Sex-Specific Effects of Cholesteryl Ester Transfer Protein (CETP) on the Perivascular Adipose Tissue.

Cholesteryl ester transfer protein (CETP) increases the atherosclerosis risk by lowering HDL-cholesterol levels. It also exhibits tissue-specific effects independent of HDL. However, sexual dimorphism of CETP effects remains largely unexplored. Here, we hypothesized that CETP impacts the perivascular adipose tissue (PVAT) phenotype and function in a sex-specific manner. PVAT function, gene and protein expression, and morphology were examined in male and female transgenic mice expressing human or simian CETP and their non-transgenic counterparts (NTg). PVAT exerted its anticontractile effect in aortas from NTg males, NTg females, and CETP females, but not in CETP males. CETP male PVAT had reduced NO levels, decreased eNOS and phospho-eNOS levels, oxidative stress, increased NOX1 and 2, and decreased SOD2 and 3 expressions. In contrast, CETP-expressing female PVAT displayed increased NO and phospho-eNOS levels with unchanged NOX expression. NOX inhibition and the antioxidant tempol restored PVAT anticontractile function in CETP males. Ex vivo estrogen treatment also restored PVAT function in CETP males. Moreover, CETP males, but not female PVAT, show increased inflammatory markers. PVAT lipid content increased in CETP males but decreased in CETP females, while PVAT cholesterol content increased in CETP females. CETP male PVAT exhibited elevated leptin and reduced Prdm16 (brown adipocyte marker) expression. These findings highlight CETP sex-specific impact on PVAT. In males, CETP impaired PVAT anticontractile function, accompanied by oxidative stress, inflammation, and whitening. Conversely, in females, CETP expression increased NO levels, induced an anti-inflammatory phenotype, and preserved the anticontractile function. This study reveals sex-specific vascular dysfunction mediated by CETP.

Transgenerational effects of maternal exposure to nicotine on structures of pituitary-gonadal axis of rats.

Smoking can lead to several diseases and cause a reduction in fertility in men and women. Among the various components of cigarettes harmful during pregnancy, nicotine stands out. It can cause a reduction in placental blood flow, compromising the development of the baby with neurological, reproductive and endocrine consequences. Thus, we aimed to evaluate the effects of nicotine on the pituitary-gonadal axis of rats exposed during pregnancy and breastfeeding (1st generation - F1), and whether the possible damage observed would reach the 2nd generation (F2). Pregnant Wistar rats received 2 mg/kg/day of nicotine throughout the entire gestation and lactation. Part of the offspring was evaluated on the first neonatal day (F1) for macroscopic, histopathological and immunohistochemical analyses of brain and gonads. Another part of the offspring was kept until 90 days-old for mating and obtainment of progenies that had the same parameters evaluated at the end of pregnancy (F2). The occurrence of malformations was more frequent and diversified in nicotine-exposed F2. Brain alterations, including reduced size and changes in cell proliferation and death, were seen in both generations of nicotine-exposed rats. Male and female gonads of F1 exposed rats were also affected. The F2 rats showed reduced cellular proliferation and increased cell death on the pituitary and ovaries, besides increased anogenital distance in females. The number of mast cells was not enough altered to indicate an inflammatory process in brain and gonads. We conclude that prenatal exposure to nicotine causes transgenerational alterations in the structures of pituitary-gonadal axis in rats.

Does maternal exposure to nicotine affect the oocyte quality and reproductive capacity in adult offspring?

Gonadal development begins in the intrauterine phase and females from most species are born with an established oocyte reserve. Exposure to drugs during gestation can compromise the offspring health, also affecting the gametes quality. Nicotine, the main component of cigarettes, is an oxidant agent capable of altering the fertility in men and women. As female gametes are susceptible to oxidative stress, this drug can damage the oolemma and affect oocyte maturation, induce errors during chromosomal segregation and DNA fragmentation. Oocyte mitochondria are particularly susceptible to injuries, contributing to the oocyte quality loss and embryonic development disruption. Thus, considering the high number of women who smoke during pregnancy, while significant events are occurring in the embryo for future fertility of offspring, we seek to verify the quality of the oocytes from adult rats exposed to nicotine during intrauterine phase and breastfeeding. Pregnant Wistar rats received nicotine by osmotic mini-pumps and the female progenies were evaluated in adulthood for oocyte quality (viability, lipid peroxidation, generation of reactive oxygen species and mitochondrial integrity) and reproductive capacity. Embryos (3dpc) and fetuses (20dpc) generated by these rats were also evaluated. The results showed that the dose of 2 mg/kg/day of nicotine through placenta and breast milk does not affect the number of oocytes and the fertility capacity of adult rats. However, it causes some morphological alterations in oocytes, mitochondrial changes, embryonic fragmentation and disruption of fetal development. The malformations in fetuses generated from these gametes can also indicate the occurrence of epigenetic modifications.

