A dominant negative mutation uncovers cooperative control of caudal Wolffian duct development by Sprouty genes.

The Wolffian ducts (WD) are paired epithelial tubules central to the development of the mammalian genitourinary tract. Outgrowths from the WD known as the ureteric buds (UB) generate the collecting ducts of the kidney. Later during development, the caudal portion of the WD will form the vas deferens, epididymis and seminal vesicle in males, and will degenerate in females. While the genetic pathways controlling the development of the UB are firmly established, less is known about those governing development of WD portions caudal to the UB. Sprouty proteins are inhibitors of receptor tyrosine kinase (RTK) signaling in vivo. We have recently shown that homozygous mutation of a conserved tyrosine (Tyr53) of Spry1 results in UB defects indistinguishable from that of Spry1 null mice. Here, we show that heterozygosity for the Spry1 Y53A allele causes caudal WD developmental defects consisting of ectopically branched seminal vesicles in males and persistent WD in females, without affecting kidney development. Detailed analysis reveals that this phenotype also occurs in Spry1+/- mice but with a much lower penetrance, indicating that removal of tyrosine 53 generates a dominant negative mutation in vivo. Supporting this notion, concomitant deletion of one allele of Spry1 and Spry2 also recapitulates the genital phenotype of Spry1Y53A/+ mice with high penetrance. Mechanistically, we show that unlike the effects of Spry1 in kidney development, these caudal WD defects are independent of Ret signaling, but can be completely rescued by lowering the genetic dosage of Fgf10. In conclusion, mutation of tyrosine 53 of Spry1 generates a dominant negative allele that uncovers fine-tuning of caudal WD development by Sprouty genes.

Hepatitis C virus micro-elimination in vulnerable populations before and during a global pandemic.

OBJECTIVE: to analyze a hepatitis C virus (HCV) microelimination strategy targeting vulnerable populations and the influence of the pandemic on its maintenance and outcomes. METHODS: in 2018, the Hepatology Unit implemented an HCV microelimination strategy for patients attending drug addiction care centers and Psychiatry Units such as the Alcoholism Treatment Unit. These centers reported suspected or confirmed cases of HCV infection directly to the hepatologists, who, after reviewing the clinical records, cite those patients if necessary. RESULTS: from June 2018 to February 2020, hepatologists were consulted on 37 anti-HCV positive patients, 31 of them were from Drug Addiction Care Centers, 5 from the Alcoholism Disorders Unit and 1 from the Department of Psychiatry. Fibrosis stage: F0-F1, 18 (50%); F2, 9 (25%); F3, 2 (4.2%); F4, 8 (20.8%). Female sex, 6 (16.7%). Required attending: Yes, 27 (73%). 25 (92.6%) went. Outpatient consultation: 10 (27%). Required treatment: 19 (51.3%). Sustained virological response: 19 (100%). Fibrosis stage of treated patients: F0-F1, 13 (68%); F2, 2 (11%); F3, 1 (5%); F4, 3 (16%). Patients from the Psychiatry Department during 2021: 11. 9 (82%) do not need to attend; 2 (18%): their situation is unknown. CONCLUSIONS: sustained communication with centers that care for populations at risk of active HCV infection is needed to detect cases, increase adherence to treatment, and rescue patients who require screening for hepatocarcinoma.

Sprouty1 Controls Genitourinary Development via its N-Terminal Tyrosine.

BACKGROUND: Studies in mice suggest that perturbations of the GDNF-Ret signaling pathway are a major genetic cause of congenital anomalies of the kidney and urinary tract (CAKUT). Mutations in Sprouty1, an intracellular Ret inhibitor, results in supernumerary kidneys, megaureters, and hydronephrosis in mice. But the underlying molecular mechanisms involved and which structural domains are essential for Sprouty1 function are a matter of controversy, partly because studies have so far relied on ectopic overexpression of the gene in cell lines. A conserved N-terminal tyrosine has been frequently, but not always, identified as critical for the function of Sprouty1 in vitro. METHODS: We generated Sprouty1 knockin mice bearing a tyrosine-to-alanine substitution in position 53, corresponding to the conserved N-terminal tyrosine of Sprouty1. We characterized the development of the genitourinary systems in these mice via different methods, including the use of reporter mice expressing EGFP from the Ret locus, and whole-mount cytokeratin staining. RESULTS: Mice lacking this tyrosine grow ectopic ureteric buds that will ultimately form supernumerary kidneys, a phenotype indistinguishable to that of Sprouty1 knockout mice. Sprouty1 knockin mice also present megaureters and vesicoureteral reflux, caused by failure of ureters to separate from Wolffian ducts and migrate to their definitive position. CONCLUSIONS: Tyrosine 53 is absolutely necessary for Sprouty1 function during genitourinary development in mice.

