LONP1 ameliorates liver injury and improves gluconeogenesis dysfunction in acute-on-chronic liver failure / 中华医学杂志(英文版)

BACKGROUND@#Acute-on-chronic liver failure (ACLF) is a severe liver disease with complex pathogenesis. Clinical hypoglycemia is common in patients with ACLF and often predicts a worse prognosis. Accumulating evidence suggests that glucose metabolic disturbance, especially gluconeogenesis dysfunction, plays a critical role in the disease progression of ACLF. Lon protease-1 (LONP1) is a novel mediator of energy and glucose metabolism. However, whether gluconeogenesis is a potential mechanism through which LONP1 modulates ACLF remains unknown.@*METHODS@#In this study, we collected liver tissues from ACLF patients, established an ACLF mouse model with carbon tetrachloride (CCl 4 ), lipopolysaccharide (LPS), and D-galactose (D-gal), and constructed an in vitro hypoxia and hyperammonemia-triggered hepatocyte injury model. LONP1 overexpression and knockdown adenovirus were used to assess the protective effect of LONP1 on liver injury and gluconeogenesis regulation. Liver histopathology, biochemical index, mitochondrial morphology, cell viability and apoptosis, and the expression and activity of key gluconeogenic enzymes were detected to explore the underlying protective mechanisms of LONP1 in ACLF.@*RESULTS@#We found that LONP1 and the expressions of gluconeogenic enzymes were downregulated in clinical ACLF liver tissues. Furthermore, LONP1 overexpression remarkably attenuated liver injury, which was characterized by improved liver histopathological lesions and decreased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in ACLF mice. Moreover, mitochondrial morphology was improved upon overexpression of LONP1. Meanwhile, the expression and activity of the key gluconeogenic enzymes were restored by LONP1 overexpression. Similarly, the hepatoprotective effect was also observed in the hepatocyte injury model, as evidenced by improved cell viability, reduced cell apoptosis, and improved gluconeogenesis level and activity, while LONP1 knockdown worsened liver injury and gluconeogenesis disorders.@*CONCLUSION@#We demonstrated that gluconeogenesis dysfunction exists in ACLF, and LONP1 could ameliorate liver injury and improve gluconeogenic dysfunction, which would provide a promising therapeutic target for patients with ACLF.

Histopathological evaluation of cirrhosis reversal / 中华肝脏病杂志

A normal liver can develop cirrhosis through long-term and repeated stimulation from various etiologies. Histological manifestations like the collapse of hepatic lobular structure (including microvascular structure) and the formation of pseudolobules can lead to portal hypertension and even decompensated cirrhosis. More and more evidence suggests that effective etiological treatment can not only delay but also reverse the progression of cirrhosis. The mechanism of cirrhosis reversal mainly includes the degradation of extracellular matrix, hepatocyte regeneration, and hepatic lobular remodeling. The "gold standard" for the evaluation of cirrhosis reversal at present is still a liver biopsy. Therefore, the histopathological evaluation of cirrhosis reversal is very important for determining the disease's prognosis, efficacy, and mechanism of exploration.

Hepatocyte apoptosis fragment product cytokeratin-18 M30 level and non-alcoholic steatohepatitis risk diagnosis: an international registry study / 中华医学杂志(英文版)

BACKGROUND@#Liver biopsy for the diagnosis of non-alcoholic steatohepatitis (NASH) is limited by its inherent invasiveness and possible sampling errors. Some studies have shown that cytokeratin-18 (CK-18) concentrations may be useful in diagnosing NASH, but results across studies have been inconsistent. We aimed to identify the utility of CK-18 M30 concentrations as an alternative to liver biopsy for non-invasive identification of NASH.@*METHODS@#Individual data were collected from 14 registry centers on patients with biopsy-proven non-alcoholic fatty liver disease (NAFLD), and in all patients, circulating CK-18 M30 levels were measured. Individuals with a NAFLD activity score (NAS) ≥5 with a score of ≥1 for each of steatosis, ballooning, and lobular inflammation were diagnosed as having definite NASH; individuals with a NAS ≤2 and no fibrosis were diagnosed as having non-alcoholic fatty liver (NAFL).@*RESULTS@#A total of 2571 participants were screened, and 1008 (153 with NAFL and 855 with NASH) were finally enrolled. Median CK-18 M30 levels were higher in patients with NASH than in those with NAFL (mean difference 177 U/L; standardized mean difference [SMD]: 0.87 [0.69-1.04]). There was an interaction between CK-18 M30 levels and serum alanine aminotransferase, body mass index (BMI), and hypertension ( P  < 0.001, P  = 0.026 and P  = 0.049, respectively). CK-18 M30 levels were positively associated with histological NAS in most centers. The area under the receiver operating characteristics (AUROC) for NASH was 0.750 (95% confidence intervals: 0.714-0.787), and CK-18 M30 at Youden's index maximum was 275.7 U/L. Both sensitivity (55% [52%-59%]) and positive predictive value (59%) were not ideal.@*CONCLUSION@#This large multicenter registry study shows that CK-18 M30 measurement in isolation is of limited value for non-invasively diagnosing NASH.

Differential therapeutic effects of Crataegus aronia and simvastatin on the hepatocyte ultrastructure in hepatic steatosis / Efectos terapéuticos diferenciales de Crataegus aronia y simvastatina sobre la estructura del hepatocito en la esteatosis hepática

Complications of fat accumulation in liver, hepatic steatosis such as liver cirrhosis and liver failure are among the common public health problems. We sought to investigate the damage to the hepatocyte ultrastructure induced by high fat diets (HFD) and compared the therapeutic effects at the cellular level of two antioxidant and lipid lowering agents; Crataegus aronia extracts and simvastatin on hepatic steatosis. Rats were either fed with HFD (model group) or low fat diets (LFD) (control group) for 15 weeks before being sacrificed and therapeutic groups started the treatment with these agents after week 11 until the sacrifice day. Harvested liver tissues were examined using transmission electron microscopy (TEM) and liver homogenates were assayed for markers of anti-oxidative stress that are known to be modulated in liver injury. TEM examinations of the model group showed a profound damage to the hepatocytes compared to the control group as demonstrated by steatosis, damaged mitochondria and vaculated cytoplasm, disrupted rough and smooth endoplasmic reticulum and nuclear membrane, dilated intercellular space between hepatocytes, and alterations in lysosomes. In addition, HFD ameliorated the anti-oxidant glutathione (GSH) and augmented the oxidative stress TBARS biomarkers. Both Crataegus aronia and simvastatin significantly reduced lipids and TBARS, and treated damage to hepatic cells, but hepatocyte structures were differentially responded to these agents. However, only Crataegus aronia induced GSH (p=0.001). We conclude that HFD-induced hepatic steatosis caused a substantial damage to the hepatocyte's ultrastructures, and Crataegus aronia and simvastatin treatments differentially reversed hepatic injuries.

