Combined precut in difficult biliary cannulation.

AIM: precut sphincterotomy refers to a variety of endoscopic techniques that are used in order to access the bile duct when conventional methods of cannulation have failed. There are not significant data (such as efficacy, safety) about the use of different techniques of precutting at the same session. We have described our experience with combined precut sphincterotomy (CPS) and we have compared our results to the use of an isolated precut. PATIENTS AND METHODS: we have performed 247 precuts of a total of 2.390 ERCPs. Patients were distributed according to the type of precut practiced: Needle-knife, transpancreatic and combined precut sphincterotomies. "Combined precut" consisted in performing first a transpancreatic sphincterotomy and, if the access was not achieved, then performing a needle-knife sphincterotomy in the same session. The data about safety and efficacy were prospectively collected. The complications were defined according to the consensus criteria. RESULTS: we performed precutting techniques in 247 patients. Needle-knife, transpancreatic, and combined precuts were performed in 125 (6.9%), 74 (4.1%) and 48 (2.6%) patients, respectively. Bile duct cannulation was successful in 48 patients (100%) in the group of combined precut, 121 patients (96.8%) in the transpancreatic group, and 67 patients (90.5%) in the needleknife group (p = 0.03). There were not differences in complications rates between the three groups. There was no pancreatitis in the combined precut group. The complications were successfully managed with conservative treatment. CONCLUSIONS: combined precut sphincterotomy seems to be a safe and successful technique in those cases of difficult bile duct cannulation.

Equity and Prevention of Cardiovascular Diseases in Latin America and the Caribbean.

Non-communicable diseases, particularly cardiovascular diseases, are the leading cause of decreased life expectancy and death in Latin America and the Caribbean. Although a lifestyle, which includes no tobacco use, good nutrition, and regular physical activity is touted as key to health, the environmental, racial, social and economic conditions, which underpin lifestyle are often ignored or considered only secondarily. Placing the main responsibility on a patient to change their lifestyle or to simply comply with pharmacological treatment ignores the specific conditions in which the individual lives. Furthermore, there are major disparities in access to both healthy living conditions as well as access to medical care. There is sufficient evidence to support advocating for policies that support healthy living, particularly healthy food choices. Progress is being made to improve the food environment with enactment of front of package nutritional labels. However, policies were enacted only after intense regional research and advocacy supporting their implementation. Government officials must rise above the pressures of commercial interests and support health-promoting policies or be exposed as self-interest groups themselves. Strong advocacy is required to persuade officials that all policies should take health into consideration both to improve lives and economies.

Suckling-induced oxytocin increase in the spinal cord of the rat.

Oxytocin (OT) is essential for parturition and milk ejection, and OT-containing fibers are present in several regions of the brain and in the spinal cord. During lactation, activation of spinal cord neurons by suckling stimulation involves deep laminae III-X including sympathetic preganglionic neurons of the intermedio-medial cell column. In the present study, experiments were designed to determine if the suckling provided by the litter increased OT levels in the spinal cord of dams, as determined by competitive immunoassay. In addition, we investigated if OT fibers reach neurons of the spinal cord that are known to respond to suckling. The OT content was higher in the hypothalamus than in the spinal cord in animals from all experimental groups. After 6 h of pup separation, OT levels decreased and suckling for 5 min induced a significant increase of OT levels in the spinal cord. Double immunostaining for Fos and OT showed OT-positive fibers adjacent to neurons that had Fos-positive nuclei, located mostly in laminae III, IV, and X. The present data support the notion that OT is released within the spinal cord in response to suckling, suggesting a role for this peptide in modulating the afferent and/or efferent responses generated by suckling.

Upregulation of GH, but not IGF1, in the hippocampus of the lactating dam after kainic acid injury.