Analytical, anthropometric and dietary factors associated with the development of fibrosis in patients with nonalcoholic fatty liver disease.

BACKGROUND: a prolonged non-alcoholic steatohepatitis (NASH) condition can lead to advanced stages of liver disease and the development of hepatocellular carcinoma. AIM: to evaluate analytical, anthropometric and dietary factors associated with the presence of fibrosis as this is the factor that most influences survival and evolution. METHODS: seventy-six patients with liver biopsy-diagnosed non-alcoholic fatty liver disease (NAFLD) were included. Biopsies were scored considering the NASH criteria of Kleiner. Analytical, anthropometric and dietary (survey) parameters were obtained. NAFLD-FS is a non-invasive fibrosis index and was assessed for each patient. Leptin, adiponectin, resistin and TNF-alpha serum levels were determined. RESULTS: fifty-six patients were male (73.7%) and the mean age was 44.5 ± 11.3 years of age (19-68). Thirty-nine (51.3%) (F1-F2: 84.6%; F3-4: 15.4%) patients had fibrosis in the liver biopsy. Seventeen females (85%) had fibrosis versus 22 males (39%), which was statistically significant by univariate analysis (p < 0.01). Patients with advanced fibrosis were older, with lower platelet counts, lower serum albumin, greater homeostatic model assessment insulin resistance (HOMA-IR), lower dietary lipids percentage, higher serum leptin levels and higher NAFLD Fibrosis Score (NAFLD-FS) values. This index had a negative predictive value of 98% and a positive predictive value of 60% for the detection of fibrosis. Variables independently associated with fibrosis (logistic regression) included male gender (protective factor) (0.09, 95% CI 0.01-0.7; p < 0.05) and HOMA-IR (1.7, 95% CI, 1.03-2.79; p < 0.05). CONCLUSIONS: gender and HOMA-IR were the only independent factors associated with fibrosis. NAFLD-FS could be considered as an accurate scoring system to rule out advanced fibrosis.

Sprouty1 is a broad mediator of cellular senescence.

Genes of the Sprouty family (Spry1-4) restrain signaling by certain receptor tyrosine kinases. Consequently, these genes participate in several developmental processes and function as tumor suppressors in adult life. Despite these important roles, the biology of this family of genes still remains obscure. Here we show that Sprouty proteins are general mediators of cellular senescence. Induction of cellular senescence by several triggers in vitro correlates with upregulation of Sprouty protein levels. More importantly, overexpression of Sprouty genes is sufficient to cause premature cellular senescence, via a conserved N-terminal tyrosine (Tyrosine 53 of Sprouty1). Accordingly, fibroblasts from knockin animals lacking that tyrosine escape replicative senescence. In vivo, heterozygous knockin mice display delayed induction of cellular senescence during cutaneous wound healing and upon chemotherapy-induced cellular senescence. Unlike other functions of this family of genes, induction of cellular senescence appears to be independent of activation of the ERK1/2 pathway. Instead, we show that Sprouty proteins induce cellular senescence upstream of the p38 pathway in these in vitro and in vivo paradigms.

Age-related differences in hepatic ischemia/reperfusion: gene activation, liver injury, and protective effect of melatonin.