Interleukin-6 and adhesion molecules VCAM-1 and ICAM-1 as biomarkers of post-acute myocardial infarction heart failure.

Acute coronary syndromes are associated with a high prevalence of complications including heart failure (HF). The aim of this study was to investigate the association of novel biomarkers with the occurrence of post-acute myocardial infarction (AMI) HF. A prospective study was conducted with patients admitted to the emergency department with ST-segment elevation myocardial infarction (STEMI). Blood and urine samples were collected for analysis of traditional and novel biomarkers, including interleukin-6, vascular cell adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1). We compared the levels of these biomarkers between patients with and without post-STEMI HF. A total of 48 patients were assessed, with a prevalence of males. Fifteen patients (31.2%) had post-STEMI HF. Patients with HF had higher mean values of IL-6, VCAM-1, and ICAM-1 compared to those who did not develop HF (57.06 vs 14.03 pg/mL, P=0.001; 1719.58 vs 1304.34 ng/mL, P=0.001; and 1594.20 vs 1158.74 ng/mL, P<0.001, respectively). The three biomarkers were shown to be good predictors of post-STEMI HF (IL-6: AUC 0.786, P=0.002; VCAM-1: AUC 0.797, P=0.001; and ICAM-1: AUC 0.825, P<0.0001), with the respective cutoff points being calculated based on the best sensitivity and specificity indexes (IL-6: 8.67 pg/mL; VCAM-1: 1501.42 ng/mL; and ICAM-1: 1262.38 ng/mL). Of the three biomarkers, only VCAM-1 and ICAM-1 had a direct linear association between them (r=0.470, P<0.0001). IL-6, VCAM-1, and ICAM-1 were associated with the development of new post-AMI HF symptoms, but only VCAM-1 and ICAM-1 correlated with each other, possibly because they have the same pathophysiological mechanism of action.

Interleukin-6 and adhesion molecules VCAM-1 and ICAM-1 as biomarkers of post-acute myocardial infarction heart failure

Acute coronary syndromes are associated with a high prevalence of complications including heart failure (HF). The aim of this study was to investigate the association of novel biomarkers with the occurrence of post-acute myocardial infarction (AMI) HF. A prospective study was conducted with patients admitted to the emergency department with ST-segment elevation myocardial infarction (STEMI). Blood and urine samples were collected for analysis of traditional and novel biomarkers, including interleukin-6, vascular cell adhesion molecule 1 (VCAM-1), and intercellular adhesion molecule-1 (ICAM-1). We compared the levels of these biomarkers between patients with and without post-STEMI HF. A total of 48 patients were assessed, with a prevalence of males. Fifteen patients (31.2%) had post-STEMI HF. Patients with HF had higher mean values of IL-6, VCAM-1, and ICAM-1 compared to those who did not develop HF (57.06 vs 14.03 pg/mL, P=0.001; 1719.58 vs 1304.34 ng/mL, P=0.001; and 1594.20 vs 1158.74 ng/mL, P<0.001, respectively). The three biomarkers were shown to be good predictors of post-STEMI HF (IL-6: AUC 0.786, P=0.002; VCAM-1: AUC 0.797, P=0.001; and ICAM-1: AUC 0.825, P<0.0001), with the respective cutoff points being calculated based on the best sensitivity and specificity indexes (IL-6: 8.67 pg/mL; VCAM-1: 1501.42 ng/mL; and ICAM-1: 1262.38 ng/mL). Of the three biomarkers, only VCAM-1 and ICAM-1 had a direct linear association between them (r=0.470, P<0.0001). IL-6, VCAM-1, and ICAM-1 were associated with the development of new post-AMI HF symptoms, but only VCAM-1 and ICAM-1 correlated with each other, possibly because they have the same pathophysiological mechanism of action.

Aorto-left ventricular tunnel: a rare cause of heart failure in the newborn.

Aorto-left ventricular tunnel is a rare congenital cardiac anomaly, consisting of a short abnormal pathway, usually from a sinus of Valsalva into the left ventricular cavity. It is usually diagnosed with echocardiography. We report a case of a newborn presenting with heart murmur and rapid progression to heart failure and left ventricular enlargement due to an aorto-left ventricular tunnel. Despite successful closure of the tunnel, the patient required a Ross procedure due to progressive aortic disease.

