Influence of creatine supplementation on bone quality in the ovariectomized rat model: an FT-Raman spectroscopy study

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Effect of caloric restriction on myenteric neuroplasticity in the rat duodenum during aging

The objective of this study was to evaluate the effects of caloric restriction (CR) on myenteric neurons in the duodenum of Wistar rats during aging. Thirty rats were divided into three groups: the C group (six-month-old animals that were fed a normal diet from weaning until six months of age), the SR group (18-month-old animals that were fed a normal diet from weaning until 18 months of age) and the CR group (18-month-old animals that were fed a 30% CR diet after six months of age). After 12 months, the animals were euthanized. Whole-mount preparations of the duodenums were either stained with Giemsa or underwent NADPH-diaphorase histochemistry to determine the general myenteric neuron population and the nitrergic neuron subpopulation (NADPH-d +), respectively. The NADPH-d-negative (NADPH-d -) neuron population was estimated based on the difference between the Giemsa-stained and NADPH-d + neurons. The neurons were counted, and the cell body areas were measured. Aging was associated with neuronal loss in the SR group, which was minimized by caloric restriction in the CR group. The density (mm(2)) of the Giemsa-stained neurons was higher in the SR group (79.09 +/- 6.25) than in the CR (92.37 +/- 11.6) and C (111.68 +/- 15.26) groups. The density of the NADPH-d + neurons was higher in the SR group (44.90 +/- 5.88) than in the C (35.75 +/- 1.6) and RC (39.14 +/- 7.02) groups. The density of NADPH-d - neurons was higher in the CR (49.73 +/- 12.08) and C (75.64 +/- 17.05) groups than in the SR group (33.82 +/- 4.5). In the C group, 32% and 68% of the Giemsa-stained myenteric neurons were NADPH-d + or NADPH-d -, respectively. With aging (SR group), the percentage of nitrergic neurons (56.77%) increased, whereas the percentage of NADPH-d - neurons (43.22%) decreased. In the CR group, the change in the percentage of nitrergic (42.37%) and NADPH-d - (57.62%) neurons was lower. As NADPH-d - neurons will be mostly cholinergic neurons, CR appears to reduce the loss of cholinergic neurons during aging. The cell body dimensions (mu m(2)) were not altered by aging or CR. Thus. CR had a protective effect on myenteric neurons during aging. (C) 2012 Elsevier B.V. All rights reserved.

Biocompatibility of different intracanal medications in rat buccal submucosa tissue

The aim of this study was to analyze the buccal tissue responses of Wistar rats to 2% chlorhexidine solution, calcium hydroxide and the association of both products. For this purpose, 30 specimens were randomly implanted in the filtrum of the four upper and lower hemiarches with a polyethylene tube containing one of the following substances: 2% chlorhexidine solution, calcium hydroxide and 2% chlorhexidine solution (test groups); calcium hydroxide and distilled water and distilled water (control groups). Ten rats each were distributed according to time interval of evaluation at 7, 15 and 30 days. The histological sections were stained with Harris hematoxylin and eosin. Analysis was per-formed with an optical microscope at x100, x200 and x400 magnifications by an expert examiner blinded to the materials. The sections were classified by scores attributed to inflammatory events and by a ranking determined according to the severity of the inflammation. The results of the inflammatory events and severity ranking were submitted to the Kruskal-Wallis test at a 0.05 level of significance. No statistically significant difference occurred among the tested materials; however, all materials showed a decreased of severity with respect to longer time intervals.

Autoradiographic investigation of effects of high-dose colchicine on dentinogenesis in rat incisors

Twelve female Wistar rats received 1.5 mg/kg of colchicine (CLC) intravenously. Control animals were similarly injected with isotonic saline solution. The animals were killed 5 h, 24 h, 3 days and 7 days after injection. Ninety minutes prior to sacrifice, all animals received an intraperitoneal injection of 3H-proline. Autoradiograms of maxillary incisors showed that CLC increased the retention of the labeled precursor in the odontoblasts. It was also shown that the odontoblasts in the different sectors of the rat incisor present different sensitivities to the CLC action.

Efeito do envelhecimento sobre o comportamento mecânico dos músculos papilares de ratos Wistar.

