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Hyponatremia is the most common electrolyte disturbance in hospitalized children, with a reported incidence of 15-30%, but its overall incidence and severity are not well known. The objective of our study was to determine the incidence, severity, and associated risk factors of community- and hospital-acquired hyponatremia on a general pediatric ward. Data of 5550 children admitted from June 2012 to December 2019 on plasma sodium and discharge diagnosis were analyzed by logistic regression model. Clinically relevant diagnostic groups were created. Hyponatremia was classified as mild, moderate, and severe. The incidence of community- and hospital-acquired hyponatremia was 15.8% and 1.4%, respectively. Most of the cases were mild (90.8%) to moderate (8.6%), with only two cases of severe community-acquired hyponatremia. There were no clinical complications in any of the hyponatremic children. Age and diagnosis at discharge were principal factors significantly correlated with hyponatremia. Community-acquired hyponatremia is more common than hospital-acquired hyponatremia in clinical practice. Severe cases of both types are rare. Children from 2 to 11 years of age presenting with infections, cardiovascular disorders, and gastrointestinal disorders are at risk of developing hyponatremia.
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Beef-calf mortality rates across tropical and subtropical Australia are high, with sub-optimal nutrition in pregnant cows being the primary risk. The nutritional deficiencies associated with calf mortality are the same as those associated with reduced milk yields. Although the highest mortality risk occurs during neonatal life, the role of inadequate milk delivery to beef neonates is not well established. This study investigated the frequency of low milk delivery in tropically adapted neonatal calves and the time for their dams to initiate full lactation in five management groups of Brahman and Droughtmaster calving cows in the dry tropics of northern Queensland, Australia. Change in calf weight in the days following birth was the primary measure of milk uptake. Plasma globulin concentration was used to indicate colostrum uptake. Across management groups, data were available on 250 calves for regression analysis of average daily gain vs. globulin and on 78 for plotting calf growth profiles. Calves had one of two growth profiles, either with immediate high growth from birth (day one) or with high growth delayed until day three. The frequency of delayed growth calves (with inadequate milk intake to gain at least 0.5 kg by day three after birth) was on average 30% across management groups, with management groups ranging 25%-50%. The frequency of calves growing ≤0.2 kg/day to day three was 15%-37%, depending on management group. The frequency of calves growing ≤0.2 kg/day to day five was 7%-20%, depending on management group. Calf globulin explained only 25% of the variation in calf average daily gain. Our study shows that a third of tropically adapted calves may experience a three-day delay to initiation of full lactation by their dams. Although study conditions were relatively benign, any additional risks with milk delivery, such as those that occur widely in tropical and subtropical northern Australia, would place such calves at risk of dehydration and mortality. Calf plasma globulin should not be used as a standalone measure of adequacy of neonatal milk delivery, especially when comparing across herds. This study demonstrates a fundamental problem of high frequency in northern Australia. The underlying risks for delayed milk delivery should be considered in the quest for practical solutions to reduce tropically adapted beef-calf mortalities.
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Alimentación Animal , Leche , Alimentación Animal/análisis , Animales , Bovinos , Calostro , Femenino , Lactancia , Parto , EmbarazoRESUMEN
BACKGROUND: This research was undertaken to evaluate Oxford Domed Lateral unicompartmental knee replacement (UKR) survival and clinical and radiological outcomes. The study also considered the influence of body mass index (BMI) on results and proposed contralateral healthy knee anatomic femorotibial angle (AFTA) as a predictor of postoperative knee alignment. METHODS: A retrospective evaluation of 41 primary Oxford Domed Lateral UKR performed in 41 patients in the same institution was undertaken on a patient group comprising of 10 men and 31 women with a mean age of 63â¯years (range: 38-81â¯years). A minimum follow-up of two years was required. RESULTS: The total revision rate was one out of 41 patients (2.4%), with a mean follow-up of 49â¯months (range: 25-84â¯months). One patient presented with a traumatic medial dislocation of the bearing after 15â¯months (resulting in an overall dislocation rate of 2.4%). The visual analogue scale (VAS) and the Oxford knee score (OKS) demonstrated significant postoperative improvement (Pâ¯<â¯0.001). Survival at five years, with revision for any reason as the endpoint, was 97.5%. No significant correlation was observed between BMI and postoperative flexion angle, radiologically measured parameters, or pre- and postoperative VAS and OKS. Significant correlation was found (râ¯=â¯0.77, Pâ¯<â¯0.001) between postoperative and contralateral healthy knee anatomic femorotibial angles. CONCLUSIONS: The Oxford Domed Lateral UKR presents a low dislocation rate and excellent mid-term clinical and functional results. Contralateral healthy knee AFTA appears to be a predictor of postoperative knee alignment.
