Cardiac mast cell proteases do not contribute to the regulation of the rat coronary vascular responsiveness to arterial delivered angiotensin I and II

Cardiac mast cells (MC) are apposed to capillaries within the heart and release renin and proteases capable of metabolizing angiotensins (Ang). Therefore, we hypothesized that mast cell degranulation could alter the rat coronary vascular responsiveness to the arterial delivered Ang I and Ang II, taking into account carboxypeptidase and chymase-1 activities. Hearts from animals that were either pretreated or not with systemic injection of the secretagogue compound 48/80 were isolated and mounted on a Langendorff apparatus to investigate coronary reactivity. The proteolytic activity of the cardiac perfusate from isolated hearts, pretreated or not with the secretagogue, toward Ang I and tetradecapeptide renin substrate was analyzed by HPLC. Coronary vascular reactivity to peptides was not affected by compound 48/80 pretreatment, despite the extensive amount of cardiac MC degranulation. Cardiac MC activation did not modify the generation of both Ang II and Ang 5-10 from Ang I by cardiac perfusate, activities that could be ascribed to MC carboxypeptidase and chymase-1, respectively. An aliskiren-resistant Ang I-forming activity was increased in perfusates from secretagogue-treated hearts. Thus, cardiac MC proteases capable of metabolizing angiotensins do not affect rat coronary reactivity to arterial delivered Ang I and II. (C) 2010 Elsevier Inc. All rights reserved.

Histological analysis of cobalt-chromium stents with and without Camouflage (R) polymer coating: experimental porcine carotid artery model

This study evaluated the arterial response to cobalt-chromium stents with and without polymer coating (Camouflage (R), Hemoteq AG, Wuerselen, Germany) implanted in pigs. Cobalt-chromium balloon-expandable stents (4 x 16 mm) were implanted in the common carotid arteries of nine pigs. Histological analysis of endothelialization, inflammation and injury was performed one month later. All stents were successfully deployed, and all but one animal survived the 30 study days. All arteries were patent. Endothelialization was nearly complete in most sections of all carotid stents in both groups. There were mild inflammatory infiltrate and mild-to-moderate injury, which were associated with the stent shafts and not significantly different between groups. Our findings suggest that, in porcine carotid arteries, the histological response to balloon-expandable cobalt-chromium stents coated with polymer (Camouflage (R), Hemoteq AG) is similar to the response to non-coated cobalt-chromium stents.

Stereological and allometric studies on neurons and axo-dendritic synapses in the superior cervical ganglia of rats, capybaras and horses

The superior cervical ganglion (SCG) in mammals varies in structure according to developmental age, body size, gender, lateral asymmetry, the size and nuclear content of neurons and the complexity and synaptic coverage of their dendritic trees. In small and medium-sized mammals, neuron number and size increase from birth to adulthood and, in phylogenetic studies, vary with body size. However, recent studies on larger animals suggest that body weight does not, in general, accurately predict neuron number. We have applied design-based stereological tools at the light-microscopic level to assess the volumetric composition of ganglia and to estimate the numbers and sizes of neurons in SCGs from rats, capybaras and horses. Using transmission electron microscopy, we have obtained design-based estimates of the surface coverage of dendrites by postsynaptic apposition zones and model-based estimates of the numbers and sizes of synaptophysin-labelled axo-dendritic synaptic disks. Linear regression analysis of log-transformed data has been undertaken in order to establish the nature of the relationships between numbers and SCG volume (V(scg)). For SCGs (five per species), the allometric relationship for neuron number (N) is N=35,067xV (scg) (0.781) and that for synapses is N=20,095,000xV (scg) (1.328) , the former being a good predictor and the latter a poor predictor of synapse number. Our findings thus reveal the nature of SCG growth in terms of its main ingredients (neurons, neuropil, blood vessels) and show that larger mammals have SCG neurons exhibiting more complex arborizations and greater numbers of axo-dendritic synapses.

