Avaliação bioquímica do extrato etanólico de própolis em ratos

Propolis is a natural product collected by honey bees containing, among other biochemical constituents, a variety of flavonoids. Propolis is a folk medicinal employed for treating various diseases. It is alleged to exhibit a broad spectrum of bioactivities. The aim of this study was to evaluate the effect of ethanolic extract of propolis (EEP) of species Plebeia droryana and Scaptotrigonea bipunctata through biochemical parameters. Rats were divided into 4 groups: (G1) untreated; (G2) ethanol treated; (G3) treated EEP of Plebeia droryana; (G4) treated of Scaptotrigonea bipunctata. The EEP (100 mg/kg b. w., daily) was administered orally to the animals, for 30 days. Treatment with EEP for two species showed reduction (p<0,05) in serum alanine aminotransferase, aspartato aminotransferase and alkaline phosphatase activity, compared to control ethanol values. The administration of EEP lowered significantly the serum levels of cholesterol (G3= 48,83±5,7 mg/dL; G4=56,91±6,5 mg/dL) and triacylglycerol (G3=45,17±4,16 mg/dL; G4=46,74± 3,90 mg/dL). The serum concentration of albumin (G3=4,16±0,6 g/dL; G4= 3,61±0,36 g/dL) increased (p<0,05) after the administration of EEP, however, it did not affect total protein and glucose concentration. The data suggest that EEP of two species caused alterations of the biochemical parameters.

Ultrastructural and Cytochemical Aspects of Metamorphosis in the Midgut of Apis mellifera L. (Hymenoptera: Apidae: Apinae)

The midgut of Apis mellifera is remodeled during metamorphosis. The epithelium and, to a lesser extent, the muscular sheath degenerate between the end of the last larval instar and the onset of pupation (prepupa).The larval epithelium is shed to the midgut lumen and digested, while a new epithelium is reconstructed from larval regenerative cells. During pupation, some reorganization still occurs, mainly in brown-eyed pupae. In pharate adult, the midgut wall shows the characteristics of adult, although some cells have pycnotic nuclei. The localization of alkaline and acid phosphatases showed that these enzymes were not involved in the reabsorption of the midgut wall.

Structural, cytochemical, immunocytochemical and ultrastructural characterization of blood granulocytes of the roadside hawk Buteo magnirostris (Gmelin, 1788) (Avian, Falconiform)

In the peripheral blood of the roadside hawk, Buteo magnirostris, the following types of granulocytic leucocytes were identified: heterophil, eosinophil and basophil. The heterophils presented acidophilic and spindle shaped granules, the eosinophils possess spherical eosinophilic granules and the basophils showed spherical and basophilic granules. The heterophils and eosinophils presented positive cytochemical reaction to glycogen and basic polyaminoacid, while the eosinophils presented sudanophilic granules, which were also positive for myeloperoxidase. The heterophils, alone, presented positivity for acid phosphatase in some granules and immunoreactivity to TGF-β1 was observed only in the cytoplasm of the eosinophils. Electron microscopy demonstrated the heterophil granules as predominantly spindle shaped, being strongly electron-dense, while the eosinophils had numerous uniformly electron-dense spherical granules and the basophils presented three different types of granules identified according to their electron-density and the aspect of their matrix.

Toxicity of hypercaloric diet and monosodium glutamate: oxidative stress and metabolic shifting in hepatic tissue.

The present study examines the effects of a hypercaloric diet on hepatic glucose metabolism of young rats, with and without monosodium glutamate (MSG) administration, and the association of these treatments with evaluating markers of oxidative stress. Male weaned Wistar rats (21 days old) from mothers fed with a hypercaloric diet or a normal diet, were divided into four groups (n=6): control (C) fed with control diet; (MSG) treated with MSG (4 mg/g) and control diet; (HD) fed with hypercaloric diet and (MSG-HD) treated with MSG and HD. Rats were sacrificed after the oral glucose tolerance test (OGTT), at 45 days of treatments. Serum was used for insulin determination. Glycogen, hexokinase(HK), glucose-6-phosphatase(G6PH), lipid hydroperoxide, superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) were determined in liver. HD rats showed hypoglycemia, hyperinsulinemia, and high hepatic glycogen, HK and decreased G6PH. MSG and MSG-HD had hyperinsulinemia, hyperglycemia, decreased HK and increased G6PH in hepatic tissue. These animals had impaired OGTT. HD, MSG and MSG-HD groups had increased lipid hydroperoxide and decreased SOD in hepatic tissue. Hypercaloric diet and monosodium glutamate administration induced alterations in metabolic rate of glucose utilization and decreased antioxidant defenses. Therefore, the hepatic glucose metabolic shifting induced by HD intake and MSG administration were associated with oxidative stress in hepatic tissue.

