTH

Zinc is known to play a relevant role in growth and development. The basic mechanisms of action of this trace element are intimately linked to the structure and action of countless enzymes involved in many different metabolic processes. In this respect, when zinc specifically acts on cartilage growth it is involved in multiple enzymatic reactions which make this a multifactorial event. Thus, we may divide the actions of zinc into three distinct types: 1) action on taste and smell acuity, appetite regulation, and food consumption and regulation; 2) action on DNA and RNA synthesis stimulating a) cell replication and differentiation of chondrocytes, osteoblasts and fibroblasts; b) cell transcription culminating in the synthesis of somatomedin-C (liver), alkaline phosphatase, collagen and osteocalcin (bone), and c) protein, carbohydrate and lipid metabolism, that is intimately related to the mechanisms of smell, taste, appetite, and food consumption and utilization; 3) action on hormonal mediation by participating in a) GH synthesis and secretion in somatomammotroph cells, b) the action of GH on liver somatomedin-C production, and c) somatomedin-C activation in bone cartilage. In addition to these multiple functions, zinc also interacts with other hormones somehow related to bone growth such as testosterone, thyroid hormones, insulin, and vitamin D-3.On the basis of the above considerations, we conclude that the integration of these mechanisms contributes to the perfect physiological functioning of bone. Tn the presence of zinc deficiency, this homeostasis is impaired, causing the weight-height deficiency detected in several species studied, the human species in particular.

SUPEROXIDE RADICAL AND TOXICITY OF ENVIRONMENTAL NICKEL EXPOSURE

Three nickel compounds were tested for pancreatic, hepatic and osteogenic damage in rats by a single i.m. injection Ni++ (7 mg kg(-1)). The nickel induced biochemical alterations included significantly increased levels of serum alkaline phosphatase in rats with NiS (75%) and NiO (50%). Amylase and aspartate transaminase were also increased, and lipoperoxide was increased in rats with NiO (5.6-fold) and NiS (3.4-fold). No serum changes were observed with NiCl2. Daily injection of Cu-Zn superoxide dismutase (SOD) conjugated with polyethylene glycol prevented the serum level changes, indicating that superoxide radical is an important intermediate in toxicity of nickel insoluble compounds.

TOXIC EFFECTS OF WATER EUTROPHICATION ON PANCREATIC, HEPATIC AND OSTEOGENIC TISSUES OF RATS

Pollution, industrial solvents, concentrations of metals and other environmental agents are widely related to biochemicals values which are used in disease diagnosis of environmental toxicity. A rat bioassay validated for the identification of toxic effects of eutrophication revealed increased serum activities of amylase, alanine transaminase (BLT) and alkaline phosphatase (ALP) in rats that received algae, filtered water and nickel or cadmium from drinking water. Serum Cu-Zn superoxide dismutase activity decreased from its basal level of 40.8 +/- 2.3 to 26.4 U/mg protein, at 7 days of algae and at 48 hr of nickel and cadmium water ingestion. The observation that lipoperoxide concentration was not altered in rats treated with filtered water, while amylase, ALT and ALP were increased in these rats and in those treated with nickel or cadmium, indicated that pancreatic, hepatic and osteogenic lesions by eutrophication were not related to superoxide radicals, and might be due to a novel toxic environmental agent found in filtered and non-filtered algae water.

Conversion of immunosuppressive monotherapy from cyclosporin A to tacrolimus reverses bone loss in rats

Tacrolimus is used for transplant patients with refractory graft rejection and those with intolerance to cyclosporin (CsA), without the disfiguring adverse effects frequently attributed to CsA therapy. Since we have shown that CsA-associated bone loss can also affect alveolar bone, the purpose of this study was to evaluate the effects of conversion of monotherapy from CsA to tacrolimus on alveolar bone loss in rats. Groups of rats were treated with either CsA (10 mg/kg/day, s.c.), tacrolimus (I mg/kg/day, s.c.), or drug vehicle for 60 and 120 days, and an additional group received CsA for 60 days followed by conversion to tacrolimus for a further 60-day period. Bone-specific alkaline phosphatase (BALP), tartrate-resistent acid phosphatase (TRAP-5b), calcium (Ca2+), interleukin (IL)-1 beta, IL-6, and tumor necrosis factor alpha (TNF-alpha) concentrations were evaluated in the serum. Analyses of bone volume, bone surface, number of osteblasts, and osteoclasts were performed. Treatment with CsA for either 60 or 120 days was associated with bone resorption, represented by lower bone volume and increased number of osteoclasts; serum BALP, TRAP-5b, IL-1 beta, IL-6, and TNF-alpha were also higher in these animals. After conversion from CsA to tacrolimus, all the altered serum markers returned to control values in addition to a significant increase of bone volume and a lower number of osteoclasts. This study shows that conversion from CsA to tacrolimus therapy leads to a reversal of the CsA-induced bone loss, which can probably be mediated by downregulation of IL-1 beta, IL-6, and TNF-alpha production.

