Replicating spine loading during functional and daily activities: An in vivo, in silico, in vitro research pipeline.

Lifestyle heavily influences intervertebral disc (IVD) loads, but measuring in vivo loads requires invasive methods, and the ability to apply these loads in vitro is limited. In vivo load data from instrumented vertebral body replacements is limited to patients that have had spinal fusion surgery, potentially resulting in different kinematics and loading patterns compared to a healthy population. Therefore, this study aimed to develop a pipeline for the non-invasive estimation of in vivo IVD loading, and the application of these loads in vitro. A full-body Opensim model was developed by adapting and combining two existing models. Kinetic data from healthy participants performing activities of daily living were used as inputs for simulations using static optimisation. After evaluating simulation results using in vivo data, the estimated six-axis physiological loads were applied to bovine tail specimens. The pipeline was then used to compare the kinematics resulting from the physiological load profiles (flexion, lateral bending, axial rotation) with a simplified pure moment protocol commonly used for in vitro studies. Comparing kinematics revealed that the in vitro physiological load protocol followed the same trends as the in silico and in vivo data. Furthermore, the physiological loads resulted in substantially different kinematics when compared to pure moment testing, particularly in flexion. Therefore, the use of the presented pipeline to estimate the complex loads of daily activities in different populations, and the application of those loads in vitro provides a novel capability to deepen our knowledge of spine biomechanics, IVD mechanobiology, and improve pre-clinical test methods.

Role of the variant rs3774261 of adiponectin gene on adiponectin levels and ratio adiponectin/leptin after biliopancreatic diversion in morbid obese subjects.

OBJECTIVE:   Single nucleotide variants (SNVs) of ADIPOQ gene on different comorbidities are related to obesity and weight loss. Despite, there are no studies evaluating the effect of rs3774261 on metabolic variables after bariatric surgery. We evaluated the effect of SNV rs3774261 of ADIPOQ gene on biochemical changes after biliopancreatic diversion surgery in morbidly obese subjects for 3 years follow-up. PATIENTS AND METHODS: One hundred and forty-nine patients (111 females/38 males) with morbid obesity (body mass index >40 kg/m2) without diabetes mellitus type 2 were enrolled. Biochemical and anthropometric evaluation were registered before and after 1, 2, and 3 years. Genotype of rs3774261 has been studied. RESULTS: Total cholesterol, LDL-cholesterol and triglyceride levels decreased in all genotype groups during the study. Although the improvement in glucose, insulin and HOMA-IR was significant in two genotypes (AA and AG); these changes were earlier in the AA genotype than in Ag and GG genotypes. Adiponectin levels increased in a significant way in subjects with AA genotype in the 3 follow-up periods (first year delta: 16.4±0.5 ng/ml; p=0.03, second year delta: 21.3±0.5 ng/ mL; p=0.02 and third year delta: 23.6±0.7 ng/mL; p=0.01) and at 3 years in subjects with AG genotype (delta: 18.3±0.4 ng/ mL; p=0.03). The ratio adiponectin/leptin increased in a significant way in subjects with AA genotype in the 3 follow-up times (first year delta: 0.40±0.1 units; p=0.02, second year delta: 0.58±0.1 units; p=0.01 and third year delta: 0.65±0.1 ng/mL; p=0.01) and at 3 years in subjects with AG genotype (delta: 0.61±0.1 ng/ mL; p=0.02). Subjects with GG genotype did not show a significant improvement in these parameters during the follow-up. CONCLUSIONS: G allele carriers of rs3774261 showed a delay in the improvement of glucose metabolism parameters, adiponectin and adiponectin/leptin ratio.

The gene variant rs2419621 of ACYL-CoA synthetase long-chain 5 gene is associated with weight loss and metabolic changes in response to a robotic sleeve gastrectomy in morbid obese subjects.