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Artroplastia de Reemplazo de Rodilla/instrumentación , Artroplastia de Reemplazo de Rodilla/métodos , Prótesis de la Rodilla , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Estudios de Seguimiento , Humanos , Articulación de la Rodilla/cirugía , Masculino , Persona de Mediana Edad , Osteoartritis de la Rodilla/cirugía , Evaluación del Resultado de la Atención al Paciente , Reoperación/estadística & datos numéricos , Estudios Retrospectivos , Escala Visual AnalógicaRESUMEN
We study transport mediated by Andreev bound states formed in InSb nanowire quantum dots. Two kinds of superconducting source and drain contacts are used: epitaxial Al/InSb devices exhibit a doubling of tunneling resonances, while, in NbTiN/InSb devices, Andreev spectra of the dot appear to be replicated multiple times at increasing source-drain bias voltages. In both devices, a mirage of a crowded spectrum is created. To describe the observations a model is developed that combines the effects of a soft induced gap and of additional Andreev bound states both in the quantum dot and in the finite regions of the nanowire adjacent to the quantum dot. Understanding of Andreev spectroscopy is important for the correct interpretation of Majorana experiments done on the same structures.
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The present study aimed to evaluate non-fiber carbohydrates (NFC) in sugarcane-based diets on rumen pH, and forage digestibility, and to describe NFC degradation curves. The study consisted of two trials. For the first trial, three rumen cannulated steers, BW of 350 ± 15 kg (mean ± SE), were assigned in a 3×3 Latin Square (LS) design. They were fed diets containing finely-ground (0.9 mm average particle size) corn (GC), steam-rolled corn (SRC), or pelleted citrus pulp (PCP). Each period had 14 d, with the first 12 for adaptation. The 13th d was for serial measurement of rumen pH, and the14th for rumen fluid collection and in vitro incubation for DM and NDF digestibility (IVDMD and IVNDFD) of bermudagrass hay (Hay), corn (CS), and sugarcane (SS) silages. In the second trial, rumen fluid of a cannulated bull, fed corn silage and a regular concentrate, was collected for in vitro digestion of NFC for multiple time points. The incubation results were used to adjust the NFC degradation curves, and calculate lag-time, feed fractions, and degradation rate. Data from first trial was analyzed in a 3×3 LS. The model for the digestibility parameters included fixed effects of forage (Feed), diets with NFC (Diet), and their interaction (Feed × Diet), and random effect of animal and period. The model for rumen pH included fixed effect of diet, time as repeated measures, animal and period as random effects. The significance was considered at probability ≤ 5% (α = 0.05). The NFC degradation curves were adjusted using the PROC NLIN procedure from SAS, and equation parameters compared using confidence intervals. There was a Diet × Time interaction on rumen pH (P = 0.04), where SRC decreased pH compared to PCP and GC diets at the time 6 h, only. There was no Feed × Diet interaction effect (P > 0.05) for any digestibility parameter. There was a Feed effect on both IVDMD and IVNDFD, either after 30 or 48 h incubation (P < 0.01). The CS had the greatest IVDMD, followed by SS and Hay, after 30 and 48 h of incubation. The CS had the greatest IVNDFD after 30 h, compared to SS and Hay. However, for IVNDFD after 48 h, CS presented the greatest mean, followed by SS and Hay. The rumen fluid from animals fed SRC decreased both IVDMD and IVNDFD (P < 0.05) of all roughages after 48 h. Results from the second trial showed that the PCP had lower Lag Time, B fraction and greater kd compared to both corn sources, and SRC had greater kd than GC. In conclusion, the SRC diet decreased rumen pH 6 h after feeding and, consequently, decreased fiber digestibility of the tropical forage sources evaluated. Although the PCP had lower lag time, and faster rate of degradation of B fraction, it did not negatively affect rumen pH or fiber digestibility of forage.(AU)