Influence of age on combined effects of cyclosporin and nifedipine on rat alveolar bone

Background: There is some evidence showing that cyclosporin A (CsA) and nifedipine (NIF) affect bone metabolism. The purpose of this work was to study the effects of CsA and NIF, given alone or concurrently, on alveolar bone of rats of different ages. Methods: Rats 15, 30, 60, and 90 days old were treated daily with 10 mg/kg body weight of CsA subcutaneously injected and/or 50 mg/kg body weight of NIF/day given orally for 60 days. Alveolar bone of the first lower molars was morphologically and stereologically evaluated in serial 5 μm bucco-lingual paraffin sections, stained with hematoxylin and eosin. Serum calcium and alkaline phosphatase levels were measured in all animals at the end of the experimental period. Results: Rats treated with CsA or NIF alone or CsA and NIF concurrently showed decreased alveolar bone density. CsA was more effective than NIF. A significant decrease in serum calcium was found only in animals treated with CsA or CsA/NIF. The results were similar regardless of age. Conclusions: These results indicate that the decrease in the alveolar bone volume in rats caused by CsA and NIF alone or concurrently is not age dependent. Furthermore, NIF (50 mg/kg) did not further increase the loss of alveolar bone volume induced by CsA (10 mg/kg).

Effects of long-term cyclosporin therapy on the periodontium of rats

Background: The treatment of cyclosporin A triggers an early bone loss and gingival overgrowth. There is a lack of studies exploring the effects of long-term cyclosporin A therapy on alveolar bone homeostasis and gingival tissue. Objective: The purpose of this study was to evaluate the effects of long-term therapy with cyclosporin A on the gingival tissue and on the alveolar bone metabolism in rats. Materials and methods: Rats were treated for 60, 120, 180 and 240 days with a daily subcutaneous injection of 10 mg/kg body weight of cyclosporin A. At the end of experimental periods, animals were killed and the serum calcium (Ca2+) and alkaline phosphatase levels were measured in all groups. After histological processing, the oral epithelium and the connective tissue, as well as volume densities of alveolar bone (Vb) and multinucleated osteoclasts (Vo), were assessed at the region of the lower first molars. Results: Significant increases in the serum alkaline phosphatase were observed in those groups that received cyclosporin A therapy. After 60 and 120 days of the treatment with cyclosporin A, evident gingival overgrowth associated with a significant increase of epithelium and connective tissue was observed, as well as a decrease of the densities of bone and an increase of densities of osteoclasts. After 180 and 240 days of the treatment, there was a reduction of the gingival overgrowth associated with significant decreases of epithelium and connective tissue, as well as an increase of bone densities and a decrease of osteoclasts. Conclusion: Within the limits of this experimental study, it can be concluded that the deleterious periodontal effects of cyclosporin A administration may be time-related side-effects. © Blackwell Munksgaard, 2004.

A comparative study of protein and enzymatic activity in venoms of some common wasps (Hymenoptera: Vespidae) from São Paulo State

The Hymenoptera Aculeata venoms, with few exceptions, have been poorly studied and characterized. Nevertheless, they have raised increasing interest due to their medical importance, since accidents with these insects are fairly frequent in Brazil and may cause severe allergic reactions. The objectives of the present work were the quantitative characterization of the main allergenic enzymes present in the venom of the species Polybia paulista, Polybia ignobilis, Polistes simillimus, and Agelaia pallipes pallipes through biochemical assays for the determination of total protein content, as well as the level of the enzymatic activity of phospholipase, hyaluronidase, acid phosphatase and esterase. These results, in addition to providing biochemical knowledge about the venom of the species in question, also supply studies that allow phylogenetic inferences among them.

Effect of Brazilian propolis on experimental hepatotoxicity in rats

Propolis (bee glue) is one of the major hive products of bees and is rich in flavonoids, which are known for their antioxidant activities. The aim of this study was to evaluate the hepatoprotective effects of the ethanolic extract of propolis (EEP) against experimental carbon tetrachloride (CCl 4)-induced liver toxicity in rats by means of biochemical indices. The animals were divided into 4 groups: GI= received mineral oil; GII= CCl 4(4mL/kg; Lp., single dose) treated; GIII= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Tetragonisca angustula, daily for 3 days and GIV= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Nannotrigonea testaceicornes, daily, for 3 days. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cholesterol and triacylglycerols were estimated after 3 days. CCl 4 caused a maximum increase (p<0,01) above biochemical parameters. As compared to CCl4 group (GII), the EEP (GIII and GIV) showed reduction in cholesterol, triacylglycerol, ALT, AST and alkaline phosphatase activity in the serum. In conclusion, these data indicate that EEP improved the dyslipidaemia, moreover, significantly attenuated increases in serum ALT and AST activities in rats with liver damage induced by carbon tetrachloride.

Enzymatic variability among venoms from different subspecies of Apis mellifera (Hymenoptera: Apidae)

The enzymatic variability was analyzed in venom extracts from bees reared in different colonies of the Africanized, A. m. ligustica and A. m. carnica subspecies. The implications of this variation focused on the biochemistry differentiation and immunogenicity of these venoms. The results showed the existence of a huge variability among the subspecies as well as among the colonies for three out of the six tested components - hyaluronidase, acid phosphatase and proteases - suggesting the utilization of these features as possible biochemical markers. Furthermore, although not statistically significant, it was found that the Africanized bee venom presented slightly higher levels of protein content and esterase activity, when compared to the other subspecies. If the esterase plays a role in the pain intensity caused by the sting, as suggested elsewhere, this might suggest a reason for a bigger algogenicity of this venom in relation to that of European bees. On the other hand, A. m. ligustica bees presented the highest levels of proteolytic and acid phosphatase activities, whose functions are not enlightened in Hymenoptera venoms. The A. m. carnica workers presented the highest hyaluronidase and the lowest acid phosphatase activity levels. The extremely variable results among colonies of the same subspecies and among subspecies, for the tested venom components, justify the absence of correlation between allergic reactions and tests with pooled venom.