Lactation embodies a natural model of morphological, neurochemical, and functional brain plasticity. In this reproductive stage, the hippocampus of the female is less sensitive to excitotoxins in contrast to nulliparity. Growth hormone (GH) and insulin-like growth factor 1 (IGF1) are known to be neuroprotective in several experimental models of brain lesion. Here, activation of the GH-IGF1 pituitary-brain axis following kainic acid (7.5 mg/kg i.p. KA) lesion was studied in lactating and nulliparous rats. Serum concentrations of GH and IGF1 were uncoupled in lactation. Compared to virgin rats, the basal concentration of GH increased up to 40% but IGF1 decreased 58% in dams, and only GH increased further after KA treatment. In the hippocampus, basal expression of GH mRNA was higher (2.8-fold) in lactating rats than in virgin rats. GH mRNA expression in lactating rats increased further after KA administration in the hippocampus and in the hypothalamus, in parallel to GH protein concentration in the hippocampus of KA-treated lactating rats (43% vs lactating control), as detected by Western blot and immunofluorescence. Except for the significantly lower mRNA concentration in the liver of lactating rats, IGF1 expression was not altered by the reproductive condition or by KA treatment in the hippocampus and hypothalamus. Present results indicate upregulation of GH expression in the hippocampus after an excitotoxic lesion, suggesting paracrine/autocrine actions of GH as a factor underlying neuroprotection in the brain of the lactating dam. Since no induction of IGF1 was detected, present data suggest a direct action of GH.

Reproductive Stage and Modulation of Stress-Induced Tau Phosphorylation in Female Rats.

Chronic stress is implicated as a risk factor for Alzheimer's disease (AD) and other neurodegenerative disorders. Although the specific mechanisms linking stress exposure and AD vulnerability have yet to be fully determined, our laboratory and others have shown that acute and repeated restraint stress in rodents leads to an increase in hippocampal tau phosphorylation (tau-P) and tau insolubility, a critical component of tau pathology in AD. Although tau phosphorylation induced by acute psychological stress is dependent on intact signaling through the type 1 corticotropin-releasing factor receptor, how sex steroids or other modulators contribute to this effect is unknown. A naturally occurring attenuation of the stress response is observed in female rats at the end of pregnancy and throughout lactation. To test the hypothesis that decreased sensitivity to stress during lactation modulates stress-induced tau-P, cohorts of virgin, lactating and weaned female rats were subjected to 30 min of restraint stress or no stress (control) and were killed 20 min or 24 h after the episode. Exposure to restraint stress induced a significant decrease in tau-P in the hippocampus of lactating rats killed 20 min after stress compared to lactating controls and virgins subjected to stress treatment. Lactating rats killed 24 hr after restraint stress exposure showed significant elevation in tau-P compared to lactating cohorts killed 20 min after stress. Levels of tau-P in these latter cohorts did not differ signficantly from control animals. Furthermore, glycogen synthase kinase (GSK)3-α levels were significantly decreased in stressed lactating animals at both timepoints. This suggests a steep, yet transient stress-induced dephosphorylation of tau, influenced by GSK3, in the hippocampus of lactating rats.

A Simple Chemical Method for Rendering Wild-Type Yeast Permeable to Brefeldin A That Does Not Require the Presence of an erg6 Mutation.

The present work aims to develop a growth medium to render a wild-type strain of Saccharomyces cerevisiae permeable to the antifungal drug Brefeldin A. In the current study, a synthetic medium containing 0.1% L-proline and supplemented with $3.0\times 10;{-3}$ % SDS is employed. When Brefeldin A is added to this medium, a wild-type strain shows increased growth sensitivity and a diminished transport of the amino acid L-leucine. Since Brefeldin A exerts its effect on the endoplasmic reticulum and the Golgi apparatus, the medium permits the study of the drug effect on the intracellular traffic of L-leucine permeases.

Lactation is a natural model of hippocampus neuroprotection against excitotoxicity.

Lactation is a temporary but complex physiological condition in which hormones and neurogenic stimulation from suckling cause maternal brain plasticity. It has been shown that lactation prevents cell damage induced by excitotoxicity in the dorsal hippocampus of the dam after peripheral administration of kainic acid (KA). The aim of this study was to determine whether lactation protects the maternal hippocampus against damage induced by intracerebral application (ICV) of KA and if lactation decreases, or only delays, this damaging effect of KA. Cell damage was assessed by Fluoro-Jade C staining in the hippocampus of virgin and lactating rats 24 or 72 h after ICV KA. Lactation prevented cell damage of the pyramidal layers of the hippocampus (CA1, CA3, and CA4), as compared to virgin rats. The longer period of KA exposure increased the difference in cell damage between these two conditions. The present results confirm that lactation is a natural model for neuroprotection, since it effectively prevents acute and chronic cell damage of the hippocampus induced by exposure to KA.