O presente estudo teve como objetivo avaliar os carboidratos não-fibrosos (CNF) em dietas à base de cana-de-açúcar sobre o pH ruminal e digestibilidade da forragem, e descrever as curvas de degradação dos CNF. O estudo foi composto de dois ensaios. No primeiro, três novilhos canulados no rúmen, com peso vivo de 350 ± 15 kg (Média ± DP), foram alocados em um quadrado latino (QL) 3×3, e alimentados com dietas contendo: milho moído (MM, tamanho de partículas 0,9 mm), laminado a vapor (MLV) ou polpa cítrica peletizada (PCP). Cada período tinha 14 d, sendo os primeiros 12 para adaptação e o 13º para a medição seriada do pH e o 14º para a coleta de líquido ruminal e incubação in vitro para digestibilidade da MS e FDN (DIVMS e DIVFDN) de feno de bermudagrass (Feno) e silagens de milho (SM) e cana (SC). No segundo ensaio, coletou-se fluido ruminal de um touro canulado, alimentado com silagem de milho e concentrado padrão, para digestão in vitro dos CNF em vários tempos. Esses resultados foram utilizados para ajustar as curvas de degradação dos CNF e calcular o tempo de colonização, frações alimentares e taxa de degradação. Os resultados do primeiro ensaio foram analisados em um QL 3×3. O modelo dos parâmetros de digestibilidade incluiu efeito fixo de forragem (Alimento), dieta com CNF (Dieta) e interação (Alimento × Dieta), e efeito aleatório de animal e período. O modelo para pH incluiu efeito fixo de Dieta, Tempo como medida repetida, animal e período como aleatórios. Foi considerada a probabilidade significativa de ≤ 5% (α = 0,05). As curvas de degradação dos CNF foram ajustadas pelo PROC NLIN do SAS, e parâmetros de equação comparados por intervalo de confiança. Houve interação Dieta × Tempo no pH ruminal (P = 0,04), onde o MLV diminuiu o pH comparado com PCP e MM apenas no tempo 6 h. Não houve interação Alimento × Dieta (P > 0,05) para nenhum parâmetro de digestibilidade. Houve efeito de Alimento sobre a DIVMS e DIVFDN, após 30 e 48 h de incubação (P < 0,01). A SM teve a maior DIVMS, seguido por SC e Feno, após 30 e 48 h de incubação. A SM teve a maior DIVFDN após 30 h, comparado com SC e Feno. No entanto, para DIVFDN após 48 h, a SM teve maior média, seguida da SC e Feno. O fluido ruminal de animais alimentados com MLV diminuiu a DIVMS e DIVFDN (P < 0.05) de todas as forragens, após 48 h. Resultados do segundo ensaio mostram que PCP diminuiu o tempo de colonização, fração B e aumentou a kd comparado com os dois milhos, e MLV apresentou maior kd que o MM. Em conclusão, a dieta com MLV diminuiu o pH ruminal no tempo 6 h e, consequentemente, diminuiu a DIVFDN das forragens avaliadas. Embora PCP tenha apresentado menor tempo de colonização e maior taxa de degradação da fração B, não afetou negativamente o pH do rúmen nem a digestibilidade da fibra das forragens. (au)
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Carbohidratos/análisis , Citrus/química , Rumiación Digestiva , Zea mays/efectos adversosRESUMEN
Molecular targeted therapy of advanced neuroendocrine tumours (NETs) of the gastroenteropancreatic (GEP) system currently encompasses approved therapy with the mammalian target of rapamycin (mTOR) inhibitor everolimus and the multi-tyrosinkinase inhibitor sunitinib. However, clinical efficacy of these treatment strategies is limited by low objective response rates and limited progression-free survival due to tumour resistance. Further novel strategies for molecular targeted therapy of NETs of the GEP system are needed. This paper reviews preclinical research models and signalling pathways in NETs of the GEP system. Preclinical and early clinical data on putative novel targets for molecular targeted therapy of NETs of the GEP system are discussed, including PI3K, Akt, mTORC1/mTORC2, GSK3, c-Met, Ras-Raf-MEK-ERK, embryogenic pathways (Hedgehog, Notch, Wnt/beta-catenin, TGF-beta signalling and SMAD proteins), tumour suppressors and cell cycle regulators (p53, cyclin-dependent kinases (CDKs) CDK4/6, CDK inhibitor p27, retinoblastoma protein (Rb)), heat shock protein HSP90, Aurora kinase, Src kinase family, focal adhesion kinase and epigenetic modulation by histone deacetylase inhibitors.