The beneficial effects of aerobic and concurrent training on metabolic profile and body composition after detraining: a 1-year follow-up in postmenopausal women.

BACKGROUND/OBJECTIVES: Aerobic and concurrent training (CT, aerobic and strength training) improves body composition and metabolic profile; however, it is not known whether these positive outcomes acquired after aerobic or CT are maintained long term (⩾6 months) after program interruption in postmenopausal women. This study investigated the changes in total and appendicular body composition, bone mineral density and metabolic profile following 16 weeks of aerobic or CT, and through 6 months and 1 year of detraining in postmenopausal women. SUBJECTS/METHODS: In total, 60 postmenopausal women were divided into the following groups: aerobic (AT), aerobic plus strength training (CT) and control group (CG), and 31 participants were assessed for the 1 year follow-up. Body composition and bone mineral density were evaluated by dual-energy X-ray absorptiometry (DXA), and total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triacylglycerol, glucose, insulin, leptin, adiponectin and plasminogen activator inhibitor-1 (PAI-1) were assessed. RESULTS: There were main effects of time for arm fat mass, arm lean mass and trunk lean mass (P<0.05). There was a statistical difference between AT and CG for leg fat mass and percentage of fat (P<0.05). After 6 months of detraining, leg lean mass decreased in relation to post-intervention, and there was a statistically significant interaction for total and appendicular lean mass (P<0.05). There were differences between CT and CG in glucose and between AT and CG in glucose and triacylglycerol (P<0.05). CONCLUSIONS: A duration of 16 weeks of aerobic or CT improved total and appendicular body composition and metabolic profile but after 6 months of detraining, leg lean mass returned to the values obtained pre-training in CT.

Reparo de lesão penetrante em parede torácica de cão, ocasionada por javali (Sus scrofa scrofa), com flape unipediculado de músculo reto abdominal / Unipediculated flap of muscle rectos abdominales to repair a penetrating injury caused by javali (Sus scrofa scrofa) on the thoracic wall of a dog

Lesões penetrantes no tórax causadas por interação animal são frequentes em cães e caracterizam-se por dano extenso e desvitalização dos tecidos moles adjacentes. Quando a musculatura local é insuficiente para a reconstrução, retalhos musculares podem ser mobilizados para reparar o defeito torácico. O presente relato tem como objetivo demonstrar uma alternativa para o reparo de defeito da parede torácica em um cão, ocasionada por interação com javali (Sus scrofa scrofa), utilizando flape unipediculado de músculo reto abdominal. Com base na literatura consultada, essa técnica reconstrutiva ainda não foi descrita. O flape de músculo reto abdominal mostrou-se uma alternativa viável no reparo de lesão extensa e infectada na parede torácica em cães.(AU)

Penetrating thoracic wounds caused by animal bites are common in dogs and are characterized by extensive trauma and adjacent soft tissue devitalization. When the local musculature is insufficient for the reconstruction, muscle flaps can be taken to repair the thoracic defect. The aim of the present report is to demonstrate an alternative to the thoracic wall defect repair in a dog which was attacked by javali (Sus scrofa scrofa), using unipediculated flap of the rectus abdominis muscle. Based on the literature, this reconstructive technique has not yet been described. The rectus abdominis muscle flap proves to be a useful alternative for the repair of extensive and infected thoracic wall lesion in dogs.(AU)

Dipeptidylpeptidase-IV activity and expression reveal decreased damage to the intrahepatic biliary tree in fatty livers submitted to subnormothermic machine-perfusion respect to conventional cold storage.

Graft steatosis is a risk factor for poor initial function after liver transplantation. Biliary complications are frequent even after normal liver transplantation. A subnormothermic machine perfusion (MP20) preservation procedure was developed by our group with high potential for reducing injury to hepatocytes and sinusoidal cells of lean and fatty livers respect to conventional cold storage (CS). We report the response of the biliary tree to CS or MP20, in lean and obese Zucker rat liver. Dipeptidylpeptidase-IV (DPP-IV), crucial for the inactivation of incretins and neuropeptides, was used as a marker. Liver morphology and canalicular network of lean livers were similar after CS/reperfusion or MP20/reperfusion. CS preservation of fatty livers induced serious damage to the parenchyma and to the canalicular activity/expression of DPP-IV whereas with MP20 the morphology and canalicular network were similar to those of untreated lean liver. CS and MP20 had similar effects on DPP-IV activity and expression in the upper segments of the intrahepatic biliary tree of fatty livers. DPP-IV expression was significantly increased after MP20 respect to CS or to the controls, both for lean and obese animals. Our data support the superiority of MP20 over CS for preserving fatty livers. Dipeptidylpeptidase-IV activity and expression reveal decreased damage to the intrahepatic biliary tree in fatty livers submitted to subnormothermic machine-perfusion respect to conventional cold storage.

