Changes in the salivary proteome of beagle dogs after weight loss.

Being overweight or obese represents an important health issue in humans and pets. The aim of this study was to investigate changes in the salivary proteome of overweight beagles after induced weight loss to better understand the physiological changes involved in this process. Five overweight/obese neutered males of pure breed beagles were evaluated. During the 3-mo period of weight loss, each animal received a strictly controlled amount of a low fat commercial diet per day. Body condition scores (BCS), body weight (BW), and serum biochemical parameters (total cholesterol, triglycerides, and C-reactive protein) were assessed weekly. Quantitative proteomics analysis by SWATH was used to evaluate the salivary proteome changes induced by weight loss treatment. BCS, BW, serum total cholesterol concentration, and abundances of 23 salivary proteins differed significantly between before and after treatment. Some of the altered protein amounts, namely of peptidyl-prolyl cis-trans isomerase, fructose-bisphosphate aldolase C, and 78-kDa glucose-regulated protein, increased after weight loss. These proteins are related with the immune system, inflammatory status, oxidative stress, and glucose metabolism. The results obtained suggest a potential use of salivary proteins in monitoring physiological changes in dogs subjected to weight loss. Moreover, the type of changes identified reinforces the postulated physiological improvements, which weight loss induces. Further research is needed to determine whether the changes observed in this study are due to weight loss, dietary changes, or a combination of both.

Physical characteristics of the eggs of red-legged partridge (Alectoris rufa) reared in captivity / Características físicas dos ovos da perdiz vermelha (Alectoris rufa) criada em cativeiro

O objetivo do presente estudo foi avaliar os efeitos da idade das fêmeas (um, dois e três anos) e do mês de postura (março, abril e maio) sobre as características físicas dos ovos da perdiz vermelha (Alectoris rufa) criada em cativeiro. O peso (W), o comprimento máximo (L) e a largura máxima (B) de 2878 ovos foram determinados diretamente, enquanto o índice de forma (B/L), o volume (V) e a superfície (S) foram calculados com base nos parâmetros determinados diretamente. A análise mostrou diferenças significativas (P<0,01) no peso dos ovos entre as diferentes idades e entre meses de postura, com menor peso nas fêmeas mais jovens. Observaram-se diferenças significativas (P<0,01) no comprimento do ovo entre as classes de idade, mas não entre os meses de postura (P>0,05). Observaram-se diferenças significativas (P<0,01) na largura máxima e no índice de forma do ovo entre as diferentes classes de idades, com valores mais elevados nas fêmeas mais velhas e no período de postura mais tardio. O volume dos ovos estimados por meio de V1= 0,51LB2e V2=0,913W foi afetado significativamente (P<0,01) pela idade e pelo mês de postura, bem como as áreas, S1=4.835W0,662, S2=4,951V10,666e S3=4,951V20,666, as quais apresentaram os mesmos efeitos.

Alterações histológicas dos rins e expressão das metalotioneínas e das proteínas de choque térmico em ratos Wistar após exposição ao fungicida tirame / Kidney histological alterations and metallothionein and heat shock protein expression in Wistar rats after fungicide thiram exposure

Avaliaram-se as alterações histológicas e a expressão das metalotioneínas (MTs) e das proteínas de choque térmico (Hsp70) nos rins de ratos Wistar após a exposição ao fungicida tirame. Os animais foram distribuídos em três grupos: grupo dieta-padrão; grupo dieta-padrão+óleo de milho; e grupo tirame. Foram encontradas diferenças significativas (P<0,05) na evolução do peso corporal entre os ratos do grupo tirame e os dos grupos controle e óleo de milho, e não foram verificadas lesões histológicas evidentes nos rins dos animais. Foram encontradas diferenças entre os animais do grupo exposto ao tirame e os dos grupos controle e óleo quanto às características histomorfométricas relativas ao corpúsculo renal - exceto para a proporção área da cápsula de Bowman:área do glomérulo - e relativas aos túbulos contorcidos proximal e distal - exceto para altura do epitélio dos túbulos distais. Nos ratos expostos ao tirame, foi observada imunomarcação positiva para as MTs, de moderada a forte, nos túbulos contorcidos da região cortical, diminuindo do córtex em direção à medula, e forte imunomarcação para as Hsp70 nas áreas do córtex e da medula, no glomérulo e nos túbulos contorcidos. Os resultados sugerem que o tirame pode ter toxicidade crônica nos mamíferos por afetar o seu crescimento e que a expressão das MTs e das Hsp70, provável resposta celular adaptativa ao estresse oxidativo causado pelo tirame, pode ser utilizada como biomarcador de exposição a este químico.