Low-intensity Treadmill Exercise-related Changes in the Rat Stellate Ganglion Neurons

Stellate ganglion (SG) represents the main sympathetic input to the heart. This study aimed at investigating physical exercise-related changes in the quantitative aspects of SG neurons in treadmill-exercised Wistar rats. By applying state-of-the-art design-based stereology, the SG volume, total number of SG neurons, mean perikaryal volume of SG neurons, and the total volume of neurons in the whole SG have been examined. Arterial pressure and heart rate were also measured at the end of the exercise period. The present study showed that a low-intensity exercise training program caused a 12% decrease in the heart rate of trained rats. In contrast, there were no effects on systolic pressure, diastolic pressure, or mean arterial pressure. As to quantitative changes related to physical exercise, the main findings were a 21% increase in the fractional volume occupied by neurons in the SG, and an 83% increase in the mean perikaryal volume of SG neurons in treadmill-trained rats, which shows a remarkable neuron hypertrophy. It seems reasonable to infer that neuron hypertrophy may have been the result of a functional overload imposed on the SG neurons by initial posttraining sympathetic activation. From the novel stereological data we provide, further investigations are needed to shed light on the mechanistic aspect of neuron hypertrophy: what role does neuron hypertrophy play? Could neuron hypertrophy be assigned to the functional overload induced by physical exercise? (C) 2008 Wiley-Liss, Inc.

Short-Term Effects of Duodenocecostomy on Body Weight, Glucose Absorption, Serum Components, and Intestinal Histopathology in Four Normal Horses

The objective of this study was to evaluate duodenocecostomy in horses performed through a ventral midline laparotomy and report its influence oil body weight, glucose absorption, serum components, and characteristics of jejunum, cecum, and large colon histology. Four horses were submitted to the duodenocecostomy technique through a ventral midline laparotomy with animals in dorsal recumbency under inhalation anesthesia, followed by abdominal exploration. A side-to-side anastomosis was performed between the duodenojejunal flexure and the base of the cecum with two simple continuous suture lines of the serosal and muscular layers. The size of the opening created was approximately 2 cm in diameter. The mucosa layer was not Sutured. After 30 days, animals were submitted to a second laparotomy to check the patency of the duodenocaecal fistula. During both laparotomy procedures, excisional biopsies of different segments of the gastrointestinal tract were performed. Information on physical examination findings, results of hematologic and histopathologic evaluations, and oral glucose absorption test were recorded. The horses did not have significant weight loss from baseline, and absorption curve of glucose did not significantly vary from baseline. Only triglycerides had significant alterations. Histologic evaluation of jejunum, cecum, and large colon did not show alterations of intestinal structure and morphology. We concluded that the proposed technique, principally in relation to the fistula size and not suturing the mucosa layer, allowed partial or total Occlusion of the fistulae without the necessity of a second surgery and avoided the permanent bypass of ingesta and weight loss.

Transient disruption of liver gap junctional intercellular communication and induction of apoptosis after administration of 1,4-bis[2-(3,5 dichloropyridyloxy)]benzene in mice

Gap junctional intercellular communication (GJIC) and connexin expression (Cx26 and Cx32) in mouse liver were studied after administration of 4-bis[2-(3,5 dichloropyridyloxy)]benzene (TCPOBOP), a phenobarbital-like enzyme inducer. Female C57BI/6 mice were administered TCPOBOP (5.8 mg/kg BW) and euthanized 0, 24, 48 and 72 hours later. Liver samples were snap frozen, or fixed in formalin, or submitted to GJIC analysis. The proliferating cell nuclear antigen (PCNA) immunohistochemistry and the Western blotting for Cx26 and Cx32 were performed. After 48 and 72 h of drug administration the liver-to-body weight ratio was increased 70% and 117% (p < 0.0001), respectively. There were temporal-dependent alterations in liver histopathology and a significant increase in cell proliferation was noted after 48h and sustained after 72h, though to a lesser extent (p < 0.0001). In addition. TCPOBOP administration induced apoptosis, which appeared to be time-dependent showing statistical significance only after 72h (p < 0.0001). Interestingly, a transient disruption by nearly 50% of GJIC capacity was detected after 48 h of drug ingestion, which recovered after 72 h (p = 0.003). These GJIC changes were due to altered levels of Cx26 and Cx32 in the livers of TCPOBOP-treated mice. We concluded that a single administration of TCPOBOP transiently disrupted the levels of GJIC due to decreased expression of connexins and increased apoptotic cell death in mouse liver. (C) 2009 Elsevier GmbH. All rights reserved.