OBJECTIVE: Genetic mechanisms have been involved in the pathogenesis of obesity and weight loss due to bariatric surgery. The aim of our work was to evaluate the effects of rs2419621 genetic variant of ACSL5 gene on weight and metabolic changes after a robotic sleeve gastrectomy. PATIENTS AND METHODS: 48 patients were enrolled. Comorbidities, biochemical and anthropometric parameters evaluation were registered before and after 3, 6 and 12 months follow up. Genotype of rs2419621 ACSL5 gene was evaluated. RESULTS: We classified the subjects with a dominant model, in two groups: those carriers T allele (TT+CT, 37.5%) and non-carriers T allele (CC, 62.5%).  We reported a statistically significant reduction of body weight, waist circumference, percentage of excess of weight loss (EWL%), blood pressure, glucose, insulin, LDL-cholesterol and triglycerides after surgery. After 12 months, delta of (EWL%; 70.1% vs. 64,2%; p=0.04), weight (40.7+4.1 kg vs. 32.5+4.8 kg; p=0.03), waist circumference (29.1+3.1 cm vs. 22.2+2.8 kg; p=0.02) and triglycerides (51.2+9.1 mg/dl vs. 32.1+8.1; p=0.02) were higher in T allele carriers than non-T allele carriers. All comorbidities improved, but the percentage of patients with hypertriglyceridemia diminished early in the 3-month follow-up in the T-allele carriers, and at 12 months, no patient with the T allele had hypertriglyceridemia. CONCLUSIONS: Our data showed that the genetic variant (rs2419621) of ACSL5 gene are associated with better improvement of adiposity and triglyceride levels in subjects with T allele, after a robotic sleeve gastrectomy.

Lupus Eritematoso Sistémico y Manifestaciones en el Sistema Nervioso Central: Un desafío diagnóstico permanente: casos Clínicos / Systemic lupus erythematosus and manifestations in the Central Nervous System: a permanent diagnostic challenge. Clinical cases

La presencia de manifestaciones neuropsiquiátricas en pacientes reumatológicos trae consigo un gran desafío diagnóstico que exige una mirada amplia, desde las bases de la medicina interna, a fin de poder orientar un estudio adecuado y el tratamiento oportuno. Junto con ello, el permanente diálogo e intercambio de miradas clínicas con otras especialidades permite tener un enfoque multidisciplinario que enriquece el abordaje de estas presentaciones complejas.

The presence of neuropsychiatric manifestations in rheumatological patients brings with it a great diagnostic challenge that requires a broad view, from the foundations of internal medicine, in order to guide the appropriate study and timely treatment of these patients. Along with this, the permanent dialogue and exchange of clinical views with other specialties allows for a multidisciplinary approach that enriches the approach to these complex presentations.

Milk production responses, rumen fermentation, and blood metabolites of dairy cows fed increasing concentrations of forage rape (Brassica napus ssp. Biennis).

The aim of this study was to determine animal performance, rumen fermentation, and health-related blood metabolites of dairy cows in mid lactation fed with increasing levels (30 and 45%) of forage rape (FR) in the diet. Twelve pregnant multiparous lactating Holstein-Friesian dairy cows were randomly allocated to 1 of 3 dietary treatments in a replicated 3 × 3 Latin square design. The experiment was divided into three 21-d periods. For the control diet, 13.0 kg (dry matter, DM) of grass silage, 3.0 kg DM of commercial concentrate, 2.7 kg of DM cold-pressed extracted canola meal, and 0.45 kg DM of solvent-extracted soybean meal were offered daily. For the other two treatments, 30 and 45% of the DM from silage, canola meal, and commercial concentrate were replaced in equal proportions with FR. Data were analyzed individually using linear and quadratic orthogonal polynomials. Ingestive behavior was altered by the inclusion of FR. We observed a linear increase in eating time at the expense of rumination time. Nevertheless, total DM intake was not affected by dietary treatments, averaging 19.5 ± 0.24 kg of DM/d. Milk yield increased linearly with increasing concentration of FR in the diet. Thus, feed efficiency of cows (kg of milk/kg of DM intake) increased linearly with the percentage of FR in the diet. Inclusion of FR in the diet had no effect on milk composition or milk sensory characteristics. Mean rumen pH of cows decreased linearly from the control to the 45% FR diet; however, dietary treatments had no effect on the daily amount of time that rumen pH was below 5.8 (252 ± 71.4), indicating no risk of subacute ruminal acidosis. Concentrations of total volatile fatty acids in the rumen and molar proportions of acetate and butyrate were increased with FR inclusion, whereas the proportion of propionate was linearly reduced. Excretion of uric acid and total purine derivatives tended to be greater for cows fed FR, which resulted in a trend toward a linear increase in estimated microbial N flow. However, N use efficiency was not affected by FR inclusion. Although differences for some hematological measures (increased white blood cell and neutrophils counts) and a quadratic response for glutamate dehydrogenase for cows fed FR in the diet (decreased with inclusion of 30% and increased with 45% in the diet) were observed, all values were within appropriate ranges for dairy cows. These results indicated that including FR to dairy cow diets, up to 45% of diet DM, improved milk production due to changes in volatile fatty acids and predicted microbial N flow and had no negative effects on dairy cow health or sensory characteristics of milk.

