Search

Background: In order to provide appropriate advice to the patient at the time of dispensing and over-the-counter (OTC) medication counseling, community pharmacists need access to current and reliable information about medicines. Brazilian pharmacists have assumed new functions such as prescribing medication, in a dependent model, based in protocols. Objective: To examine the practice of community pharmacists in a Brazilian State, focusing on OTC recommendation. Method: A cross-sectional survey of community pharmacists in a state of Brazil was conducted from October 2013 to January 2014, with data collection through a pre-piloted self-administered anonymous survey via Survey Monkey® platform. Following ethical approval, the online instrument was sent to 8,885 pharmacists registered in Parana State, Brazil, focusing on professionals working in community pharmacies. The questionnaire assessed the community pharmacy setting, the search for information, the knowledge of the evidence-based practice, the important factors to consider when recommending an OTC medicine, and the pharmacist prescribing. Responses were imported into SPSS® (version 22.0) for analysis. Nonparametric tests were used to assess the association between responses and demographic information with a significance level less than 5% (p

The use of recombinant bovine somatotropin (rbST) leads to an increase in variable amounts of milk production in buffalo, but there is a lack of information on the influence of rbST on their metabolism. This study looked at the effects of a single 500 mg dose of rbST on the lipid profile, liver and kidney function, and physical, chemical, and cellular constitution of milk in 14 buffalo over 14 days, from the 100th day of lactation, compared with 14 animals in a control group. From the first day after rbST, there was a rise in beta-hydroxybutyrate (β-HBO), possibly due to higher dry matter intake or the biotransformation of NEFA into β-HBO. The treatment did not influence blood glucose, non-esterified fatty acids (NEFAs), triglycerides, cholesterol, total protein, albumin, AST, GGT, bilirubin, urea, or creatinine levels. In 71.3% of the buffalo, there was a gradual increase in milk production, with the maximal response occurring in the first week followed by a gradual decrease, whilst in 21.4%, the increase in production occurred between 7 and 10 days. Only 7.1% of the animals did not respond. On the 3rd, 5th, 7th, and 10th days after treatment, an increase was found in daily milk production between the two groups equal to 1.04, 1.52, 1.42, and 1.06 L, respectively. In relative terms, this means an increase in milk production, respectively, of 15.1%, 21.0%, 19.8%, and 15.1%. The constitution of the milk showed no difference in the amounts of fat, lactose, total solids, or somatic cell count; however, on the third day after rbST administration, there was a decrease in protein. Notably, from the fifth day, the protein values showed no statistical difference. It can be concluded that the use of rbST in buffalo from the 100th day of lactation is metabolically safe since the treatment neither caused imbalances in fat metabolism nor overloaded the liver or renal function, and the changes in milk composition were transient and limited to a decrease in milk protein.

The protein composition in goat milk undergoes changes throughout the different lactation periods, displaying distinct characteristics that are influenced by the dynamic nature of protein composition and concentration during the transition from colostrum secretion to mature milk. To evaluate the dynamics of whey proteins of Saanen goats during the colostral phase and the first month of lactation, 110 milk samples from 11 healthy mammary halves of seven Saanen goats were selected through a clinical evaluation. Whey was obtained by rennet coagulation of the mammary secretion. The biuret method determined total protein concentration, and their fractions were identified by 12% dodecyl sulfate-polyacrylamide gel electrophoresis. Maximum concentrations of all protein fractions were observed in the first 12 h of lactation, reducing throughout the study. Modification of the protein predominance was also observed. The transition from colostrum secretion to milk occurred 5 or 7 d postpartum.