PURPOSE--To evaluate the effects of age on mechanical performance of rat myocardium. METHODS--Left ventricular papillary muscles were isolated from male Wistar rats at 1, 3, 6 and 12 months of age. Muscles were studied isometrically and isotonically, stimulated at 0.2 Hz, perfused with Krebs-Henseleit solution having an external calcium concentration of 2.52 mM, and maintained at 28 degrees C. RESULTS--Peak isometric developed tension was significantly higher in 1 month than 3, 6 and 12 months. Peak rate of isometric tension rise decreased substantially between 1, 3 and 12 months. Time to peak isometric developed tension showed a significant increase of both 3 and 12 months of age. Time to half relaxation increased significantly from 3 to 6 and from 3 to 12 months. Maximum rate of tension decline decreased from 3 to 6 and from 3 to 12 months. No difference in resting tension was noted among any group. Isotonically, peak shortening and time to peak shortening increased from 1 to 3 months of age. Time to half re-lengthening increased from 3 to 6 and from 3 to 12 months of age. No difference in peak shortening velocity, peak relaxation velocity and relative change in muscle length was noted among any groups. CONCLUSION--The maturation affects the mechanical performance of cardiac muscle.

Alcohol-induced alterations in the palate epithelium of the rat fetus.

Maternal alcoholism (ethanol and sugar cane brandy) during gestation induces delayed cellular growth and differentiation in fetal rat palate epithelium, with increased nuclear, cytoplasmic and cellular volumes, increased epithelial and keratin thickness and decreased cellular numerical density.

Ultrastructural study of the coagulating gland of Wistar rats submitted to experimental chronic alcohol ingestion

Morphological changes of coagulating gland of Wistar rats submitted to the experimental chronic alcoholism were noted. Ultrastructurally, it was observed reduction of the granular endoplasmic reticulum cisternae and nuclei with basal position and irregular shape. The epithelial cells presented pycnotic nuclei and the mitochondrial cristae disappeared, characteristic of cellular degeneration.

Circadian time-dependent effects of fencamfamine on inhibition of dopamine uptake and release in rat striatal slices

Fencamfamine (FCF) is a central stimulant that facilitates central dopaminergic transmission through inhibition of dopamine uptake and enhanced release of the transmitter. We evaluated the changes in the inhibition of uptake and the release of striatal [ 3H]-dopamine at 9:00 and 21:00 h, times corresponding to maximal and minimal behavioral responses to FCF, respectively. Adult male Wistar rats (200-250 g) maintained on a 12-h light/12-h dark cycle (lights on at 7:00 h) were used. In the behavioral experiments the rats (N = 8 for each group) received FCF (3.5 mg/kg, ip) or saline at 9:00 or 21:00 h. Fifteen minutes after treatment the duration of activity (sniffing, rearing and locomotion) was recorded for 120 min. The basal motor activity was higher (28.6 ± 4.2 vs 8.4 ± 3.5 s) after saline administration at 21:00 h than at 9:00 h. FCF at a single dose significantly enhanced the basal motor activity (38.3 ± 4.5 vs 8.4 ± 3.5 s) and increased the duration of exploratory activity (38.3 ± 4.5 vs 32.1 ± 4.6 s) during the light, but not the dark phase. Two other groups of rats (N = 6 for each group) were decapitated at 9:00 and 21:00 h and striata were dissected for dopamine uptake and relase assays. The inhibition of uptake and release of [ 3H]-dopamine were higher at 9:00 than at 21:00 h, suggesting that uptake inhibition and the release properties of FCF undergo daily variation. These data suggest that the circadian time-dependent effects of FCF might be related to a higher susceptibility of dopamine presynaptic terminals to the action of FCF during the light phase which corresponds to the rats' resting period.

Ultrastructural study of the lateral lobe of the prostate of Wistar rats submitted to experimental chronic alcohol ingestion

Alcoholism is considered today one of the most serious social and medical problems. Different investigators have demonstrated that chronic exposure to alcohol causes morphological and physiological changes in the testes and in the accessory sex glands, in the prostate in particular. In the present study, experimental rats were divided into groups receiving sugar cane brandy diluted to 30° G.L. (30%, v/v) and Purina ration ad libitum for 60, 120, 180, 240 and 300 days, respectively. At the end of each period the animals were sacrificed and the lateral lobe of the prostate was collected for histological examination. The results showed atrophied epithelium, accumulation of lipidic droplets and rupture of the microvilli that line the cell surface facing the lumen of cells of the secretory epithelium of the lateral lobe of the prostate.

Ultrastructural changes on the epithelial cells of uterine tubes of Wistar rats after chronic ethanol ingestion

The present paper describes the morphological alterations of the epithelial layer of the uterine tubes of rats submitted to experimental chronic alcoholism using anatomical, histological, ultrastructural and morphometric methods. Sixty adult rats (Rattus norvegicus albinus) at the same age (3 months) and with a mean body weight of 228 g were divided into two groups. The control group received solid diet (Purina rat chow) and tap water ad libitum. The alcoholic group received the same solid diet and was allowed to drink only sugar cane brandy dissolved in 30° Gay Lussac (v/v). After periods of 90, 180 and 270 days of treatment animals at normal estrus were anaesthetised with ethyl ether, weighed and sacrificed. Subsequently, the uterine tubes were dissected, weighed and prepared for TEM and SEM methods. The final mean body weights were similar in the control and alcoholic groups. The morphometric analysis showed no difference between control and alcoholic epithelial height. The alcoholic animals showed ultrastructural alterations: intense lipid droplet and lysosomes accumulation, dilated rough endoplasmic reticulum cisternae and vacuolization in both periods of treatment. It was concluded that alcohol acts as a toxin on the epithelial layer of the uterine tubes of rats.

