Enzymes involved in l-carnitine biosynthesis are expressed by small intestinal enterocytes in mice: Implications for gut health

BackgroundCarnitine is essential for mitochondrial β-oxidation of long-chain fatty acids. Deficiency of carnitine leads to severe gut atrophy, ulceration and inflammation in animal models of carnitine deficiency. Genetic studies in large populations have linked mutations in the carnitine transporters OCTN1 and OCTN2 with Crohn's disease (CD), while other studies at the same time have failed to show a similar association and report normal serum carnitine levels in CD patients.MethodsIn this report, we have studied the expression of carnitine-synthesizing enzymes in intestinal epithelial cells to determine the capability of these cells to synthesize carnitine de novo. We studied expression of five enzymes involved in carnitine biosynthesis, namely 6-N-trimethyllysine dioxygenase (TMLD), 4-trimethylaminobutyraldehyde dehydrogenase (TMABADH), serine hydroxymethyltransferase 1 and 2 (SHMT1 and 2) and γ-butyrobetaine hydroxylase (BBH) by real-time PCR in mice (C3H strain). We also measured activity of γ-BBH in the intestine using an ex vivo assay and localized its expression by in situ hybridization.ResultsOur investigations show that mouse intestinal epithelium expresses all five enzymes required for de novo carnitine biosynthesis; the expression is localized mainly in villous surface epithelial cells throughout the intestine. The final rate-limiting enzyme γ-BBH is highly active in the small intestine; its activity was 9.7 ± 3.5 pmol/mg/min, compared to 22.7 ± 7.3 pmol/mg/min in the liver.ConclusionsWe conclude that mouse gut epithelium is able to synthesize carnitine de novo. This capacity to synthesize carnitine in the intestine may play an important role in gut health and can help explain lack of clinical carnitine deficiency signs in subjects with mutations with OCTN transporters.     

Enhanced proliferation in colorectal epithelium of patients with type 2 diabetes correlates with β-catenin accumulation

Aimsβ-Catenin accumulation promotes proliferation. However, the correlation between proliferation of colorectal epithelium and β-catenin in type 2 diabetes mellitus (DM) patients remains unclear.MethodsColorectal epithelium samples from distal ends of colorectal adenocarcinomas without histological aberrances were divided into two groups: DM patients with type 2 DM for more than 1 year (n = 27) and non-DM patients without hyperglycemia (n = 20). Samples from patients without colorectal epithelial disease or hyperglycemia served as a control group (n = 6). Proliferative index was calculated as the percentage of proliferating cell nuclear antigen positive cells. Wnt/β-catenin signaling was assessed immunohistochemically and phosphorylation of β-catenin was assessed by immunofluorescence.ResultsCompared with the non-DM or control group, the proliferative index and expression of lactate dehydrogenase A and Wnt/β-catenin signaling were significantly higher in the DM group (all p < 0.01). The proliferative index correlated positively with β-catenin expression (Spearman correlation coefficient = 0.55; p < 0.01). Reduced phosphorylation at serine 33/37 and increased phosphorylation at serine 675 of β-catenin were detected in the DM group (all p < 0.01).ConclusionsEnhanced proliferation, accompanied by increased aerobic glycolysis, was detected in colorectal epithelium of patients with diabetes. β-Catenin accumulation with altered phosphorylation correlated with the proliferative changes.     

Discontinuation of hormone replacement therapy in young GH-treated hypopituitary women increases liver enzymes

ObjectiveHypopituitarism, often characterized by hypogonadism, is associated with central obesity, increased cardiovascular and endocrine morbidity and mortality. In Turner syndrome, which is also characterized by hypogonadism liver enzymes are often elevated, but readily suppressed by a short course of hormone replacement therapy (HRT). We investigated the effect of HRT on liver enzymes, lipid levels and measures of insulin sensitivity 26 in hypopituitary women.DesignWe studied 26 hypopituitary women (age 38.8 ± 11.0 (mean ± SD years), BMI 27.4 ± 5.1 kg/m²) during HRT and 28 days off therapy.MethodsWe measured liver enzymes, fasting levels of lipids, insulin and glucose as well as adiponectin and leptin levels. Body composition was assessed by means of anthropometry and bioimpedance.ResultsAlanine transaminase (ALT) and aspartate transaminase (AST) increased after discontinuation of HRT (ALT; treated: 22.3 ± 11.5 vs. untreated: 27.1 ± 11.1 (U/L) (P < 0.02); AST; treated: 20.4 ± 6.1 vs. untreated: 24.6 ± 8.9 (U/L) (P < 0.002)), whereas other liver function tests remained unchanged. Measures of insulin sensitivity and fasting lipids were also unaffected by HRT, whereas leptin levels decreased with cessation of HRT (leptin; treated: 23 (8–71) vs. untreated: 20 (8–64) (μg/L) (P < 0.0005)).ConclusionShort time discontinuation of HRT in young hypopituitary women increased liver enzymes, whereas measures of insulin sensitivity and lipid levels remained unchanged. We speculate that the estrogen component of HRT has beneficial effects on hepatic metabolism through various pathways. Further studies including liver imaging and with a time-dependent design are needed to clarify the role of HRT on liver enzyme levels, metabolic variables and liver fat content.     

