Effects of increased doses of lysine in a rumen-protected form on plasma amino acid concentration and lactational performance of dairy cows fed a lysine-deficient diet.

The objective of these studies was to determine the effects of feeding a novel rumen-protected Lys (RP-Lys) product on plasma AA, lactational performance, and Lys bioavailability. To evaluate RP-Lys on lactation performance a corn-based diet (42.5% of corn silage and 21.9% of corn and corn by-products, on DM basis) was formulated to be Lys deficient but adequate in Met, energy, and metabolizable protein. Thirty-six lactating Holstein cows were fed either a Lys-deficient control diet (CON) with no added RP-Lys, or diets containing 0.3% of RP-Lys (0.3RP-Lys) or 0.6% of RP-Lys (0.6RP-Lys) for 8 wk. There were no effects on dry matter intake (mean ± SD; 26.1 ± 0.58 kg/d), milk yield (37.9 ± 0.72 kg/d), or milk composition to the RP-Lys supplementation. No effect was observed on plasma AA concentrations except for His. Plasma His was linearly reduced by Lys feeding (42.6, 41.2, 30.0 ± 4.09 µM, for CON, 0.3RP-Lys, and 0.6RP-Lys, respectively). Calculated efficiency of Lys utilization decreased linearly with RP-Lys supplementation. In the companion study, 3 rumen-cannulated lactating dairy cows were used in a 3 × 3 Latin square design to assess the bioavailability of the RP-Lys. Free Lys (HCl-Lys), RP-Lys, and water were administered separately by postruminal bolus dosing. The Lys bioavailability was assessed by the ratio of area under the curve of Lys plasma concentration for RP-Lys compared with HCl-Lys and discounted for the area under the curve for water bolus dose. The estimated bioavailability of the RP-Lys was 24.4% ± 4.61. In summary, increased supplemental doses of Lys had no effect on Lys plasma concentration and lactational performance when fed to dairy cows on a corn-based diet, although altered Lys as % of essential AA was observed. However, the lack of effects should be considered in light of the lower-than-expected bioavailability of the RP-Lys.

Mitochondrial genome analysis in penile carcinoma.

Penile cancer is a rare neoplasm that seems to be linked to socio-economic differences. Mitochondrial genome alterations are common in many tumors types and are reported as regulating oxidative metabolism and impacting tumorigenesis. In this study, we evaluate for the first time the mitochondrial genome in penile carcinoma (PeCa), aiming to evaluate heteroplasmy, mitochondrial DNA (mtDNA) mutational load and mtDNA content in Penile tumors. Using next generation sequencing (NGS), we sequenced the mitochondrial genome of 13 penile tumors and 12 non-neoplastic tissue samples, which allowed us to identify mtDNA variants and heteroplasmy. We further evaluated variant's pathogenicity using Mutpred predictive software and calculated mtDNA content using quantitative PCR. Mitochondrial genome sequencing revealed an increase number of non-synonymous variants in the tumor tissue, along with higher frequency of heteroplasmy and mtDNA depletion in penile tumors, suggesting an increased mitochondrial instability in penile tumors. We also described a list of mitochondrial variants found in penile tumor and normal tissue, including five novel variants found in the tumoral tissue. Our results showed an increased mitochondrial genome instability in penile tumors. We also suggest that mitochondrial DNA copy number (mtDNAcn) and mtDNA variants may act together to imbalance mitochondrial function in PeCa. The better understanding of mitochondrial biology can bring new insights on mechanisms and open a new field for therapy in PeCa.

Effects of treatment of preweaning dairy calves with recombinant bovine somatotropin on immune responses and somatotropic axis.

Weaning may be associated with negative energy balance and body weight loss when calves are still immunologically immature, predisposing them to infectious diseases. The aim of the present experiment was to investigate the effects of treatment of preweaning dairy calves with recombinant bovine somatotropin (rbST) on the somatotropic axis, selected immune parameters, and hematology of calves around weaning. Thirty-six Holstein female calves were randomly assigned to receive 1.5 to 1.8 mg of rbST (Posilac, Elanco Animal Health, Greenfield, IN) per kilogram of body weight or to receive injections of saline (saline solution 0.9%, Valley Vet Supply, Marysville, KS) every 7 d from 21 to 63 d of life. Calves were fed milk replacer ad libitum from birth to 38 d of age (d -11), when progressive weaning started, and calves were weaned at 49 d of age (d 0). Calves were weighed at birth and weekly from 21 to 63 d of age, when wither height also was measured. Calves were vaccinated with 0.5 mg of ovalbumin on study d -28 and -7. Blood samples were collected on d -28, -25, -21, -11, 0, 3, 7, and 14. Polymorphonuclear leukocytes were isolated and challenged ex vivo with Escherichia coli to determine phagocytosis and oxidative burst capacity. Additionally, expression of cluster of differentiation (CD)62L and CD18 by granulocyte, lymphocyte, and CD14+ monocyte were determined. Blood samples were also used to determine hematological parameters and concentrations of growth hormone, insulin-like growth factor-1, insulin, glucose, fatty acids, ß-hydroxybutyrate, haptoglobin, and anti-ovalbumin IgG. Calves treated with rbST had greater concentrations of growth hormone and insulin-like growth factor-1 from d -25 to 14 than control calves, whereas insulin, fatty acid, and ß-hydroxybutyrate concentrations did not differ. On d -11, glucose concentration was greater for rbST-treated calves. Treatment did not affect polymorphonuclear lymphocyte phagocytosis and oxidative burst, but intensity of expression of CD62L and CD18 by granulocytes tended to be increased by rbST treatment. Treatment did not affect the concentration of anti-ovalbumin IgG in serum. Haptoglobin concentration was reduced in rbST treated calves on d 3 and we noted a tendency for hematocrit to be lower in rbST-treated calves. Treatment did not affect body weight, wither height, and average daily gain, despite the fact that rbST-treated calves had lower daily milk replacer intake. The relatively minor improvements in immune responses resulting from rbST treatment of weaning calves may not be sufficient to reduce the incidence of infectious diseases.