Long-term nitric oxide inhibition and chronotropic responses in rat isolated right atria

The long-term administration of nitric oxide synthesis inhibitors induces arterial hypertension accompanied by left ventricular hypertrophy and myocardial ischemic lesions. Because the enhancement of sympathetic drive has been implicated in these phenomena, the current study was performed to determine the potency of β-adrenoceptor agonists and muscarinic agonists on the spontaneous rate of isolated right atria from rats given long-term treatment with the nitric oxide inhibitor N(ω)-nitro-L-arginine methyl ester (L-NAME). Atrial lesions induced by long-term treatment with L-NAME were also evaluated. Long-term L-NAME treatment caused a time-dependent, significant (P<0.05) increase in tail-cuff pressure compared with control animals. Our results showed that the potency of isoproterenol, norepinephrine, carbachol, and pilocarpine in isolated right atria from rats given long-term treatment with L-NAME for 7, 15, 30, and 60 days was not affected as compared with control animals. Addition of L-NAME in vitro (100 μmol/L) affected neither basal rate nor chronotropic response for isoproterenol and norepinephrine in rat heart. Stereological analysis of the right atria at 15 and 30 days revealed a significant increase on amount of fibrous tissues in L-NAME- treated groups (27±2.3% and 28±1.3% for 15 and 30 days, respectively; P<0.05) as compared with the control group (22±1.1%). Our results indicate that nitric oxide does not to interfere with β-adrenoceptor-mediated and muscarinic receptor-mediated chronotropic responses.

Apoptosis in the early developing periodontium of rat molars

Development of the periodontium involves a series of complex steps that result in the formation of root dentine, cementum, bone and fibres of the ligament. These precisely controlled and timed events require the participation of the enamel organ derived epithelial cells of Hertwig's (HRS) and ectomesenchymal cells of the dental follicle. These events involve rapid turnover of the tissues and cells, including disappearance of epithelial cells of HRS. Thus, it seemed likely to us that programmed cell death (apoptosis) may play a role in the development of the periodontium. Fragments of first molars, obtained from 14- and 29-day-old rats, were fixed in glutaraldehyde-formaldehyde and processed for light and electron microscopy. For the TUNEL method for detection of apoptosis, specimens were fixed in 4% formaldehyde and embedded in paraffin. Results confirmed that epithelial cells of HRS maintain a close relationship with the forming dentine root, and that they may become trapped in the dentino-cemental junction. Some of the epithelial cells exhibited ultrastructural features which are consistent with the interpretation that they were undergoing programmed cell death, i.e. apoptosis. Periodontal fibroblast-like cells showed typical images of apoptosis and engulfed apoptotic bodies. TUNEL positive structures were present in all corresponding regions. It seems therefore that apoptosis of epithelial cells of HRS and fibroblast-like cells of the periodontal ligament constitutes an integral part of the developmental process of the tissues of the periodontium. (C) 2000 Wiley-Liss, Inc.

Collagen fiber reorganization in the rat ventral prostate following androgen deprivation: A possible role for smooth muscle cells

BACKGROUND. Stroma plays an essential role in glandular function in different systems. In the prostate, it is responsible for the development and maintenance of the differentiated state of the epithelium. The marked reduction in the epithelial compartment of the prostate gland following castration is followed by a similarly important reorganization of the stroma. In this work, we characterized the reorganization of collagen fibers in the ventral prostate of castrated rats. METHODS. Histochemical tests and immunohistochemistry for type I and III collagens plus confocal microscopy of triple-labeled (collagen III, actin, and DNA) tissue sections were employed. RESULTS. We showed that collagen fibers are composed of type I and type III collagens and that they are progressively concentrated around the epithelial structures (ducts and acini) and become increasingly undulated and folded. Double-labeling of collagen fibers and F-actin demonstrated that smooth muscle cells (SMC) are intimately associated with collagen fibers. CONCLUSIONS. The results demonstrated a marked reorganization of the collagen fibers, and suggest an active role of the SMC in the reorganization of the fibrillar components of the stroma. (C) 2000 Wiley-Liss, Inc.