Uptake and retention of 65Zn in lithium-treated rat liver: Role of zinc

AimTo evaluate the effects of zinc on the biokinetics of ⁶⁵Zn in rat and its distribution in various organs and in subcellular compartment following lithium therapy.MethodsFemale wistar rats received either lithium treatment at a dose of 1.1 g/kg in diet, zinc alone at a dose of 227 mg/L in drinking water, and combined lithium plus zinc for duration of four months.ResultsAfter four months of lithium treatment, liver enzymes increased significantly (glutamic oxaloacetic transaminase, +66.73%; glutamic pyruvic transaminase, +63.70%; alkaline phosphatase, +40.28%; p ≤ 0.001); zinc supplementation to lithium-treated rats significantly reduced liver enzymes (glutamic oxaloacetic transaminase, −13.11%; glutamic pyruvic transaminase, −21.78%; alkaline phosphatase, −11.77%; p ≤ 0.001). The biological half-lives of ⁶⁵Zn showed an initial fast component (Tb₁) and a slower component (Tb₂). A significant increase in Tb₂ (38.82%, p ≤ 0.001) in liver was observed following lithium treatment, which significantly decreased following zinc treatment (21.71%, p ≤ 0.001). A significant decrease in the uptake of ⁶⁵Zn (53.93%, p ≤ 0.01) in liver was observed and in nuclear (p ≤ 0.01), mitochondrial (p ≤ 0.01), and microsomal (52.67%, p ≤ 0.001) fractions. A significant increase in the uptake of ⁶⁵Zn (82.92%, p ≤ 0.05) in liver microsomal fraction (34.09%, p ≤ 0.001) was observed in lithium-treated rats receiving zinc supplementation.ConclusionThe study suggests that zinc has the potential to regulate the biokinetics of ⁶⁵Zn and its subcellular distribution in rat liver following lithium therapy.     

Phosphorylation of eIF2α mitigates endoplasmic reticulum stress and hepatocyte necroptosis in acute liver injury

Introduction and objectivesNecroptosis and endoplasmic reticulum (ER) stress has been implicated in acute and chronic liver injury. Activated eukaryotic initiation factor 2 alpha (eIF2α) attenuates protein synthesis and relieves the load of protein folding in the ER. In this study, we aimed to analyze the impact of eIF2α phosphorylation on hepatocyte necroptosis in acute liver injury.Materials and methodsMale BALB/c mice were injected with tunicamycin or d-galactosamine, and LO2 cells were incubated with tunicamycin to induce acute liver injury. 4-Phenylbutyric acid (PBA) and salubrinal were used to inhibit ER stress and eIF2α dephosphorylation, respectively. We analyzed the eIF2α phosphorylation, ER stress, and hepatocyte necroptosis in mice and cells model.ResultsTunicamycin or d-galactosamine significantly induced ER stress and necroptosis, as well as eIF2α phosphorylation, in mice and LO2 cells (p < 0.05). ER stress aggravated tunicamycin-induced hepatocyte necroptosis in mice and LO2 cells (p < 0.05). Elevated eIF2α phosphorylation significantly mitigated hepatocyte ER stress (p < 0.05) and hepatocyte necroptosis in mice (34.37 ± 3.39% vs 22.53 ± 2.18%; p < 0.05) and LO2 cells (1 ± 0.11 vs 0.33 ± 0.05; p < 0.05). Interestingly, tumor necrosis factor receptor (TNFR) 1 protein levels were not completely synchronized with necroptosis. TNFR1 expression was reduced in d-galactosamine-treated mice (p < 0.05) and cells incubated with tunicamycin for 12 and 24 h (p < 0.05). ER stress partially restored TNFR1 expression and increased necroptosis in tunicamycin-incubated cells (p < 0.05).ConclusionsThese results imply that ER stress can mediate hepatocyte necroptosis independent of TNFR1 signaling and elevated eIF2α phosphorylation can mitigate ER stress during acute liver injury.     