Expression of hepatic genes related to energy metabolism during the transition period of Holstein and F1 Holstein-Gir cows.

The aim of this study was to investigate the expression of genes encoding enzymes and other factors involved with carbohydrate and lipid metabolism in the liver of 2 genetic groups of dairy cows during the transition period. We analyzed the expression of glucose-6-phosphatase (G6PC), cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C), methylmalonyl-CoA mutase (MUT), ß-hydroxybutyrate dehydrogenase-2 (BDH2), acetyl-CoA carboxylase (ACC), carnitine palmitoyltransferase-2 (CPT2), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), glucose transporter-2 (SLC2A2), and the transcription factor peroxisome proliferator-activated receptor α (PPARA). Blood concentrations of glucose, nonesterified fatty acids, and ß-hydroxybutyrate were also determined. Liver biopsies and blood samples were taken at d 15 prepartum and at d 6, 21, 36, 51, and 66 postpartum from Holsteins (n = 6) and F1 Holstein-Gir (n = 6) cows. Cows were kept under the same prepartum and postpartum management conditions. The results showed that the expression of G6PC, PEPCK-C, BDH2, ACC, CPT2, HMGCR, SLC2A2, and PPARA genes did not differ between genetic groups. Except for PEPCK-C, no interaction between genetic groups and the experimental period was observed. Within both groups of cows, G6PC and PEPCK-C gene expression decreased when comparing prepartum gene expression with 21 and 36 DIM, and increased in d 51 postpartum. MUT mRNA levels differed between the 2 genetic groups and displayed a significant increase after d 36 postpartum, whereas mRNA levels of HMGCR tended to increase when comparing d 21 and 36 to d 51 postpartum. Glucose concentrations also differed between genetic groups, being significantly higher in the plasma of F1 Holstein-Gir cows than in Holstein cows, but no differences were found within each group during the analysis period. ß-Hydroxybutyrate and nonesterified fatty acid concentrations did not differ between genetic groups, but displayed increased levels from prepartum to d 6 and 21 postpartum. Our results indicated that expression in the liver of genes involved with glucose and fatty acid metabolism were similar in both groups of cows and significant differences were observed between the 2 groups in the expression of MUT, a gene involved in propionate metabolism.

Incremental amounts of ground flaxseed decrease milk yield but increase n-3 fatty acids and conjugated linoleic acids in dairy cows fed high-forage diets(1).

The objective of this study was to investigate the effect of incremental amounts of ground flaxseed (GFX) on milk yield and concentrations and yields of milk components, milk fatty acids (FA) profile, ruminal metabolism, and nutrient digestibility in dairy cows fed high-forage diets. Twelve multiparous Jersey cows averaging (mean ± SD) 112±68d in milk and 441±21kg of body weight and 8 primiparous Jersey cows averaging 98±43d in milk and 401±43kg of body weight were randomly assigned to treatment sequences in a replicated 4×4 Latin square design. Each period lasted 21d with 14d for diet adaptation and 7d for data and sample collection. Treatments were fed as a total mixed ration (63:37 forage-to-concentrate ratio) with corn meal and soybean meal replaced by incremental levels (i.e., 0, 5, 10, or 15% diet dry matter) of GFX. The ruminal molar proportions of acetate and butyrate decreased linearly with GFX supplementation, whereas the ruminal molar proportion of propionate increased linearly resulting in decreased acetate-to-propionate ratio. Apparent total-tract digestibilities of nutrients either decreased (dry matter) or tended to decrease (organic matter, neutral detergent fiber, acid detergent fiber) linearly in cows fed GFX. Milk yield decreased linearly in cows fed increasing amounts of GFX, which is explained by the linear reduction in dry matter intake. Except for the concentrations of milk protein and urea N, which decreased linearly with GFX supplementation, no other changes in the concentration of milk components were observed. However, yields of milk protein and fat decreased linearly with GFX supplementation. The linear decrease in the yields of milk fat and protein are explained by reduced milk yield, whereas that in milk urea N is explained by decreased crude protein intake. No treatment effects were observed for plasma urea N and nonesterified fatty acids, serum cortisol, and body weight change. Milk odd- and branched-chain FA and saturated FA decreased linearly with GFX supplementation. Milk trans-11 18:1, α-linolenic acid, cis-9,trans-11 18:2, and the sum of n-3 FA all increased linearly and quadratically, whereas the milk ratio of n-6 to n-3 decreased linearly in cows fed GFX. Overall, compared with the control diet (0% GFX), the diet with 15% GFX supplementation resulted in the lowest milk yield but highest milk proportions and yields (data not shown) of cis-9,trans-11 18:2 and n-3 FA.