RAS Mutation Decreases Overall Survival After Optimal Cytoreductive Surgery and Hyperthermic Intraperitoneal Chemotherapy of Colorectal Peritoneal Metastasis: A Modification Proposal of the Peritoneal Surface Disease Severity Score.

BACKGROUND: Cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) are currently the most accepted treatment for peritoneal metastases from colorectal cancer. Restrictive selection criteria are essential to obtain the best survival benefits for this complex procedure. The most widespread score for patient selection, the peritoneal surface disease severity score (PSDSS), does not include current biological factors that are known to influence on prognosis. We investigated the impact of including RAS mutational status in the selection criteria for these patients. METHODS: We studied the risk factors for survival by multivariate analysis using a prospective database of consecutive patients with carcinomatosis from colorectal origin treated by CRS and HIPEC in our unit from 2009 to 2017. The risk factors obtained were validated in a multicentre, international cohort, including a total of 520 patients from 15 different reference units. RESULTS: A total of 77 patients were selected for local análisis. Only RAS mutational status (HR: 2.024; p = 0.045) and PSDSS stage (HR: 2.90; p = 0.009) were shown to be independent factors for overall survival. Early PSDSS stages I and II associated to RAS mutations impaired their overall survival with no significant differences with PSDSS stage III overall survival (p > 0.05). These results were supported by the international multicentre validation. CONCLUSIONS: By including RAS mutational status, we propose an updated RAS-PSDSS score that outperforms PSDSS alone providing a quick and feasible preoperative assessment of the expected overall survival for patients with carcinomatosis from colorectal origin undergone to CRS + HIPEC.

Anti TNF como inductor de lupus cutáneo: caso clínico / Anti TNF as inducer of cutaneous lupus: clinical case

El mayor acceso a las terapias biológicas para el tratamiento de múltiples enfer-medades autoinmune trae consigo el mayor riesgo de padecer eventos adversos relacionados al uso de estos2,4. Presentamos un caso clínico de una paciente con diagnóstico de artritis reumatoide en tratamiento con ANTI TNF

The greater access to biological therapies for the treatment of multiple autoim-mune diseases brings with it the greatest risk of suffering adverse events related to the use of these (2,4). We present a clinical case of a patient diagnosed with rheumatoid arthritis in treatment with ANTI TNF

The association of the rs670 variant of APOA1 gene with insulin resistance and lipid profile in morbid obese patients after a biliopancreatic diversion surgery.

OBJECTIVE: A common G-to-A transition (rs670) of the Apoprotein subtype 1 APOA1 gene has been evaluated. The presence of the A allele has been related with increased activity. We investigated the role of this genetic variant (rs670) on lipoprotein levels and anthropometric parameters after biliopancreatic diversion (BPD) surgery in morbid obese patients. PATIENTS AND METHODS: Sixty-three patients with morbid obesity without diabetes mellitus type 2 were enrolled. Biochemical and anthropometric evaluation were registered before and after one, two and three years follow-up. RESULTS: Genotype distribution was 73% (n=46) GG, 25.6% (n=16) GA and 1.4% (n=1) AA for the rs670 polymorphism. Percent excess weight loss, anthropometric and biochemical parameters improved in both groups (GG vs. GA±AA). The decrease of fasting insulin levels at 1 years (delta: -3.7±1.4 mUI/L vs. -2.9±1.2 mUI/L; p=0.02), 2 years (delta: -4.8±0.3 mUI/L vs. -4.0±0.2 mUI/L; p=0.01) and 3 years (delta: -6.7±3.1 mUI/L vs. -3.9±2.1 mUI/L; p=0.03) was higher in A allele carriers than in non carriers. The improvement of HOMA-IR levels at 1 years (delta: -3.7±1.4 mUI/L vs. -2.9±1.2 mUI/L; p=0.02), 2 years (delta: -4.8±0.3 mUI/L vs. -4.0±0.2 mUI/L; p=0.01) and 3 years (delta: -6.7±3.1 mUI/L vs. -3.9±2.1 mUI/L; p=0.03) was also higher in A allele carriers than non-carriers. Finally, the increase of HDL-cholesterol levels at 1 years (delta: 2.2±0.6 mg/dl vs. -1.2±0.2 mg/dl; p=0.001), 2 years (delta: 2.5±0.4 mg/dl vs. 0.3±0.1 mg/dl; p=0.01) and 3 years (delta: 2.4±0.6 mg/dl vs. 0.4±2.3 mg/dl; p=0.02) was higher in A allele carriers than non-carriers. CONCLUSIONS: This variant of the APOA1 gene showed important effects on HDL-cholesterol, HOMA-IR and insulin resistance after DBP for 3 years.