Mensuração de hormônios andrógenos, estrógeno, fosfatase ácida prostática (PAP) e antígeno prostático específico (PSA) em cães adultos com próstata normal e hiperplásica

Prostatic diseases have been a common problem in middle age and older intact male dogs. Among these, benign prostatic hyperplasia (BHP) is the most frequent, age-related and hormonal-dependent condition of human and canine prostate. Blood samples were collected from 37 male intact dogs, tree years old dogs or more to determine androgens, estrogen, prostatic acid phosphatase (PAP) and prostatic specific antigen (PSA) according to histopathological aspects. Low levels of estrogen and high levels of prostatic specific antigen (PSA) were founded in dogs with BHP, respectively. Seric and urinary PAP levels were high in dogs with hyperplasia.

Localização histoquímica de algumas hidrolases no epidídimo do gerbilo

This study mainly showed that alkaline phosphatase expression had been present in the proximal regions of the epididymidis ductus of the gerbil which comprised the initial segment and proximal caput. The reactivities of acid phosphatase and ATPase were strong in the proximal and distal regions of the epididymidis ductus at the level of the apical cytoplasm and epithelium, except at the corpus level, a very thin isthmus located between the caput and cauda epididymidis, and as a general rule a low enzymatic reactive region of the epididymis of gerbil. SDH revealed also low activities in all the regions and regional structures of the duct, except into the luminal content formed by storaged spermatozoa, prior on the cauda level. The enzymes presented in the epididymis were correlated to some histophysiological roles such as the enzymatic mediation of endocytosis, secretion, absorption and active transport concerning to phosphatases and ATPase and a possible mitochondrial role of SDH could occur at the spermatozoa level in which the middle pieces were formed by a great amount of mitochondria.

Proposal of a new technique for bile duct reconstruction after iatrogenic injury. Study in dogs and review of the literature

Purpose: Interposition of a jejunal tube between the common bile duct and duodenum. Methods: Five adult mongrel dogs of both sexes, weighing on average 22.3 kg (18 to 26.5 kg), were used. Obstructive jaundice was induced by ligation of the distal common bile duct. After one week, a 2.5-cm long jejunal tube was fabricated from a segment of the loop removed 15 cm from the Treitz angle and interposed between the common bile duct and duodenum. Results: The animals presented good clinical evolution and no complications were observed. After 6 weeks, complete integration was noted between the bile duct mucosa, tube and duodenum and a significant reduction in total bilirubin and alkaline phosphatase was observed when compared to the values obtained one week after ligation of the common bile duct. Conclusion: The jejunal tube interposed between the dilated bile duct and duodenum showed good anatomic integration and reduced total bilirubin and alkaline phosphatase levels in the animals studied.

The materno-fetal interface in llama (Lama guanicoe glama)

Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5μm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

Comparative morphology of the ileum of three species of Cephalotes (formicidae, myrmicinae)

The histology (H.E.), histochemistry (carbohydrates, proteins, collagen, and calcium), enzyme profile (ATPase and Acid phosphatase), and ultramorphology of the ileum of Cephalotes atratus, C. clypeatus and C. pusillus were studied in order to obtain some information about the similarities of these species. The relationship among these three species, as well as the results obtained for the wall, and the contents of this portion of the digestive tract were discussed. In C. clypeatus the ileum is somewhat smaller than that of C. atratus, despite the similarities in body size. The ATPase results showed 2 different regions: the anterior region responsible for absorbing material from the haemolymph and the posterior region for absorbtion from the ileum lumen. Other histochemical analyses failed to show any differences.

Efeito do alumínio no conteúdo de poliaminas livres e atividade da fosfatase ácida durante o crescimento de brotações de Eucalyptus grandis × E. urophylla cultivadas in vitro

This paper is aiming to evaluate the effects of different levels of the aluminum on the growth of the Eucalyptus grandis x E. urophylla shoots cultivated in vitro. Evaluations were carried out on pH and chemicals modifications of the culture medium by Geochem program, polyamines contents (putrescine, espermidine e espermine) and acid phosphatase ativity on the shoots. The trial had a totally randomized design with four treatments and four replicates. The treatments were: 0.0, 6.75,13.50 and 27 mg.L -1 of AlCl 3.6H 2. Evaluations were carried out on the 4th, 8th, 12th, 16th, 20th, 24th and 28th day of culture. The addition of the aluminium, in all concentrations, affected the culture medium ionic equilibrium, the morphology of the shoots, reduced the pH on the medium, induced an increase in polyamines content and higher acid phosphatase activity.