Kinetic characterization of hypophosphatasia mutations with physiological substrates

We have analyzed 16 missense mutations of the tissue-nonspecific AP (TNAP) gene found in patients with hypophosphatasia. These mutations span the phenotypic spectrum of the disease, from the lethal perinatal/infantile forms to the less severe adult and odontohypophosphatasia. Site-directed mutagenesis was used to introduce a sequence tag into the TNAP cDNA and eliminate the glycosylphosphatidylinositol (GPI)-anchor recognition sequence to produce a secreted epitope-tagged TNAP (setTNAP). The properties of GPI-anchored TNAP (gpiTNAP) and setTNAP were found comparable. After introducing each single hypophosphatasia mutation, the setTNAP and mutant TNAP cDNAs were expressed in COS-1 cells and the recombinant flagged enzymes were affinity purified. We characterized the kinetic behavior, inhibition, and heat stability properties of each mutant using the artificial substrate p-nitrophenylphosphate (pNPP) at pH 9.8. We also determined the ability of the mutants to metabolize two natural substrates of TNAP, that is, pyridoxal-5'-phosphate (PLP) and inorganic pyrophosphate (PPi), at physiological pH. Six of the mutant enzymes were completely devoid of catalytic activity (R54C, R54P, A94T, R206W, G317D, and V365I), and 10 others (A16V, A115V, A160T, A162T, E174K, E174G, D277A, E281K, D361V, and G439R) showed various levels of residual activity. The A160T substitution was found to decrease the catalytic efficiency of the mutant enzyme toward pNPP to retain normal activity toward PPi and to display increased activity toward PLP. The A162T substitution caused a considerable reduction in the pNPPase, PPiase, and PLPase activities of the mutant enzyme. The D277A mutant was found to maintain high catalytic efficiency toward pNPP as substrate but not against PLP or PPi. Three mutations ( E174G, E174K, and E281K) were found to retain normal or slightly subnormal catalytic efficiency toward pNPP and PPi but not against PLP. Because abnormalities in PLP metabolism have been shown to cause epileptic seizures in mice null for the TNAP gene, these kinetic data help explain the variable expressivity of epileptic seizures in hypophosphatasia patients.

Response of rat bone marrow cells to commercially pure titanium submitted to different surface treatments

Objectives. Alterations in the commercially pure titanium (cpTi) surface may be undertaken to improve its biological properties. The aim of this study is to investigate the biocompatibility of cpTi submitted to different surface treatments.Methods. The cpTi surfaces were prepared so that machined and blasted surfaces, either acid etched or not, were compared using rat bone marrow cells cultured to differentiated into osteoblast. For attachment evaluation, cells were cultured for 4 and 24 h. Cell morphology was evaluated after 3 days. After 7, 14, and 21 days cell proliferation was evaluated. Total protein content and alkaline phosphatase (ALP) activity were evaluated after 14 and 21 days. For bone-like nodule formation, cells were cultured for 21 days. Data were compared by analysis of variance.Results. Cell attachment, cell morphology, cell proliferation, and ALP activity were not affected by surface treatments. Total. protein content was reduced by blasted and acid etched surface. Bone-Like nodule formation was significantly reduced by blasted, acid etched, and a combination of both blasted and acid etched surfaces.Conclusions. Based on these results, it can be suggested that cpTi surfaces that were submitted only to machining treatment favor the final event of osteoblastic differentiation of the rat bone marrow cells, evidenced by increased bone-Like nodule formation. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

PHYLOGENETIC STUDIES OF SOME SPECIES OF THE GENUS COFFEA .2. NUMERICAL-ANALYSIS OF ISOENZYMATIC DATA

Thirteen species of Coffea were studied for five enzymes systems, including alpha and beta esterase, alkaline phosphatase, acid phosphatase, malate dehydrogenase and acid dehydrogenase. Three coefficients of similarity: Simple Matching, Jaccard and Ochiai and three different clustering methods: Single Linkage, Complete Linkage and Unweighted Pair Group, using Arithmetic Averages (UPGMA) were used to analyse the data.The phylogenetic relationships among the twelve diploid species and between them and the tetraploid species C. arabica showed that similarity among species of the same subsection is not always greater than among species of different subsections. In addition, although there are several similarity groups in common, established by isoenzymatic polymorphism, morphological characteristics, chemical data, crossability and geographic distribution, there is no common trend among the phylogenetic relationships as indicated by all these different evaluating procedures.