Las complicaciones de la acumulación de grasa en el hígado, la esteatosis hepática como la cirrosis hepática y la insuficiencia hepática se encuentran entre los problemas comunes de salud pública. Se intentó investigar el daño a la ultraestructura de los hepatocitos inducido por la dieta alta en grasas (DAG) y se compararon los efectos terapéuticos a nivel celular de dos antioxidantes y agentes hipolipemiantes; Extracto de Crataegus aronia y simvastatina sobre esteatosis hepática. Las ratas fueron alimentadas con DAG (grupo modelo) o dieta baja en grasa (DBG) (grupo control) durante 15 semanas antes de sacrificarse y los grupos terapéuticos comenzaron el tratamiento con estos agentes después de la semana 11 hasta el día del sacrificio. Se examinaron los tejidos hepáticos usando microscopía electrónica de transmisión (MET) y se analizaron homogeneizados de hígado para marcadores de estrés anti-oxidativo, que se sabe están modulados en la lesión hepática. Los exámenes MET del grupo DAG mostraron un grave daño de los hepatocitos en comparación con el grupo control, demostrado por esteatosis, daño mitocondrial y citoplasma vacío, retículo endoplásmico rugoso y liso y membrana nuclear, el espacio intercelular dilatado entre hepatocitos y alteraciones en los lisosomas. Además, DAG mejoró el anti-oxidante glutatión (GSH) y aumentó el estrés oxidativo TBARS biomarcadores. Tanto Crataegus aronia como simvastatina redujeron significativamente los lípidos y TBARS, trataron el daño a las células hepáticas, pero las estructuras de hepatocitos respondieron diferencialmente a estos agentes. Sin embargo, sólo Crataegus aronia indujo GSH (p = 0,001). Concluimos que la esteatosis hepática inducida por HFD causó un daño sustancial a la ultraestructura del hepatocito y los tratamientos de Crataegus aronia y simvastatina diferenciaron las lesiones hepáticas.

Management Strategies for Liver Fibrosis

Abstract: Liver fibrosis resulting from chronic liver injury are major causes of morbidity and mortality worldwide. Among causes of hepatic fibrosis, viral infection is most common (hepatitis B and C). In addition, obesity rates worldwide have accelerated the risk of liver injury due to nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH). Also liver fibrosis is associated with the consumption of alcohol, or autoimmune hepatitis and chronic cholangiophaties. The response of hepatocytes to inflammation plays a decisive role in the physiopathology of hepatic fibrosis, which involves the recruitment of both pro- and anti-inflammatory cells such as monocytes and macrophages. As well as the production of other cytokines and chemokines, which increase the stimulus of hepatic stellate cells by activating proinflammatory cells. The aim of this review is to identify the therapeutic options available for the treatment of the liver fibrosis, enabling the prevention of progression when is detected in time.

Effect of cigarette smoking on the structure of hepatocytes: TEM study / Efecto del tabaquismo sobre la estructura de los hepatocitos: estudio MET

The liver is one of the major organs that is indirectly affected by cigarette smoke. The aim of this project is to define the histologic and ultrastructural changes in normal liver cells after exposing animals to cigarette smoke. Thirty albino rats were exposed to cigarette smoke for 90 days, followed by morphologic examination of their livers under light microscope and electron microscope. The liver cells of cigarette smoke exposed rats showed mild swelling with increased eosinophilia. Ultrastructural examination of these cells demonstrated cytoplasm with highly proliferated and crowded mitochondria. There were many electron dense mitochondria. These mitochondria were pleomorphic in shape compared to mitochondrias of control rats. Also, loss of mitochondrial cristae and widening of the intermembranous space was noticed. It is concluded that smoking exerts cellular damage and oxidative stress on normal liver cells resulting in ultrastructural changes.

El hígado es uno de los principales órganos indirectamente afectado por el humo del cigarrillo. El objetivo de este trabajo fue definir los cambios histológicos y ultraestructurales de las células normales del hígado después de exponer a los animales al humo del cigarrillo. Treinta ratas albinas fueron expuestas al humo de cigarrillo durante 90 días, seguido de un examen morfológico de los hígados bajo microscopio de luz y microscopio electrónico. Las células hepáticas de las ratas expuestas al humo de cigarrillo mostraron una leve inflamación con un aumento de la eosinofilia. En el examen ultraestructural de estas células se observó el citoplasma mitocondrial altamente proliferado y saturado. Se observó gran cantidad de mitocondrias electrón-densas y éstas presentaban forma pleomórfica en comparación con las mitocondrias del grupo control. Además, se observó pérdida de las crestas mitocondriales y ensanchamiento del espacio intermembranoso. Se concluye que el tabaquismo ejerce daño celular y estrés oxidativo en las células hepáticas normales, lo que resulta en la aparición de cambios ultraestructurales.