Reduced welfare and productivity of dairy goats have often been associated with poor claw health, especially conditions such as claw overgrowth and deformations. It is known that periodic claw trimmings have prophylactic and therapeutic effects on these problems, and this study aimed to evaluate if the additional use of an angle grinder to finish trimming overgrown and deformed goat claws, after the usual trimming using hoof shears, could provide further changes in these claws. For this, twelve Saanen goats (57.29 ± 11.15 kg of body weight, 3.08 ± 1.78 years old) were selected by presence of severe claw overgrowth, and absence of claw alterations of other nature. Their claws were trimmed in two steps, first using hoof shears and then using an angle grinder. Morphometric, baropodometric, and conformational aspects of all claws were assessed before claw trimming and after each trimming step. To analyse the effects of the trimming steps in each claw, the Tukey's test was used on parametric data, with 5% probability, and descriptive statistics were used on non-parametric data. Although this is a small pilot study, results suggest that using an angle grinder after the use of hoof shears, could further reduce heel length and sole width of claws, as well as reduce the number of deformed claws. The incorporation of the second trimming tool, could also further increase the frequency with which the point of maximum pressure was found in the toes, rather then in the heels of the claws as seen in deformed claws., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

The aim of the present study is to evaluate the erythrogram and iron serum profiles of neonatal calves born spontaneously or born by elective cesarean section with or without dexamethasone induction. The research was performed on 38 newborn Nellore calves. Three groups of calves were assigned according to the type of birth: calves born by spontaneous vaginal calving ( n = 10), calves born by elective cesarean section without inducing labor ( n = 14), and calves born by elective cesarean section with labor induction with dexamethasone ( n = 14). Blood samples to assess red blood cell count (RBC), hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), concentration of mean corpuscular hemoglobin (MCHC), serum iron (SFe), total capacity to bind iron to transferrin (TIBIC), and transferrin saturation index (TSI) were performed at calving (0, 3, 6, and 12 h of life) and on 1, 2, 3, 5, 7, 10, 15, and 30 days of life. Regardless of the experimental group (calves born spontaneously, or born by elective cesarean section with or without dexamethasone induction), in the first day of life there was a decrease in the number of red blood cells, hemoglobin rates, and values of the globular volume. In the period of the first 10 days of life, animals from spontaneous vaginal delivery quickly recovered values of erythrocytes, hemoglobin, and packed cell volume, whereas animals born by elective C-section (induced and uninduced) did not recover as quickly in their rates of hemoglobin and packed cell volume values. In calves born by elective C-section (induced and uninduced), it was observed in their period between 10 and 30 days of life that the MCV and MCH were reduced by passing the presenting microcytic hypochromic when compared with calves obtained by spontaneous vaginal delivery. In the period between 10 and 30 days of life, the levels of SFe and TSI in animals born by elective C-section (induced and uninduced) are significantly lower. The differences in the erythrogram values between Nellore calves born spontaneously and those by elective C-section with or without induction must be considered consequent to the process of neonatal adaptation to extrauterine life. Iron supplementation in the first month of life in calves from cesarean could be recommended to prevent anemia of this iron deficiency.

Several studies have described the effects of human tumor necrosis factor alpha (hTNF-α) on Schistosoma mansoni. hTNF-α affects the worm's development, metabolism, egg-laying, gene expression and protein phosphorylation. The available data on the influence of hTNF-α on egg-laying in S. mansoni are controversial, but understanding the mechanism of egg-laying regulation in this species is essential in combating schistosomiasis. We characterized the effects of in vitro treatment of S. mansoni adult worms with different doses of hTNF-α (5, 20 and 40 ng/ml) for 5 days. We explored the effects on egg-laying rate, glucose levels, ATP metabolism, and messenger RNA (mRNA) expression levels of lactate dehydrogenase, glucose transporters and the parasite gene which acts as an hTNF-α receptor, SmTNFR. hTNF-α influenced egg-laying in a time- and dose-dependent manner: at a dose of 40 ng/ml, egg-laying increased on day 2 and decreased on days 3 and 4; at 20 ng/ml, egg-laying decreased on day 3; while at 5 ng/ml, egg-laying decreased on day 4. The total number of eggs produced was not affected by the different treatments, but the egg-laying dynamics were: the median egg-laying time decreased significantly with treatment, and egg developmental stages and size were also affected. At 5 and 20 ng/ml hTNF-α, lactate production diminished on day 3 up to day 5, while glucose uptake increased on day 5. At 40 ng/ml, glucose uptake diminished on day 1 up to day 3, while ATP accumulation was detected on day 5. No significant changes in mRNA expression were detected in any of the treatments. We found that crosstalk involving hTNF-α and parasite signaling plays a role in the fine-scale regulation of the worm's metabolism and physiology, and points to new strategies for disease control., (© 2022. The Author(s).)