BACKGROUND: Ischemia/reperfusion (I/R) causes functional and structural damage to liver cells, this being more pronounced with increasing age of the tissue. Melatonin is a pineal indole that has been shown to play an important role as a free radical scavenger and anti-inflammatory molecule. MATERIAL AND METHODS: The age-dependent responses to I/R were compared in 2-mo-old and 14-mo-old male Wistar rats. After 35 min of hepatic ischemia followed by 36 h of reperfusion, rats were sacrificed. Sham-operated control rats underwent the same protocol without real vascular occlusion. Animals were intraperitoneally injected with 10 mg/kg melatonin 24 h before the operation, at the time of surgery, and 12 and 24 h after it. The tissues were submitted to histopathologic evaluation. The levels of ALT and AST were analyzed in plasma. The expression of TNF-α, IL-1ß, IL-10, MCP-1, IFN-γ, iNOS, eNOS, Bad, Bax, Bcl2, AIF, PCNA, and NFKB1 genes were detected by RT-PCR in hepatic tissue. RESULTS: I/R was associated with significant increases in the expression of pro-inflammatory and pro-apoptotic genes in liver. Older rats submitted to I/R were found to respond with increased liver damage as compared with young rats, with serum ALT and AST levels significantly higher than in young animals. Mature rats also showed more evident increases in expression of pro-inflammatory cytokines (IL-1ß, MCP-1, and IFN-γ) as well as a decrease in the mRNA expression of IL-10 as compared with young animals. Pro-apoptotic genes (Bax, Bad, and AIF) were significantly enhanced in liver after I/R, without differences between young and mature animals. However, the expression of Bcl2 gene did not show any change. Melatonin treatment was able to lower the expression of pro-inflammatory cytokines and pro-apoptotic genes and to improve liver function, as indicated by normalization of plasma AST and ALT levels and by reduction of necrosis and microsteatosis areas. CONCLUSIONS: Melatonin treatment was able to reduce the I/R-stimulated pro-inflammatory and pro-apoptotic genes in the rat liver. Since older animals showed a more marked increase in inflammation and in liver injury, the treatment was more effective in those subjects.

The practical utility of non-invasive indices in metabolic hepatic steatosis.

BACKGROUND: Metabolic hepatic steatosis (metHS) is the most frequent cause of chronic liver disease in our environment. The "gold standard" for its diagnosis continues to be liver biopsy, but this is an invasive technique, is not risk-free, and has great interobserver variability, so noninvasive diagnostic methods are necessary. OBJECTIVE: To determine the diagnostic accuracy of non-invasive methods based on clinical and analytical data compared to liver biopsy, and to analyse their concordance with each other in the overall cohort and in subpopulations at risk of metHS. METHODS: Prospective observational study of 245 patients aged 19-80 years diagnosed with metHS by liver biopsy. Steatosis indices were calculated: FLI (Fatty Liver Index), LAP (Liver Accumulation Product), HSI-(Hepatitis Score Index) and fibrosis indices: Non-alcoholic fatty liver disease fibrosis score (NFS), fibrosis-4 index (FIB-4) and Hepamet Fibrosis Score (HFS). RESULTS: The non-invasive steatosis indices showed high sensitivity, and those of fibrosis, high specificity. To assess steatosis, FLI was the most sensitive index in all subpopulations (89-97%), except in women. To assess fibrosis, HFS offers maximum sensitivity in diabetics (86.7%) and is the index with the highest negative predictive value overall. The COR curves for non-invasive indices in steatosis and fibrosis compared to liver biopsy showed greater areas under the curve for the fibrosis indices, with NFS and HFS offering greater diagnostic accuracy (area > 0.8, p < 0.05). HFS also offers high diagnostic sensitivity in the diabetic population. CONCLUSIONS: Non-invasive indices of steatosis are more sensitive and those of fibrosis more specific than liver biopsy. NFS and HFS offer the highest diagnostic accuracy, with HFS having the highest negative predictive value.

Beneficial effects of melatonin on cardiological alterations in a murine model of accelerated aging.

This study investigated the effect of aging-related parameters such as inflammation, oxidative stress and cell death in the heart in an animal model of accelerated senescence and analyzed the effects of chronic administration of melatonin on these markers. Thirty male mice of senescence-accelerated prone (SAMP8) and 30 senescence-accelerated-resistant mice (SAMR1) at 2 and 10 months of age were used. Animals were divided into eight experimental groups, four from each strain: two young control groups, two old untreated control groups, and four melatonin-treated groups. Melatonin was provided at two different dosages (1 and 10 mg/kg/day) in the drinking water. After 30 days of treatment, the expression of inflammatory mediators (tumor necrosis factor-alpha, interleukin 1 and 10, NFkBp50 and NFkBp52), apoptosis markers (BAD, BAX and Bcl2) and parameters related to oxidative stress (heme oxygenases 1 and 2, endothelial and inducible nitric oxide synthases) were determined in the heart by real-time reverse transcription polymerase chain reaction (RT-PCR). Inflammation, as well as, oxidative stress and apoptosis markers was increased in old SAMP8 males, when compared to its young controls. SAMR1 mice showed significantly lower basal levels of the measured parameters and smaller increases with age or no increases at all. After treatment with melatonin, these age-altered parameters were partially reversed, especially in SAMP8 mice. The results suggest that oxidative stress and inflammation increase with aging and that chronic treatment with melatonin, a potent antioxidant, reduces these parameters. The effects were more marked in the SAMP8 animals.