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This research aimed to evaluate the effects of the beta-agonist zilpaterol hydrochloride (ZH) on carcass traits, subprimal yield, meat quality, palatability traits, and gene expression in Nellore heifers. Zilpaterol increased Longissimus lumborum area and did not change back fat thickness, meat color, and cooking loss. Heifers fed ZH had greater hindquarter weight and carcass percentage. Muscles from hindquarter were heavier for animals fed ZH. Forequarter (% of carcass) decreased and brisket did not change with ZH supplementation. There were no differences between treatments for steak aroma, beef flavor, and off-flavor. However, tenderness and juiciness were reduced by ZH, depending on postmortem aging. Zilpaterol increased Calpain-1, Calpain-2, and calpastatin mRNA expression, with no effect of day of slaughter or ZH×Day interaction. In conclusion, ZH supplementation improved hypertrophy, meat production, and debone yield in Nellore heifers, which led to decreased tenderness and to increased mRNA expression in the calpain-calpastatin system.
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Proteínas de Unión al Calcio/metabolismo , Calpaína/metabolismo , Carne Roja , Compuestos de Trimetilsililo/farmacología , Animales , Proteínas de Unión al Calcio/genética , Calpaína/genética , Bovinos , Color , Comportamiento del Consumidor , Culinaria , Aditivos Alimentarios/farmacología , Calidad de los Alimentos , Expresión Génica , Humanos , Concentración de Iones de Hidrógeno , ARN Mensajero , GustoRESUMEN
No disponible
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Femenino , Humanos , Masculino , Especificaciones Sanitarias/métodos , /organización & administración , /normas , Técnicas de Laboratorio Clínico/normas , Sistemas de Información en Laboratorio Clínico/organización & administración , Sistemas de Información en Laboratorio Clínico/normas , Ciencia del Laboratorio Clínico/organización & administración , Ciencia del Laboratorio Clínico/normas , AlgoritmosAsunto(s)
Análisis Químico de la Sangre/normas , Ensayos de Aptitud de Laboratorios/organización & administración , Garantía de la Calidad de Atención de Salud/normas , Sociedades Científicas/normas , Urinálisis/normas , Comités Consultivos , Consenso , Humanos , Ensayos de Aptitud de Laboratorios/normas , Guías de Práctica Clínica como Asunto , Garantía de la Calidad de Atención de Salud/organización & administración , Control de Calidad , EspañaRESUMEN
The aim of this study was to evaluate the use of real-time polymerase chain reaction (qPCR) combined with DNA extraction directly from composite milk and bulk tank samples for detection and enumeration of Streptococcus agalactiae (SAG) causing subclinical mastitis. Dilutions of sterile reconstituted skim milk inoculated with SAG ATCC 13813 were used to establish a standard curve (cfu/mL) for the qPCR assay targeting SAG. The analytical sensitivity and repeatability of the qPCR assay were determined. Bulk tank (BTM; n = 38) and composite milk samples (CM; n = 26) collected from lactating cows with positive isolation of SAG were submitted to the qPCR protocol and SAG plate counting, with results from both methods compared. Amplification of DNA was not possible in two out of 64 samples, indicating that qPCR was able to detect SAG in 96 and 97% of BTM and CM samples, respectively. The inter-assay coefficient of variation was <5%, showing that the technique had adequate repeatability. The qPCR protocol can be a high-throughput and rapid diagnostic assay to accurately detect SAG from BTM and CM samples compared with conventional microbiological culture method. However, the evaluated qPCR protocol is not accurate for enumerating SAG in milk samples, probably due to quantification of DNA of non-viable cells.