Metritis in dairy cows is classified as a puerperal disease that affects all layers of the uterine epithelium and occurs during the first 21 days after parturition. The objectives of this study were to identify potential predictive biomarkers of metritis, to monitor the biochemical profile of pre-metritic dairy cows, and to evaluate the reproductive performance of dairy cows with metritis. Fifty-five cows without metritis and fifteen cows that showed clinical signs of metritis were enrolled in this study. Blood samples were collected at -21, -14 and - 7 days before calving and at parturition and assayed for urea, creatinine, aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), total cholesterol, fibrinogen, total calcium, β-hydroxybutyrate (BHB) and non-esterified fatty acids (NEFA) concentrations. Serum fibrinogen on 21, 14 and 7 days prepartum and at parturition was the strongest discriminator between metritic and control cows. Moreover, metritic cows had lower (P < 0.05) concentrations of urea, creatinine, cholesterol and calcium and higher (P < 0.05) serum AST, GGT activities, and serum fibrinogen, BHB and NEFA concentrations when compared to control cows. The number of days to first ovulation, insemination, services per pregnancy and days to pregnancy were greater (P < 0.05) for metritic cows than the control group, and the follicle size was smaller (P < 0.05) for metritic cows than the control group. Our results indicate alterations in the biochemical profile of dairy cows with metritis and the negative impact on the reproductive performance of metritic cows raised in tropical conditions. In addition, this study showed that the serum fibrinogen could be used as screening biomarkers to indicate cows that might have metritis during the postpartum period., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: Rumenostomy is a useful procedure commonly performed in cattle for medical treatment of domestic ruminants with forestomach diseases. Methylene blue (MB)-mediated antimicrobial photodynamic therapy (APDT) has been broadly investigated to treat infected wounds., Aim: The aim of the study was to evaluate the effectiveness of MB-mediated APDT (MB-APDT) combined with chlorhexidine and zinc oxide ointment on wound healing process after rumenostomy., Methods: Fourteen Nelore bulls were subjected to rumenostomy procedure. Animals were randomly divided into MB-APDT (MB associated with a red diode laser performed immediately after surgery and repeated on days 3, 5, 7 and 10) and control groups. Daily care included topical cleaning with chlorhexidine 2% followed by topical zinc oxide ointment. Animals were followed-up until the 28th day., Results: Wounds presented a better post-surgical profile in MB-APDT group when it was compared with the control group. In MB-APDT group, it was also possible to observe less pain on palpation of wounds borders, less edema and inflammatory exudate. Additionally, animals from MB-APDT group were faster discharged from the cattle care facility., Conclusion: Our results support the use of MB-APDT for the post-surgical management of rumenostomy. This pilot study ratifies the use of APDT in cattle and also suggests that it could be performed for other surgical procedures as a complementary approach or an alternative for topical administration of antibiotics., (Copyright © 2020. Published by Elsevier B.V.)

The neonatal period is a challenging phase for calves, and during this phase constant adaptations are required. The aim of the present study was to evaluate the invasive hemodynamics with the Swan-Ganz catheter in neonate calves to understand adaptive changes during the first 30 days of life. A prospective and observational study was conducted with 10 Holstein calves. Assessments of the right atrial pressure (RAP), right ventricular pressure (RVP), pulmonary artery pressure (PAP), pulmonary capillary pressure (PW), cardiac output (CO), heart rate (HR), pulmonary vascular resistance (PVR), and blood gas levels were performed. The analyses of PAP, PVR, PW, HR, sO 2 , and arterial blood gases differed ( p < 0.05) between the evaluated periods. Our results indicated transient pulmonary artery hypertension during the process of extrauterine adaptation during the first 30 days of life. This hypertension must be considered as physiological and consequent to the neonatal adaptation process.