Histopathological differences in patients with biopsy-proven non-alcoholic fatty liver disease with and without type 2 diabetes.

BACKGROUND AND AIMS: Prevalence of non-alcoholic fatty liver disease (NAFLD) in developed countries is 30% in the general population and 50% in patients with type 2 diabetes mellitus (T2DM). The aim of this study was to compare the severity of NAFLD, as assessed by liver biopsy and using the non-invasive index NAFLD Fibrosis Score (NFS), in subjects with and without T2DM. PATIENTS AND METHODS: The study sample consisted of 217 patients with biopsy-proven NAFLD. Anthropometric assessments, laboratory tests, histological criteria established by the Non-alcoholic Steatohepatitis Clinical Research Network (NASH CRN), and the NFS were recorded. RESULTS: Patients with T2DM (n=36; 16.5%) had higher HOMA-IR values (6.3±3.6 vs. 3.3±2.4; p<0.0001), GGT levels (125.2±102.3 vs. 82.5±70.6IU/l; p<005), and NFS index (-0.6±0.2 vs. -1.8±0.1; p<0.001) than subjects with no T2DM. Patients with T2DM were found higher rates of NASH (72.2% vs. 48.6%; p<0.05), advanced steatosis (80.6% vs. 63%; p<0.05), and liver fibrosis (75% vs. 43.1%, p<0.05) than patients with no T2DM. Patients with T2DM also had higher NFS values (-0.6±1.2 vs. -1.8±1.8: p=0.01). A logistic regression analysis adjusting for age, gender and BMI showed a significant independent association between NASH and presence of T2DM (OR=4.2: 95% CI: 1.4-12.1; p=0.007). A second model adjusting for the same covariates showed T2DM to be an independent factor associated to advanced fibrosis (OR=4.1; 95% CI: 1.7-9.7). CONCLUSION: Patients with T2DM have more advanced degrees of NAFLD and advanced fibrosis as assessed by liver biopsy and the NFS index. Particular attention should be paid to the study and monitoring of NASH in patients with T2DM.

Protective effect of resveratrol against inflammation, oxidative stress and apoptosis in pancreas of aged SAMP8 mice.

Aging is a physiological state in which a progressive decline in organ functions is accompanied by the development of age-related diseases. Resveratrol supplementation has been shown to exert anti-inflammatory and antioxidant effects in various mammalian models of aging. Senescence-accelerated mice (SAM) are commonly used as animal models to investigate the aging process. In the present study, the effects of inflammation, oxidative stress and apoptosis in pancreas of two different types of SAM (SAMR1 or resistant to aging, and SAMP8 or prone to aging) have been analysed, as well as the effect of resveratrol administration (5mg/kg/day) on these parameters in the SAMP8 strain. mRNA expressions of sirtuin 1 and FoxO factors were found to be decreased with aging in SAMP8 mice. An increase in inflammatory status and nuclear-factor kappa B (NFκB) protein expression was also observed in old mice, together with a decrease of anti-apoptotic markers and antioxidant-enzyme activity. Resveratrol administration was able to increase sirtuin 1 mRNA expression, as well as decreasing NFκB expression and reducing the proinflammatory and prooxidant status associated with age. In conclusion, resveratrol was able to modulate the inflammatory, oxidative and apoptotic status related to aging, thereby exerting a protective effect on pancreas age-induced damage.

Immunochromatographic test for detection of adenovirus from respiratory samples: is it a real solution for pediatric emergency department?