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Carga Bacteriana/métodos , Leche/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Streptococcus agalactiae/aislamiento & purificación , Animales , Infecciones Asintomáticas , Bovinos , Mastitis Bovina/microbiología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Streptococcus agalactiae/genéticaRESUMEN
Resveratrol, a modulator of several signaling proteins, can exert off-target effects involving the peroxisome proliferator-activated receptor (PPAR) transcription factors. However, evidence for the direct interaction between this polyphenol and PPARs is lacking. Here, we addressed the hypothesis that resveratrol and its metabolites control aspects of PPAR transcriptional activity through direct interaction with PPARs. Bioaffinity chromatographic studies with the immobilized ligand-binding domains (LBDs) of PPARγ and PPARα and isothermal titration calorimetry allowed the binding affinities of resveratrol, resveratrol 3-O-glucuronide, resveratrol 4-O-glucuronide, and resveratrol 3-O-sulfate to both PPAR-LBDs to be determined. Interaction of resveratrol, resveratrol 3-O-glucuronide, and resveratrol 4-O-glucuronide with PPARγ-LBD occurred with binding affinities of 1.4, 1.1, and 0.8 µM, respectively, although only resveratrol bound to the PPARα-LBD with a binding affinity of 2.7 µM. Subsequently, X-ray crystallographic studies were carried out to characterize resveratrol binding to the PPARγ-LBD at the molecular level. The electron density map from the crystal structure of the complex between PPARγ-LBD and resveratrol revealed the presence of one molecule of resveratrol bound to the LBD of PPARγ, with the ligand occupying a position close to that of other known PPARγ ligands. Transactivation assays were also performed in HepG2 cells, with the results showing that resveratrol was not a PPAR agonist but instead was able to displace rosiglitazone from PPARγ and Wy-14643 from PPARα with IC50 values of (27.4±1.8) µM and (31.7±2.5) µM, respectively. We propose that resveratrol acts as a PPAR antagonist through its direct interaction with PPARγ and PPARα.
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Receptores Activados del Proliferador del Peroxisoma/antagonistas & inhibidores , Receptores Activados del Proliferador del Peroxisoma/metabolismo , Estilbenos/metabolismo , Estilbenos/farmacología , Sitios de Unión , Cristalografía por Rayos X , Relación Dosis-Respuesta a Droga , Células Hep G2 , Humanos , Modelos Moleculares , Receptores Activados del Proliferador del Peroxisoma/química , Pirimidinas/farmacología , Resveratrol , Rosiglitazona , Relación Estructura-Actividad , Tiazolidinedionas/farmacología , Células Tumorales CultivadasRESUMEN
Coagulase-negative staphylococci (CoNS) are among the main pathogens causing bovine intramammary infection (IMI) in many countries. However, one of the limitations related to the specific diagnosis of CoNS is the lack of an accurate, rapid, and convenient method that can differentiate the bacterial species comprising this group. The aim of this study was to evaluate the ability of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to accurately identify CoNS species in dairy cow IMI. In addition, the study aimed to determine the frequency of CoNS species causing bovine IMI. A total of 108 bacterial isolates were diagnosed as CoNS by microbiological cultures from two milk samples collected from 21 dairy herds; the first sample was collected at the cow level (i.e., 1,242 composite samples from all quarters), while the second sample was collected at the mammary quarter level (i.e., 1,140 mammary samples collected from 285 cows). After CoNS isolation was confirmed by microbiological culture for both samples, all CoNS isolates (n=108) were genotypically differentiated by PCR restriction fragment length polymorphism (RFLP) analysis of a partial groEL gene sequence and subjected to the MALDI-TOF MS identification procedure. MALDI-TOF MS correctly identified 103 (95.4%) of the CoNS isolates identified by PCR-RFLP at the species level. Eleven CoNS species isolated from bovine IMI were identified by PCR-RFLP, and the most prevalent species was Staphylococcus chromogenes (n=80; 74.1%). In conclusion, MALDI-TOF MS may be a reliable alternative method for differentiating CoNS species causing bovine IMI.