The synthetic peptide PnPP-19 induces peripheral antinociception via activation of NO/cGMP/KATP pathway: Role of eNOS and nNOS.

BACKGROUND: and purpose: The peptide PnPP-19, derived from the spider toxin PnTx2-6 (renamed as δ-CNTX-Pn1c), potentiates erectile function by activating the nitrergic system. Since NO has been studied as an antinociceptive molecule and PnPP-19 is known to induce peripheral antinociception, we intended to evaluate whether PnPP-19 could induce peripheral antinociception through activation of this pathway. EXPERIMENTAL APPROACH: Nociceptive thresholds were measured by paw pressure test. PGE2 (2 µg/paw) was administered intraplantarly together with PnPP-19 and inhibitors/blockers of NOS, guanylyl cyclase and KATP channels. The nitrite concentration was accessed by Griess test. The expression and phosphorylation of eNOS and nNOS were determined by western blot. KEY RESULTS: PnPP-19 (5, 10 and 20 µg/paw) induced peripheral antinociception in rats. Administration of NOS inhibitor (L-NOarg), selective nNOS inhibitor (L-NPA), guanylyl cyclase inhibitor (ODQ) and the blocker of KATP (glibenclamide) partially inhibited the antinociceptive effect of PnPP-19 (10 µg/paw). Tissue nitrite concentration increased after PnPP-19 (10 µg/paw) administration. Expression of eNOS and nNOS remained the same in all tested groups, however the phosphorylation of nNOS Ser852 (inactivation site) increased and phosphorylation of eNOS Ser1177 (activation site) decreased after PGE2 injection. Administration of PnPP-19 reverted this PGE2-induced effect. CONCLUSIONS AND IMPLICATIONS: The peripheral antinociceptive effect induced by PnPP-19 is resulting from activation of NO-cGMP-KATP pathway. Activation of eNOS and nNOS might be required for such effect. Our results suggest PnPP-19 as a new drug candidate to treat pain and reinforce the importance of nNOS and eNOS activation, as well as endogenous NO release, for induction of peripheral antinociception.

PnPP-19, a spider toxin peptide, induces peripheral antinociception through opioid and cannabinoid receptors and inhibition of neutral endopeptidase.

BACKGROUND AND PURPOSE: The synthetic peptide PnPP-19 has been studied as a new drug candidate to treat erectile dysfunction. However, PnTx2-6, the spider toxin from which the peptide was designed, induces hyperalgesia. Therefore, we intended to investigate the role of PnPP-19 in the nociceptive pathway. EXPERIMENTAL APPROACH: Nociceptive thresholds were measured by paw pressure test. PnPP-19 was administered intraplantarly alone or with selective cannabinoid or opioid receptor antagonists. The hydrolysis of PnPP-19 by neutral endopeptidase (NEP) (EC 3.4.24.11), an enzyme that cleaves enkephalin, was monitored by HPLC and the cleavage sites were deduced by LC-MS. Inhibition by PnPP-19 and Leu-enkephalin of NEP enzyme activity was determined spectrofluorimetrically. KEY RESULTS: PnPP-19 (5, 10 and 20 µg per paw) induced peripheral antinociception in rats. Specific antagonists of µ opioid receptors (clocinnamox), δ opioid receptors (naltrindole) and CB1 receptors (AM251) partly inhibited the antinociceptive effect of PnPP-19. Inhibition of fatty acid amide hydrolase by MAFP or of anandamide uptake by VDM11 enhanced PnPP-19-induced antinociception. NEP cleaved PnPP-19 only after a long incubation, and Ki values of 35.6 ± 1.4 and 14.6 ± 0.44 µmol·L(-1) were determined for PnPP-19 and Leu-enkephalin respectively as inhibitors of NEP activity. CONCLUSIONS AND IMPLICATIONS: Antinociception induced by PnPP-19 appears to involve the inhibition of NEP and activation of CB1, µ and δ opioid receptors. Our data provide a greater understanding of the antinociceptive effects of PnPP-19. This peptide could be useful as a new antinociceptive drug candidate.