The aim of the present study was to evaluate the biochemical profile and reproductive performance in postpartum dairy cows with clinical metritis. The research was carried out using performed on 213 pluriparous lactating cows from two dairy herds in southeast Brazil. Dairy cows were assigned into a group of healthy cows (n = 147) and cows with metritis (n = 66). Blood samples to assess serum albumin, cholesterol, fibrinogen, calcium, gamma-glutammyltransferase (GGT), aspartate aminotransferase (AST) and β-hydroxybutyrate (BHBA) concentrations were performed on day 8 ± 2 postpartum. Metritic cows had lower (p < .05) serum albumin, cholesterol and calcium concentration, and higher (p < .05) GGT and AST activity, and BHBA and fibrinogen concentrations when compared to the healthy group. The calving-to-conception interval and services per pregnancy were higher in metritic cows (p < .01) than in healthy cows and the conception rate at first service was lower in metric group (p < .01). Variable importance in projection plots demonstrated that cholesterol was the main discriminator between metritic and healthy cows. Our results indicated alterations in the biochemical profile and impaired reproduction performance in metritic cows., (© 2020 Wiley-VCH GmbH.)

Background: The TNF signaling pathway is involved in the regulation of many cellular processes (such as apoptosis and cell proliferation). Previous reports indicated the effect of human TNF-α on metabolism, physiology, gene expression and protein phosphorylation of the human parasite Schistosoma mansoni and suggested that its TNF receptor was responsible for this response. The lack of an endogenous TNF ligand reinforced the idea of the use of an exogenous ligand, but also opens the possibility that the receptor actually binds a non-canonical ligand, as observed for NGFRs., Methods: To obtain a more comprehensive view, we analyzed platyhelminth genomes deposited in the Wormbase ParaSite database to investigate the presence of TNF receptors and their respective ligands. Using different bioinformatics approaches, such as HMMer and BLAST search tools we identified and characterized the sequence of TNF receptors and ligand homologs. We also used bioinformatics resources for the identification of conserved protein domains and Bayesian inference for phylogenetic analysis., Results: Our analyses indicate the presence of 31 TNF receptors in 30 platyhelminth species. All platyhelminths display a single TNF receptor, and all are structurally remarkably similar to NGFR. It suggests no events of duplication and diversification occurred in this phylum, with the exception of a single species-specific duplication. Interestingly, we also identified TNF ligand homologs in five species of free-living platyhelminths., Conclusions: These results suggest that the TNF receptor from platyhelminths may be able to bind canonical TNF ligands, thus strengthening the idea that these receptors are able to bind human TNF-α. This also raises the hypothesis that an endogenous ligand was substituted by the host ligand in parasitic platyhelminths. Moreover, our analysis indicates that death domains (DD) may be present in the intracellular region of most platyhelminth TNF receptors, thus pointing to a previously unreported apoptotic action of such receptors in platyhelminths. Our data highlight the idea that host-parasite crosstalk using the TNF pathway may be widespread in parasitic platyhelminths to mediate apoptotic responses. This opens up a new hypothesis to uncover what might be an important component to understand platyhelminth infections.