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Enfermedades de los Bovinos/microbiología , Coagulasa/genética , Mastitis Bovina/microbiología , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Infecciones Estafilocócicas/microbiología , Staphylococcus/genética , Animales , Bovinos , Femenino , Leche/microbiologíaRESUMEN
Sperm cryopreservation is widely used for both research and reproduction purposes, but its effect on sperm DNA damage remains controversial. Sperm DNA fragmentation (SDF) has become an important biomarker to assess male infertility. In particular, the differentiation between single- and double-stranded DNA fragmentation (ssSDF and dsSDF) has clinical implications for male infertility where ssSDF is associated with reduced fertility, whereas dsSDF is associated with increased risk of miscarriage. In this study, semen samples from 30 human males have been analysed in both fresh and cryopreserved using the alkaline and neutral Comet assays. Results show an increase of about 10% of ssSDF, assessed by the alkaline Comet assay, regardless of the male fertility status. Neutral Comet analysis of dsSDF does not show any statistical increase when comparing fresh and cryopreserved samples in any of the patient groups. Results support previous reports that oxidative stress is the major effector in DNA damage during sample cryopreservation, as, on one hand, ssSDF has previously been related to oxidative damage and, on the other hand, we have not found any effect on dsSDF. Therefore, there might be a slight risk of decreased fertility after using a freezed sample, but no evidence for increased miscarriage risk from cryopreserved spermatozoa should be expected.
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Criopreservación , Fragmentación del ADN , Preservación de Semen/efectos adversos , Espermatozoides/citología , Ensayo Cometa , Humanos , Infertilidad Masculina , Masculino , Estrés Oxidativo , Semen , Análisis de SemenRESUMEN
Sperm DNA fragmentation (SDF) is becoming an important test to assess male infertility. Several different tests are available, but no consensus has yet been reached as to which tests are most predictive of infertility. Few publications have reported a comprehensive analysis comparing these methods within the same population. The objective of this study was to analyze the differences between the five most common methodologies, to study their correlations and to establish their cut-off values, sensitivity and specificity in predicting male infertility. We found differences in SDF between fertile donors and infertile patients in TUNEL, SCSA, SCD and alkaline Comet assays, but none with the neutral Comet assay. The alkaline COMET assay was the best in predicting male infertility followed by TUNEL, SCD and SCSA, whereas the neutral COMET assay had no predictive power. For our patient population, threshold values for infertility were 20.05% for TUNEL assay, 18.90% for SCSA, 22.75% for the SCD test, 45.37% for alkaline Comet and 34.37% for neutral Comet. This work establishes in a comprehensive study that the all techniques except neutral Comet are useful to distinguish fertile and infertile men.
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Fragmentación del ADN , Infertilidad Masculina/diagnóstico , Análisis de Semen/métodos , Espermatozoides/citología , Cromatina/metabolismo , Ensayo Cometa , Humanos , Etiquetado Corte-Fin in Situ , Infertilidad Masculina/genética , MasculinoRESUMEN
O artigo aborda o significado que toma o termo fé no campo da psicanálise, tal como introduzido por Bion no contexto de seu pensamento a partir do livro Transformações. Para penetrar a penumbra de significados do termo, a autora se dirige à concepção religiosa de fé, usando subsídios do pensamento de Søren Kierkegaard. O artigo procede a uma aproximação dos dois vértices numa ilustração clínica do conceito de fé.
This paper explores the meaning of faith in the field of psychoanalysis, as introduced by Bion in the context of his thoughts since 1965, in his book Tansformations. The author searches the religious concept of faith in the thoughts of Søren Kierkegaard and tries to approximate the two vertices through a clinical illustration of the concept of faith.