Effects of milk replacer feeding strategies on performance, ruminal development, and metabolism of dairy calves.

Performance, rumen development, and metabolism of male Holstein calves (n=54) were evaluated according to 1 of 3 liquid feeding strategies: 4 L of milk replacer (MR)/d until 60 d old (4 L-60 d), 6L of MR/d until 29 d old and 4 L/d from 30 to 60 d (6L-29 d/4 L-60 d), or 6L of MR/day until 60 d old (60 d-6L). Water and starter were provided ad libitum. Intakes of MR and starter were monitored daily and body weight (BW) weekly. Blood samples for glucose and insulin concentrations and ruminal content samples for volatile fatty acids and ruminal ammonia concentrations were collected at 15, 30, 45, 60, 75, and 90 d of age. Six calves on each treatment were euthanized at 30, 60, and 90 d of age. Empty weights of forestomach and abomasum, papillae length, and mitotic index were measured. Average MR intake/d (expressed as a percent of BW) was greater for calves receiving 6L/d than for those receiving 4 L/d until 30 d of age. Calves on the 6 L-29 d/4 L-60 d had the smallest MR intake from 30 to 60 d old, followed by the 4 L-60 d and 6L-60 d treatments. Starter intake (kg of dry matter/day) did not differ between groups. It increased from 0.065 kg/d in the first month to 0.386 kg/d in the second month, and to 2.065 kg/d after weaning. Weight gain was greater for calves fed more MR in the first month, but no difference was observed during the second month. After weaning, 6L-60 d calves had greater rate of weight gain than others and were heavier at 30, 60, and 90 d of age. Weight of empty forestomachs, ruminal pH, and ammoniac nitrogen concentration were not different among groups. Propionate concentration was lower for 6 L-60 d calves, but acetate and butyrate concentrations were not influenced by MR feeding strategy. Calves fed more MR until d 30 had greater ruminal epithelium mitotic index. The different MR feeding strategies did not influence papillae length or ruminal epithelium thickness. Lesions such as ruminal parakeratosis or hyperkeratosis were not observed. The MR feeding strategy did not affect glucose concentration, but insulin was higher in 6L-60 d calves than in the other groups. Glucose concentration increased with age, whereas insulin decreased until 45 d old, and then started to increase until 90 d. In conclusion, MR feeding strategy did not influence ruminal development. Feeding calves 6 L of MR/d over 60 d resulted in greater rate of weight gain without negative effects on starter intake or forestomach development. The weight advantage that 6L-60 d calves obtained preweaning was maintained until 90 d of age.

Gene expression study related with the intrinsic pathway of apoptosis in bladder cancer by real-time PCR technique.

We examined the expression of anti-apoptotic genes (XIAP and Bcl-2) and apoptotic genes (cytochrome c, caspase-9, Apaf-1) in tissue samples of patients with superficial bladder cancer. Thirty-two bladder cancer tissue samples (8 papillary urothelial neoplasm of low malignant potential, 10 low-grade, and 14 high-grade) and 8 normal bladder tissue samples from necropsy were used for the study of gene expression by real-time PCR analysis. Analysis of the expression of apoptotic gene constituents of an apoptosome demonstrated an increase in Apaf-1 expression in the three tumor grades when compared with the control (P < 0.01, P < 0.05, and P < 0.01), low expression of caspase-9 in all groups (P < 0.05), and an increase in cytochrome c expression in all tumor grades in relation to the control, although without statistically significant difference. The expression of anti-apoptotic genes revealed an increase in XIAP expression in all tumor grades in relation to the control, although without statistically significant difference, and low expression of Bcl-2 in all tumor grades and the control (P < 0.05). The results proved that there is low evidence of apoptotic activity by the intrinsic pathway, demonstrated by the low expression of caspase-9 and considerable increase in XIAP expression, which may render these genes potential therapeutic targets in bladder cancer treatment.

Pan-cancer genomic analysis shows hemizygous PTEN loss tumors are associated with immune evasion and poor outcome.