Prenatal lipopolysaccharide reduces motor activity after an immune challenge in adult male offspring

Prenatal lipopolysaccharide (LPS) exposure causes reproductive, behavioral and neurochemical injuries in both the mother and pups. Previous investigations by our group showed that prenatal LPS administration (100 mu g/kg, i.p.) on gestational day 9.5 impaired the male offspring`s social behavior in infancy and adulthood. In the present study, we investigated whether these social behavioral changes were associated with motor activity impairment. Male rat pups treated prenatally with LPS or not were tested for reflexological development and open field general activity during infancy. In adulthood, animals were tested for open field general activity, haloperidol-induced catalepsy and apomorphine-induced stereotypy; striatal dopamine levels and turnover were also measured. Moreover, LPS-treated or untreated control pups were challenged with LPS in adulthood and observed for general activity in the open field. In relation to the control group, the motor behavior of prenatally treated male pups was unaffected at basal levels, both in infancy and in adulthood, but decreased general activity was observed in adulthood after an immune challenge. Also, striatal dopamine and metabolite levels were decreased in adulthood. In conclusion, prenatal LPS exposure disrupted the dopaminergic system involved with motor function, but this neurochemical effect was not accompanied by behavioral impairment, probably due to adaptive plasticity processes. Notwithstanding, behavioral impairment was revealed when animals were challenged with LPS, resulting in enhanced sickness behavior. (C) 2010 Elsevier B.V. All rights reserved.

Pfaffia paniculata (Brazilian ginseng) roots decrease proliferation and increase apoptosis but do not affect cell communication in murine hepatocarcinogenesis

Pfaffia paniculata (Brazilian ginseng) roots and/or its extracts have shown anti-neoplastic, chemopreventive, and anti-angiogenic properties. The aim of this work was to investigate the chemopreventive mechanisms of this root in Mice Submitted to the infant model of hepatocarcinogenesis, evaluating the effects oil cellular proliferation, apoptosis. and intercellular communication. Fifteen-day-old BALB/c male mice were given, i.p., 10 mu g/g of the carcinogen N-nitrosodiethylamine (DEN). Animals were separated into three groups at weaning and were given different concentrations of powdered P. paniculata root (0%, 2%, or 10%) added to commercial food for 27 weeks. Control group (CT) was not exposed to the carcinogen and was given ration without the root. After euthanasia, the animals` liver and body weight were measured. Liver fragments were sampled to Study intercellular communication, molecular biology, and histopathological analysis. Cellular proliferation was evaluated by immunohistochemistry for PCNA, apoptosis was evaluated by apoptotic bodies count and alkaline cornet technique, and inter-cellular communication by diffusion of lucifer yellow dye, immunofluorescence, western blot and real-time PCR for connexins 26 and 32. Chronic treatment with powdered P. paniculata root reduced cellular proliferation and increased apoptosis in the 2%, group. Animals in the 10% group had an increase in apoptosis with chronic inflammatory process. Intercellular communication showed no alterations in any of the groups analyzed. These results Indicate that chemopreventive effects of P. paniculata are related to the control of cellular proliferation and apoptosis, but not to cell communication and/or connexin expression, and are directly Influenced by the root concentration. (C) 2009 Elsevier GmbH. All rights reserved.