The aims of this study were to characterize the biochemical profile, the reproductive performance and to identify potential predictive biomarkers of disease state of dairy cows with hyperketonemia, lipomobilization and hypocalcemia raised in tropical conditions in southeastern Brazil. Dairy cows (n = 50) were divided into a group of healthy cows (n = 14), cows with lipomobilization (n = 14), cows with hypocalcemia (n = 11), and a group of cows with hyperketonemia (n = 11). Evaluation of body condition score (BCS), body weight (BW) and blood samples was performed on 21, 14, 7, 4 and 2 days before calving, parturition, 1, 7, 14, 21, 30, 45 and 60 days post-partum and milk production was recorded on days 7, 14, 21, 30, 45 and 60 after parturition. Blood samples were assayed for aspartate aminotransferase (AST), gamma-glutammyltransferase (GGT), albumin, total protein, globulin, fibrinogen, total cholesterol, triglyceride, urea and creatinine concentrations. The biochemical profile, BCS, BW, milk production and reproductive performance differed (p < .05) among the groups. Our findings indicate changes in the biochemical profile of dairy cows with metabolic diseases and impaired production and fertility of dairy cows in this group. Variable importance in projection plots demonstrated that cholesterol, urea, total protein, albumin and fibrinogen in the serum were the strongest discriminators between cows with hypocalcemia and healthy cows; and AST, cholesterol, urea and triglycerides for cows with hyperketonemia and healthy cows; and cholesterol, urea, triglycerides, total protein and fibrinogen for lipomobilization and healthy cows, which might be useful as predictive biomarkers of the disease state., (© 2020 Blackwell Verlag GmbH.)

Background: Peripartum is a challenging phase for the health of cows., Objectives: This study analysed the haematological profile of Holstein cows during peripartum., Method: Blood samples were collected on days 18, 12, 8, 5 and 2 before calving, at parturition, and on days 1, 7, 14, 21, 30, 45 and 60 postpartum. Analyses of red blood cell (RBC) count, haemoglobin concentration, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and RBC distribution width were performed; body condition score (BCS) and milk yield were evaluated., Results: Red blood cell (the highest value was 6.10 × 1012/L at parturition and the lowest recorded value was 5.03 × 1012/L 21 days after parturition), haemoglobin and haematocrit (the highest values were 10.48 g/dL and 33.47% at parturition, respectively and the lowest values were 8.28 g/dL and 26.13% on day 30 after parturition, respectively); BCS (the highest and the lowest values were 3.50 points and 2.73 points on days 18 before parturition and 45 after parturition, respectively) and milk production (the lowest and the highest values were 21.48 L and 27.02 L on days 7 and 45 after parturition, respectively) were significantly different (p 0.05) during the peripartum period. Of the total cows (n = 48), 41.7% had RBC, haemoglobin and haematocrit below the reference intervals during at least one collection point during the postpartum period., Conclusion: This study demonstrated that dairy cows included in this investigation suffered alterations in select haematological variables during the postpartum period.

Climate change is seen as a significant threat to the sustainability of livestock production systems in many parts of the world, particularly in tropical regions. Extreme meteorological events can result in catastrophic production and death of livestock. Heat waves in particular can push vulnerable animals beyond their survival threshold limits. However, there is little information about buffalo responses to sudden changes in the thermal environment, specifically the heat waves. This study aimed to quantify the thermoregulatory and blood biochemical responses of heat-acclimatized buffaloes to a simulated heat wave. The experiment was designed in a climatic chamber with two periods of 4 days each. Twelve heat acclimated buffalo heifers aged 18 months were used. The climatic chamber environment was set as follows: 4-day period (P1) simulating the same weather conditions of a summer in humid tropical climate used as a baseline, with daily cycle with Ta and RH at 27 ± 1 °C and 76% from 0600 h to 1900 h and 24 ± 1 °C and 80% from 1900 h to 0600 h, and 4-day period (P2), simulating a daily heat wave cycle, from 0600 h to 1900 h with Ta and RH kept at 36 °C and 78% and from 1900 h to 0600 h, 27 °C and 74%. All animals were subject to both treatments and data were analyzed by a repeated measure analysis of variance, with post-hoc pooling comparison performed by Tukey's test. In P2, there was observed a significant increase in respiratory frequency ( p < 0.01), found four times in P1. The sweating rates were quite high in both periods; still, there were significant increases in P2 compared to P1 ( p < 0.01) (4931 and 3201 g/m 2 /h, respectively). A slight but significant increase in rectal temperature was observed during the day ( p < 0.01), with a rising until 1900 h. The simulated heat wave in P2 did not affect the values of the erythrogram or leukogram, excluding the significant reduction in K + ( p < 0.05). The low heat storage and the subsequent fast and full recovery of the thermal balance late afternoon appear to be related to the high sweating rate values. The massive sweating rate emphasizes its relevance in the maintenance of buffalo homeothermy. The absence of changes in hematological parameters has revealed the considerable physiological resilience of buffaloes toward simulated heat waves.