Diallel analysis of corn for special use as corn grits: determining the main genetic effects for corn gritting ability.

Corn grits are used for various purposes such as flakes, snacks, livestock feed, hominy, extruded products, beer, etc. The grit size proportion varies according to the hybrid, and thus, once the use of the grits is linked to the particle size, determining the genetic effects is essential to develop hybrids for any specific use. For this purpose a complete diallel series of crosses, involving eight parents, was performed near Maringá, PR, Brazil. The objective of this study was to evaluate the general (GCA) and specific (SCA) combining abilities of 28 progeny for selection of hybrids for breeding programs and extraction of inbred lines for hybrid development. The response variables, such as plant height, ear insertion height, crop stand, grain yield, and grits, small grits and bran production, were gauged and appraised for each of the 28 progeny. The trait effects and GCA were significant for all response variables, while for SCA, only grain yield and crop stand showed significance (P < 0.05), according to Griffing (1955) analysis. A significant weak negative partial correlation was found between grain yield and grits conversion. In relation to the hybrid selection for breeding programs, the parent IAC Nelore was highly recommended for recurrent selection and the hybrids IPR 119 x HT 392 and IAC Nelore x HD 332 for the extraction of pure lines for hybrid development.

Diferentes níveis de incorporação de farinha de resíduos de jundiás (Rhamdiaquelen) cultivados na dieta / Different levels of jundiá waste meal in the diet

O estudo foi desenvolvido no Laboratório de Piscicultura do Departamento de Zootecnia da Universidade Federal de Santa Maria, com duração de oito semanas, de abril a junho de 2011. O objetivo foi definir o melhor nível de incorporação do resíduo de peixes na alimentação de juvenis de jundiá, em sistema de recirculação d'água, utilizando-se 450 alevinos de jundiás, com peso médio inicial de 7,2±0,27g. Foram testadas cinco diferentes dietas, em três repetições, com níveis diferentes (0; 3,75; 7,5; 15; e 30 por cento) de incorporação de farinha de carcaça de jundiás com vísceras (FCJCV). As dietas continham, aproximadamente, 37 por cento de PB e 3200kcal de EM/kg de dieta. Foram coletados dados de ganho em peso diário, sobrevivência, fator de condição, taxa de crescimento específico, conversão alimentar aparente, rendimento de carcaça, taxa de eficiência proteica e deposição de proteína e lipídio corporal. O maior ganho de peso foi para o nível 30 por cento de inclusão de FCJCV, com peso final de 45,47g e ganho em peso diário de 0,68g/dia. A dieta com nível de 30% de inclusão de FCJCV apresentou melhores valores para taxa de crescimento específico (3,34 por cento/dia), conversão alimentar aparente (1,05: 1), biomassa final 1.105,45g, deposição proteica corporal (64,9mg/dia), taxa de eficiência proteica (1,9) e coeficiente de retenção proteica (1,96 por cento). Concluiu-se que a utilização de farinha de resíduos de processamento de jundiá no nível de 30% de inclusão na dieta foi a que apresentou maior eficiência no desenvolvimento de juvenis de jundiá...

The study was carried out at the Fish Farming Laboratory of the Animal Science Department, Federal University of Santa Maria, during 8 weeks, from April to June 2011. The objective of this study was to define the best level of incorporation of waste in food fish of juvenile Jundiá in water recirculation system using 450 fingerlings of jundiá with initial average weight of 7.2±0.27g. We tested 5 different diets in three repetitions with different levels of incorporation of carcasse jundiá with viscera meal (FCJCV) (0, 3.75, 7.5, 15, and 30 percent). The diets contained 37 percent CP and 3200kcal/kg of ME. Data were collected from daily weight gain, survival, condition factor, specific growth rate, feed conversion, carcass yield, protein efficiency ratio and deposition of body protein and lipid. The weight gain was greater with the inclusion of 30 percent FCJCV with final weight of 45.47g and daily weight gain of 0.68g/day. The diet with inclusion of 30 percent FCJCV showed better values for specific growth rate (3.34 percent/day), feed conversion ratio (1.05: 1), final biomass (1105.45g), body protein deposition (64.9mg/day), protein efficiency ratio (1.9) and protein retention coefficient (1.96 percent). In conclusion, the use of processing waste meal of jundiá in the level of 30 percent in the diet showed the highest efficiency in the development of juvenile jundiá...