The effect of liver test abnormalities on the prognosis of COVID-19

IntroductionCOVID-19 caused by the SARS-CoV-2 continues to spread rapidly across the world. In our study, we aim to investigate the relationship between the liver enzymes on admission (AST, ALT, ALP, GGT) and severity of COVID-19. We evaluated course of disease, hospital stay, liver damage and mortality.Materials and methodsOur study included 614 patients who were hospitalized with the diagnosis of COVID-19 between 03.16.20 and 05.12.20. Patients with liver disease, hematological and solid organ malignancy with liver metastases were excluded, resulting in 554 patients who met our inclusion criteria. We retrospectively evaluated liver transaminase levels, AST/ALT ratio, cholestatic enzyme levels and R ratio during hospital admission and these were compared in terms of morbidity, mortality and clinical course.ResultsMean age of 554 subjects were 66.21 ± 15.45 years, 328 (59.2%) were men. The mean values of liver enzymes on admission were AST (36.2 ± 33.6 U/L), ALT (34.01 ± 49.34 U/L), ALP (78.8 ± 46.86 U/L), GGT (46.25 ± 60.05 U/L). Mortality rate and need for intensive care unit were statistically significant in subjects that had high ALT–AST levels during their admission to the hospital (p = 0.001). According to the ROC analysis AST/ALT ratio was a good marker of mortality risk (AUC = 0.713: p = 0.001) and expected probability of intensive care unit admission (AUC = 0.636: p = 0.001). R ratio, which was used to evaluate prognosis, showed a poor prognosis rate of 26.5% in the cholestatic injury group, 36.1% in the mixed pattern group and 30% in the hepato-cellular injury group (p 0.001).ConclusionsALT–AST elevation and AST/ALT ratio >1 was associated with more severe course and increased mortality in COVID-19.     

Serum osteopontin levels as a predictor of portal inflammation in patients with nonalcoholic fatty liver disease

BackgroundOsteopontin is a secreted phosphorylated glycoprotein that is expressed by a variety of cell types and that mediates numerous and diverse biological functions.Osteopontin knockout mice are protected from obesity-induced hepatic steatosis. In the present study, we sought to investigate whether serum osteopontin concentrations are associated with liver histology in patients with nonalcoholic fatty liver disease.MethodsSerum levels of osteopontin were measured by enzyme-linked immunosorbent assay in 179 well-characterized patients with nonalcoholic fatty liver referred for liver histology and 123 control subjects.ResultsSerum osteopontin levels were markedly higher in patients with nonalcoholic fatty liver disease than in controls (p < 0.001). Multivariable analysis showed that osteopontin levels were strongly and independently associated with both portal inflammation (β = 0.294, p < 0.01) and serum aminotransferase levels (aspartate aminotransferase: β = 0.295, p < 0.01; alanine aminotransferase; β = 0.285, p < 0.01).ConclusionIn summary, these data demonstrate that serum levels of osteopontin are elevated in nonalcoholic fatty liver disease and are a significant independent predictor of portal inflammation in this clinical entity.     

Lipid accumulation is ahead of epithelial-to-mesenchymal transition and therapeutic intervention by acetyl-CoA carboxylase 2 silence in diabetic nephropathy

ObjectiveThe study investigated the relationship between epithelial-to-mesenchymal transition (EMT) and lipotoxicity in diabetic nephropathy as well as the protective effect of acetyl-CoA carboxylase 2 (ACC2) silence.MethodsHigh glucose (30 mmol/L) cultured human proximal tubular epithelial cells (HK-2 cells) were used. Triglyceride content, fatty acid β-oxidation rate, malonyl CoA content, and marker proteins of EMT, including E-cadherin (E-cad), α-smooth muscle actin (α-SMA) and transforming grow factor-β (TGF-β), were assessed. Silence of ACC2 was achieved by ACC2-shRNA lentivirus transfection.ResultsIn cultured human proximal tubular cells, high glucose induced fatty acid deposit before phenotypical and morphological changes of EMT. At 48 h, more triglyceride content, more malonyl CoA content and lower fatty acid β-oxidation rate were detected. However, increased expression of TGF-β, accompanied by loss of E-cad and acquisition of α-SMA, was observed at 98 h but not at 48 h. The silence of ACC2 in HK-2 cells led to restored cell morphology with less lipid deposition and less malonyl-CoA content, which resulted from faster β-oxidation rate.ConclusionThe progress of lipotoxicity participates in the development of diabetic nephropathy in early stage before EMT. The manipulation of lipid metabolism might act as a promising therapeutic intervention for diabetic nephropathy.     