A non-hepatotropic parasite infection increases mortality in the acetaminophen-induced acute liver failure murine model: possible roles for IL-5 and IL-6

We evaluated the effects of a non-hepatotropic parasite infection (Taenia crassiceps) on the outcome of acetaminophen-induced acute liver failure in mice. Uninfected and T. crassiceps infected mice orally received either 300 mg/kg acetaminophen or water as vehicle (n = 5 per group). Survival analysis, hepatocyte necrosis, alanine aminotransferase (ALT) levels, CYP2E1 protein, interleukin (IL-) 5, and IL-6 were assessed for all groups. All infected mice died within 16 h after exposure to acetaminophen (Tc+APAP group), whereas only one-third of uninfected animals exposed to acetaminophen (APAP group) died. Uninfected (Control group) and infected (Tc group) mice that received the vehicle showed no liver damage. Tc+APAP mice exhibited massive liver necrosis characterised by marked balloning degeneration of hepatocytes and higher serum ALT compared to Control, Tc, and APAP animals. Liver tissue from Tc+APAP mice also displayed increased expression of CYP2E1 protein and higher mRNA and protein levels of IL-5 and IL-6 compared to the other groups. These findings suggest that non-hepatotropic parasite infections may increase mortality following acute liver failure by promoting hepatocyte necrosis via IL-5 and IL-6-dependent CYP2E1 overproduction. This study identifies new potential risk factors associated with severe acute liver failure in patients.

Comparative immunohistochemical assessment of the effect of repetitive anesthesia with isoflurane and sevoflurane on rat liver / Avaliação imunohistoquímica comparativa do efeito da anestesia repetitiva com isoflurano e sevoflurano sobre o fígado de rato

Abstract Background and objectives: Inhalation anesthetics are used in human, as well as veterinary medical practice. In the present study we investigated the effect of isoflurane and sevoflurane on rat hepatocytes. Methods: A total of 40 Wistar female rats were used in this study. Animals were divided in groups of 5 rats. Groups IM, SM served as control groups. Groups I1, I2, I3 were used to study isoflurane and S1, S2, S3 for sevoflurane study. They were anesthetized 3 times, for 2 h long, at 2 days interval with a concentration of: 1.5% isoflurane (I1, I2, I3) and 2% sevoflurane (S1, S2, S3). The oxygen supply throughout the anesthesia was 1 L O2/min. Groups IM, IS, I1, S1 were sacrificed immediately after the last anesthesia. Groups I2, S2 were sacrificed 6 h after the last anesthesia, and groups I3, S3, 24 h post-anesthesia. Liver samples were harvested to highlight caspase-3 in apoptotic hepatocytes. Results: Following isoflurane administration, there were less than 1% cells in apoptosis highlighted in rat livers from groups IM, I1 and I2. At 24 h post-anesthesia (group I3), a small number of apoptotic hepatocytes was highlighted (around 3.23% cells in apoptosis), with a strictly periacinar disposition, randomly distributed in a small number of hepatic lobules. After sevoflurane administration, less than 1% apoptotic hepatocytes were identified at all control moments throughout the study. Conclusions: The results suggest that the anesthetics do not present a considerable hepatotoxicity. The comparative assessment of the two anesthetics shows that sevoflurane is superior to isoflurane.

Resumo Justificativa e objetivos: Anestésicos inalatórios são usados em humanos e também na prática médica veterinária. No presente estudo investigamos o efeito de isoflurano e sevoflurano em hepatócitos de rato. Métodos: Foram usados neste estudo 40 ratos Wistar fêmeas. Os animais foram divididos em grupos de cinco. Os grupos IM e SM serviram como controle. Os grupos I1, I2 e I3 foram usados para o estudo de isoflurano e os grupos S1, S2 e S3 para o estudo de sevoflurano. Os ratos foram anestesiados três vezes, durante duas horas em intervalos de dois dias, com uma concentração de 1,5% de isoflurano (I1, I2, I3) e 2% de sevoflurano (S1, S2, S3). O fornecimento de oxigênio durante a anestesia foi de 1 L O2/min. Os grupos IM, IS, I1 e S1 foram sacrificados imediatamente após a última anestesia. Os grupos I2 e S2 foram sacrificados seis horas após a última anestesia e os grupos I3 e S3 foram sacrificados 24 horas após a anestesia. Amostras dos fígados foram colhidas para ressaltar a caspase-3 em hepatócitos apoptóticos. Resultados: Após a administração de isoflurano, havia menos de 1% das células em apoptose em destaque nos fígados dos ratos dos grupos IM, I1 e I2. Às 24 horas após a anestesia (grupo I3), um pequeno número de hepatócitos apoptóticos foi destacado (3,23% de células em apoptose), com uma disposição estritamente periacinar, distribuídos aleatoriamente em um pequeno número de lóbulos hepáticos. Após a administração do sevoflurano, menos de 1% de hepatócitos apoptóticos foi identificado em todos os momentos de controle ao longo do estudo. Conclusões: Os resultados sugerem que os anestésicos não apresentam uma hepatotoxicidade considerável. A avaliação comparativa dos dois anestésicos mostra que sevoflurano é superior ao isoflurano.

Diferentes níveis protéicos na dieta de girinos de rã-touro (Lithobates catesbeianus) e seus efeitos sobre o tecido hepático / Different protein levels in the diet of bullfrog tadpoles (Lithobates catesbeianus) and its effects on the liver tissue

Para verificação da ocorrência de alterações morfológicas no fígado, foram coletados dez animais de cada dieta, no 30º dia (1º período experimental) e no 60º dia (2º período experimental), sendo esse, o último dia do período experimental e início do clímax da metamorfose. Os girinos foram distribuídos em delineamento experimental inteiramente casualizado (DIC) com três tratamentos: dieta experimental com 32,68% de proteína bruta e duas dietas comerciais com 37,92% e 57,53% de proteína bruta. Os órgãos coletados foram fixados em solução de Bouin e depois submetidos às práticas da rotina histotecnológica e coradas com hematoxilina - Eosina (HE). Na avaliação microscópica do fígado foram encontradas diversas alterações morfológicas como: Desorganização e vacuolização de hepatócitos, infiltrado inflamatório mononuclear periportal, multifocal e difuso, infiltrado inflamatório eosinofílico, congestão, hemorragia, hemólise e necrose. As alterações hepáticas encontradas neste estudo sugerem que as dietas utilizadas para os girinos de rã-touro, não atenderam suas necessidades dietéticas, afetando a homeostasia dos mesmos, comprometendo assim, sua sanidade.(AU)

To verify the occurrence of liver morphological changes, bullfrog tadpoles were fed three diets: an experimental diet with 32.68% crude protein and two commercial diets with 37.92% and 57.53% crude protein. Tadpoles were distributed into a completely randomized design (CRD) with three treatments and four replications. Liver samples were collected twice, on the 30th and 60th day of the experiment. Tadpole livers were fixed in Bouin's solution, then subjected to routine histotechnology procedures, and stained with hematoxylin - eosin (HE). Microscopic evaluation of liver tissue showed several morphological changes like disorganization and vacuolization of hepatocytes; periportal, multifocal and diffuse mononuclear inflammatory infiltrate, eosinophilic inflammatory infiltration, congestion, hemorrhage, hemolysis and necrosis. The liver changes found in this study suggest that diets used for bullfrog tadpoles, did not meet their dietary needs, what affected their homeostasis, thus compromising their health.(AU)

Can the intermittent training generate alterations on the liver tissue of rats submitted to a hyperlipidic diet? / ¿El entrenamiento intermitente puede generar alteraciones sobre el tejido hepático de ratas sometidas a una dieta alta en grasa?