Genetic gains in the UENF-14 popcorn population with recurrent selection.

The popcorn breeding program of Universidade Estadual do Norte Fluminense Darcy Ribeiro aims to provide farmers a cultivar with desirable agronomic traits, particularly with respect to grain yield (GY) and popping expansion (PE). We evaluated full-sib families from the seventh cycle of recurrent selection and estimated the genetic progress with respect to GY and PE. Eight traits were evaluated in 200 full-sib families that were randomized into blocks with two replicates per set in two contrasting environments, Campos dos Goytacazes and Itaocara, located in north and northwest Rio de Janeiro State, respectively. There were significant differences between sets in families with respect to all traits evaluated, which indicates genetic variability that may be explored in future cycles. Using random economic weights in the selection of superior progenies, the Mulamba and Mock index showed gains for PE and GY of 5.11 and 7.78%, respectively. Significant PE and GY increases were found when comparing the evolution of mean values of these two parameters that were assessed at cycles C0-C6 and predicted for C7. Thus, an advanced-cycle popcorn cultivar with genotypic superiority for the main traits of economic interest can be made available to farmers in Rio de Janeiro State.

Preliminary validation of an instrument to assess social support and tuberculosis stigma in patients' families.

SETTING: Ribeirão Preto, São Paulo, Brazil. OBJECTIVE: To develop and validate a preliminary instrument for assessing social support and tuberculosis (TB) stigma in families of TB patients. DESIGN: A literature review on social support and TB stigma was used to generate the theoretical domains for the instrument. A focus group was then conducted with TB patients and their families to revise the domains. Reviewers were invited to judge the appropriateness of the items in the instrument. A cross-sectional survey was carried out among 110 family members to assess the factorial structure using principal component analysis and confirmatory factor analysis to assess construct validity. Reliability was assessed in terms of internal consistency using Cronbach's alpha. RESULTS: After semantic validation and a pilot study, 23 items were selected for the scale. Examination of the factorial structure of the 16 items that were factorable using principal component analysis led to the extraction of two factors. The 16-item instrument was assessed for construct validity with confirmatory factor analysis, which confirmed a model with four items for each dimension. CONCLUSION: The study analysed the psychometric properties of an instrument that is still in its preliminary stages. Other studies on a similar scale in the Brazilian setting are required.

Contexte : Ribeirão Preto, Brésil.Objectif : Elaborer et valider un instrument préliminaire afin d'évaluer le soutien social et la stigmatisation liée à la tuberculose (TB) dans les familles des patients.Schéma : Une revue de littérature relative au soutien social et à la stigmatisation liée à la TB a permis de générer les domaines théoriques de l'instrument. Un groupe focal a ensuite été réalisé avec des patients tuberculeux et leurs familles afin de revoir ces domaines. Les réviseurs ont été invités à juger le caractère approprié des éléments de l'instrument. Une enquête traversable auprès de 110 membres des familles a été effectuée afin d'évaluer la structures factorielles grâce à l'analyse du composant principal, avec une analyse du facteur de confirmation qui a permis de vérifier la validité de la construction. La fiabilité a été évaluée par la cohérence interne grâce au coefficient α de Cronbach.Résultats : Vingt-trois éléments composaient l'échelle après la validation sémantique et l'étude pilote. L'exploration de la structure factorielle des 16 items qui étaient favorables grâce à l'analyse du composant principal a extrait deux facteurs. L'instrument à 16 items a été évalué en termes de validité de la construction avec une analyse du facteur de confirmation qui a confirmé un modèle à quatre items pour chaque dimension.Conclusion : L'étude a démontré les propriétés psychométriques d'un instrument encore au stade préliminaire. Cette échelle doit être testée à travers d'autres études dans le contexte brésilien.