Renoprotective effect of a novel selective PPARα modulator K-877 in db/db mice: A role of diacylglycerol-protein kinase C-NAD(P)H oxidase pathway

ObjectiveSeveral clinical studies have shown the beneficial effects of peroxisome proliferator-activated receptor α (PPARα) agonists on diabetic nephropathy. However, the molecular mechanism is not fully understood. Here we show that K-877, a novel selective PPARα modulator, ameliorates nephropathy in db/db mice via inhibition of renal lipid content and oxidative stress.Methods and resultsK-877 (0.5 mg/kg/day) was administered to db/db mice for 2 or 12 weeks. Short-term treatment did not affect body weight or plasma glucose levels in db/db mice, but attenuated albuminuria, along with improvement of plasma lipid profiles, lipid content including total diacylglycerol (DAG) levels, protein kinase C (PKC) activity, NAD(P)H oxidase-4 expression, and oxidative stress markers, all of which were significantly increased in diabetic kidneys. It increased phosphorylation of 5′-AMP activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC), and expression of several genes mediating fatty acid β-oxidation. In addition, long-term treatment ameliorated renal mesangial expansion in db/db mice and improved glycemic control.ConclusionsK-877 administration ameliorates diabetic nephropathy, at least in part, via inhibition of renal lipid content and oxidative stress. The underlying mechanism may be mediated by modulating the renal AMPK-ACC pathway, subsequent acceleration of fatty acid β-oxidation and inhibition of fatty acid synthesis, and thus inhibition of the DAG-PKC-NAD(P)H oxidase pathway, in addition to its systemic effect including improvement of the plasma lipid profile and glycemic control.     

Influence of β2-adrenoceptor gene polymorphisms on β2-adrenoceptor expression in human lung

BackgroundThe aim of the present study was to establish whether polymorphisms, especially those within the promoter region, of the β₂-adrenoceptor gene (ADRB2) influence β₂-adrenoceptor expression in human lung.MethodsThe density of β-adrenoceptors in human lung tissue (n = 88) was determined by saturation binding using the radioligand, iodinated cyanopindolol. Discrimination of β₁- and β₂-adrenoceptors was determined using the highly selective β₁-adrenoceptor antagonist, CGP20712A. Genotype was determined at 5 positions of ADRB2 previously reported as polymorphic. Potential influences of single nucleotide polymorphisms (SNPs) within the promoter region (?367, ?47) and coding block (46, 79, 491) of ADRB2 on β₂-adrenoceptor expression were investigated.ResultsThe density of β₂-adrenoceptors was variable among the 88 lung preparations studied ranging from 17 to 177 fmol/mg protein (mean ± S.E.M., 72 ± 4 fmol/mg protein). There was no influence of genotype on β₂-adrenoceptor expression for any of the polymorphisms studied except at position 491. The polymorphism at position 491C > T, leading to a change from thr to ile at amino acid 164, is uncommon. Preparations genotyped as heterozygous (126 ± 15 fmol/mg protein; n = 5) expressed significantly (P = 0.0005) higher levels of β₂-adrenoceptor than those that were homozygous (69 ± 4 fmol/mg protein; n = 83).ConclusionWith the exception of position 491, these data indicate that polymorphisms of ADRB2 do not influence β₂-adrenoceptor expression in human lung.     