Effects of type II pyrethroid cyhalothrin on rat innate immunity: A flow cytometric study

Synthetic type II pyrethroids induce anxiety, immunosuppresion or, alternatively, immunostimulatory effects in laboratory animals. Macrophages and neutrophils are known to be key elements in cellular immune responses. The present study was designed to investigate the in vivo effects of cyhalothrin (1.0 and 3.0 mg/kg/once daily for 7 days) on macrophage and neutrophil activities, using a flow cytometry method. Results showed that cyhalothrin treatment decreased the percentage and intensity of phagocytosis performed by macrophages, but did not alter these parameters in neutrophils: and also decreased basal neutrophil oxidative burst and increased S. aureus-induced neutrophil oxidative burst, but did not alter these responses in macrophages. The present results are discussed in the light of a possible indirect action of cyhalothrin on macrophage and neutrophil activities via hypothalamic pituitary adrenal (HPA) axis activation. A possible direct effect of cyhalothrin on macrophage and neutrophil activities is also considered. (C) 2008 Elsevier B.V. All rights reserved.

Insulin Affects Tissue Organization and the Kinetics of Epithelial Cell Death in the Rat Ventral Prostate After Castration

Prostate growth and physiology are regulated by steroid hormones and modulated by multiple endocrine factors We investigated the action of insulin on the tissue organization and kinetics of epithelial cells in the rat ventral prostate (VP) in response to castration up to 120 hours after surgery by using an acute protocol of alloxan induced diabetes Diabetes caused a reduction in volume density (Vv(o)/) and volume of the epithelium The effects of castration on the epithelium were accelerated in the diabetic animals as determined by changes in V(o)/, and volume The smooth muscle cells became atrophic and apparently relaxed in response to castration in contrast to the spinous aspect observed in nondiabetic castrated rats Counting of apoptotic nuclei in the epithelium showed the classical apoptosis peak at 72 hours in nondiabetic rats and an advance of the apoptosis peak to 48 hours after castration in diabetic rats Insulin restored the time of the peak to 72 hours These results were confirmed after immunostaining for cleaved caspase 3 and suggest a survival and antiapoptotic effect on VP epithelial cells in both the presence and absence of androgen stimulation This idea is supported by the observation that insulin also reduced the overall rate of apoptosis at all experimental points analyzed before and after castration

Validation of body condition score as a predictor of subcutaneous fat in Nelore (Bos indicus) cows

The aim of the present study was to determine the relationship among body weight (BW), body condition score (BCS) and rump fat thickness (RFAT) measured by ultrasonography, and validate the relationship between BCS and RFAT over the time. Two hundred sixty and six Nelore cows had their BW, BCS and RFAT evaluated at five different moments during the production cycle: M1) weaning: M2) parturition, M3) 42 days post-partum; M4) 82 days postpartum and M5) 112 days post-partum. A BCS value was attributed for each cow following a I to 5 points scale. Ultrasonographic images for RFAT measurement were obtained using a 3.5 MHz linear transducer. Images were immediately analyzed as soon as they were formed and frozen. Body condition scores and ultrasound measurements were collected on the same day by a single trained technician. The relationship between BCS and RFAT values was investigated by regression models. The analysis of similarity among the five obtained models was performed using the proc MIXED from SAS and the correlations among variables were analyzed with proc CORR from SAS. The BCS was able to predict RFAT in Nelore cows in all different moments evaluated. Also, it was shown that BCS presented high correlation (r=0.82 to 0.93) and relationship (R(2) = 0.73 to 0.92) with RFAT. However, both BCS and RFAT showed low correlation (r=0.37 to 0.50) and relationship (R(2) = 0.13 to 0.25) with BW. The BCS classification by visual method using a 1 to 5 point scale, was able to predict the RFAT in Nelore cows over the time. (C) 2008 Elsevier B.V. All rights reserved.

Genotype effects on body temperature in dairy cows under grazing conditions in a hot climate including evidence for heterosis