Regulation of cell death receptor S-nitrosylation and apoptotic signaling by Sorafenib in hepatoblastoma cells.

Nitric oxide (NO) plays a relevant role during cell death regulation in tumor cells. The overexpression of nitric oxide synthase type III (NOS-3) induces oxidative and nitrosative stress, p53 and cell death receptor expression and apoptosis in hepatoblastoma cells. S-nitrosylation of cell death receptor modulates apoptosis. Sorafenib is the unique recommended molecular-targeted drug for the treatment of patients with advanced hepatocellular carcinoma. The present study was addressed to elucidate the potential role of NO during Sorafenib-induced cell death in HepG2 cells. We determined the intra- and extracellular NO concentration, cell death receptor expression and their S-nitrosylation modifications, and apoptotic signaling in Sorafenib-treated HepG2 cells. The effect of NO donors on above parameters has also been determined. Sorafenib induced apoptosis in HepG2 cells. However, low concentration of the drug (10nM) increased cell death receptor expression, as well as caspase-8 and -9 activation, but without activation of downstream apoptotic markers. In contrast, Sorafenib (10 µM) reduced upstream apoptotic parameters but increased caspase-3 activation and DNA fragmentation in HepG2 cells. The shift of cell death signaling pathway was associated with a reduction of S-nitrosylation of cell death receptors in Sorafenib-treated cells. The administration of NO donors increased S-nitrosylation of cell death receptors and overall induction of cell death markers in control and Sorafenib-treated cells. In conclusion, Sorafenib induced alteration of cell death receptor S-nitrosylation status which may have a relevant repercussion on cell death signaling in hepatoblastoma cells.

Increased milk protein synthesis in response to exogenous growth hormone is associated with changes in mechanistic (mammalian) target of rapamycin (mTOR)C1-dependent and independent cell signaling.

The objective of this study was to determine if increased milk protein synthesis observed in lactating dairy cows treated with growth hormone (GH) was associated with mechanistic (or mammalian) target of rapamycin complex 1 (mTORC1) regulation of downstream factors controlling nucleocytoplasmic export and translation of mRNA. To address this objective, biochemical indices of mammary growth and secretory activity and the abundance and phosphorylation status of mTORC1 pathway factors were measured in mammary tissues harvested from nonpregnant lactating dairy cows 6 d after treatment with a slow-release formulation of GH or saline (n=4/group). Treatment with GH increased mammary parenchymal weight and total protein content and tended to increase ribosome number and cell size, whereas protein synthetic efficiency, capacity, and cell number were unchanged. Cellular abundance of the mTORC1 components mTOR and (phosphorylated) mTOR(Ser2448) increased, as did complex eukaryotic initiation factor 4E:eukaryotic initiation factor 4E binding protein 1 (eIF4E:4EBP1), whereas no change was observed for mTORC1-downstream targets 4EBP1, 4EBP1(Ser65), p70/p85(S6K) and p70(S6K)Thre389/p85(S6K)Thre412. Changes in activation were not observed for any of the targets measured. These results indicate that GH treatment influences signaling to mTORC1 but not downstream targets involved in the nucleocytoplasmic export and translation of mRNA. Increased eIF4E:4EBP1 complex formation indicates involvement of the mitogen-activated protein kinase (MAPK) pathway. Abundance of MAPK pathway components eIF4E, eIF4E(Ser209), eIF4E:eIF4G complex, MAP kinase-interacting serine/threonine-protein kinase 1 (MKNK1), MKNK1(Thr197202), and ribosomal protein S6 kinase, 90kDa, polypeptide 1 (RPS6KA1) increased significantly in response to GH, whereas relative activation of the proteins was unchanged. Expression of IGFBP3 and IGFBP5 increased, that of IGF1R decreased, and that of IGF1 remained unchanged in response to GH. PatSearch analysis of the milk caseins αS1-casein, αS2-casein, and ß-casein, MAPK signaling target RPS6KA1, and proliferation gene IGFBP3 mRNA indicated that all contained putative eIF4E-sensitivity elements. In response to GH, these genes were all upregulated, suggesting that increased abundance of eIF4E and eIF4E(Ser209) plays a role in mediating their nucleocytoplasmic export. We propose that, in response to GH, the IGF1-IGF1R-MAPK signaling cascade regulates eIF4E-mediated nucleocytoplasmic export and translation of mRNA, whereas mTOR controls cell renewal, cell turnover, and rRNA transcription through an alternative signaling cascade.

