Impact of Adherence to the Mediterranean Diet on Antioxidant Status and Metabolic Parameters in NAFLD Patients: A 24-Month Lifestyle Intervention Study.

BACKGROUND: The Mediterranean Diet (MedDiet) is recognized as a healthy dietary pattern. Non-alcoholic fatty liver disease (NAFLD) is characterized by the excessive accumulation of fat in the liver. OBJECTIVES: To assess the antioxidant status in erythrocytes, plasma, and peripheral blood mononuclear cells (PBMCs) of NAFLD patients following a 24-month lifestyle intervention based on the MedDiet. Adult patients (n = 40; aged 40-60 years) diagnosed with NAFLD by magnetic resonance imaging were divided into two groups based on their adherence to the MedDiet. Consumption was assessed using a validated 143-item semiquantitative Food Frequency Questionnaire. Anthropometrics, biochemistry parameters, intrahepatic fat contents (IFC), antioxidants, and inflammatory biomarkers were measured in plasma and erythrocytes before and after the intervention. RESULTS: After the intervention, body mass index (BMI) and plasma levels of total cholesterol, low-density lipoprotein cholesterol (LDL-chol), triglycerides, malondialdehyde (MDA), and cytokeratin-18 (CK18) decreased, and high-density lipoprotein cholesterol (HDL-chol) increased. Participants with high adherence to MedDiet showed lower IFC, hepatic enzyme (AST, ALT, and GGT), glycemia, oxidase LDL (oxLDL) plasma levels, and erythrocyte MDA levels. Higher antioxidant activity (erythrocyte catalase-CAT, superoxide dismutase-SOD, glutathione peroxidase-GPx, glutathione reductase-GRd, and total glutathione-GSH as well as PBMCs-CAT gene expression) was observed in these patients, along with a reduction of PBMCs reactive oxygen species production and Toll-like receptor 4 (TLR4) expression. Inverse associations were observed between adherence to the MedDiet and BMI, glycemia, AST, IFC, and CK18 plasma levels and oxLDL, CAT, SOD, and GRd activities in erythrocytes. A significant linear regression was observed between adherence to the MedDiet and antioxidant score. CONCLUSIONS: Adherence to the MedDiet is associated with improved plasma and PBMC antioxidant and inflammatory biomarker profiles and high antioxidant defences in erythrocytes.

Effect of a Six-Month Lifestyle Intervention on the Physical Activity and Fitness Status of Adults with NAFLD and Metabolic Syndrome.

(1) Background: Physical inactivity has been linked to NAFLD, and exercise has been reported as useful to reduce intrahepatic fat content in NAFLD. (2) Objectives: To assess the physical activity (PA) and fitness status after a six-month lifestyle intervention (diet and PA) in adults with NAFLD and metabolic syndrome (MetS). (3) Design: Prospective cohort analysis of data obtained between baseline and six-year parallel-group randomized trial (n = 155, aged 40-60 years old, with MetS and NAFLD). Participants were randomized into three nutritional and PA intervention groups: Conventional diet (CD); MedDiet-high meal frequency (MD-HMF); MedDiet-physical activity (MD-PA). (4) Methods: PA and fitness status were assessed using a validated Minnesota questionnaire, ALPHA-FIT test battery, accelerometers, and functional fitness score. Information related to age, gender, education level, marital status, socioeconomic status, smoking habit, and alcohol consumption were also obtained. (5) Results: The CD group had higher improvement in standing handgrip than the MD-HMF group. The MD-PA group did more modified push-up repetitions than the CD group. The MD-PA and CD groups showed higher sitting handgrip than the MD-HMF group. The MD-HMF group showed the highest decrease in aerobic capacity. The MD-PA group showed lower light intensity PA/day than the CD and MD-HMF groups. The MD-PA group showed higher moderate intensity PA than the CD and MD-HMF groups. The CD group reported more METs per day than the MD-HMF group. (6) Conclusions: Lifestyle six-month intervention with diet and regular PA improved functional fitness in middle-aged patients with NAFLD and MetS. Aerobic capacity improved in patients who followed a Mediterranean diet and regular training sessions at six months.

Albuminuria Is Associated with Hepatic Iron Load in Patients with Non-Alcoholic Fatty Liver Disease and Metabolic Syndrome.