In tumors, somatic mutations of the PTEN suppressor gene are associated with advanced disease, chemotherapy resistance, and poor survival. PTEN loss of function may occur by inactivating mutation, by deletion, either affecting one copy (hemizygous loss) leading to reduced gene expression or loss of both copies (homozygous) with expression absent. Various murine models have shown that minor reductions in PTEN protein levels strongly influence tumorigenesis. Most PTEN biomarker assays dichotomize PTEN (i.e. presence vs. absence) ignoring the role of one copy loss. We performed a PTEN copy number analysis of 9793 TCGA cases from 30 different tumor types. There were 419 (4.28%) homozygous and 2484 (25.37%) hemizygous PTEN losses. Hemizygous deletions led to reduced PTEN gene expression, accompanied by increased levels of instability and aneuploidy across tumor genomes. Outcome analysis of the pan-cancer cohort showed that losing one copy of PTEN reduced survival to comparable levels as complete loss, and was associated with transcriptomic changes controlling immune response and the tumor microenvironment. Immune cell abundances were significantly altered for PTEN loss, with changes in head and neck, cervix, stomach, prostate, brain, and colon more evident in hemizygous loss tumors. These data suggest that reduced expression of PTEN in tumors with hemizygous loss leads to tumor progression and influences anticancer immune response pathways.

Targeted uptake of folic acid-functionalized polymeric nanoparticles loading glycoalkaloidic extract in vitro and in vivo assays.

Solanum lycocarpum fruits contain two major glycoalkaloids (GAs), solamargine (SM) and solasonine (SS). These compounds are reported as cytotoxic. However, they have poor water solubility and low bioavailability. To overcome these disadvantages and getting an efficient formulation the current study aimed to develop, characterize, and test the effectiveness of a nanotechnology-based strategy using poly(D,L-lactide) (PLA) nanoparticles functionalized with folate as delivery system of glycoalkaloidic extract (AE) for bladder cancer therapy. The strategic of adding folic acid into nanoformulations can increase the selectivity of the compounds to the cancer cells reducing the side effects. Our results revealed the successful preparation of AE-loaded folate-targeted nanoparticles (NP-F-AE) with particle size around 177 nm, negative zeta potential, polydispersity index <0.20, and higher efficiency of encapsulation for both GAs present in the extract (>85 %). To investigate the cellular uptake, the fluorescent dye coumarin-6 was encapsulated into the nanoparticle (NP-F-C6). The cell studies showed high uptake of nanoparticles by breast (MDA-MB-231) and bladder (RT4) cancer cells, but not for normal keratinocytes cells (HaCaT) indicating the target uptake to cancer cells. The cytotoxicity of nanoparticles was evaluated on RT4 2D culture model showing 2.16-fold lower IC50 than the free AE. Furthermore, the IC50 increased on the RT4 spheroids compared to 2D model. The nanoparticles penetrated homogeneously into the urotheliumof porcine bladder. These results showed that folate-conjugated polymeric nanoparticles are potential carriers for targeted glycoalkaloidic extract delivery to bladder cancer cells.

Hypoenhancing prostate cancers on dynamic contrast-enhanced MRI are associated with poor outcomes in high-risk patients: results of a hypothesis generating study.

PURPOSE: To investigate the association of hypoenhancement on dynamic Contrast enhanced (DCE) with prostate cancer patient outcomes. MATERIAL AND METHODS: This was a single-institution retrospective Institutional Review Board (IRB)-approved cohort study of 54 men who had prostate Magnetic Resonance Imaging (MRI) within 6 months of cancer diagnosis between 01/2012 to 03/2014. Two readers independently identified the dominant MRI-lesions utilizing Prostate Imaging-Reporting and Data System-version2- guidelines. These lesions were classified as hypoenhancing or hyperenhancing, compared to normal peripheral zone using quantitative DCE analysis. The t test for unequal sample sizes and the two-sample Wilcoxon rank-sum tests were used to compare groups. Logistic regression determined if DCE characteristics predict the development of metastases or prostate cancer death. RESULTS: Time-to-progression was significantly shorter for hypoenhancing tumors (6.2 vs. 24.8 months, p = 0.05). Men with these lesions had a higher odds of having poor outcome (univariate logistic regression, odds ratio (OR) 6.79, 95% confidence interval (CI) 1.45-31.72, p = 0.02; multivariate analysis, OR 2.05, 95% CI 0.30-13.72, p = 0.47). Hypoenhancing tumors were larger (33.1 vs. 19.1 mm, p < 0.001) and more likely to be intermediate (Gleason scores 3 + 4 and 4 + 3) and high-grade (Gleason scores ≥ 4 + 4) prostate cancers (p = 0.05). Men in the hypoenhancing group had a higher mean prostate-specific antigen (PSA) value (87.6 vs. 24.8 ng/dL, p = 0.01) and PSA density (1.54 vs. 0.72, p = 0.03). The mean Ktrans and kep of hypoenhancing lesion were lower when compared to hyperenhancing lesions (p = 0.03 and p = 0.04). Ve values did not differ (p = 0.25). CONCLUSION: Men with hypoenhancing prostate cancers may have a worse prognosis than men with hyperenhancing tumors.