The histological alterations and the expression of metallothionein (MTs) and heat shock protein (Hsp70) in the kidney of Wistar rats after thiram fungicide exposure were evaluated. Animals were distributed into three groups: standard diet group, standard diet + corn oil group and thiram group. Significant differences were found (P<0,05) in the evolution of body weight between rats in the thiram group and those in the control and corn oil groups, and no histological lesions were evident in the animals' kidneys. Differences were found among animals in the group exposed to thiram and the control and oil groups regarding histomorphometric characteristics of the renal corpuscle - except for the proportion in the area of Bowman's capsule: glomerulus area - and regarding the height of the epitelium in the distal tubules. In rats exposed to thiram, a positive moderate to strong immunoexpression was observed for MTs, in the cortical convulated tubules decreasing the cortex towards the medulla, and a strong immunoexpression for Hsp70 in the cortex and medulla areas, in the glomerulus and convulated tubules. The results suggest that thiram may have chronic toxicity in mammals affecting their growth, and that the expression of MTs and Hsp70, a probable cellular adaptive response to the oxidative stress caused by thiram, may be used as a biomarker of exposure to this chemical.

Efeitos da semicarbazida no esqueleto de ratos Wistar em crescimento / Effects of semicarbazide on skeleton of growing Wistar rats

The effects of semicarbazide hydrochloride on skeletal tissues were evaluated using Wistar rats. In the animals submitted to the administration of semicarbazide, radiological examination showed bone and articular alterations with growth plates enlargement. Histological exams showed irregular patterns and proliferation of chondrocytes and excessive accumulation of extracellular matrix. Immunohistochemical exams showed PCNA positive reaction in pre- and hypertrofic chondrocytes. These results confirm that semicarbazide induces important changes in skeletal tissues and that has potential utility in experimental osteolathryrism.

Effect of dithiocarbamate thiram on Wistar rat growth plate and articular cartilage / Efeito do ditiocarbamato tirame sobre a placa de crescimento e cartilagem articular de ratos Wistar

Avaliou-se o efeito do tirame, ditiocarbamato largamente utilizado na agricultura como antifúngico e repelente de roedores, na ossificação endocondral de mamíferos, usando, como modelo, ratos Wistar. Não foram observadas lesões na cartilagem articular, nem nas placas de crescimento, o que pode ser atribuído à dose utilizada e à duração do ensaio. A diminuição da altura da placa de crescimento nos animais aos quais foi administrado o tirame parece traduzir o atraso verificado no crescimento em geral, e não um efeito específico na cartilagem, uma vez que as diferentes zonas da placa epifisária mantiveram as proporções dos animais do grupo-controle. Embora não tenham sido verificados, no presente trabalho, os efeitos registrados para outras espécies nos tecidos cartilaginosos, sugere-se a avaliação dos efeitos crónicos do tirame no crescimento e no desenvolvimento dos ossos longos em mamíferos.

Effect of condensed tannin ingestion in sheep and goat parotid saliva proteome.