BACKGROUND: Increased albuminuria is associated with increased serum ferritin, insulin resistance, and non-alcoholic fatty liver disease (NAFLD). Liver iron accumulation is also related to hyperferritinemia, insulin resistance, and NAFLD; however, there is no evidence on its relationship with albuminuria. AIMS: To assess the relationship between hepatic iron load and urine albumin-to-creatinine ratio (UACR) in patients with metabolic syndrome (MetS) and NAFLD. METHODS: In total, 75 MetS and NAFLD patients (aged 40-60 years, BMI 27-40 kg/m2) were selected from a cohort according to available data on hepatic iron load (HepFe) by magnetic resonance imaging (MRI). Subjects underwent anthropometric measurements, biochemistry testing, and liver MRI. Increased albuminuria was defined by UACR. RESULTS: UACR correlated with NAFLD, HepFe, triglycerides, serum ferritin, fasting insulin, insulin resistance (calculated using the homeostatic model assessment for insulin resistance-HOMA-IR- formula), and platelets (p < 0.05). Multiple regression analysis adjusted for gender, age, eGFR, HbA1c, T2DM, and stages of NAFLD, found that HepFe (p = 0.02), serum ferritin (p = 0.04), fasting insulin (p = 0.049), and platelets (p = 0.009) were associated with UACR (R2 = 0.370; p = 0.007). UACR, liver fat accumulation, serum ferritin, and HOMA-IR increased across stages of HepFe (p < 0.05). Patients with severe NAFLD presented higher HepFe, fasting insulin, HOMA-IR, and systolic blood pressure as compared to patients in NAFLD stage 1 (p < 0.05). CONCLUSION: Hepatic iron load, serum ferritin, fasting insulin, and platelets were independently associated with albuminuria. In the context of MetS, increased stages of NAFLD presented higher levels of HepFe. Higher levels of HepFe were accompanied by increased serum ferritin, insulin resistance, and UACR. The association between iron accumulation, MetS, and NAFLD may represent a risk factor for the development of increased albuminuria.

Oxidative Stress and Pro-Inflammatory Status in Patients with Non-Alcoholic Fatty Liver Disease.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is characterized by excessive fat accumulation, especially triglycerides, in hepatocytes. If the pathology is not properly treated, it can progress to nonalcoholic steatohepatitis (NASH) and continue to fibrosis, cirrhosis or hepatocarcinoma. OBJECTIVE: The aim of the current research was to identify the plasma biomarkers of liver damage, oxidative stress and inflammation that facilitate the early diagnosis of the disease and control its progression. METHODS: Antioxidant and inflammatory biomarkers were measured in the plasma of patients diagnosed with NAFLD (n = 100 adults; 40-60 years old) living in the Balearic Islands, Spain. Patients were classified according to the intrahepatic fat content (IFC) measured by magnetic resonance imaging (MRI). RESULTS: Circulating glucose, glycosylated haemoglobin, triglycerides, low-density lipoprotein-cholesterol, aspartate aminotransferase and alanine aminotransferase were higher in patients with an IFC ≥ 2 of NAFLD in comparison to patients with an IFC of 0 and 1. The plasma levels of catalase, irisin, interleukin-6, malondialdehyde, and cytokeratin 18 were higher in stage ≥2 subjects, whereas the resolvin D1 levels were lower. No differences were observed in xanthine oxidase, myeloperoxidase, protein carbonyl and fibroblast growth factor 21 depending on liver status. CONCLUSION: The current available data show that the severity of NAFLD is associated with an increase in oxidative stress and proinflammatory status. It may be also useful as diagnostic purpose in clinical practice.

The use of adipose mesenchymal stem cells and human umbilical vascular endothelial cells on a fibrin matrix for endothelialized skin substitute.