Fatores de influência sobre o desempenho reprodutivo em vacas leiteiras / [Influence factors on reproductive performance in dairy cows]

O objetivo deste estudo foi avaliar os efeitos das estações do parto e da inseminação, da retenção de placenta, da natimortalidade e da ordem de parto sobre o desempenho reprodutivo de vacas leiteiras. Utilizou-se a regressão logística, em que as variáveis dependentes foram concepção à primeira inseminação pós-parto e percentual de vacas prenhes no rebanho aos 100, 150 ou 200 DEL e as variáveis independentes foram a estação do parto e a estação da inseminação, a retenção de placenta, a natimortalidade e a ordem de parto. A estação do parto e a estação da inseminação influenciaram o desempenho reprodutivo, reduzindo a fertilidade das vacas nas estações quentes do ano. O atraso na primeira inseminação aumentou a concepção na primeira inseminação, mas reduziu o percentual de vacas prenhes ao longo da lactação. O desempenho reprodutivo da vaca leiteira foi afetado por vários fatores, relacionados à vaca, ao ambiente e ao manejo a ela imposto.(AU)

The objective of this study was to evaluate the effects of the calving season, insemination, placenta retention, stillbirth and birth order on the reproductive performance of dairy cows. Logistic regression was used, where the dependent variables were conception at the first postpartum insemination and the percentage of cows pregnant in the herd at 100, 150 and 200 DEL. The independent variables were delivery season and insemination season, placenta retention, stillbirth and delivery order. The calving season and the insemination season influenced reproductive performance, and in the hot seasons of the year the fertility of the cows was reduced. The delay in the first insemination increased conception in the first insemination but reduced the percentage of pregnant cows during lactation. The reproductive performance of the dairy cow was affected by several factors, related to the cow, the environment and the management imposed on it.(AU)

Comparison of PI-RADS 2, ADC histogram-derived parameters, and their combination for the diagnosis of peripheral zone prostate cancer.

PURPOSE: The purpose of this study was to compare the PI-RADS V2 scores, ADC histogram-derived parameters, and their combination for the diagnosis of clinically significant peripheral zone prostate cancer (PCa). MATERIALS AND METHODS: The IRB approved this retrospective study of 47 men who underwent 1.5 Tesla endorectal prostate magnetic resonance imaging (MRI). Informed consent was waived. Two readers identified and scored MRI lesions using PI-RADS V2. Their mean, median, 10th, 25th, 75th percentile ADC values, and normalized ratio were also calculated. Multilevel logistic regression and receiver-operating characteristic (ROC) curve analyses assessed their diagnostic performance. Clinically significant PCa was defined as tumor volume over 0.5 cc and Gleason grade of 4 or 5 on prostatectomy. RESULTS: The area under the ROC curve (A z) of the overall and diffusion-weighted imaging (DWI) PI-RADS V2 scores were 0.69 and 0.84 (reader-1), and 0.68 and 0.73 (reader-2). The A z of ADC parameters ranged from 0.68 to 0.75 for both readers. Compared to other predictors, DWI PI-RADS V2 yielded the highest A z for identification of significant cancer; but, except for reader-1 75th percentile ADC, the differences were not statistically significant (P > 0.05). Adding ADC parameters to PI-RADS V2 scores did not improve their diagnostic ability. CONCLUSION: DWI PI-RADS V2 score may a better predictor of clinically significant PCa than the overall PI-RADS V2 score, but its diagnostic performance was not significantly improved by the addition of objective ADC value measurements.

Effects of bovine mammary gland biopsy and increased milking frequency on post-procedure udder health, histology, and milk yield.

Sixteen cows in early lactation were randomly distributed into two groups in order to evaluate the effects of mammary biopsies and increased milking frequency on tissue characteristics, post-biopsy udder health and histology. One group was milked twice a day (2×) starting on the 2nd day after calving, until 28 days in milk (DIM). The other group was milked four times a day (4×) from two to 21 DIM, and twice a day (2×) from 22 to 28 DIM. On days 2, 7, 14, 21, and 28 postpartum, one fragment of secretory tissue was collected from one mammary quarter at a time. Collections were alternated between the four mammary quarters per collection day. A total of 80 mammary tissue samples were collected. Qualitative and quantitative analyses of the tissues were conducted by histologic examination. Animal health was assessed by observation of feed intake behavior immediately after biopsy, and weight and body condition score before and one week after biopsy. Udder health was assessed daily from calving to 60 DIM with California Mastitis Test (CMT) and by noting alterations in the milk such as blood, milk clots, blood clots, clinical signs of mastitis. Milk composition and somatic cell count (SCC) were analyzed before and after the biopsies. Milk production was evaluated before biopsy, on the day of biopsy, and after the biopsy. An average of 10 fields at 40× magnification was obtained from each sample. There were no evident changes in mammary morphology as a result of milking two or four times/day at any of the evaluated time points. Biopsy wounds healed rapidly without infection. Intramammary bleeding and CMT alterations were observed in 96% and 75% of the biopsied mammary quarters, respectively. Clinical mastitis was diagnosed in 12% of the biopsied quarters. Different milking frequencies had no effect on the frequency and duration of post-biopsy alterations. Milk production decreased after biopsies done on days 2 for 2× and 4× groups, but it returned to pre-biopsy values within 1 day. Milk composition and SCC were affected transiently. Increased milking frequency did not influence udder health. Post-biopsy recovery was rapid and the procedure proved effective without damaging the cows' health.