Rapid diagnosis of adenoviral respiratory infections is required in order to decide optimal treatment strategies. The adenovirus antigen immunochromatography Adeno Respiratory Card Letitest (Leti diagnostics, Barcelona, Spain), was evaluated versus the shell-vial culture and multiplex PCR (Clart Pneumovir Version 3.0, Genomica, Madrid, Spain), in nasopharyngeal washes and oropharyngeal swabs specimens from subjects with respiratory tract infections. Between April 2011 and November 2012, 224 patients were included. The IC Adeno Respiratory Card Letitest was positive for 77.9% (74 of 95) of patients diagnosed at bedside. Using multiplex-PCR as the reference standard, the overall sensitivity was 77.9% and the specificity was 73.6%. Taking shell-viral culture as the reference method, the Adeno Respiratory Card Letitest (Leti diagnostics, Barcelona, Spain) sensitivity and specificity values were 80.0% and 60.9%, respectively. Using RT-PCR (Clart Pneumovir Version 3.0, Genomica, Madrid, Spain) as the reference standard, the viral culture sensitivity was 53.2% and the specificity was 100%. The Adeno Respiratory Card Letitest (Leti diagnostics, Barcelona, Spain) assay could be a simple and rapid method for antigenic detection of adenovirus in pediatric respiratory samples although it would be necessary to improve the specificity and sensitivity of the test.

Melatonin can improve insulin resistance and aging-induced pancreas alterations in senescence-accelerated prone male mice (SAMP8).

The aim of the present study was to investigate the effect of aging on several parameters related to glucose homeostasis and insulin resistance in pancreas and how melatonin administration could affect these parameters. Pancreas samples were obtained from two types of male mice models: senescence-accelerated prone (SAMP8) and senescence-accelerated-resistant mice (SAMR1). Insulin levels in plasma were increased with aging in both SAMP8 and SAMR1 mice, whereas insulin content in pancreas was decreased with aging in SAMP8 and increased in SAMR1 mice. Expressions of glucagon and GLUT2 messenger RNAs (mRNAs) were increased with aging in SAMP8 mice, and no differences were observed in somatostatin and insulin mRNA expressions. Furthermore, aging decreased also the expressions of Pdx-1, FoxO 1, FoxO 3A and Sirt1 in pancreatic SAMP8 samples. Pdx-1 was decreased in SAMR1 mice, but no differences were observed in the rest of parameters on these mice strains. Treatment with melatonin was able to decrease plasma insulin levels and to increase its pancreatic content in SAMP8 mice. In SAMR1, insulin pancreatic content and plasma levels were decreased. HOMA-IR was decreased with melatonin treatment in both strains of animals. On the other hand, in SAMP8 mice, treatment decreased the expression of glucagon, GLUT2, somatostatin and insulin mRNA. Furthermore, it was also able to increase the expression of Sirt1, Pdx-1 and FoxO 3A. According to these results, aging is associated with significant alterations in the relative expression of pancreatic genes associated to glucose metabolism. This has been especially observed in SAMP8 mice. Melatonin administration was able to improve pancreatic function in old SAMP8 mice and to reduce HOMA-IR improving their insulin physiology and glucose metabolism.

Melatonin treatment protects liver of Zucker rats after ischemia/reperfusion by diminishing oxidative stress and apoptosis.

Fatty livers occur in up to 20% of potential liver donors and increase cellular injury during the ischemia/reperfusion phase, so any intervention that could enable a better outcome of grafts for liver transplantation would be very useful. The effect of melatonin on liver ischemia/reperfusion injury in a rat model of obesity and hepatic steatosis has been investigated. Forty fa/fa Zucker rats were divided in 4 groups. 3 groups were subjected to 35 min of warm hepatic ischemia and 36 h of reperfusion. One experimental group remained untreated and 2 were given 10mg/kg melatonin intraperitoneally or orally. Another group was sham-operated. Plasma ALT, AST and hepatic content of ATP, MDA, hydroxyalkenals, NOx metabolites, antioxidant enzyme activity, caspase-9 and DNA fragmentation were determined in the liver. The expression of iNOS, eNOS, Bcl2, Bax, Bad and AIF were determined by RT-PCR Melatonin was effective at decreasing liver injury by both ways as assessed by liver transaminases, markers of apoptosis, of oxidative stress and improved liver ATP content. Melatonin administration decreased the activities or levels of most of the parameters measured in a beneficial way, and our study identified also some of the mechanisms of protection. We conclude that administration of melatonin improved liver function, as well as markers of pro/antioxidant status and apoptosis following ischemia/reperfusion in obese rats with fatty liver. These data suggest that this substance could improve outcome in patients undergoing liver transplantation who receive a fatty liver implant and suggest the need of clinical trials with it in liver transplantation.