Studies indicate that increasing physical activity and decreasing levels of fat in the liver help to decrease the risk of morbidity from liver conditions. This study aimed to evaluate the effects of an eight week intermittent training program on the liver tissue of rats subjected to a hyperlipidic diet. The study consisted of 30 male Wistar rats, divided into the following groups: Sedentary Control (SC) Exercise Control (EC) (fed on standard feed and water) Sedentary Obese (SO) and Exercise Obese (EO) (fed on bacon, ham, sausage, biscuits, soda and standard feed), which performed intermittent training through electrically stimulated jumps, with three sets of 12 repetitions, three times per week for eight weeks. At the end of the training period, the animals were euthanized, and their livers removed for histological processing and hematoxylin and eosin staining (HE). Soon afterwards, caryometric analysis of the hepatocyte nuclei was performed. From the presented results it can be seen that the hepatocyte nuclei of the obese animals were smaller in relation to those of the control animals, therefore, exercise combined with an appropriate diet proved to be efficient in not causing alterations in the hepatocyte nuclei, conserving normal cell function and reducing the chances of the appearance of tissue damage. Furthermore, exercise in isolation cannot be considered a protective factor against the alteration of the liver cells.

Estudios indican que el aumento de la actividad física y la disminución de los niveles de grasa en el hígado ayudan a reducir el riesgo de morbilidad por enfermedades hepáticas. El objetivo fue evaluar el efecto de un programa de entrenamiento intermitente de ocho semanas en el tejido hepático de ratas con dieta alta en grasa. Treinta ratas Wistar machos fueron divididas en grupos Control Sedentario (SC), Control Ejercicio (CE) (con fuente de alimentación que consiste en la ración estándar y agua), Obeso Sedentario (OS) y Obeso Ejercicio (OE) (con alimentación compuesta por tocino, mortadela, salchichas, galletas, refrescos y ración estándar), sometidas a un entrenamiento intermitente a través de saltos mediante estimulación eléctrica con tres series de 12 repeticiones, tres veces por semana durante ocho semanas. Al final del período de entrenamiento, los animales fueron sacrificados; se extrajeron sus hígados para el procesamiento histológico y tinción con hematoxilina y eosina (HE). Luego se realizó el análisis cariométrico de los núcleos de los hepatocitos. Se observó que los núcleos de los hepatocitos fueron menores en los animales obesos en comparación con los núcleos de hepatocitos de los animales de control, pues el ejercicio combinado con una dieta adecuada se mostró eficiente para que causar alteraciones en los núcleos de hepatocitos, y esa combinación puede retener la función normal de las células y disminuir las posibilidades de la aparición de daños en los tejidos. Además, el ejercicio aislado no puede ser considerado como un factor de protección contra la alteración las células del hígado.

Morfometría nuclear de hepatocitos esteatósicos y esteatósicos estimulados con laser infrarrojo / Nuclear morphometric of the steatotic hepatocytes and stimulated steatotisc with infrared laser

Un total de 24 ratas hembras de 4 meses de vida con peso aproximado de 250 g recibieron una solución de alcohol 40 % disuelto en agua lo cual derivó en una esteatosis alcohólica multivesicular. A 12 de estas ratas esteatósicas se le aplicó estimulaciones de láser infrarrojo con dosis de 8 J/cm2 durante 15 días consecutivos. Posteriormente las ratas fueron sacrificadas y se extrajeron muestras tanto de hígado esteatósico como del estimulado para enseguida ser procesadas para microscopía electrónica de transmisión. De hepatocitos esteatósicos y esteatósicos estimulados se obtuvieron microfotografías electrónicas de transmisión con aumentos finales de 9.500 X, las cuales fueron sometidas a estudios morfométricos para determinar tanto el número de poros nucleares como de fracciones volumétricas de los siguientes componentes celulares: Areas celular y nuclear, fracciones volumétricas de núcleos y nucléolos, eu y heterocromatina. De igual manera se determinó la relación nucleo-citoplasmática de ambos tipos celulares. Del análisis de los resultados entre hepatocitos alcohólicos y alcohólicos estimulados se visualiza que existen notables diferencias en todos los componentes celulares cuantificados. Se concluye que los efectos de la estimulación con lásr infrarrojo provoca en los hepatocitos una drástica transformación en su ultraestructura y en su morfología, situación que se traduciría, por ende, en una variación funcional, representando de esta manera el efecto que dicha estimulación provoca en los hepatocitos.

A total of 24 female rats, aged 4 months and weighing approximately 250 g and they given a solution of 40 % alcohol dissolved in water, leading to alcoholic multivesicular steatosis and 12 of rats was given and infrared laser with dose of 8 J/cm2 during 15 d. The rats were then killed and samples of steatosis and stimulated and were taken and processed for examination by transmission electron microscope. Transmission electron microscope microphotographs steatotic hepatocytes and stimulated steatotic were obtained with final magnification of 9,500 X. They were subjected to morphometric studies to determine the number of nuclear pores and volumetric fractions and areas the following components: cellular and nuclear area, volumetric fractions of nucleus, nucleolus, eu and heterochromatin, nucleocytoplamic ratio of each cell type was determined. Analysis of the results between alcoholic hepatocytes and stimulate alcoholic shows that noticeable differences exist in all the cell components quantified. It is concluded that the effects of the stimuli of laser infrared provoke in the hepatocytes, a drastic transformation of their ultrastructure and morphology. This finally leads to functional variations, representing the effects produced by this stimulate in the hepatocytes.