Basal GLP-1 levels in morbidly obese patients following biliopancreatic diversion surgery.

BACKGROUND: Previous studies addressing the changes of glucagon-like peptide-1 (GLP-1) concentrations in morbidly obese patients after bariatric surgery have demonstrated conflicting results. The aim of the present study was to investigate the changes in serum GLP-1 levels 9 months after biliopancreatic diversion in morbidly obese patients without diabetes mellitus. METHODS: A sample of 40 morbidly obese patients without diabetes mellitus was enrolled. Biochemical and anthropometrical evaluations were conducted at basal and 9 months after surgery. RESULTS: The mean patient age was 46.6 ± 13.1 years, and the mean preoperative body mass index (BMI) was 47.1 ± 18.1. A significant decrease in BMI, weight, waist circumference, fat mass, glucose, LDL cholesterol, total cholesterol, and triglyceride levels was observed after 9 months. Serum basal GLP-1 levels did not change after surgery (0.65 ± 0.18 ng/ml vs. 0.66 ± 0.17 ng/ml; n.s.). Postsurgical correlation analysis showed a negative association between basal GLP-1 and HDL cholesterol (r = -0.57; p < 0.01). CONCLUSIONS: Fasting GLP-1 concentrations did not change after massive weight loss with biliopancreatic diversion in morbidly obese patients without diabetes mellitus.

Lys656Asn polymorphism of leptin receptor, leptin levels and insulin resistance in patients with non alcoholic fatty liver disease.

BACKGROUND: Some studies have pointed to a role of leptin and insulin resistance in pathogenesis of non alcoholic fatty liver disease (NAFLD). The aim of our study was to investigate the influence of Lys656Asn polymorphism LEPR gene on the histological changes, insulin resistance and leptin levels in overweight patients. MATERIAL AND METHODS: A population of 76 patients with NAFLD was recruited in a cross sectional study. A biochemical analysis of serum was measured. Genotype of LEPR gene Lys656Asn was studied. RESULTS: Nineteen patients (25%) had the genotype Lys656Asn and 4 patients genotype Asn656Asn (mutant type group) and 53 patients (69.7%) Lys656Lys (wild type group). Body mass index, weight, fat mass, waist circumference, waist to hip ratio, glucose levels and HOMA-IR were higher in mutant than wild type group. LEPR polymorphism is in any way related with liver lesions. The multivariate analysis adjusted by age, sex, BMI and genotype showed an independently association of lobular inflammation 4.19 (CI95%: 1.37-12.77), portal inflammation 1.97 (CI95%: 1.05-3.74) and steatosis 9.23 (CI95%: 1.47-57.83) with HOMA. Liver steatosis was associated with leptin levels (1.09 (CI95%: 1.06-1.18)), too. CONCLUSION: Lys656Asn polymorphism of LEPR gene is associated with obesity parameters, insulin resistance and glucose levels in patients with NAFLD. In logistic regression analysis, only insulin resistance was associated with portal inflammation), lobular inflammation and steatosis; liver steatosis was related with leptin levels, too.

Influence of G1359A polimorphysm of the cannabinoid receptor gene (CNR1) on insulin resistance and adipokines in patients with non alcoholic fatty liver disease.