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Humanos , Psicoanálisis , ReligiónRESUMEN
O artigo aborda o significado que toma o termo fé no campo da psicanálise, tal como introduzido por Bion no contexto de seu pensamento a partir do livro Transformações. Para penetrar a penumbra de significados do termo, a autora se dirige à concepção religiosa de fé, usando subsídios do pensamento de Søren Kierkegaard. O artigo procede a uma aproximação dos dois vértices numa ilustração clínica do conceito de fé.(AU)
This paper explores the meaning of faith in the field of psychoanalysis, as introduced by Bion in the context of his thoughts since 1965, in his book Tansformations. The author searches the religious concept of faith in the thoughts of Søren Kierkegaard and tries to approximate the two vertices through a clinical illustration of the concept of faith.(AU)
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Humanos , Psicoanálisis , ReligiónRESUMEN
The peroxisome proliferator-activated receptors (PPARs) belong to the nuclear receptor superfamily. In the last years novel PPARs ligands have been identified and these include PPARα/γ dual agonists. To rapidly identify novel PPARs dual ligands, a robust binding assay amenable to high-throughput screening toward PPAR isoforms would be desirable. In this work we describe a parallel assay based on the principles of frontal affinity chromatography coupled to mass spectrometry (FAC-MS) that can be used to characterize dual agonists. For this purpose the ligand binding domain of PPARα receptor was immobilized onto the surface of open tubular capillaries to create new PPAR-alpha-OT columns to be used in parallel with PPAR-gamma-OT columns. The two biochromatographic systems were used in both ranking and Kd experiments toward new ureidofibrate-like dual agonists for subtype selectivity ratio determination. In order to validate the system, the Kd values determined by frontal analysis chromatography were compared to the affinity constants obtained by ITC experiments. The results of this study strongly demonstrate the specific nature of the interaction of the ligands with the two immobilized receptor subtypes.
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Cromatografía de Afinidad/instrumentación , Cromatografía de Afinidad/métodos , Espectrometría de Masas/métodos , PPAR alfa/química , PPAR gamma/química , Calorimetría , Descubrimiento de Drogas/métodos , Proteínas Inmovilizadas/agonistas , Proteínas Inmovilizadas/química , Proteínas Inmovilizadas/metabolismo , Ligandos , PPAR alfa/agonistas , PPAR alfa/metabolismo , PPAR gamma/agonistas , PPAR gamma/metabolismo , Unión Proteica , Reproducibilidad de los ResultadosRESUMEN
Objetivou-se com este estudo avaliar o efeito da época de corte sobre a composição bromatológica e digestibilidade in vitro de genótipos de cana-de-açúcar, comparar a composição bromatológica e digestibilidade in vitro das frações colmo e folhas, e estabelecer possíveis correlações entre composição química e digestibilidade in vitro da planta inteira de genótipos da cana-de-açúcar. Nove genótipos foram colhidos em maio e setembro de 2006. Em maio, somente a fração planta inteira foi analisada, em setembro foram separados em colmos, folhas ou planta inteira para determinação da composição bromatológica, sacarose (POL) e digestibilidade in vitro. Colmos apresentaram menor teor de fibra em detergente neutro (FDN) e lignina na MS, e maior digestibilidade in vitro da matéria seca (DIVMS) do que folhas. Em contrapartida, apresentaram menor digestibilidade in vitro da FDN (DIVFDN), maior teor de lignina na FDN e menor concentração de proteína bruta (PB). Houve redução no teor e digestibilidade da FDN com o avanço da maturidade, enquanto que DIVMS, POL e lignina aumentaram. A DIVMS se mostrou negativamente correlacionada com FDN e FDN/POL, porém não houve correlação entre DIVFDN e FDN ou FDN/POL. Conclui-se que o avanço da maturidade promoveu redução na DIVFDN e aumento na DIVMS, não existindo correlação genética entre acúmulo de açúcar e digestibilidade in vitro da fibra. Os dados deste artigo indicam não ser esperado que a seleção de genótipos com maior DIVFDN do colmo altere o teor de açúcar do mesmo.