Marco de referencia: Ribeirão Preto, en Brasil.Objetivo: Validar una herramienta preliminar de evaluación del apoyo social y los estigmas relacionados con la tuberculosis (TB) en las familias de los pacientes.Método: Mediante una recensión bibliográfica sobre el apoyo social y los estigmas relacionados con la TB se definieron los dominios teóricos de la herramienta. Luego, en un grupo de opinión conformado por pacientes con diagnóstico de TB y sus familiares se verificaron los dominios. Se invitaron revisores con el objeto de juzgar la idoneidad de los elementos de la escala. Se llevó a cabo una encuesta transversal a 110 miembros de las familias de pacientes con el propósito de evaluar la estructura factorial, mediante un análisis de componentes principales y se aplicó un análisis factorial confirmatorio a fin de apreciar la validez conceptual de la escala. Mediante un análisis de la consistencia interna se examinó la fiabilidad de la escala con medición del coeficiente α de Cronbach.Resultados: Tras la validación semántica y el estudio preliminar, la escala estaba compuesta por 23 elementos. Se exploró la estructura factorial de los 16 elementos factorizables mediante el análisis de componentes principales y se extrajeron dos factores. La validez conceptual del instrumento de 16 elementos se verificó mediante un análisis factorial confirmatorio, el cual ratificó un modelo con cuatro elementos en cada dimensión.Conclusión: El estudio puso en evidencia las propiedades psicométricas de un instrumento que se encuentra en fases preliminares de desarrollo. Es necesario poner a prueba esta escala en otros estudios en el contexto del Brasil.

Subnormothermic machine perfusion for non-heart-beating donor liver grafts preservation in a Swine model: a new strategy to increase the donor pool?

We previously reported that subnormothermic machine perfusion (sMP; 20°C) is able to improve the preservation of livers obtained from non-heart-beating donors (NHBDs) in rats. We have compared sMP and standard cold storage (CS) to preserve pig livers after 60 minutes of cardiac arrest. In the sMP group livers were perfused for 6 hours with Celsior at 20°C. In the CS group they were stored in Celsior at 4°C for 6 hours as usual. To simulate liver transplantation, both sMP- and CS-preserved livers were reperfused using a mechanical continuous perfusion system with autologus blood for 2 hours at 37°C. At 120 min after reperfusion aspartate aminotransferase levels in sMP versus CS were 499 ± 198 versus 7648 ± 2806 U/L (P < .01); lactate dehydrogenase 1685 ± 418 versus 12998 ± 3039 U/L (P < .01); and lactic acid 4.78 ± 3.02 versus 10.46 ± 1.79 mmol/L (P < .01) respectively. The sMP group showed better histopathologic results with significantly less hepatic damage. This study confirmed that sMP was able to resuscitate liver grafts from large NHBD animals.

Dipeptidylpeptidase--IV, a key enzyme for the degradation of incretins and neuropeptides: activity and expression in the liver of lean and obese rats.

Given the scarcity of donors, moderately fatty livers (FLs) are currently being considered as possible grafts for orthotopic liver transplantation (OLT), notwithstanding their poor tolerance to conventional cold preservation. The behaviour of parenchymal and sinusoidal liver cells during transplantation is being studied worldwide. Much less attention has been paid to the biliary tree, although this is considered the Achille's heel even of normal liver transplantation. To evaluate the response of the biliary compartment of FLs to the various phases of OLT reliable markers are necessary. Previously we demonstrated that Alkaline Phosphatase was scarcely active in bile canaliculi of FLs and thus ruled it out as a marker. As an alternative, dipeptidylpeptidase-IV (DPP-IV), was investigated. This ecto-peptidase plays an important role in glucose metabolism, rapidly inactivating insulin secreting hormones (incretins) that are important regulators of glucose metabolism. DPP-IV inhibitors are indeed used to treat Type II diabetes. Neuropeptides regulating bile transport and composition are further important substrates of DPP-IV in the enterohepatic axis. DPP-IV activity was investigated with an azo-coupling method in the liver of fatty Zucker rats (fa/fa), using as controls lean Zucker (fa/+) and normal Wistar rats. Protein expression was studied by immunofluorescence with the monoclonal antibody (clone 5E8). In Wistar rat liver, DPP-IV activity and expression were high in the whole biliary tree, and moderate in sinusoid endothelial cells, in agreement with the literature. Main substrates of DPP-IV in hepatocytes and cholangiocytes could be incretins GLP-1 and GIP, and neuropeptides such as vasoactive intestinal peptide (VIP) and substance P, suggesting that these substances are inactivated or modified through the biliary route. In lean Zucker rat liver the enzyme reaction and protein expression patterns were similar to those of Wistar rat. In obese rat liver the patterns of DPP-IV activity and expression in hepatocytes reflected the morphological alterations induced by steatosis as lipid-rich hepatocytes had scarce activity, located either in deformed bile canaliculi or in the sinusoidal and lateral domains of the plasma membrane. These findings suggest that bile canaliculi in steatotic cells have an impaired capacity to inactivate incretins and neuropeptides. Incretin and/or neuropeptide deregulation is indeed thought to play important roles in obesity and insulin-resistance. No alteration in enzyme activity and expression was found in the upper segments of the biliary tree of obese respect to lean Zucker and Wistar rats. In conclusion, this research demonstrates that DPP-IV is a promising in situ marker of biliary functionality not only of normal but also of fatty rats. The approach, initially devised to investigate the behaviour of the liver during the various phases of transplantation, appears to have a much higher potentiality as it could be further exploited to investigate any pathological or stressful conditions involving the biliary tract (i.e., metabolic syndrome and cholestasis) and the response of the biliary tract to therapy and/or to surgery.