Beneficial effect of melatonin treatment on age-related insulin resistance and on the development of type 2 diabetes.

Abstract This paper will review the effect of aging on glucose metabolism and insulin resistance in pancreas and in peripheral tissues and how melatonin administration could affect these parameters. In SAMP8 mice insulin levels in plasma were found to be increased together with enhanced HOMA-IR values, whereas insulin content in pancreas showed a decrease with aging. Aging in SAMP8 mice was also associated with a significant increase in the relative expression of both protein and mRNA of different pro-inflammatory mediators. Furthermore, aging was associated with a decrease in the expression of Pdx-1, FoxO 1 and FoxO 3A and Sirt 1 in pancreas SAMP8 samples. Melatonin administration was able to reduce these age-related alterations, decreasing plasma insulin levels and increasing its pancreatic content in SAMP8 mice. HOMA-IR was decreased with melatonin treatment in all animals. Conversely, in SAMP8 mice, melatonin treatment decreased the expression of glucagon, GLUT2, somatostatin and insulin. Furthermore it was also able to increase the expression of Sirt 1, Pdx-1 and FoxO 3A. The present study has shown that aging is associated with significant alterations in the relative expression of pancreatic genes involved in both insulin secretion and glucose metabolism and that these are associated with an increase in inflammation and oxidative stress. Melatonin administration was able to reduce oxidative stress and inflammation and thus to improve pancreatic function in old mice. By doing so, insulin resistance is diminished and plasma insulin is reduced, enhancing insulin pancreatic content and reducing plasma glucose levels and HOMA index.

Factores analíticos, antropométricos y dietéticos asociados al desarrollo de fibrosis en pacientes con enfermedad por hígado graso no alcohólico / Analytical, anthropometric and dietary factors associated with the development of fibrosis in patients with nonalcoholic fatty liver disease

Antecedentes: la esteatohepatitis no alcohólica (EHNA) mantenida en el tiempo puede conducir a estadios avanzados de enfermedad hepática y al desarrollo de hepatocarcinoma. Objetivos: evaluar los factores analíticos, antropométricos y dietéticos asociados a la presencia de fibrosis hepática, evento que más influye en supervivencia y evolución. Métodos: fueron estudiados setenta y seis pacientes diagnosticados de enfermedad por hígado graso no alcohólica mediante biopsia. Las biopsias fueron clasificadas según el NAS-score (Kleiner). Se obtuvieron parámetros analíticos, antropométricos y dietéticos y se calculó el índice no invasivo NAFLD Fibrosis Score (NFLD-FS). Se determinaron los niveles séricos de leptina, adiponectina, resistina y TNF-alfa. Resultados: cincuenta y seis pacientes eran hombres (73,7%), con una edad media de 44,5 ± 11,3 años (19-68). Pacientes con fibrosis en biopsia: 39 (51,3%) (F1-F2: 84,6%; F3-4: 15,4%). Univariante: 17 mujeres (85%) presentaban fibrosis, frente a 22 hombres (39%) (p = 0,000). Los pacientes con fibrosis avanzada tenían mayor edad, menor recuento de plaquetas, menor albúmina sérica, mayor resistencia a la insulina (homeostatic model assessment insulin resistance, HOMA-IR), menor ingesta de lípidos, mayor nivel de leptina sérica y valores más altos de NAFLD-FS. Este índice presenta para detectar fibrosis avanzada un valor predictivo negativo del 98% y un valor predictivo positivo del 60%. Variables asociadas de forma independiente a la presencia de fibrosis (regresión logística): sexo masculino (factor protector) (0,09, IC 95%, 0,01-0,7; p < 0,05) y HOMA-IR (1,7, IC 95% 1,03-2,79; p < 0,05). Conclusiones: el sexo y el HOMA-IR son los únicos factores independientes que se asociaron a la presencia de fibrosis hepática en biopsia. El NAFLD-FS es un buen marcador no invasivo para descartar la presencia de fibrosis avanzada