The objectives of this study were to evaluate the effect of time of harvest on chemical composition and in vitro digestibility of sugarcane genotypes, to compare chemical composition and in vitro digestibility of the stem and leaf fractions, and to determine possible correlations between chemical composition and in vitro digestibility of the whole plant in sugarcane genotypes. Nine genotypes were harvested in May and September of 2006. In May, only the whole-plant fraction was analyzed, in September the genotypes were separated in stems, leaves or whole-plant for determination of chemical composition, sucrose (POL) and in vitro digestibility. Stems had lower neutral detergent fiber (NDF) and lignin in the DM, and greater in vitro dry matter digestibility (IVDMD) than leaves. However, stems had lower in vitro NDF digestibility (IVNDFD), higher lignin in the NDF and lower crude protein (CP). The NDF and IVNDFD were reduced with advanced maturity, while IVDMD, POL and lignin were increased. IVDMD was negatively correlated with NDF and NDF/POL, however there was no correlation between IVFDND and NDF or NDF/POL. It can be concluded that with the advance in maturity the IVNDFD was reduced and IVDMD was increased, and there was no genetic correlation between accumulation of sugar and in vitro fiber digestibility. Data from this study indicate that it is not expected that selection of genotypes with greater stem IVNDFD would alter the sugar content of the plant.
Asunto(s)
Saccharum/química , Factores de Tiempo , Digestión , GenotipoRESUMEN
Objetivou-se com este estudo avaliar o efeito da época de corte sobre a composição bromatológica e digestibilidade in vitro de genótipos de cana-de-açúcar, comparar a composição bromatológica e digestibilidade in vitro das frações colmo e folhas, e estabelecer possíveis correlações entre composição química e digestibilidade in vitro da planta inteira de genótipos da cana-de-açúcar. Nove genótipos foram colhidos em maio e setembro de 2006. Em maio, somente a fração planta inteira foi analisada, em setembro foram separados em colmos, folhas ou planta inteira para determinação da composição bromatológica, sacarose (POL) e digestibilidade in vitro. Colmos apresentaram menor teor de fibra em detergente neutro (FDN) e lignina na MS, e maior digestibilidade in vitro da matéria seca (DIVMS) do que folhas. Em contrapartida, apresentaram menor digestibilidade in vitro da FDN (DIVFDN), maior teor de lignina na FDN e menor concentração de proteína bruta (PB). Houve redução no teor e digestibilidade da FDN com o avanço da maturidade, enquanto que DIVMS, POL e lignina aumentaram. A DIVMS se mostrou negativamente correlacionada com FDN e FDN/POL, porém não houve correlação entre DIVFDN e FDN ou FDN/POL. Conclui-se que o avanço da maturidade promoveu redução na DIVFDN e aumento na DIVMS, não existindo correlação genética entre acúmulo de açúcar e digestibilidade in vitro da fibra. Os dados deste artigo indicam não ser esperado que a seleção de genótipos com maior DIVFDN do colmo altere o teor de açúcar do mesmo.
The objectives of this study were to evaluate the effect of time of harvest on chemical composition and in vitro digestibility of sugarcane genotypes, to compare chemical composition and in vitro digestibility of the stem and leaf fractions, and to determine possible correlations between chemical composition and in vitro digestibility of the whole plant in sugarcane genotypes. Nine genotypes were harvested in May and September of 2006. In May, only the whole-plant fraction was analyzed, in September the genotypes were separated in stems, leaves or whole-plant for determination of chemical composition, sucrose (POL) and in vitro digestibility. Stems had lower neutral detergent fiber (NDF) and lignin in the DM, and greater in vitro dry matter digestibility (IVDMD) than leaves. However, stems had lower in vitro NDF digestibility (IVNDFD), higher lignin in the NDF and lower crude protein (CP). The NDF and IVNDFD were reduced with advanced maturity, while IVDMD, POL and lignin were increased. IVDMD was negatively correlated with NDF and NDF/POL, however there was no correlation between IVFDND and NDF or NDF/POL. It can be concluded that with the advance in maturity the IVNDFD was reduced and IVDMD was increased, and there was no genetic correlation between accumulation of sugar and in vitro fiber digestibility. Data from this study indicate that it is not expected that selection of genotypes with greater stem IVNDFD would alter the sugar content of the plant.