Spirulina enhanced the skeletal muscle protein in growing rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Free activity in the cage associated with body weight and restoration of bone structural and mechanical properties in growing rats after hindlimb unloading

We investigated the importance of daily free activity in the cage and body weight gain during the recovering of bone structural and mechanical properties in growing rats after hindlimb unloading. Eight-week-old male Wistar rats were randomly divided into control (CG, n=24) and suspended (SG, n=24) groups. Animals from SG underwent a four-week hindlimb unloading period by tail-suspension. Animals from CG and those from SG after release were kept in collective cages and sacrificed at the age of 12, 16 and 20 weeks. Both femurs were removed and its area, bone mineral density (BMD), resistance to failure and stiffness were determined. Four-week hindlimb unloading decreased (p < 0.05) body weight (CG, 373.00 +/- 9.47 vs. SG, 295.86 +/- 9.19 g), BMD (CG, 0.19 +/- 0.01 vs. SG, 0.15 +/- 0.01 g/cm(2)), bone resistance to failure (CG, 147.75 +/- 5.05 vs. SG, 96.40 +/- 5.95 N) and stiffness (CG, 0.38 +/- 0.01 vs. SG, 0.23 +/- 0.02 N/m). Eight weeks of free activity in cage recovered (p > 0.05) the body weight (CG, 472.75 +/- 14.11 vs. SG, 444.75 +/- 18.91 g), BMD (CG, 0.24 +/- 0.01 vs. SG, 0.22 +/- 0.01 g/cm(2)), bone resistance to failure (CG, 195.73 +/- 10.06 vs. SG, 178.45 +/- 8.48 N) and stiffness (CG, 0.56 +/- 0.02 vs. SG, 0.47 +/- 0.03 N/m) of SG animals. Body weight correlated strongly with bone structural and mechanical properties (p < 0.0001). In conclusion, free activity in the cage associated with body weight gain restored bone structural and mechanical properties in growing rats after hindlimb unloading.

Decreased rate of protein synthesis, caspase-3 activity, and ubiquitin-proteasome proteolysis in soleus muscles from growing rats fed a low-protein, high-carbohydrate diet

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Arraste via fecal de nutrientes da ingestão produzido por bagaço de mandioca hidrolisado

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeitos a longo prazo de alterações de dieta numa fase precoce da vida sobre o crescimento posterior: estudo experimental em ratos jovens

Pós-graduação em Pediatria - FMB

Decrease in the number and apoptosis of alveolar bone osteoclasts in estrogen-treated rats

Bone is a mineralized tissue that is under the influence of several systemic, local and environmental factors. Among systemic factors, estrogen is a hormone well known for its inhibitory function on bone resorption. As alveolar bone of young rats undergoes continuous and intense remodeling to accommodate the growing and erupting tooth, it is a suitable in vivo model for using to study the possible action of estrogen on bone. Thus, in an attempt to investigate the possibility that estrogen may induce the death of osteoclasts, we examined the alveolar bone of estrogen-treated rats.Fifteen, 22-d-old female rats were divided into estrogen, sham and control groups. The estrogen group received estrogen and the sham group received corn oil used as the dilution vehicle. After 8 d, fragments containing alveolar bone were removed and processed for light microscopy and transmission electron microscopy. Sections were stained with hematoxylin and eosin and tartrate-resistant acid phosphatase (TRAP)-an osteoclast marker. Quantitative analysis of the number of TRAP-positive osteoclasts per mm of bone surface was carried out. For detecting apoptosis, sections were analyzed by the Terminal deoxynucleotidyl transferase-mediated dUTP Nick-End Labeling (TUNEL) method; TUNEL/TRAP combined methods were also used.The number of TRAP-positive osteoclasts per mm of bone surface was significantly reduced in the estrogen group compared with the sham and control groups. TRAP-positive osteoclasts exhibiting TUNEL-positive nuclei were observed only in the estrogen group. In addition, in the estrogen group the ultrastructural images revealed shrunken osteoclasts exhibiting nuclei with conspicuous and tortuous masses of condensed chromatin, typical of apoptosis.Our results reinforce the idea that estrogen inhibits bone resorption by promoting a reduction in the number of osteoclasts, thus indicating that this reduction may be, at least in part, a consequence of osteoclast apoptosis.