Saliva appears as a defence mechanism, against potential negative effects of tannins, in some species of animals which have to deal with these plant secondary metabolites in their regular diets. This study was carried out to investigate changes in parotid saliva protein profiles of sheep (Ovis aries) and goats (Capra hircus), induced by condensed tannin ingestion. Five Merino sheep and five Serpentina goats were maintained on a quebracho tannin enriched diet for 10 days. Saliva was collected through catheters inserted on parotid ducts and salivary proteins were separated by two-dimensional gel electrophoresis. Matrix-assisted Laser desorption ionization - time of flight (MALDI-TOF) and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to identify the proteins whose expression levels changed after tannin consumption. Although no new proteins appeared, quebracho tannin consumption increased saliva total protein concentration and produced changes in the proteome of both species. While some proteins were similarly altered in both species parotid salivary protein profile, sheep and goats also presented species-specific differences in response to tannin consumption.

Polymeric piezoelectric actuator substrate for osteoblast mechanical stimulation.

Bone mass distribution and structure are dependent on mechanical stress and adaptive response at cellular and tissue levels. Mechanical stimulation of bone induces new bone formation in vivo and increases the metabolic activity and gene expression of osteoblasts in culture. A wide variety of devices have been tested for mechanical stimulation of cells and tissues in vitro. The aim of this work was to experimentally validate the possibility to use piezoelectric materials as a mean of mechanical stimulation of bone cells, by converse piezoelectric effect. To estimate the magnitude and the distribution of strain, finite numerical models were applied and the results were complemented with the optical tests (Electronic Speckle Pattern Interferometric Process). In this work, osteoblasts were grown on the surface of a piezoelectric material, both in static and dynamic conditions at low frequencies, and total protein, cell viability and nitric oxide measurement comparisons are presented.

Sheep and goat saliva proteome analysis: a useful tool for ingestive behavior research?

Sheep and goats differ in diet selection, which may reflect different abilities to deal with the ingestion of plant secondary metabolites. Although saliva provides a basis for immediate oral information via sensory cues and also a mechanism for detoxification, our understanding of the role of saliva in the pre-gastric control of the intake of herbivores is rudimentary. Salivary proteins have important biological functions, but despite their significance, their expression patterns in sheep and goats have been little studied. Protein separation techniques coupled to mass spectrometry based techniques have been used to obtain an extensive comprehension of human saliva protein composition but far fewer studies have been undertaken on animals' saliva. We used two-dimensional electrophoresis gel analysis to compare sheep and goats parotid saliva proteome. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) and liquid chromatography tandem mass spectrometry (LC-MS/MS) were used to identify proteins. From a total of 260 sheep and 205 goat saliva protein spots, 117 and 106 were identified, respectively. A high proportion of serum proteins were found in both salivary protein profiles. Major differences between the two species were detected for proteins within the range of 25-35 kDa. This study presents the parotid saliva proteome of sheep and goat and highlights the potential of proteomics for investigation relating to intake behavior research.

Salivary amylase induction by tannin-enriched diets as a possible countermeasure against tannins.

Tannins are characterized by protein-binding affinity. They have astringent/bitter properties that act as deterrents, affecting diet selection. Two groups of salivary proteins, proline-rich proteins and histatins, are effective precipitators of tannin, decreasing levels of available tannins. The possibility of other salivary proteins having a co-adjuvant role on host defense mechanisms against tannins is unknown. In this work, we characterized and compared the protein profile of mice whole saliva from animals fed on three experimental diets: tannin-free diet, diet with the incorporation of 5% hydrolyzable tannins (tannic acid), or diet with 5% condensed tannins (quebracho). Protein analysis was performed by one-dimensional gel electrophoresis combined with Matrix-Assisted Laser Desorption Ionization-Time of Flight mass spectrometry to allow the dynamic study of interactions between diet and saliva. Since abundant salivary proteins obscure the purification and identification of medium and low expressed salivary proteins, we used centrifugation to obtain saliva samples free from proteins that precipitate after tannin binding. Data from Peptide Mass Fingerprinting allowed us to identify ten different proteins, some of them showing more than one isoform. Tannin-enriched diets were observed to change the salivary protein profile. One isoform of alpha-amylase was overexpressed with both types of tannins. Aldehyde reductase was only identified in saliva of the quebracho group. Additionally, a hypertrophy of parotid salivary gland acini was observed by histology, along with a decrease in body mass in the first 4 days of the experimental period.