Supplementation with different non-fiber carbohydrate sources in dairy cow diets with high or low rumen-undegradable protein content / Suplementação com diferentes fontes de carboidratos não fibrosos em dietas para vacas leiteiras com alto ou baixo teor de proteína não degradável no rúmen

The objective for this study was to evaluate the supplementation with different non-fibrous carbohydrate (NFC) sources for grazing dairy cows in diets with high or low concentrations of rumen undegradable protein (RUP). Twelve multiparous cows averaging 30kg/d of milk production and 45±23 days in milk in a 4x4 Latin square design with 2 × 2 factorial arrangement were used. The difference in NFC content of the diets was achieved using ground corn (GC) or dried citrus pulp (DCP). Dietary RUP was altered by the partial replacement of soybean meal (LRUP) by roasted whole soybean and corn gluten meal (HRUP). Greater dry matter intake (DMI) was observed in DCP treatments compared to GC treatments (19.2 and 17.7kg/day of DM, respectively). Milk fat concentration was lower for LRUP treatments (3.4%) when compared to HRUP (3.6%). The milk protein concentration was greater for GC treatments (2.9%) in relation to DCP treatments (2.8%). Lower milk ureic nitrogen was observed when DCP diets were fed compared to those with GC (17.8 and 21.2mg/dL, respectively). The use of DCP as CNF source increased dry matter intake as well as milk fat concentration, although no differences in milk production were observed for RUP level.(AU)

Objetivou-se avaliar a suplementação com diferentes fontes de carboidratos não fibrosos (CNF) em vacas manejadas a pasto em dietas com alta ou baixa concentração de proteína não degradável no rúmen (PNDR). Doze vacas multíparas com média de 30kg leite/ dia e 45±23 dias em lactação foram utilizadas em delineamento quadrado latino 4x4 com arranjo fatorial 2 × 2. As fontes de CNF foram milho moído (MM) e polpa cítrica seca (PC). A PNDR da dieta foi alterada pela substituição parcial do farelo de soja da dieta de baixa PNDR (BPNDR) por grão de soja tostado e farelo de glúten de milho dieta de alta PNDR (APNDR). Foi observado maior consumo de matéria seca (CMS) nos tratamentos com PC quando comparado aos tratamentos com MM (19,2 e 17,7kg / dia de MS, respectivamente). O teor de gordura do leite foi menor para os tratamentos com BPNDR (3,4%) em comparação aos tratamentos com APNDR (3,6%). O teor de proteína do leite foi maior nos tratamentos com MM (2,9%) em relação aos tratamentos com PC (2,8%). Foram observados menores teores de nitrogênio ureico no leite dos animais alimentados com PC em relação aos com MM (17,8 e 21,2mg / dL, respectivamente). O uso de PC como fonte de CNF aumentou o consumo de matéria seca, bem como o teor de gordura do leite, embora não tenham sido observadas diferenças na produção de leite relacionada a alteração nos teores de PNDR.(AU)

Combination therapy using oral alpha-blockers and intracavernosal injection in men with erectile dysfunction.

OBJECTIVES: Intracavernosal injection with a combination of agents (ie, phentolamine plus papaverine or alprostadil) has been used in an effort to increase efficacy and reduce side effects compared with single agents. The purpose of this pilot study was to determine the potential role of oral alpha-blockers in combination with intracavernosal therapy in men with erectile dysfunction, for whom intracavernosal therapy alone failed. METHODS: Thirty-eight consecutive men with moderate to severe erectile dysfunction on the basis of history and examination and with minimal or no therapeutic response to intracavernosal alprostadil injection therapy were evaluated. All patients received daily doxazosin titrated to 4 mg over 3 weeks in combination with intracavernosal therapy as needed for 12 weeks. Efficacy was assessed at 4, 8, and 12 weeks after doxazosin titration using the 1 5-item, self-administered International Index of Erectile Function (IIEF) and a global efficacy question (GEQ: Did treatment improve your erections?). RESULTS: For the group, the mean baseline IIEF score before therapy was 29.7+/-9.8. After intracavernosal therapy (mean dose 34.7+/-7.3 microg), IIEF improved to 36.1+/-1 1.4 (17.7%). After addition of doxazosin, IIEF improved to 48.6+/-13.4, 46.4+/-10.9, and 51.5+/-14.3 at 4, 8, and 12 weeks, respectively (P < 0.01). The GEQ response improved from 25.7% at baseline to 81.4% at 12 weeks. Overall 22 (57.9%) of 38 patients with the combined regimen had a significant (more than 60% improvement in IIEF) therapeutic response. CONCLUSIONS: The addition of an oral alpha-blocker may have a beneficial effect in patients with erectile dysfunction for whom intracavernosal therapy alone fails. The synergistic effects of vascular dilation and blockade of sympathetic inhibition may explain this response. The potential role of alpha-blockade in synergy with other agents designed to treat erectile dysfunction remains to be determined.