BACKGROUND: Considering the evidence that endogenous cannabinoid system plays a role in metabolic aspects of body weight and metabolic syndrome components such as non alcoholic fatty liver disease (NAFLD). The aim of our study was to investigate the influence of this polymorphism on insulin resistance, liver histological changes, anthropometric parameters and adipocytokines in patients with NAFLD. MATERIAL AND METHODS: A population of 71 patients with NAFLD was recruited in a cross sectional study. A biochemical analysis of serum was measured. Genotype of G1359A polymorphism of CB1 receptor gene CB1 receptor was studied. Forty one patients (36.9%) had the genotype G1359G (wild type group) and twenty nine (26.1%) patients G1359A or A1359A (mutant type group). RESULTS: Twenty four 24 patients (32,3%) had a Brunt grade > 4 and 12 patients (17%) had a significative fibrosis (F > = 2). HOMA values were higher in wild type group than mutant type group. Adiponectin and visfatin levels were higher in mutant type group. Moreover, TNF-alpha and resistin levels were higher in wild type group than mutant type group. Patients with mutant genotype showed less frequently elevated levels of AST. AST > 40 UI/L was detected in 28.5% of patients in the mutant vs. 53% of patients with wild genotype, p < 0.05. Patients with mutant type group presented a percentage of Brunt grade > = 4 less frequently than patients with wild type group (28.5%vs 7.1%). CONCLUSION: A variant of the polymorphism G1359A CBR1 is associated with lower levels of HOMA, TNF-alpha, resistin and higher levels of adiponectin than patients with the wild variant of this polymorphism. Besides, patients with A allele variant shown lower Brunt grade in liver biopsy.

Factors affecting mortality following myeloablative cord blood transplantation in adults: a pooled analysis of three international registries.

A retrospective analysis was conducted to examine factors affecting early mortality after myeloablative, single-unit cord blood transplantation (CBT) for hematological malignancies in adolescents and adults. Data were collected from the three main CBT registries pooling 514 records of unrelated, single, unmanipulated, first myeloablative allogeneic CBTs conducted in North America or Europe from 1995 to 2005, with an HLA match ≥ 4/6 loci, in patients aged 12-55. Overall 100-day, 180-day and 1-year survival (Kaplan-Meier method) were 56, 46 and 37%, respectively, with no significant heterogeneity across registries. Multivariate analysis showed cell dose < 2.5 × 107/kg (odds ratio (OR) 2.76, P < 0.0001), older age (P = 0.002), advanced disease (P = 0.02), positive CMV sero-status (OR 1.37 P = 0.11), female gender (OR 1.43, P = 0.07) and limited CBT center experience (< 10 records contributed, OR 2.08, P = 0.0003) to be associated with higher 100-day mortality. A multivariate model predictive of 1-year mortality included similar prognostic factors except female gender. Transplant year did not appear as a significant independent predictor. This is the first analysis to pool records from three major CBT registries in the United States and Europe. In spite of some differences in practice patterns, survival was remarkably homogeneous. The resulting model may contribute to better understanding factors affecting CBT outcomes.

Covered metal stents for the treatment of biliary complications after orthotopic liver transplantation.

BACKGROUND: Biliary complications, a major source of morbidity after orthotopic liver transplantation (OLT), are increasingly being treated by endoscopic retrograde cholangiopancreatography (ERCP). Endoscopic management has been shown to be superior to percutaneous therapy and surgery. Covered self-expandable metal stents (CSEMSs) may be an alternative to the current endoscopic standard treatment with periodic plastic stent replacement. OBJECTIVE: To assess the safety and efficacy of temporary CSEMS insertion for biliary complications after OLT. METHODS: From November 2001 to December 2009, the 242 OLT performed in 226 patients included 67 cases that developed post-OLT leaks or strictures (29.6%), excluding ischemic biliary complications. CSEMSs were used in 22 patients (33%), 18 male and 4 female, with an overall median age of 55 years (range, 29-69). In-house OLT patients underwent an index ERCP at 26 days (range, 8-784) after OLT. Their records were reviewed to determine ERCP findings, technical success, and clinical outcomes. RESULTS: ERCP with sphincterotomy was performed in all 22 patients, revealing 18 with biliary strictures alone (82%), 3 with strictures and leaks (14%), and 1 with strictures and choledocholithiasis (4%). All strictures were anastomotic. All patients had 1-2 plastic stents inserted across the anastomosis (11 had prior balloon dilation); stones were successfully removed, for an initial technical success rate of 100% (22/22). CSEMSs, were placed at the second ERCP in 14 patients, at the third in 7, and at the fourth in 1. With a median follow-up of 12.5 months (range, 3-25) after CSEMS removal, 21/22 patients (95.5%) remain stricture free and one relapsed, requiring repeat CSEMS insertion. Four patients experienced pain after CSEMS insertion. At CSEMS removal, migration was noted in 5 cases, into either the distal duodenum (n=4) or the proximal biliary tree (n=1), and embedding was seen in 1 case. There were no serious complications; no patients needed hepatojejunostomy. CONCLUSIONS: ERCP is a safe first-line approach for post-OLT biliary complications. It was highly successful in a population with anastomotic leaks and strictures. The therapeutic role of ERCP to manage biliary complications after OLT in the long term is not well known. In our experience, the high rate (close to 95%) of efficacy and its relative safety allowed us to use CSEMS to manage refractory biliary post-OLT strictures. CSEMS insertion may preclude most post-OLT hepatojejunostomies.