We compared diurnal patterns of vaginal temperature in lactating cows under grazing conditions to evaluate genotype effects on body temperature regulation. Genotypes evaluated were Holstein, Jersey, Jersey x Holstein and Swedish Red x Holstein. The comparison of Holstein and Jersey versus Jersey x Holstein provided a test of whether heterosis effects body temperature regulation. Cows were fitted with intravaginal temperature recording devices that measured vaginal temperature every 15 min for 7 days. Vaginal temperature was affected by time of day (P < 0.0001) and genotype x time (P < 0.0001) regardless of whether days in milk and milk yield were used as covariates. Additional analyses indicated that the Swedish Red x Holstein had a different pattern of vaginal temperatures than the other three genotypes (Swedish Red x Holstein vs others x time; P < 0.0001) and that Holstein and Jersey had a different pattern than Jersey x Holstein [(Holstein + Jersey vs Jersey x Holstein) x time, P < 0.0001]. However, Holstein had a similar pattern to Jersey [(Holstein vs Jersey) x time, P > 0.10]. These genotype x time interactions reflect two effects. First, Swedish Red x Holstein had higher vaginal temperatures than the other genotypes in the late morning and afternoon but not after the evening milking. Secondly, Jersey x Holstein had lower vaginal temperatures than other genotypes in the late morning and afternoon and again in the late night and early morning. Results point out that there are effects of specific genotypes and evidence for heterosis on regulation of body temperature of lactating cows maintained under grazing conditions and suggest that genetic improvement for thermotolerance through breed choice or genetic selection is possible.

Ultrastructure of Motor Nerve Terminals in the Anterior Third of Wistar Rat Tongue

The nerve terminals of intrinsic muscular fibers of the tongue of adult wistar rats was studied by using silver impregnation techniques, transmission electron microscopy (TEM), and high resolution scanning electron microscopy (HRSEM) to observe the nerve fibers and their terminals. Silver impregnation was done according to Winkelman and Schmit, 1957. For TEM, small blocks were fixed in modified Karnovsky solution, postfixed in 1% buffered osmium tetroxide solution, and embedded in Spurr resin. For HRSEM, the parts were fixed in 2% osmium tetroxide solution with 1/15 M sodium phosphate buffer (pH 7.4) at 4 degrees C for 2 h, according to the technique described by Tanaka, 1989. Thick myelinated nerve bundles were histologically observed among the muscular fibers. The intrafusal nerve fiber presented a tortuous pathway with punctiform terminal axons in clusters contacting the surface of sarcolemma. Several myelinated nerve fibers involved by collagen fibers of the endoneurium were observed in HRSEM in three-dimensional aspects. The concentric lamellae of the myelin sheath and the axoplasm containing neurofilaments interspersed among the mitochondria were also noted. In TEM, myofibrils, mitochondria, rough endoplasmic reticulum, Golgi`s apparatus, and glycogen granules were observed in sarcoplasm. It is also noted that the sarcomeres constituted by myofilaments with their A, I, and H bands and the electron dense Z lines. In areas adjacent to muscular fibers, there were myelinated and unmyelinated nerve fibers involved by endoneurium and perineurium. In the region of the neuromuscular junction, the contact with the sarcolemma of the muscular cell occurs forming several terminal buttons and showing numerous evaginations of the cell membrane. In the terminal button, mitochondria and numerous synaptic vesicles were observed. Microsc. Res. Tech. 72:464-470, 2009. (C) 2009 Wiley-Liss. Inc.

Laser phototherapy effect on protein metabolism parameters of rat salivary glands

The aim of this study was to evaluate the effects of infrared diode laser phototherapy (LP) on tissues of the submandibular gland (SMG) and parotid gland (PG). Wistar rats were randomly divided into experimental (A and B) and control (C) groups. A diode laser, 808 nm wavelength, in continuous wave mode, was applied to the PG, SMG and sublingual gland in the experimental groups on two consecutive days. The doses were 4 J/cm(2) and 8 J/cm(2), and total energy was 7 J and 14 J, respectively. The power output (500 mW) and power density (277 mW/cm(2)) were the same for both experimental groups. In order to visualize the area irradiated by the infrared laser, we used a red pilot beam (650 nm) with 3 mW maximum power for the experimental groups. For the control group, the red pilot beam was the only device used. The SMG and PG were removed after 1 week of the first irradiation. Total protein concentration, amylase, peroxidase, catalase and lactate dehydrogenase assays were performed, as well as histological analysis. Statistical tests revealed significant increase in the total protein concentration for groups A and B in the parotid glands (P < 0.05). Based on the results of this study, LP altered the total protein concentration in rats` parotid glands.