Alcohol and HBV synergistically promote hepatic steatosis

Background and aimsHepatitis virus and alcohol are the main factors leading to liver damage. Synergy between hepatitis B virus (HBV) and alcohol in promoting liver cell damage and disease progression has been reported. However, the interaction of HBV and ethanol in hepatic steatosis development has not been fully elucidated.MethodsEight-week-old male C57BL/6 mice were treated with or without HBV, ethanol, or the combination of HBV and ethanol (HBV + EtOH), followed by a three-week high-fat diet (HFD) regimen. Liver histology, serum biomarkers, and liver triglyceride levels were analysed. Furthermore, a meta-analysis of the effects of alcohol and HBV on hepatic steatosis in populations was performed.ResultsHepatic steatosis was significantly more severe in the HBV + EtOH group than in the other groups. The serum alanine aminotransferase, aspartate aminotransferase and liver triglyceride levels in the HBV + EtOH group were also significantly higher than those in the other groups. The HBeAg and HBsAg levels in the HBV + EtOH group were significantly higher than those in the pair-fed HBV-infected mice. In addition, the meta-analysis showed that alcohol consumption increased the risk of hepatic steatosis by 43% in HBV-infected patients (pooled risk ratio (RR) = 1.43, P < 0.01).ConclusionsAlcohol and HBV synergistically promote high-fat diet-induced hepatic steatosis in mice. In addition, alcohol consumption increases the risk of hepatic steatosis in HBV-infected patients.     

Sonographic features suggestive of amyloidosis in hemodialysis patients: Relations to serum beta2-microglobulin

Aim of the workTo determine sonographic features suggestive of amyloidosis in hemodialysis patients complaining of shoulder pain, and to study their relations to serum beta2-microglobulin (β2M).Patients and methodsClinical examination, skeletal survey, musculoskeletal ultrasonography of the shoulder joints, and serum β2M were done for 32 patients with end stage renal disease, who were regular on hemodialysis.ResultsSerum β2M levels were markedly raised in all patients, and increased with increasing duration of dialysis (r = 0.91, p < 0.001). Twenty-five patients had a non homogeneous thickening of the supraspinatus tendon >7 mm, and thickening of the biceps tendon >4 mm, 30 had synovial deposits, 27 had subdeltoid bursa effusion, 25 had thickened subacromial bursa, 7 had supraspinatus tendon tear, and 17 had bony erosions. Serum β2M levels significantly correlated with thickened supraspinatus tendon (>7 mm) and supraspinatus tendon tear (r = 0.41, p = 0.03 and r = 0.42, p = 0.01 respectively). Long time on hemodialysis was the significant independent determinant for supraspinatus tendon tear and humeral head erosions (p = 0.001 for each).ConclusionElevated serum β2M levels and sonographic features suggestive of dialysis-related amyloidosis (DRA) were found in all hemodialyzed patients complaining of shoulder pain either with or without clinical and/or radiological features suggestive of DRA. So, for diagnosis of DRA, sonographic features should correspond to the presence of clinically or radiologically evident β2M amyloid, and we should exclude other causes of non-amyloid changes.     

Serum sex steroids and steroidogenesis-related enzyme expression in skeletal muscle during experimental weight gain in men

ObjectivesLow-circulating testosterone is associated with development of type 2 diabetes in obese men. In this study, we examined the effects of experimental overfeeding and weight gain on serum levels of sex hormones and skeletal muscle expression of steroidogenic enzymes in healthy men with (FH+) and without (FH–) a family history of type 2 diabetes.MethodsFollowing a 3-day lead in energy balanced diet, FH+ (n = 9) and FH– men (n = 11) were overfed by 5200 kJ/day (45% fat) for 28 days. Body weight, fasting glucose, insulin, sex steroid, sex hormone binding globulin (SHBG) levels, insulin sensitivity (hyperinsulinaemic-euglycaemic clamp) and body fat (DXA) were assessed in all individuals at baseline and day 28, and sex steroidogenesis-related enzyme expression in vastus lateralis biopsies was examined in a subset (n = 11).ResultsBody weight, fat mass and fasting insulin levels were increased by overfeeding (P < 0.01) and insulin was increased significantly more in FH+ men (P < 0.01). Serum sex hormone binding globulin (SHBG) and 5α-dihydrotestosterone (DHT) were reduced with overfeeding (P < 0.05), and serum testosterone and DHT were reduced to a greater extent in FH+ men (P < 0.05). Overfeeding reduced mRNA expression of 3β-hydroxysteroid dehydrogenase (HSD) and 17βHSD (P ≤ 0.007), independently of group. 5α-Reductase (SRD5A1) mRNA expression was not changed overall, but a time by group interaction was observed (P = 0.04).ConclusionOverfeeding reduced SHBG and muscle expression of enzymes involved in the formation of testosterone in skeletal muscle. Men with a family history of T2DM were more susceptible to deleterious outcomes of overfeeding with greater reductions in serum testosterone and DHT and greater increases in markers of insulin resistance, which may contribute to increased risk of developing type 2 diabetes.     