Altered alkaline phosphatase activity in obese Zucker rats liver respect to lean Zucker and Wistar rats discussed in terms of all putative roles ascribed to the enzyme.

Biliary complications often lead to acute and chronic liver injury after orthotopic liver transplantation (OLT). Bile composition and secretion depend on the integrated action of all the components of the biliary tree, starting from hepatocytes. Fatty livers are often discarded as grafts for OLT, since they are extremely vulnerable to conventional cold storage (CS). However, the insufficiency of donors has stimulated research to improve the usage of such marginal organs as well as grafts. Our group has recently developed a machine perfusion system at subnormothermic temperature (20°C; MP20) that allows a marked improvement in preservation of fatty and even of normal rat livers as compared with CS. We sought to evaluate the response of the biliary tree of fatty liver to MP20, and a suitable marker was essential to this purpose. Alkaline phosphatase (AlkP, EC 3.1.3.1), frequently used as marker of membrane transport in hepatocytes and bile ducts, was our first choice. Since no histochemical data were available on AlkP distribution and activity in fatty liver, we have first settled to investigate AlkP activity in the steatotic liver of fatty Zucker rats (fa/fa), using as controls lean Zucker (fa/+) and normal Wistar rats. The AlkP reaction in Wistar rats was in accordance with the existing data and, in particular, was present in bile canaliculi of hepatocytes in the periportal region and midzone, in the canals of Hering and in small bile ducts but not in large bile ducts. In lean ZR liver the AlkP reaction in Hering canals and small bile ducts was similar to Wistar rat liver but hepatocytes had lower canalicular activity and besides presented moderate basolateral reaction. The difference between lean Zucker and Wistar rats, both phenotypically normal animals, could be related to the fact that lean Zucker rats are genotypically heterozygous for a recessive mutated allele. In fatty liver, the activity in ductules and small bile ducts was unchanged, but most hepatocytes were devoid of AlkP activity with the exception of clusters of macrosteatotic hepatocytes in the mid-zone, where the reaction was intense in basolateral domains and in distorted canaliculi, a typical pattern of cholestasis. The interpretation of these data was hindered by the fact that the physiological role of AlkP is still under debate. In the present study, the various functions proposed for the role of the enzyme in bile canaliculi and in cholangiocytes are reviewed. Independently of the AlkP role, our data suggest that AlkP does not seem to be a reliable marker to study the initial step of bile production during OLT of fatty livers, but may still be used to investigate the behaviour of bile ductules and small bile ducts.

Decreased apoptosis in fatty livers submitted to subnormothermic machine-perfusion respect to cold storage.

Machine perfusion at subnormothermic temperature (20°C), MP20, was developed by Vairetti et al. and showed to afford a better preservation of fatty livers respect to traditional cold storage (CS) in terms of enzyme release into the perfusate and bile, glycogen stores, energy charge and oxidative stress. Here we investigated whether it also caused decreased cell death by apoptosis. Fatty and lean Zucker rats were submitted to MP20 or CS for 6 h and reperfused normothermically for 2 h. Apoptotic cells were revealed by immunohistochemistry of activated caspase-3 and M30 (new epitope on CK18 degraded by caspase-3) and by the TUNEL assay. Portal pressure was also determined. A statistically significant reduction of hepatocyte apoptosis, but especially of sinusoidal cells was determined for fatty livers submitted to MP20 respect to CS. Portal pressure was significantly lower after MP20 respect to CS. The reduction of sinusoidal cell death by apoptosis without need for anti-apoptotic therapies appears particularly positive since apoptotic sinusoidal cells hinder microcirculation in the sinusoids and are thrombogenic. These results further confirm the potential of MP20 for preserving fatty livers that would be otherwise discarded as grafts, and thus for increasing the donor pool for liver transplantation.