Intermittent alpha-blocker therapy in the treatment of men with lower urinary tract symptoms.

OBJECTIVES: To determine the safety and efficacy of intermittent alpha-blocker therapy in men with lower urinary tract symptoms (LUTS) in a prospective study. Alpha-blockers have been demonstrated to be safe and effective in the treatment of men with LUTS. To date, the role of varying dosing regimens in responding patients has not been well studied. METHODS: Men with LUTS were entered into this prospective open label, parallel, randomized trial. In phase 1, patients were treated with alfuzosin, 2.5 mg three times daily for 3 months. In phase 2, those patients who had a significant therapeutic response were randomized into one of the following three groups: (1) maintenance of alfuzosin; (2) alfuzosin every other day; and (3) discontinuation of alfuzosin (ie, no treatment). Patients were followed up for a total of 6 months. Parameters of evaluation included the International Prostate Symptom Score (IPSS), global satisfaction, peak urinary flow rate (Qmax), and adverse events. RESULTS: At 3 months, there were 79 patients who were categorized as having obtained a therapeutic response: IPSS decreased to 7.6 +/- 3.2 and Qmax increased to 11.3 +/- 2.9 mL/s. After randomization, IPSS was 7.1 +/- 2.9 and 6.5 +/- 2.5 for group 1; 6.5 +/- 3.2 and 6.7 +/- 2.1 for group 2; and 11.4 +/- 4.8 and 12.3 +/- 4.9 for group 3 at 3 and 6 months, respectively. Qmax was 12.7 +/- 4.8 and 11.7 +/- 5.2 mL/s for group 1; 12.2 +/- 3.9 and 11.9 +/- 3.7 mL/s for group 2; and 9.7 +/- 2.5 and 9.3 +/- 2.1 mL/s for group 3 at 3 and 6 months, respectively. Global satisfaction at 6 months was the same for groups 1 and 2. There were no differences in adverse events among the three groups. CONCLUSIONS: In men with LUTS who responded to alfuzosin, changing the dosing regimen from daily to once every other day resulted in similar efficacy and safety at 3 and 6 months. By contrast, complete cessation of alfuzosin resulted in recurrence of both symptoms and impaired urinary flow. These data provide evidence that in responding patients, intermittent alpha-blocker therapy may be a reasonable therapeutic regimen. The role of intermittent alpha-blocker therapy using other agents, as well as in a large cohort of men with LUTS, remains to be determined.

Safety and efficacy of sildenafil in postmenopausal women with sexual dysfunction.

OBJECTIVES: Sildenafil has been demonstrated to be safe and effective in the treatment of men with erectile dysfunction. The role of sildenafil in treating women with sexual dysfunction has heretofore not been reported. The purpose of this preliminary study was to ascertain the response of postmenopausal women with self-described sexual dysfunction treated with sildenafil for 3 months. METHODS: Thirty-three consecutive postmenopausal women with sexual dysfunction based on history were entered in this open-label, nonrandomized study. All patients received 50 mg of sildenafil. Efficacy was assessed at weeks 4, 8, and 12 using a newly developed 9-item, self-administered Index of Female Sexual Function (IFSF) and a global efficacy question ([GEQ] Did treatment improve your sexual function?). The IFSF quantifies the domains of desire, quality of sexual intercourse, overall satisfaction with sexual function, orgasm, lubrication, and clitoral sensation. RESULTS: Of the group, 30 women (91 %) completed the study and were available for follow-up at 3 months. Mean baseline IFSF score before therapy was 24.8+/-9.8. Mean usage of sildenafil was 3.1+/-1.4 times per week for the duration of the study. The IFSF score improved to 29.5+/-7.6, 30.3+/-8.5, and 31.4+/-10.4 at 4, 8, and 12 weeks, respectively (P = 0.25). Mean scores for questions 2 (lubrication), 8 (orgasm), and 9 (clitoral sensation) improved by 23.2%, 7.4%, and 31.3%, respectively, at 12 weeks. Seven women (21%) noted improvement on the GEQ. Overall, only 6 (18.1%) of 33 patients had a significant (more than 60% improvement in IFSF score) therapeutic response. Clitoral discomfort and "hypersensitivity" occurred in 7 women (21%), 3 of whom withdrew from the study. Other side effects, which did not result in withdrawal from the study, included headache (n = 5), dizziness (n = 4) and dyspepsia (n = 3). CONCLUSIONS: The data suggest that sildenafil is well tolerated in postmenopausal women with sexual dysfunction. Overall sexual function did not improve significantly, although there were changes in vaginal lubrication and clitoral sensitivity. The role of sildenafil in treating sexual dysfunction in various cohorts of women remains to be determined.