The main aim of the current study was to assess the prevalence of anaemia in Holstein dairy cows during the puerperium, and the haematological and biochemical profile of dairy cows with and without anaemia. The study was conducted in seven dairy herds in São Paulo State, Brazil. The evaluated sample comprised a total of 336 Holstein cows. Blood samples were collected at postpartum day 25 ± 3. Haematological analysis included white blood cell, red blood cell and platelet count, haematocrit value, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. The biochemical profile encompassed cholesterol, non-esterified fatty acids, β-hydroxybutyrate, albumin, globulin, fibrinogen, calcium and total bilirubin concentrations. The prevalence of anaemia was 16.3% in all herds, and this was not affected by clinical diseases, milk production, parity and body score condition. Moreover, anaemic cows had lower red blood cell count, haematocrit, haemoglobin, serum cholesterol and calcium concentrations and higher white blood cell and platelet counts, mean corpuscular haemoglobin, red cell distribution width, non-esterified fatty acids, β-hydroxybutyrate, fibrinogen and globulin concentrations when compared with non-anaemic cows. The results indicate changes in energy balance and an inflammatory process in anaemic cows.

Purulent vaginal discharge (PVD) is a prevalent uterine disease of dairy cows during the puerperium that affects the milk production and affects the profitability of farms. The objectives of this study were to evaluate the biochemical profile, the body condition score, the milk production of cows with PVD and the effects PVD on reproductive performance. A total of 338 Holstein dairy cows aged from 3 to 5 years, from three commercial dairy farms, from Brazil, were used. Blood samples were collected within 25 ± 3 days post-partum from Holstein dairy cows without PVD (control cows, n = 242) and cows with PVD (n = 96), based on scoring of the vaginal discharge. The body condition score and milk production were recorded on the day of sampling. The biochemical profile encompassed albumin, urea, gamma-glutamyl transferase, calcium, fibrinogen and cholesterol concentrations. The number of services per pregnancy was lower (p < 0.01), and the number of days until first insemination and the median time to pregnancy were higher in cows with PVD (p < 0.01) when compared with control cows. Milk production and body condition score were lower (p < 0.01) in cows with PVD than in control group. Cows with PVD had lower (p < 0.05) serum albumin, urea, calcium and cholesterol concentrations, and higher serum gamma-glutamyl transferase activity and fibrinogen concentration than cows without PVD. Our results show that cows with PVD have changes in the biochemical profile and negative effects on production and reproduction performance., (© 2019 Blackwell Verlag GmbH.)

The objective of this study was to evaluate the effects of different levels of NDF in high-concentrate diets on performance and aspects ruminal, blood, and carcass parameters of finishing lambs. Twenty-four lambs, with initial BW of 24 ± 3 kg, were randomly divided into individual pens, in a completely randomized design, comprising three treatments with eight repetitions each. The treatments consisted of dietary levels of 15%, 20%, and 25% of NDF, based on DM. The diets were composed of corn, soybean meal, minerals, and corn silage and were offered twice a day, with daily control of what was offered and refused, to determine dry matter intake (DMI). The animals were weighed weekly and slaughtered after 50 days of confinement. Performance evaluations, blood parameters, carcass, and ruminal parameters were evaluated. Lambs fed with lower dietary fiber had better carcass yield (P < 0.005). Blood count, blood biochemical parameters, rumen short-chain fatty acid production and protozoan population were not affected by the dietary NDF level. It was concluded that in high-concentrate diets with 15% NDF resulting in better weight gain of the animals in the confinement, good carcass yields without the animal having altered health and ruminal parameters.