Background: a prolonged non-alcoholic steatohepatitis (NASH) condition can lead to advanced stages of liver disease and the development of hepatocellular carcinoma. Aim: to evaluate analytical, anthropometric and dietary factors associated with the presence of fibrosis as this is the factor that most influences survival and evolution. Methods: seventy-six patients with liver biopsy-diagnosed non-alcoholic fatty liver disease (NAFLD) were included. Biopsies were scored considering the NASH criteria of Kleiner. Analytical, anthropometric and dietary (survey) parameters were obtained. NAFLD-FS is a non-invasive fibrosis index and was assessed for each patient. Leptin, adiponectin, resistin and TNF-alpha serum levels were determined. Results: fifty-six patients were male (73.7%) and the mean age was 44.5 ± 11.3 years of age (19-68). Thirty-nine (51.3%) (F1-F2: 84.6%; F3-4: 15.4%) patients had fibrosis in the liver biopsy. Seventeen females (85%) had fibrosis versus 22 males (39%), which was statistically significant by univariate analysis (p < 0.01). Patients with advanced fibrosis were older, with lower platelet counts, lower serum albumin, greater homeostatic model assessment insulin resistance (HOMA-IR), lower dietary lipids percentage, higher serum leptin levels and higher NAFLD Fibrosis Score (NAFLD-FS) values. This index had a negative predictive value of 98% and a positive predictive value of 60% for the detection of fibrosis. Variables independently associated with fibrosis (logistic regression) included male gender (protective factor) (0.09, 95% CI 0.01-0.7; p < 0.05) and HOMA-IR (1.7, 95% CI, 1.03-2.79; p < 0.05). Conclusions: gender and HOMA-IR were the only independent factors associated with fibrosis. NAFLD-FS could be considered as an accurate scoring system to rule out advanced fibrosis

Histopathological differences in patients with biopsy-proven non-alcoholic fatty liver disease with and without type 2 diabetes / Diferencias histopatológicas en pacientes con diabetes mellitus tipo 2 y no diabéticos con enfermedad hepática grasa no alcohólica diagnosticada con biopsia

Background and aims: Prevalence of non-alcoholic fatty liver disease (NAFLD) in developed countries is 30% in the general population and 50% in patients with type 2 diabetes mellitus (T2DM). The aim of this study was to compare the severity of NAFLD, as assessed by liver biopsy and using the non-invasive index NAFLD Fibrosis Score (NFS), in subjects with and without T2DM. Patients and methods: The study sample consisted of 217 patients with biopsy-proven NAFLD. Anthropometric assessments, laboratory tests, histological criteria established by the Non-alcoholic Steatohepatitis Clinical Research Network (NASH CRN), and the NFS were recorded. Results: Patients with T2DM (n=36; 16.5%) had higher HOMA-IR values (6.3±3.6 vs. 3.3±2.4; p<0.0001), GGT levels (125.2±102.3 vs. 82.5±70.6IU/l; p<005), and NFS index (−0.6±0.2 vs. −1.8±0.1; p<0.001) than subjects with no T2DM. Patients with T2DM were found higher rates of NASH (72.2% vs. 48.6%; p<0.05), advanced steatosis (80.6% vs. 63%; p<0.05), and liver fibrosis (75% vs. 43.1%, p<0.05) than patients with no T2DM. Patients with T2DM also had higher NFS values (−0.6±1.2 vs. −1.8±1.8: p=0.01). A logistic regression analysis adjusting for age, gender and BMI showed a significant independent association between NASH and presence of T2DM (OR=4.2: 95% CI: 1.4-12.1; p=0.007). A second model adjusting for the same covariates showed T2DM to be an independent factor associated to advanced fibrosis (OR=4.1; 95% CI: 1.7-9.7). Conclusion: Patients with T2DM have more advanced degrees of NAFLD and advanced fibrosis as assessed by liver biopsy and the NFS index. Particular attention should be paid to the study and monitoring of NASH in patients with T2DM