Changes in c-Fos and NOS expression in the PVH of lactating rats in response to excitotoxicity and stress.

This study investigated how lactation modified the expression patterns of Fos and nitric oxide synthase in the hypothalamic paraventricular nucleus (PVH) induced by excitotoxicity and stress. Kainic acid or egg white treatment weakly activated Fos expression in the PVH of lactating in comparison to diestrus or ovariectomized (OVX) rats. Labels for NADPH-diaphorase and nNOS revealed a different distribution pattern in the PVH depending on the physiological condition and challenge. The present results confirm that lactation attenuates the PVH activational response to stress and excitotoxicity, and both stimuli induced nitric oxide expression in the PVH of diestrus, lactating, and OVX rats.

Fos expression induced by milk ingestion in the caudal brainstem of neonatal rats.

Prominent Fos expression in the nucleus of the solitary tract (NTS) related to feeding has been reported in the brainstem of adult animals. In this study, we used a Fos-guided immunohistochemical approach to determine the brainstem areas activated specifically in response to milk ingestion in rat pups at two different ages. Rats at 9 or 18 days postpartum were isolated from the mother for a 6-h period, after which they were returned to the mother for a suckling period of either 5 or 90 min and then perfused at 90 min after the beginning of suckling. Control groups were sacrificed before or after the 6-h-deprivation period and showed little or no Fos-ir. In contrast, a 90-min-suckling episode after 6 h of deprivation induced strong Fos-ir in the caudal regions of the NTS and in the spinal nucleus of the trigeminal (SPV). Moderate expression was observed in the rostral NTS and in the nucleus raphé obscurus. In rat pups that suckled for only 5 min, the main area activated was the SPV. Fos immunostaining was detected in only 1% of the catecholaminergic neurons from the NTS after milk ingestion. The experimental design employed here allowed us to distinguish brainstem areas activated by milk ingestion from those activated by suckling action in rat pups. In contrast to adult rats, catecholaminergic neurons from the caudal NTS seem to contribute little to the regulation of feeding at this age.

Neuroprotective effects of lactation against kainic acid treatment in the dorsal hippocampus of the rat.

Marked hippocampal changes in response to excitatory amino acid agonists occur during pregnancy (e.g. decreased frequency in spontaneous recurrent seizures in rats with KA lesions of the hippocampus) and lactation (e.g. reduced c-Fos expression in response to N-methyl-d,l-aspartic acid but not to kainic acid). In this study, the possibility that lactation protects against the excitotoxic damage induced by KA in hippocampal areas was explored. We compared cell damage induced 24 h after a single systemic administration of KA (5 or 7.5 mg/kg bw) in regions CA1, CA3, and CA4 of the dorsal hippocampus of rats in the final week of lactation to that in diestrus phase. To determine cellular damage in a rostro-caudal segment of the dorsal hippocampus, we used NISSL and Fluorojade staining, immunohistochemistry for active caspase-3 and TUNEL, and we observed that the KA treatment provoked a significant loss of neurons in diestrus rats, principally in the pyramidal cells of CA1 region. In contrast, in lactating rats, pyramidal neurons from CA1, CA3, and CA4 in the dorsal hippocampus were significantly protected against KA-induced neuronal damage, indicating that lactation may be a natural model of neuroprotection.