In recent years, the reconstruction of human skin by tissue engineering represents a clinical challenge and has offered a therapeutic alternative. Avascular engineered skin equivalents have been available for several years and used to treat wounds due to burns, nonhealing ulcers, and surgical excisions. They are constituted by different types of cultured cells included in a three-dimensional structure that permits cellular proliferation to create tissue substitutes. The major drawback of these artificial skin substitutes is their lack of blood supply, since the endurance and cell proliferation of the substitute depend on an adequate oxygen and nutrient supply and on toxin removal. These functions are served by the vascular system. We have produced a new model of endothelialized skin substitute that promotes the formation of capillary-like structures by seeding human umbilical vein endothelial cells (HUVECs) with dermal fibroblasts and human adipose-derived mesenchymal stem cells (hADMSCs) in a fibrin matrix. Dermal fibroblasts and hADMSCs produce extracellular matrix that stimulates cellular growth and proliferation. hADMSCs secrete significant quantities of angiogenic and antiapoptotic factors (vascular endothelial growth factor and hepatocyte growth factor), which induce in vitro differentiation of these cells into endothelial cells promoting angiogenesis and participating in tissue repair and skin regeneration processes. We obtained the artificial skin substitute with similar structure to native skin, including dermis and epidermis. We demonstrated that endothelial cells (CD31 and von Willebrand factor positive) proliferated and organized themselves into capillary-like structures within the fibrin matrix. The epidermis showed a complete epithelization by squamous cells (AE1/AE3 cytokeratin positive) with intracytoplasmic keratohyalin granules, hyperkeratosis, and parakeratosis. We have established a novel artificial skin substitute that facilitates the formation of capillary-like structures that may provide a novel therapeutic approach to different skin defects and prove to be a useful tool for regenerative medicine.

Assessment of ovarian cycles in the African elephant (Loxodonta africana) by measurement of salivary progesterone metabolites.

Monitoring ovarian cycles through hormonal analysis is important in order to improve breeding management of captive elephants, and non-invasive collection techniques are particularly interesting for this purpose. However, there are some practical difficulties in collecting proper samples, and easier and more practical methods may be an advantage for some institutions and/or some animals. This study describes the development and validation of an enzymeimmunoassay (EIA) for progestins in salivary samples of African elephants, Loxodonta africana. Weekly urinary and salivary samples from five non-pregnant elephant cows aged 7-12 years were obtained for 28 weeks and analyzed using EIA. Both techniques correlated positively (r = 0.799; P < 0.001), and the cycle characteristics obtained were identical. The results clearly show that ovarian cycles can be monitored by measuring progestins from salivary samples in the African elephant. This is a simple and non-invasive method that may be a practical alternative to other sampling methods used in the species.

Profile of patients with hepatic hydatid disease not treated surgically.

BACKGROUND: hepatic hydatid disease (HHD) is still an important health problem in certain areas of Spain where it is endemic. The treatment of HHD is usually surgical but certain patients are found to be ineligible after assessment for surgery (asymptomatic disease, comorbidity, patient refusal, or other). MATERIAL AND METHODS: description of patients assessed in the Department of Surgery for hepatic hydatid disease. RESULTS: in a group of 70 patients with HHD, 27 patients were not treated surgically (mean age: 72.7 years [range: 47-97], 14 women [51.8%]). The number of cysts presented by these patients was 33, with 1.22 cyst/patient (range: 1-4). The cyst size was 5.5 cm (range: 2.1-12.5 cm). The cysts, according to the WHO classification, were CE1: 3 patients, CE3B: 5 patients, CE4: 10 patients and CE5: 9 patients. The form of presentation was: symptomatic in 9 patients, although only 6 were attributable to HHD (22%) and asymptomatic in 18 patients. In these cases, imaging was performed for study of tumor extension in 6 patients and diverse medical reasons in 12. Only two therapeutic interventions were performed: endoscopic retrograde cholangiopancreatography (ERCP) with insertion of a bile duct stent, and puncture-aspiration-injection-re-aspiration (PAIR), both in patients who did not wish to undergo surgery.Ten patients had surgical indications: CE1 (3 patients), CE3B (5 patients), CE4 (1 patient), and CE5 (1 patient). The reasons why the patients did not undergo surgical treatment were: refusal (9 patients) and advanced neoplasm (1 patient). Surgery was judged necessary in 5 patients. In the mean follow-up period of 17 months (range: 1-37), no surgery was performed. CONCLUSIONS: there were various causes for not performing surgical intervention of HHD after medical evaluation: asymptomatic patients, older patients, patients with multiple pathologies and oncologic patients. Usually, they were patients who voluntarily chose not to undergo surgery.