Ultrastructure of fetal mice hepatocytes exposed in utero to diazepam / Ultraestructura de hepatocitos fetales de ratón expuestos in utero a diazepam

Previous research has shown that fetal mice hepatic cells from females treated with diazepam (Valium) during pregnancy depict cytoplasmic and nuclear modifications when observed with photonic microscope. The purpose of this work is to investigate if diazepam administered subcutaneously (SC) to pregnant mice females induces ultraestructural alterations in the cytoplasmic organelles and nucleus to fetal hepatocytes. Transmission electron microscopy observations of fetal hepatocytes from pregnant females treated with a single daily dose of diazepam 2.7 mg/kg/bw/SC administered from 6th to 15th days of gestation revealed that they frequently presented disorganized and dilated rough endoplasmic reticulum cisterns, membranous elements, abundant Golgi complex and glycogen granules, around large vacuoles. The voluminous nucleus shows atypical distribution of chromatin. These alterations could modify the hepatocyte's physiology and probably persist after birth.

Estudios previos muestran que las células hepáticas de fetos de ratón, de hembras tratadas con diazepam (Valium) durante la gestación, presentan modificaciones citoplásmicas y nucleares que se pueden observar con el microscopio fotónico, por lo que el propósito de este trabajo es determinar si el diazepam administrado por vía subcutánea (SC) a hembras gestantes de ratón, induce alteraciones ultraestructurales de los organelos citoplásmicos y del núcleo de los hepatocitos fetales. En los fetos de ratón del grupo experimental de hembras gestantes, tratadas con dosis únicas diarias de 2,7 mg/kg de peso corporal administradas por vía SC del 6° al 15° día de la gestación, se observó con el microscopio electrónico de transmisión que los hepatocitos fetales presentaban con frecuencia retículo endoplásmico rugoso desorganizado, con cisternas dilatadas; había elementos membranosos y complejo de Golgi abundante, al igual que gránulos de glucógeno que rodeaban a grandes vacuolas. Los núcleos eran voluminosos, con la cromatina distribuida atípicamente. Estas alteraciones podrían modificar la fisiología de los hepatocitos y probablemente persistan después del nacimiento.

Depression as a Clinical Determinant of Dependence and Low Quality of Life in Elderly Patients with Cardiovascular Disease / Depressão como Determinante Clínico de Dependência e Baixa Qualidade de Vida em Idosos Cardiopatas

Background: The aging process promotes a progressive increase in chronic-degenerative diseases. The effect of these diseases on the functional capacity has been well recognized. Another health parameter concerns “quality of life related to health”. Among the elderly population, cardiovascular diseases stand out due to the epidemiological and clinical impact. Usually, these diseases have been associated with others. This set of problems may compromise both independence and quality of life in elderly patients who seek cardiologic treatment. These health parameters have not been well contemplated by cardiologists. Objective: Evaluating, among the elderly population with cardiovascular disease, which are the most relevant clinical determinants regarding dependence and quality of life. Methods: This group was randomly and consecutively selected and four questionnaires were applied: HAQ, SF-36, PRIME-MD e Mini Mental State. Results: The study included 1,020 elderly patients, 63.3% women. The group had been between 60 and 97 years-old (mean: 75.56 ± 6.62 years-old). 61.4% were independent or mild dependence. The quality of life total score was high (HAQ: 88.66 ± 2.68). 87.8% of patients had a SF-36 total score > 66. In the multivariate analysis, the association between diagnoses and high degrees of dependence was significant only for previous stroke (p = 0.014), obesity (p < 0.001), lack of physical activity (p = 0.016), osteoarthritis (p < 0.001), cognitive impairment (p < 0.001), and major depression (p < 0.001). Analyzing the quality of life, major depression and physical illness for depression was significantly associated with all domains of the SF-36. Conclusion: Among an elderly outpatient cardiology population, dependence and quality of life clinical determinants are not cardiovascular comorbidities, especially the depression. .

Fundamento: Com o envelhecimento, a prevalência de doenças crônico-degenerativas sofreu aumento progressivo. A repercussão dessas doenças sobre a capacidade funcional foi reconhecida. Outro parâmetro de saúde é a “qualidade de vida relacionada à saúde”. Na população idosa, as doenças cardiovasculares destacam-se pelo impacto epidemiológico e clínico. Elas, geralmente, vêm associadas a outras afecções. Esse conjunto de problemas pode comprometer a independência e a qualidade de vida do idoso que busca tratamento cardiológico. Objetivo: Avaliar, em uma população de idosos cardiopatas, quais são os determinantes clínicos mais relevantes de dependência e de qualidade de vida. Métodos: O grupo foi selecionado aleatória e consecutivamente, sendo aplicados quatro questionários: HAQ, SF-36, PRIME‑MD e Mini Exame do Estado Mental. Resultados: Incluiu-se 1020 idosos, 63,3% mulheres. O grupo tinha em média 75,56 ± 6,62 anos. 61,4% mostrou-se independente ou com dependência leve. O escore de qualidade de vida foi elevado (HAQ: 88,66 ± 2,68). 87,8% dos pacientes apresentou escore total do SF-36 ≥ 66. À análise multivariada, a associação entre os diagnósticos e graus elevados de dependência foi significante apenas para acidente vascular cerebral prévio (p = 0,014), obesidade (p < 0,001), sedentarismo (p = 0,016), osteoartrite (p < 0,001), déficit cognitivo (p < 0,001), e depressão maior (p < 0,001). Ao analisarmos a qualidade de vida, a depressão maior e a depressão por doença física associou-se significativamente com todos os domínios do SF-36. Conclusão: Em uma população de idosos cardiopatas, os determinantes clínicos mais relevantes de prejuízos para dependência e qualidade de vida foram as comorbidades não cardiovasculares, particularmente a depressão. .