Interstitial thermometry in men undergoing electrovaporization of the prostate.

BACKGROUND AND OBJECTIVES: Transurethral electrovaporization of the prostate (TVP) has been utilized increasingly in the therapeutic management of benign prostatic hyperplasia (BPH). The temperature profile within both prostatic and periprostatic tissue has been presented as a parameter of safety in various preliminary studies. This prospective study measured interstitial temperature during TVP in both the prostate and the surrounding tissue of 18 men. METHODS: These 18 men undergoing TVP had three interstitial thermocouple probes placed under ultrasound guidance. Probes were positioned in the rectal wall and at the 5 and 7 o'clock position of the prostate capsule. A fourth probe was placed within 1 mm of the area of vaporization to determine "lesion" temperature. Temperature was measured at baseline and at 15-minute intervals as TVP was performed utilizing the VaporTrode at 240 to 280 W with a Valley Lab Force 40 generator. RESULTS: The maximum temperature variability was 1.9 degrees C. The temperature within 0.5 mm of the area of vaporization was >100 degrees C. These results were independent of the temperature of the irrigating solution. CONCLUSIONS: These results provide compelling evidence that high vaporization temperatures do not affect surrounding prostatic tissue. In theory, the risk of injury to either the rectum or the neurovascular bundle during TVP should be extremely low.

Cyclin D1 expression in prostate carcinoma.

The purpose of this study was to investigate the relationship between cyclin D1 expression and clinicopathological parameters in patients with prostate carcinoma. We assessed cyclin D1 expression by conventional immunohistochemistry in 85 patients who underwent radical prostatectomy for prostate carcinoma and 10 normal prostate tissue samples retrieved from autopsies. We measured nuclear immunostaining in the entire tumor area and based the results on the percentage of positive tumor cells. The preoperative prostate-specific antigen (PSA) level was 8.68±5.16 ng/mL (mean±SD). Cyclin D1 staining was positive (cyclin D1 expression in >5% of tumor cells) in 64 cases (75.4%) and negative (cyclin D1 expression in ≤5% of tumor cells) in 21 cases (including 15 cases with no immunostaining). Normal prostate tissues were negative for cyclin D1. Among patients with a high-grade Gleason score (≥7), 86% of patients demonstrated cyclin D1 immunostaining of >5% (P<0.05). In the crude analysis of cyclin D1 expression, the high-grade Gleason score group showed a mean expression of 39.6%, compared to 26.9% in the low-grade Gleason score group (P<0.05). Perineural invasion tended to be associated with cyclin D1 expression (P=0.07), whereas cyclin D1 expression was not associated with PSA levels or other parameters. Our results suggest that high cyclin D1 expression could be a potential marker for tumor aggressiveness.

PTEN loss in Gleason grade 7 prostate tumors exhibits intratumoral heterogeneity and is associated with unfavorable pathological features

Background: PTEN loss is observed in 20­30% of prostate cancers and is associated with a poor outcome, but clinical details of the impact of this biomarker are unclear for intermediate grade tumors. Methods: We investigated 43 radical prostatectomy-derived grade 7 prostate tumors from the Clinics Hospital of Ribeirão Preto. Tissue microarray (TMA) blocks were constructed and PTEN copy number status was determined for all patients through fluorescence in situ hybridization (FISH). To determine the presence of PTEN protein loss in our study cohort, we performed immunohistochemistry (IHC) in TMA sections. We then developed an automated algorithm in HALO™ to identify regions of PTEN protein loss in whole prostate scanned sections from ten patients with known PTEN deletion status by FISH. Clinical analyses were conducted to determine the associations between PTEN loss and patient outcome. All statistical analyses were conducted in R v3.4.3 with P-values below 0.05 being considered statistically significant. Results: In this study of 43 grade 7 tumors, we found PTEN deletions by FISH in 18.9% of tumors, and PTEN protein loss by IHC in 16.3% of tumors. Both techniques were highly concordant and complementary. Clinical analysis demonstrated that PTEN deletion by FISH was significantly associated with positive margin invasion (P = 0.04) and Gleason score upgrade (P = 0.001). Digital image analysis of ten representative tumors demonstrated distinct intratumoral heterogeneity for PTEN protein loss in four tumors. Conclusions: This study shows that PTEN loss in Gleason grade 7 tumors can be heterogeneous and that a systematic analysis of this biomarker using a combination of FISH, IHC, and digital imaging may identify patients with a greater risk of poor outcome (AU)