Antecedentes y objetivos: La prevalencia de la enfermedad hepática grasa no alcohólica (NAFLD) en los países desarrollados es del 30% de la población general y del 50% de los pacientes con diabetes mellitus tipo 2 (DM2). El objetivo de este estudio fue comparar la gravedad de NAFLD evaluado por biopsia hepática y con un índice no invasivo NAFLD Fibrosis Score (NFS) en sujetos con DM2 frente a pacientes no diabéticos. Pacientes y métodos: Este estudio se llevó a cabo entre 217 pacientes con diagnostico mediante biopsia de NAFLD. Se registraron la valoración antropométrica, pruebas de laboratorio, criterios histológicos establecidos por la Red de Investigación Clínica de Esteatohepatitis No Alcohólica (NASH) y NFS. Resultados: Los pacientes con DM2 (n=36; 16,5%) tuvieron más HOMA-IR (6,3±3,6 vs. 3,3±2,4; p<0,0001), GGT (125,2±102,3 vs. 82,5±70,6UI/L); p<0,05) e índice NFS (−0,6±0,2 vs. −1,8±0,1; p<0,001) que los sujetos sin DM2. Los pacientes con DM2 presentaron mayor porcentaje de EHNA (72,2 vs. 48,6%; p<0,05), grado avanzado de esteatosis (80,6 vs. 63%; p<0,05) y fibrosis hepática (75 vs. 43,1%; p<0,05) que los pacientes sin DM2. Los pacientes con DM2 presentaron también valores más altos de NFS (−0,6±1,2 vs. −1,8±1,8; p=0,01). El análisis de regresión logística ajustado por edad, sexo e IMC mostró asociación significativa independiente entre la esteatohepatitis y la presencia de DM2 (OR=4,2; IC 95%: 1,4-12,1; p=0,007). Un segundo modelo ajustado por las mismas covariables mostró que la DM2 fue un factor independiente asociado a la fibrosis avanzada (OR=4,1; IC 95%: 1,7-9,7). Conclusión: Los pacientes con DM2 tienen grados más avanzados de NAFLD y fibrosis avanzada evaluados mediante biopsia hepática y el índice NFS. Debe prestarse especial atención al estudio y seguimiento de la esteatohepatitis en pacientes con DM2

Effect of chronic melatonin administration on several physiological parameters from old Wistar rats and SAMP8 mice.

UNLABELLED: The effect of melatonin administration on age-induced alterations in hepatocytes, central nervous system, immune system, and skin are reviewed. Twenty-two-month-old Wistar rats and SAMP8 (senescence prone) mice of 10 months of age were used as experimental models. Wistar rats were analyzed untreated or after the chronic administration of melatonin at a dose of 1 mg/kg/day in the drinking water for 10 weeks. At the end of the treatment period, the various parameters were investigated. Results were compared with those of 2-month-old controls. In hepatocytes, aging induced a significant increase in oxidative stress, inflammation, and apoptosis when compared to young animals. Melatonin administration significantly ameliorated all these age-related changes. The impairment of the cardiovascular system with aging appears to contribute to the increased morbidity and mortality of the aged subjects. The process was investigated in SAMP8 mice of 10 months of age. Melatonin was provided for 30 days at two different dosages (1 mg/kg/day and 10 mg/kg/day), also in the drinking water. After treatment, the expression of inflammatory mediators (tumor necrosis factor-α, interleukin 1 and 10, NFκBp50 and NFκBp52), apoptosis markers (BAD, BAX, and Bcl2), and parameters related to oxidative stress (heme oxygenases 1 and 2, endothelial and inducible nitric oxide synthases) were determined in the heart. Inflammation as well as oxidative stress and apoptosis markers were increased in old SAMP8 males, as compared to young controls. After treatment with melatonin, these age-altered parameters were partially reversed. The results suggest that oxidative stress and inflammation increase with aging and that chronic treatment with melatonin is able to reduce these parameters. In the skin, a reduction of epidermal thickness together with a marked increase of the hypodermis with great fat accumulation was observed in old rats, together with an increase in caspase 3, 8 of nucleosomes and LPO and a reduction in Bcl2 levels in the cultured keratinocytes. Melatonin treatment was able to reduce the fat content of the hypodermis and to increase Bcl2 and reduce nucleosomes, caspases, and LPO in keratinocytes. CONCLUSION: Melatonin administration exerts a beneficial effect against age-induced changes in several physiological parameters in Wistar rats and SAMP 8 mice.