Effects of C358A missense polymorphism of the endocannabinoid degrading enzyme fatty acid amide hydrolase on weight loss and cardiovascular risk factors 1 year after biliopancreatic diversion surgery.

BACKGROUND: Bariatric surgery is the most effective long-term treatment of morbid obesity and also results in a reduction of obesity-associated co-morbidities. We investigated the role of the polymorphism (C358A) of the fatty acid amide hydrolase gene on the clinical outcomes 1 year after biliopancreatic diversion in morbidly obese patients. METHODS: A total of 67 morbidly obese patients (body mass index >40 kg/m(2)) underwent biliopancreatic diversion. Their weight, blood pressure, basal glucose, triglycerides, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol were measured at the baseline visit and at each follow-up visit. The frequency of the metabolic co-morbidities was recorded at each visit. RESULTS: Of the 67 patients, 46 (68.7%) had genotype C358C (wild-type group) and 21 (10.3%) had genotype C358A (mutant-type group). In the wild- and mutant-type groups, the body mass index, weight, waist circumference, systolic blood pressure, and glucose, total cholesterol, low-density lipoprotein cholesterol, and triglyceride concentrations decreased, without statistical significance between the 2 groups. The initial percentage of weight loss at 9 months and 1 year of follow-up was greater in the mutant-type group (9 months, 22.1% versus 28.8%, P <.05; and 1 year, 28.3% versus 36.4%, P <.05). CONCLUSION: The allele A358 of fatty acid amide hydrolase was associated with a better initial percentage of excess weight loss 9 and 12 months after biliopancreatic diversion.

Role of -55CT polymorphism of UCP3 gene on non alcoholic fatty liver disease and insulin resistance in patients with obesity.

BACKGROUND AND AIMS: Some studies have pointed to a role of UCP3 in the regulation of biochemical and fat parameters in overweight patients. The aim of our study was to investigate the influence of -55CT polymorphism of UCP3 gene (rs1800849) on histological changes and insulin resistance in patients with non-alcoholic fatty liver disease (NAFLD). MATERIAL AND METHODS: A population of 39 patients with NAFLD was recruited in a cross sectional study. The inclusion criterion was the presence of biopsy-proven NAFLD. A biochemical analysis of serum (lipid profile, and adipocytokines) was measured. An anthropometric analysis was assessed, too. Genotype of UCP3 gene -55CT was studied. RESULTS: Nine patients (23%) had the genotype 55CC (mutant type group) and 30 patients (77%) 55CT (wild type group).TT genotype was not detected. Insulin levels and HOMA were higher in mutant type group (insulin: 17.7 +/- 10.9 mUI/L vs 11.9 +/- 4.7 mUI/L/; p < 0.05) and (HOMA: 3.2 +/- 1.8 vs 4.5 +/- 2.8; p < 0.05). Adiponectin levels were lower in mutants type group (36.5 +/- 28.1 ug/ml vs 21.5 +/- 18.6 ug/ml:p < 0.05). Moderate-severe inflammation and moderate-severe steatosis were more frequent in mutant type group, with higher levels of insulin and lower levels of adiponectin than mild stages. CONCLUSION: -55CT genotype is associated with high insulin resistance and low adiponectin levels than -55CC genotype. Patients with -55CT genotype have more frequently moderate-severe steatosis and inflammation than -55CC genotype.