La restriction énergétique réduit le stress oxydant de l’aorte et du cœur et corrige le risque athérogène chez le rat rendu obèse

Aim of the studyThe aim was to see if 40% of caloric restriction can improve in rats, the oxidative stress induced by a high fat diet on liver, heart and aorta, and have a positive impact on the lipoproteins-cholesterol contents.MethodsMale Wistar rats (n = 12) consumed at weaning obesegenic diet. The obese rats were divided into two groups (n = 6) each consumed for 4 weeks a normocaloric diet (1,60 MJ) or a calorie restricted diet (40% of energy of the standard diet [0.96 MJ]). A control group (n = 6) was fed a standard diet during the experiment.ResultsCaloric restriction decreased cholesterol and low-density lipoproteins-cholesterol, whereas the high-density lipoproteins-cholesterol was elevated. Levels of cholesterol in the aorta and heart were lowered in the group that consumed caloric restriction compared to normocaloric diet. The values of thiobarbituric acid reactive substances and hydroperoxides were lowered in the aorta and the heart. The caloric restriction compared to normocaloric diet increased positively the superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase activities in the heart and those of catalase and superoxide dismutase in aorta.ConclusionIn obese rats, caloric restriction may play a role in the antioxidant defense of cardiovascular system by reducing proatherogenic cholesterol and lipid peroxidation and increasing the antioxidant activity of some enzymes, which was in favor of reduction of cardiometabolic risk associated with obesity.     

Pro-inflammatory Interleukin-18 and Caspase-1 serum levels in liver failure are unaffected by MARS treatment

BackgroundThe pro-inflammatory cytokine IL-18 and its activator Caspase-1 are involved in acute liver failure and acute-on-chronic-liver-failure. In acute liver failure and acute-on-chronic-liver-failure, the MARS system has been used to support liver function. Enhancement of IL-18, as seen in other extracorporeal-support systems like hemodialysis might thus have mitigated beneficial effects of the MARS system in acute hepatic failure.Patients and methodsWe measured serum concentrations of IL-18 and Caspase-1 in 10 patients with acute liver failure and 10 patients suffering from acute-on-chronic-liver-failure, who were all treated with MARS. Thirteen patients suffering from chronic hepatic failure and 15 healthy individuals served as controls. Data are given as mean with 95% CI.ResultsBaseline IL-18 serum concentrations were significantly increased in acute liver failure and acute-on-chronic-liver-failure patients as compared to chronic hepatic failure (P = 0.0039 and P = 0.0011, respectively) and controls (P = 0.0028 and P = 0.0014, respectively). Caspase-1 serum concentrations were as well significantly elevated in the acute liver failure and acute-on-chronic-liver-failure groups as compared to chronic hepatic failure patients (P = 0.0039 and P = 0.0232, respectively) and controls P < 0.0001 and P < 0.0007, respectively). IL-18 and Caspase-1 did not change significantly during MARS treatment in acute liver failure and acute-on-chronic-liver-failure patients.ConclusionsMARS had no effect on IL-18 and Caspase-1 serum concentrations in acute liver failure and acute-on-chronic-liver-failure, providing no evidence of harmful effects by the increase of these potentially hepatocidal cytokines.     