Percepção dos idosos jovens e longevos gaúchos quanto aos espaços públicos em que vivem / The perception of the young and long-lived elderly ‘Gauchos’ (from the State of Rio Grande do Sul, Brazil) about the public spaces they live in Resumo

Em 2050, o número de brasileiros residindo em áreas urbanas passará dos 200 milhões e 29% da população será composta por idosos. Os idosos longevos possuem 80 anos ou mais, os idosos jovens são aqueles que apresentam idade entre 60 e 79 anos. O objetivo foi verificar a diferença da percepção de idosos jovens e longevos do Rio Grande do Sul quanto ao ambiente urbano em que vivem. Estudo de base populacional, observacional, descritivo, retrospectivo, com paradigma de análise quantitativa, analisou dados da pesquisa Perfil dos Idosos do RS, realizado pelo Instituto de Geriatria e Gerontologia da PUCRS em parceria com a Escola de Saúde Pública do RS. A amostra foi composta por 6913 questionários respondidos por idosos de 59 cidades. A análise dos dados foi realizada com cada um dos grupos etários e as variáveis independentes foram testadas pelo Qui-Quadrado, sendo o nível de significância menor que 0,05. Como resultado, a percepção de dificuldades, como poucos bancos, falta de faixas de segurança, tempo de sinal muito curto para pedestres, degraus muito altos e mau cheiro dos banheiros públicos, foi maior entre os idosos jovens. Os idosos longevos perceberam menos esses fatores, porém, referiram que frequentam menos os ambientes comunitários.

By 2050, the number of Brazilians living in urban areas will be over 200 million and 29% of the population will be elderly. The long-lived elderly are 80 or more years old and the young elderly are between 60 and 79 years of age. The scope of this article was to verify the difference in perception between the young elderly and the long-lived elderly from Rio Grande do Sul (RS) about the urban environment they live in. This is a population-based, observational, descriptive, retrospective study with a quantitative analysis paradigm. Data was analyzed from Elderly Profile research in RS conducted by the Geriatric and Gerontological Institute of PUCRS in partnership with the RS School of Public Health. The sample consisted of 6913 questionnaires answered by the elderly from 59 cities. Data analysis was performed for each age group and independent variables were processed using the Chi-square test, with p under 0.05. Results showed that the perception of difficulties such as a lack of park benches and safety strips, short traffic light times for pedestrians, high steps and bad-smelling public toilets was greater among the young elderly. The long-lived elderly noticed these facts less, though they admitted that they frequent community environments less often.

Instrumento para avaliação da qualidade do Time de Resposta Rápida em um hospital universitário público / Instrument for assessing the quality of the Rapid Response Team at a university public hospital / Instrumento para la evaluación de la calidad del Equipo de Respuesta Rápida en un hospital universitario público

O artigo descreve a construção de um questionário para avaliação da qualidade do atendimento de um Time de Resposta Rápida, em Hospital Universitário de Londrina-PR, fundamentado no modelo conceitual de Donabedian (estrutura-processoresultado). A coleta de dados ocorreu no mês de março de 2012 e o processo de adequação do questionário foi desenvolvido com a aplicação da Técnica Delphi com a participação de 15 especialistas. Ao término do estudo obteve-se um questionário com 37 enunciados, sendo alcançado índice de concordância final na pesquisa com valores superiores a 80% para todos os conceitos. Espera-se que as contribuições do grupo de especialistas tornem o instrumento confiável e seja aplicado em outros serviços semelhantes. Aplicações futuras deste instrumento poderão trazer subsídios para melhor avaliação da qualidade dos serviços de equipes de Resposta Rápida.

The paper describes the construction of a questionnaire to assess the quality of care of a Rapid Response Team at the University Hospital of Londrina, based on the conceptual model of Donabedian (structure-process-outcome). Data collection occurred in March 2012 and the process of adjusting the questionnaire was developed with the application of the Delphi technique involving 15 experts. At the end of the study the questionnaire contained 37 statements, achieving final compliance level higher than 80% in all concepts. It is hoped that the contributions of the expert group produce a more reliable questionnaire to be applied in other similar services. Future applications of this instrument may provide information to better assess the quality of teams of Rapid Response services.

El artículo describe la construcción de un cuestionario para la evaluación de la calidad de la atención de un Equipo de Respuesta Rápida en un Hospital Universitario de Londrina, Paraná, basado en el modelo conceptual de Donabedian (estructura-procesoresultado). La recolección de datos ocurrió durante el mes de marzo de 2012 y el proceso de ajuste del cuestionario fue desarrollado por medio de la Técnica Delphi con la participación de 15 especialistas. Al término del estudio se obtuvo un cuestionario con 37 enunciados, alcanzándose un índice de concordancia final en la investigación con valores superiores al 80% para todos los conceptos. Se espera que las contribuciones del grupo de especialistas afiancen la confiabilidad del instrumento y que el cuestionario sea utilizado en otros servicios semejantes. Las aplicaciones futuras podrán traer subsidios para evaluar mejor la calidad de los servicios de los equipos de Respuesta Rápida.

Análisis morfométrico de hepatoesteatosis microvesicular alcohólica de ratas / Morphometric analysis of alcoholic microvesicular hepatic steatosis in rats

Un total de 24 ratas hembras de 4 meses de vida con peso aproximado de 250 gramos fueron divididas en dos grupos de animales rotulados como A y B. El grupo A se mantuvo con pellet y agua ad libitum sirviendo como controles mientras que el grupo B conservaba el pellet y recibía una solución de alcohol 40% disuelto en agua lo cual derivó en una esteatosis alcohólica multivesicular. Ambos grupos se mantuvieron en estas condiciones por 60 días. Posteriormente las ratas fueron sacrificadas y se extrajeron muestras tanto de hígado normal-control como de hígado graso para enseguida ser procesadas para microscopía electrónica de transmisión. De hepatocitos normales y esteatósicos se obtuvieron microfotografías electrónicas de transmisión con aumentos finales de 9.500X, las cuales fueron sometidas a estudios morfométricos para determinar fracciones volumétricas de los siguientes componentes celulares: Retículo endoplasmático rugoso, mitocondrias, inclusiones lipídicas y de glicógeno, eu y heterocromatina. De igual manera se cuantificaron las áreas celulares y nucleares. Del análisis de los resultados entre hepatocitos normales y alcohólicos se visualiza que existen notables diferencias en todos los componentes celulares cuantificados. Se concluye que los efectos de la ingesta diaria de alcohol provoca en los hepatocitos una esteatosis microvesicular que genera una drástica transformación en su ultraestructura y en su morfología, situación que se traduciría, por ende, en una variación funcional, representando de esta manera el efecto que esta droga provoca en los hepatocitos.