The bovine whey consists of more than 200 different types of proteins, of which β-lactoglobulin, α-lactalbumin, serum albumin, immunoglobulins and lactoferrin predominate. However, their concentrations are not stable due to the existence of protein dynamics during a transition from colostrum secretion to mature milk. To evaluate the dynamics of whey proteins of Jersey cows during a colostral phase and first month of lactation and an influence of the number of lactations, 268 milk samples from 135 Jersey cows were selected through a clinical evaluation. Whey was obtained by rennet coagulation of the mammary secretion. The concentration of total proteins was determined by the biuret method and their fractions were identified by 12% dodecyl sulfate-polyacrylamide gel electrophoresis (12% SDS-PAGE). Maximum concentrations of all protein fractions were observed in the first 12 h of lactation, reducing over the course of the study. Modification of the protein predominance was also observed. The transition from colostrum secretion to milk occurred between 24 and 72 h postpartum. There was an influence of the number of lactations on the dynamics of whey proteins, indicating that multiparous cows had better immunological and nutritional quality when compared to primiparous cows.

The activity of the ubiquitin proteasome system (UPS) and the level of oxidative stress contribute to the transition from compensated cardiac hypertrophy to heart failure in hypertension. Moreover, aerobic exercise training (AET) is an important therapy for the treatment of hypertension, but its effects on the UPS are not completely known. The aim of this study was to evaluate the effect of AET on UPS's activity and oxidative stress level in heart of spontaneously hypertensive rats (SHR). A total of 53 Wistar and SHR rats were randomly divided into sedentary and trained groups. The AET protocol was 5×/week in treadmill for 13 weeks. Exercise tolerance test, non-invasive blood pressure measurement, echocardiographic analyses, and left ventricle hemodynamics were performed during experimental period. The expression of ubiquitinated proteins, 4-hydroxynonenal (4-HNE), Akt, phospho-Akt(ser473), GSK3β, and phospho-GSK3β(ser9) were analyzed by western blotting. The evaluation of lipid hydroperoxide concentration was performed using the xylenol orange method, and the proteasomal chymotrypsin-like activity was measured by fluorimetric assay. Sedentary hypertensive group presented cardiac hypertrophy, unaltered expression of total Akt, phospho-Akt, total GSK3β and phospho-GSK3β, UPS hyperactivity, increased lipid hydroperoxidation as well as elevated expression of 4-HNE but normal cardiac function. In contrast, AET significantly increased exercise tolerance, decreased resting systolic blood pressure and heart rate in hypertensive animals. In addition, the AET increased phospho-Akt expression, decreased phospho-GSK3β, and did not alter the expression of total Akt, total GSK3β, and ubiquitinated proteins, however, significantly attenuated 4-HNE levels, lipid hydroperoxidation, and UPS's activity toward normotensive group levels. Our results provide evidence for the main effect of AET on attenuating cardiac ubiquitin proteasome hyperactivity and oxidative stress in SHR rats.

Recombinant coagulation factor IX must be produced in mammalian cells because FIX synthesis involves translational modifications. Human cell culture-based expression of human coagulation factor IX (hFIX) is expensive, and large-scale production capacity is limited. Transgenic animals may greatly increase the yield of therapeutic proteins and reduce costs. In this study, we used a lentiviral system to obtain transgenic cells and somatic cell nuclear transfer (SCNT) to produce transgenic animals. Lentiviral vectors carrying hFIX driven by 3 bovine β-casein promoters were constructed. Bovine epithelial mammary cells were transduced by lentivirus, selected with blasticidin, plated on extracellular matrix, and induced by lactogenic hormones; promoter activity was evaluated by quantitative PCR. Transcriptional activity of the 5.335-kb promoter was 6-fold higher than the 3.392- and 4.279-kb promoters, which did not significantly differ. Transgenic bovine fibroblasts were transduced with lentivirus carrying the 5.335-kb promoter and used as donor cells for SCNT. Cloned transgenic embryo production yielded development rates of 28.4%, similar to previous reports on cloned non-transgenic embryos. The embryos were transferred to recipient cows (N = 21) and 2 births of cloned transgenic cattle were obtained. These results suggest combination of the lentiviral system and cloning may be a good strategy for production of transgenic cattle.