Apoptotic bone cells may be engulfed by osteoclasts during alveolar bone resorption in young rats

The alveolar bone is a suitable in vivo physiological model for the study of apoptosis and interactions of bone cells because it undergoes continuous, rapid and intense resorption/remodelling, during a long period of time, to accommodate the growing tooth germs. The intensity of alveolar bone resorption greatly enhances the chances of observing images of the extremely rapid events of apoptosis of bone cells and also of images of interactions between osteoclasts and osteocytes/osteoblasts/bone lining cells. To find such images, we have therefore examined the alveolar bone of young rats using light microscopy, the TUNEL method for apoptosis, and electron microscopy. Fragments of alveolar bone from young rats were fixed in Bouin and formaldehyde for morphology and for the TUNEL method. Glutaraldehyde-formaldehyde fixed specimens were processed for transmission electron microscopy. Results showed TUNEL positive round/ovoid structures on the bone surface and inside osteocytic lacunae. These structures - also stained by hematoxylin - were therefore interpreted, respectively, as osteoblasts/lining cells and osteocytes undergoing apoptosis. Osteoclasts also exhibited TUNEL positive apoptotic bodies inside large vacuoles; the nuclei of osteoclasts, however, were always TUNEL negative. Ultrathin sections revealed typical apoptotic images - round/ovoid bodies with dense crescent-like chromatin - on the bone surface, corresponding therefore to apoptotic osteoblasts/lining cells. Osteocytes also showed images compatible with apoptosis. Large osteoclast vacuoles often contained fragmented cellular material. Our results provide further support for the idea that osteoclasts internalize dying bone cells; we were however, unable to find images of osteoclasts in apoptosis. (C) 2001 Harcourt Publishers Ltd.

Effects of leucine supplemented diet on intestinal absorption in tumor bearing pregnant rats

Background: It is known that amino acid oxidation is increased in tumor-bearing rat muscles and that leucine is an important ketogenic amino acid that provides energy to the skeletal muscle.Methods: To evaluate the effects of a leucine supplemented diet on the intestinal absorption alterations produced by Walker 256, growing pregnant rats were distributed into six groups. Three pregnant groups received a normal protein diet (18% protein):pregnant (N), tumor-bearing (WN), pair-fed rats (Np). Three other pregnant groups were fed a diet supplemented with 3% leucine (15% protein plus 3% leucine):leucine (L), tumor-bearing (WL) and pair-fed with leucine (Lp). Non pregnant rats (C), which received a normal protein diet, were used as a control group. After 20 days, the animals were submitted to intestinal perfusion to measure leucine, methionine and glucose absorption.Results: Tumor-bearing pregnant rats showed impairment in food intake, body weight gain and muscle protein content, which were less accentuated in WL than in WN rats. These metabolic changes led to reduction in both fetal and tumor development. Leucine absorption slightly increased in WN group. In spite of having a significant decrease in leucine and methionine absorption compared to L, the WL group has shown a higher absorption rate of methionine than WN group, probably due to the ingestion of the leucine supplemented diet inducing this amino acid uptake. Glucose absorption was reduced in both tumor-bearing groups.Conclusions: Leucine supplementation during pregnancy in tumor-bearing rats promoted high leucine absorption, increasing the availability of the amino acid for neoplasic cells and, mainly, for fetus and host utilization. This may have contributed to the better preservation of body weight gain, food intake and muscle protein observed in the supplemented rats in relation to the non-supplemented ones.

Root surface treatment with vitamin C in tooth replantation: microscopic study in rats

AIM: Late tooth replantation is a worthy procedure, especially in growing patients, being the best option for tooth avulsion. This study evaluated the effects of root surface treatment with 2% acidulated phosphate sodium fluoride and effervescent vitamin C (Redoxon) in late replantation. METHODS: Twenty rat teeth (Rattus norvegicus, albinus, Wistar) were extracted and left on a table for 6 h. Then, the dental papilla and enamel organ were sectioned and the pulp was removed through the apex. After removal of the periodontal ligament with a blade, the animals were divided into 2 groups: Group I teeth were immersed in 2% acidulated phosphate sodium fluoride solution for 10 min and obturated with Ca(OH)(2) paste. Group II teeth were immersed in effervescent vitamin C solution (Redoxon 2 g) for 10 min and obturated as for Group I. After these procedures, teeth were replanted and animals were killed after 60 days. RESULTS: The study revealed a larger amount of replacement resorption in Group I and larger amount of ankylosis in Group II, with statistically significant difference and absence of inflammatory resorption. CONCLUSION: The substances used for root surface treatment were unable to prevent replacement resorption and ankylosis, which are expected when the periodontal ligament has been lost.