A total of 24 female rats, aged 4 months and weighing approximately 250 g, were divided into two groups, called A and B. The group A animals were kept on pellets and water ad libitum and served as controls, while group B animals were fed pellets and given a solution of 40% alcohol dissolved in water, leading to alcoholic multivesicular steatosis. Both groups were kept under these conditions for 60 days. The rats were then euthanized and samples of normal-control and fatty liver were taken and processed for examination by transmission electron microscope. Transmission electron microscope microphotographs of normal and steatotic hepatocytes were obtained with final magnification of 9,500 X. They were subjected to morphometric studies to determine the volumetric fractions of the following cell components: Rough Endoplasmic Reticulum, mitochondria, lipid and glycogen inclusions, and eu- and heterochromatin. In addition, the cell and nucleus areas were quantified and the nucleo cytoplasmic ratio of each cell type was determined. Analysis of the results between normal and alcoholic hepatocytes shows that noticeable differences exist in all the cell components quantified. It is concluded that the effects of the daily consumption of alcohol provoke microvesicular steatosis in the hepatocytes, generating a drastic transformation of their ultrastructure and morphology. This finally leads to functional variations, representing the effects produced by this drug in the hepatocytes.

Mollusc C-reactive protein crosses species barrier and reverses hepatotoxicity of lead in rodent models.

Achatina fulica C-reactive protein (ACRP) reversed the toxic effects of lead nitrate both in vivo in mice and in vitro in rat hepatocytes restoring the basal level of cell viability, lipid peroxidation, reduced glutathione and superoxides. Cytotoxicity was also significantly ameliorated in rat hepatocytes by in vitro pre-treatments with individual subunits (60, 62, 90 and 110 kDa) of ACRP. Annexin V-Cy3/CFDA dual staining showed significant reduction in the number of apoptotic hepatocytes pre-treated with ACRP. ACRP induced restoration of mitochondrial membrane potential was remarkable. ACRP pre-treatment prevented Pb-induced apoptosis mediated by caspase activation. The antagonistic effect of ACRP may be due to scavenging of reactive oxygen species which maintained the homeostasis of cellular redox potential as well as reduced glutathione status. The results suggest that ACRP crosses the species barrier and it may be utilized as a viable exogenous agent of cytoprotection against heavy metal related toxicity.

[Characteristics of hepatocyte rough endoplasmic reticulum single cationic channel in streptozocin-induced diabetic rats]

The role of ion channels and particularly cationic channels in the pathogenesis of various diseases are being considered carefully. The diabetes mellitus is a common disease which is initiated by ion channel disturbances. This study was done to determine the characteristics of hepatocyte rough endoplasmic reticulum single cationic channel in Streptozocin-induced diabetic rats. This experimental study was done on 10 male adult Wistar rats and animals were randomly allocaied into diabetic and control groups. Diabetes induced by STZ [65 mg/kg/bw] intraperitounally. Rough endoplasmic reticulum vesicles were extracted following rat liver excision, homogenization and ultracentrifuging. The bilayer membrane formation was prepared by painting phosphatidylcholine on 250 micro M aperture in between Cis and Trans sides. The RER vesicles incorporation was performed through gentle and delicate touch of membrane using a dentistry needle. The Pclamp9 software was used for ion channel activity characteristic analysis. The cationic channel current amplitude did not change significantly in voltages more than+3o mV but their open probability [Po] decreased in diabetic group [P<0.05]. More severe changes in channel activity were seen in potentials less than the reverse potential. In addition to significant increase of channel Po [P<0.05], also, the channel unitary currents were significantly decreased [P<0.05]. The mean current amplitude and channel open probability in voltage+40 mV were 17 +/- 2.14 pA and 0.68 +/- 0.01 in control group respectively, whereas, the values of these parameters reached to 18.5 +/- 2.5 and 0.26 +/- 0.03, respectively. In voltage-10 mV, the values of mean current amplitude and Po were-22.3 +/- 2.14 pA and<0.1 in control group, respectively but the values changed to-13.1 +/- 0.08 and 0.62 +/- 0.03 in diabetic group. It seems that RER cationic channel is involved in metabolic changes which cause by diabetes mellitus and this disease can cause probably a channel gating kinetic and behavior change by inducing metabolic stresses

Características estereológicas del hígado de rata (Rattus norvergicus) sometidas a menopausia inducida por ovariectomía / Stereologic characteristics of the liver of rats (Rattus norvergicus) submitted to ovariectomy-induced menopause

Durante la menopausia ocurren cambios hormonales, como disminución de estradiol y aumento de las hormonas folículo estimulante y luteinizante. En mujeres premenopáusicas la fibrosis y esteatosis hepática es menor que en mujeres postmenopáusicas, mejorando cuando se administra terapia hormonal de reemplazo. El objetivo del estudio fue determinar aspectos cuantitativos del hígado de ratas sometidas a un modelo experimental de menopausia inducida por ovariectomía. Utilizamos 10 ratas Sprague-Dawley (Rattus norvergicus) adultas, sanas. Las ratas fueron pesadas y divididas en dos grupos (ovariectomía bilateral=Grupo Ovx y control=Grupo Sham). A los 60 días fueron sacrificadas y retirado el hígado, obteniéndose un trozo de cada hígado. Se realizaron 5 cortes de 4 um y se tiñeron con HE. Se analizaron 5 campos por lámina. Se usó el test M42. Los parámetros estereológicos considerados fueron: Nv, Vv, Sv y número total (Nt) de hepatocitos y sinusoides hepáticos. El peso promedio del grupo Ovx y Sham fue 300,8 g y 285,8 g respectivamente. La Nv de hepatocitos de ratas sometidas a cirugías Sham (CS) y Ovx (CO) fue de 6,89 x105/mm3 y 7,70 x105/mm3, respectivamente. La Vv de hepatocitos CS y CO fue de 71,92 por ciento y 80,48 por ciento, respectivamente. La Sv de hepatocitos CS y CO fue de 260,61 mm2/mm3 y 273,64 mm2/mm3, respectivamente. El volumen promedio de hígados CS fue de 8,40 mm3 y el Nt de hepatocitos promedio fue 5,79 x 106. El volumen promedio de los hígados CO fue 9,28 mm3 y el Nt de 7,15 x 106. Respecto a los sinusoides, la Nv en el hígado CS y CO fue de 2,81 x105/mm3 y 2,64 x105/mm3, respectivamente. La Vv de sinusoides en CS y CO fue de 16,74 por ciento y 16,46 por ciento, respectivamente. La Sv de sinusoides en ratas CS y CO fue de 49,40 mm2/mm3 y 45,21 mm2/mm3, respectivamente. El Nt de sinusoides CS y CO fue en promedio 2,37 x 106 y 2,45 x 106, respectivamente. La ausencia de estrógeno puede generar fibrosis y esteatosis hepática como lo señala...