Análisis de la mortalidad en enfermedades Hemato-oncológicas malignas en pediatría en hospitales públicos de Argentina / Analysis of Mortality due to Hemato-Oncological Pediatrics Malignancies in Public Hospitals in Argentina

INTRODUCCIÓN: En Argentina, la mortalidad por enfermedades malignas en edad pediátrica ocupa un lugar relevante y sus causas todavía no han sido estudiadas en el país. OBJETIVO: Analizar las tasas, causas y etapas de los fallecimientos relacionados con neoplasias en centros públicos seleccionados, desde enero de 2000 a diciembre de 2010. MÉTODOS: Se analizaron las historias clínicas de los pacientes fallecidos por cáncer en centros registrados en el Registro Oncopediátrico Hospitalario Argentino (ROHA) y en los registros individuales de los servicios de Hemato-Oncología. Se clasificaron las causas de mortalidad, la etapa en la cual se produjo el óbito y su relación con el tratamiento o con la patología de base. Se pesquisaron las causas de comorbilidad y las demoras en el diagnóstico y tratamiento. RESULTADOS: En 13 centros se analizó exitosamente un promedio >70...

INTRODUCTION: In Argentina, the mortality of pediatric malignant diseases occupies an important place causes have not yet been studied in the country. OBJECTIVE:To analyze mortality rates, causes and moment of death related to neoplasias in selected public centers from January 2000 until December 2010. METHODS: The analysis was conducted in clinical records of patients who died due to cancer. The cases were registered in the Argentine Hospital Oncopediatric Registry (ROHA)and by different registries belonging to hemato-oncological departments. Mortality causes were classified according to the phase of therapy when the event occurred and the relation shipof death with the treatment or underlying disease. Causes of comorbility and delays in diagnosis/treatment were also analyzed. RESULTS: In 13 centers, more than 70...

Efecto de iones amonio sobre la entrada de L-leucina en Saccharomyces cerevisiae: represíon e inhibicíon of transport systems / Effect of ammoniun ions on the uptake of L-leucine: repression and inhibition of transport systems

En Saccharomyces cerevisiae la entrada de L-leucina es mediada por la permeasa general de aminoácidos, GAP, y dos sistemas cinéticamente caracterizados, uno de alta afinidad y baja velocidad, S1, para concentraciones externas de L-leucina 0,05-01mM y otro de baja afinidad, alta velocidad, S2, para concentraciones externas 1.0mM. En células crecidas en medios suplementados con amonio, como única fuente de nitrógeno, los valores de entrada e incorporación son menores que en células crecidas en medios suplementados con L-prolina. En condiciones de represión de la GAP por iones amonio, la entrada de L-leucina es mediada por los sistemas S1 y S2. Los dos sistemas son parcialmente inhibidos por efecto de iones amonio. En condiciones de depresion de la GAP, por crecimiento en L-prolina, la entrada de L-leucina es mediada por los sistemas S1 y GAP, bajas concentraciones externas de L-leucina mediada por centraciones externas. El amonio inhibe en mayor extensión la entrada de L-leucina mediada por La GAP

Determinación del pH intracelular por distribución de ácido benzoico en S. cerevisiae, transporte de aminoácidos y gradiente de protones / Determination of internal pH of S. cerevisiae from the distribution of benzoic acid: amino acids transport and proton gradient

El (pHi) de Saccjaromyces cerevisiae, cepa silvestre y su mutante rhose determinó por el método de distribución intra-extracelilar del ácido 14C-benzoico. Los valores de pHi obtenidos (con un pH externo de 4,5) variarón con la depa de levadura y dependieron de las condiciones metabólicas de las mismas. En células de la cepa silvestre, energizada por incubación previa con glucosa 5 mM, el pHi osció entre 6,15-6,40 y el gradiente de protones a través de la membrana deltapH entre 1,65-1,90 ó -97 a -112 mV. Esos valores fueron mayores que los de células ayunadas: pHi 5,90, deltapH 1,40 ó -82 mV. En esas dos condiciones metabólicas, los valores en la mutante rho- fueron algo menores que en la cepa silvestre; levadura rho- energizada pHi 6,05, deltapH 1,55 ó -91 mV; levadura rho- ayunada pHi 5,70, deltapH 1,20 ó -71 mV. Los protonófros DNP y PCP produjeron una disminución del pHi del deltapH y una inhibición de la entrada de L-leucina por los sistemas S1, alta afinidad y baja velocidad y S2, baja afinidad y alta velocidad. Los valores de la disminución del deltapH y la inhibición del transporte de L-leucina obtenidos indican que no hay una relación estricta entre el deltapH y el proceso de transporte