Moderate physical activity from childhood contributes to metabolic health and reduces hepatic fat accumulation in adult rats

Background: Obesity, oxidative stress and inflammation, by triggering insulin resistance, may contribute to the accumulation of hepatic fat, and this accumulation by lipotoxicity can lead the organ to fail. Because obesity is growing at an alarming rate and, worryingly, in a precocious way, the present study aimed to investigate the effects of moderate physical training performed from childhood to adulthood on liver fat metabolism in rats. Methods. Twenty rats that were 28days old were divided into two groups: control (C) and trained (T). The C Group was kept in cages without exercise, and the T group was submitted to swimming exercise for 1hour/day, 5days/week from 28 to 90days of age (8weeks) at 80% of the anaerobic threshold determined by the lactate minimum test. At the end of the experiment, the body weight gain, insulin sensitivity (glucose disappearance rate during the insulin tolerance test), concentrations of free fatty acids (FFA) and triglycerides (TG) and hepatic lipogenic rate were analyzed. For the statistical analysis, the Student t-test was used with the level of significance preset at 5%. Results: The T group showed lower body weight gain, FFA concentrations, fat accumulation, hepatic lipogenic rate and insulin resistance. Conclusion: The regular practice of moderate physical exercise from childhood can contribute to the reduction of obesity and insulin resistance and help prevent the development of accumulation of hepatic fat in adulthood. © 2013de Moura et al; licensee BioMed Central Ltd.

Changes in cells's secretory organelles and extracellular matrix during endochondral ossification in the mandibular condyle of the growing rat

The mandibular condyle from 20-day-old rats was examined in the electron microscope with particular attention to intracellular secretory granules and extracellular matrix. Moreover, type II collagen was localized by an immunoperoxidase method. The condyle has been divided into five layers: (1) the most superficial, articular layer, (2) polymorphic cell layer, (3) flattened cell layer, (4) upper hypertrophic, and (5) lower hypertrophic cell layers. In the articular layer, the cells seldom divide, but in the polymorphic layer and upper part of the flattened cell layer, mitosis gives rise to new cells. In these layers, cells produce two types of secretory granules, usually in distinct stacks of the Golgi apparatus; type a, cylindrical granules, in which 300-nm-long threads are packed in bundles which appear lucent after formaldehyde fixation; and type b, spherical granules loaded with short, dotted filaments. The matrix is composed of thick banded lucent fibrils in a loose feltwork of short, dotted filaments. The cells arising from mitosis undergo endochondral differentiation, which begins in the lower part of the flattened cell layer and is completed in the upper hypertrophic cell layer; it is followed by gradual cell degeneration in the lower hypertrophic cell layer. The cells produce two main types of secretory granules: type b as above; and type c, ovoid granules containing 300-nm-long threads associated with short, dotted filaments. A possibly different secretory granule, type d, dense and cigar-shaped, is also produced. The matrix is composed of thin banded fibrils in a dense feltwork. In the matrix of the superficial layers, the lucency of the fibrils indicated that they were composed of collagen I, whereas the lucency of the cylindrical secretory granules suggested that they transported collagen I precursors to the matrix. Moreover, the use of ruthenium red indicated that the feltwork was composed of proteoglycan; the dotted filaments packed in spherical granules were similar to, and presumably the source of, the matrix feltwork. The superficial layers did not contain collagen II and were collectively referred to as perichondrium. In the deep layers, the ovoid secretory granules displayed collagen II antigenicity and were likely to transport precursors of this collagen to the matrix, where it appeared in the thin banded fibrils. That these granules also carried proteoglycan to the matrix was suggested by their content of short dotted filaments. Thus the deep layers contained collagen II and proteoglycan as in cartilage; they were collectively referred to as the hyaline cartilage region.

Femoral Biomechanic and microtomography from male rats submitted to dietary restricition supplemented with sucrose

Food restriction reduces body weight and influence bone mass and also is correlated with bone mineral density (BMD). Mechanisms have been proposed for the loss of BMD after body weight reduction, including reduced energy intake. Growing 8 wk-old Wistar male rats were randomly divided into Control and Calorie restriction associated with sucrose 30% (CRS). These animals were subjected to intermittent food restriction during 8 weeks and had free access to tap water and sucrose30% in distilled water. The rats were euthanized at the end of week 8, blood collected from abdominal aorta artery, femurs cleaned of adherent soft tissues, scanned using dual energy X-ray absorptiometry, structural and material properties determined by three-point bending testing in the mid-diaphyseal region, bone surface tested in a microhardness tester and microstructure was assessed in a microcomputer tomography. In CRS animals body weight decreased significantly relative to the Control animals. There was a clear option for high-sucrose beverage in CRS animals. No difference was observed in biochemical, densitometric and biomechanical analyzes. Results from micro CT showed only significant difference in connectivity of trabecular bone. It has been suggested that rats submitted to food restriction consumed sugar not because of its inherent palatability, but in order to alter their macronutrient balance and animals need to meet energy demands in high-sucrose.