Dispensação farmacêutica: uma análise dos conceitos legais em relação à prática profissional / Pharmaceutical dispensing: an analysis of legal concepts in relation to professional practice

O âmbito profissional do farmacêutico sofreu mudanças ao longo dos anos, provocando a descaracterização das farmácias comunitárias em virtude da acentuada concepção mercantil que vem apresentando e do distanciamento do farmacêutico na relação direta com a comunidade. Dessa forma, a dispensação de medicamentos se tornou um ato mecânico, desprovido dos cuidados necessários para a assistência à saúde, havendo, contudo, uma tendência de incorporação de novas práticas. Nesse sentido, ao estudar os conceitos de dispensação explicitados nas legislações sanitárias e profissionais brasileiras, esse artigo tem como objetivo analisá-los em relação ao contexto atual. Trata-se de um estudo teórico, realizado com a técnica da análise de conteúdo. Os resultados mostram que apesar das necessidades e novas demandas apontadas no cenário farmacêutico, ocorreram poucas mudanças nos conceitos de dispensação nas legislações estudadas, apontando a necessidade de se reafirmar a dispensação como atividade inserida no contexto da assistência à saúde.

The pharmacist?s professional scope of practice has changed over the years, causing a loss of the special role of community pharmacies, due to the strongly market-oriented conception that is emerging and the distancing of the pharmacist from a direct relationship with the community. Thus, the dispensing of drugs has become a mechanical act, devoid of the care necessary for health provision, there being, however, a tendency to incorporate new practices. Therefore, by studying the concepts of dispensing spelled out in the health laws and by health professionals in Brazil, the aim of this article is to analyze them in relation to the current context. This is a theoretical study, carried out by the technique of content analysis. The results show that despite the new demands and requirements noted in the pharmaceutical scenario, there have been few changes in the concepts of dispensing in the laws examined, indicating a need to reaffirm dispensing as an activity that has a place within the context of health care.

Multiple trichoepitheliomas--a novel mutation in the CYLD gene.

BACKGROUND: Trichoepitheliomas are benign neoplasms with follicular differentiation. They may present as a solitary lesion or as multiple lesions. Multiple trichoepitheliomas are inherited in an autosomal dominant pattern within families, with both variable penetrance and expressivity. Recent investigations support that mutations in CYLD, the gene affected in familial cylindromatosis as well as in Brooke-Spiegler syndrome, are also responsible for multiple trichoepitheliomas. OBJECTIVE: The authors report the case of a 9-year-old African girl with multiple facial trichoepitheliomas in whom a mutation in the CYLD gene was hypothesised. MATERIALS AND METHODS: After genomic DNA extraction from the peripheral blood, a molecular analysis of the CYLD gene was performed by PCR, DHPLC and automated sequencing. RESULTS: A novel heterozygous mutation in exon 18 of the CYLD gene (c.2449delT) was identified, with a deletion of one nucleotide resulting in a premature translational termination codon at amino acid position 831 on the affected allele (p.Cys817Valfs X15). CONCLUSIONS: The predominating tumours define the classification of these three entities. Nevertheless, studies suggest that they can simply represent phenotypic variations of the same disease spectrum, sharing common genetic mutations.

Kluyveromyces lactis but not Pichia fermentans used as adjunct culture modifies the olfactory profiles of Cantalet cheese.

Yeasts are commonly detected in cheese. Two yeast species, Kluyveromyces lactis and Pichia fermentans, were isolated at high populations from raw-milk Cantalet cheese, a French Protected Denomination of Origin hard cheese. To investigate the interest of these 2 species as adjunct cultures to promote flavor development of Cantalet cheese, they were added at 10(5) cfu/mL to microfiltered milk. The global microbiological, biochemical, and flavor changes induced by the presence of the yeasts in cheese were determined. Adjunct yeasts were present at 10(6) cfu/g in curd, declined to 10(4) to 10(5) cfu/g in cheese, and did not influence gross composition, content of free amino acids, or content of free fatty acids. By using 8-way gas chromatography-olfactometry in parallel with gas chromatography-mass spectrometry, 30 odorous compounds of Cantalet cheese were identified. The olfactory profiles of K. lactis cheeses contained significantly greater levels of 8 odorous compounds (ethanol, ethyl hexanoate, 4 aldehydes, and 2 branched-chain acids) compared with the control and P. fermentans cheeses. Sensory analysis of cheeses flavor discriminated K. lactis cheeses on only 2 attributes (acetaldehyde and alcohol odors). This study shows that yeast contribution is species-specific and that K. lactis, at a population of 10(6) viable cells/g, can influence Cantalet cheese flavor.