During menopause, hormonal changes such as decreased estradiol and increased follicle stimulating and luteinizing hormones occur. In premenopausal women fibrosis and hepatic steatosis is less than in postmenopausal women and improves when hormone replacement therapy is administered. The aim of this study was to determine quantitative aspects of the liver of rats undergoing an experimental model of ovariectomy-induced menopause. We used 10 Sprague-Dawley (Rattus norvergicus) healthy adults. The rats were weighed and divided into two groups (bilateral oophorectomy = Group Ovx and control = Group Sham). The animals were sacrificed at 60 days and the liver removed, obtaining a piece of each liver. Five cuts were made of 4 microns and stained with HE. We analyzed five fields per slide. M42 test was used. Stereological parameters considered were as follows: Nv, Vv, Sv and total number (Tn) of hepatocytes and sinusoids. The average weight of Ovx and Sham group was 300.8 g and 285.8 g, respectively. The Nv of hepatocytes of rats subjected to sham surgery (CS) and Ovx (CO) was 6.89 and 7.70 x105/mm3 x105/mm3, respectively. The Vv of CS and CO hepatocytes was 71.92 percent and 80.48 percent, respectively. SV and CO, CS hepatocytes was 260.61mm2/mm3 and 273.64 mm2/mm3 , respectively. The average volume of CS livers was 8.40 mm3 and hepatocyte Tn averaged 5.79 x 106. The average volume was 9.28 mm3 CO livers and Tn of 7.15 x 106. Regarding the sinusoids, the Nv in the liver CS and CO was 2.81 x105/mm3 and 2.64 x105/mm3, respectively. The Vv of sinusoids in CS and CO was 16.74 percent and 16.46 percent, respectively. The sinusoids in rats Sv CS and CO was 49.40 mm2/mm3 and 45.21 mm2/mm3, respectively. The sinusoids Tn CS and CO averaged 2.37 x 106 and 2.45 x 106, respectively. The lack of estrogen can cause fibrosis and hepatic steatosis as noted by other authors, but also changes in the organization and the proportion of the components of the liver, factors to ...

Histopathological effects of aloe barbadensis and soybean Oil on rat liver / Efectos histopatológicos de aloe barbadensis y aceite de soya en hígado de rata

Aloe Barbadensis, which is a species of Aloe vera, is a popular plant used by the common people and in alternative medicine. This study aimed to analyze the effects of Aloe Barbadensis and soybean oil on liver. For this study Wistar Albino female rats were taken and divided into 3 equal groups; the first group was the control group wherein no treatment was applied, second group in which the dissolved form of A. barbadensis in the soybean oil was applied (25 mg/day), and the third group which only soybean oil was applied (500 mg/day). Biopsy materials were taken from the lobus dexter of the livers of the rats and analyzed with light microscope after the necessary standard processing of histologic slides. Group I demonstrated normal structural characteristics of rat liver. In Group II and Group III, we observed nuclear enlargement, mild increase in chromatin and hydropic degeneration and binucleation in some hepatocytes. Liver histology demonstrated congestion in portal veins, sinusoids and the central veins. Merely in Group III, portal venous congestion and in Group II sinusoidal congestion was evident parenchyma of the liver. Additionally in Group III liver histology demonstrated plasmocyte infiltration in portal areas. Our study showed that using soybean with Aloe Barbadensis is synergystic and increasing each others effects. However we didn't observe mononeuclear infiltrations in Group II, these show antinflammatory effects of Aloe Barbadensis. It is determined that, depending on the used dose of Aloe Barbadensis, the toxic effect can change. If Aloe Barbadensis used very high doses it can have toxic effect on hepatocytes.

Aloe Barbadensis, una especie de Aloe vera, es una planta popular usada por el común de las personas y también en la medicina alternativa. El estudio tuvo como objetivo analizar los efectos del Aloe Barbadensis y aceite de soya en el hígado. Para el estudio se emplearon ratas Wistar hembras Albino y se dividieron en 3 grupos: grupo control I sin tratamiento; grupo II A. barbadensis disuelta en aceite de soja (25 mg / día), y grupo III tratado sólo con aceite de soja (500 mg / día). Fueron extraídas biopsias del lóbulo derecho del hígado de las ratas y luego se analizaron con microscopio de luz. En el grupo I el hígado de las ratas era normal. En los grupos II y III, se observó aumento del tamaño nuclear, leve aumento de la cromatina y degeneración hidrópica y binucleación en algunos hepatocitos. La histología hepática mostró la congestión en las venas porta, sinusoides y las centrales. En el grupo III, la congestión venosa portal y en el Grupo II la congestión sinusoidal fue evidente. Además, el Grupo III reveló infiltración de plasmocitos en áreas portales. El uso de soja con Aloe Barbadensis es sinergista y aumenta cada uno de otros efectos. Infiltraciones mononucleares en el grupo III determinan la reacción inflamatoria. Sin embargo, no observamos infiltración mononuclear en el Grupo II, éste mostró efectos antiinflamatorios de la Aloe Barbadensis. Esto determina que, dependiendo de la dosis usada de Aloe Barbadensis, los efectos tóxicos pueden cambiar. Si es usado en altas dosis Aloe Barbadensis puede producir efectos tóxicos en los hepatocitos.