The spectrum of β-thalassemia mutations in Gaza Strip,Palestine

Backgroundβ-Thalassemia is a disorder caused by mutations at the hemoglobin β-gene (HBB) locus. Its most important manifestation, the major form, is characterized by severe hypochromic and hemolytic anemia and is inherited in an autosomal recessive mode. In Gaza Strip, Palestine 0.02% of the population has been identified as β-thalassemia major.Design and methodsAn assessment of mutations was performed in 49 transfusion dependent patients with β-thalassemia major and in 176 β-thalassemia carriers diagnosed with a mean erythrocyte cell volume (MCV) < 80 fl and a proportion of HbA₂ > 3.5%. In addition 39 individuals suspicious for β-thalassemia carrier status due to a reduced MCV (< 80 fl) but a normal HBA₂ were screened.ResultsBy screening with three hybridization assays a proportion of 80% of the thalassemic chromosomes from patients and carriers was identified to carry five different mutations of the hemoglobin (Hb) β-gene. Subsequent DNA sequencing confirmed these and revealed further 9% of the chromosomes to be affected by other mutations. In addition six chromosomes from suspicious carriers were detected to carry β-thalassemia mutations. Of the 15 different HBB mutations identified the variant IVS-I-110 G > A was the most frequent mutation identified in 34% of the thalassemic chromosomes, followed by IVS-I-1 G > A, IVS-I-6 T > C, Codon 39 C > T, and Codon 37 G > A. Three novel HBB variants were discovered by direct sequencing of the gene: 5′ UTR-50 (?/G), 5′ UTR-43 C > T, and IVS-II-26 T > G.ConclusionsThe spectrum of HBB mutations described is of the Mediterranean type whereby the allele frequencies of the most common mutations differ from those, which were previously described for the population of the Gaza Strip and other Palestinian populations. The data presented may promote the introduction of molecular testing to the Palestinian premarital screening program for β-thalassemia in Gaza Strip, which will improve the screening protocol and genetic counseling in the future.     

Predictive accuracy of serum hyaluronic acid as a non-invasive marker of fibrosis in a cohort of multi-transfused Egyptian children with β-thalassaemia major

Background and study aimLiver disease remains a major cause of morbidity and mortality in patients with β-thalassaemia major (β-TM); therefore, its identification at an early stage is of great significance. Serum hyaluronic acid (HA) is considered as a non-invasive marker that appears early before pathological changes occur. We aim to determine the predictive accuracy of HA in detecting and staging hepatic fibrosis in β-TM patients.Patients and methods30 Egyptian children with β-TM, and 15 age and sex-matched controls were studied. All had abdominal ultrasonography (US), measurement of serum amino-transferases (ALT, AST); hepatitis C, B and human immunodeficiency viruses (HCV, HBV, HIV) sero-markers, serum ferritin and HA. Liver biopsy was done for patients and fibrosis was scaled using Metavir scoring system and liver iron concentration (LIC) was measured.ResultsTwenty patients (67.7%) had sero-markers of HCV, none had HBV or HIV. Serum HA was significantly higher in patients (90.78 ± 28.79 ng/ml) compared to controls (21.1 ± 13.24 ng/ml) with p < 0.05. No difference between HCV infected and non-infected patients was detected. Positive significant correlation was detected between serum HA and stages of fibrosis by histopathology and US. No correlation was found between serum HA and age, sex, weight, height, haemoglobin level, platelet count, AST, serum ferritin, necro-inflammatory grade, and LIC.ConclusionsSerum HA is a valuable non-invasive marker that may contribute to the assessment of liver fibrosis in multi-transfused children and adolescents with β-TM, irrespective of concomitant HCV infection.     

Immediate postoperative Fibrosis-4 predicts postoperative liver failure for patients with hepatocellular carcinoma undergoing curative surgery

BackgroundPostoperative liver failure remains the main complication and predominant cause of hepatectomy-related mortality for patients undergoing liver resection.AimOur aim is to investigate whether immediate postoperative Fibrosis-4 could predict postoperative liver failure.MethodsWe retrospectively enrolled 1353 consecutive hepatocellular carcinoma patients undergoing radical resection. The characteristics and clinical outcomes were compared between patients with high and low immediate postoperative Fibrosis-4. Risk factors for hepatic failure were evaluated by univariate and multivariate analysis.ResultsUsing a receiver operating characteristic curve, immediate postoperative Fibrosis-4 showed good prediction ability for postoperative liver failure (AUROC = 0.647, P < 0.001). With the optimal cut-off value of 5.9, the high postoperative Fibrosis-4 group (Fibrosis-4 < 5.9) had higher postoperative complication (39.1% vs 28.6%, P < 0.001), mortality (2.8% vs 0.6%, P < 0.001) and liver failure (13.9% vs 6.2%, P < 0.001). In addition, patients with high Fibrosis-4 had worse and delayed recovery of liver function. By univariate and multivariate analysis, Fibrosis-4, as well as liver removed volume, total bilirubin and albumin was identified as independent risk factor for postoperative liver failure.ConclusionsImmediate postoperative Fibrosis-4 showed good prediction ability for postoperative liver failure, and required measure should be taken to prevent liver failure when high postoperative Fibrosis-4 appeared.     

Hyperdynamic circulatory syndrome in a mouse model transgenic for SerpinB3

Introduction and objectivesSerpinB3 is a cysteine protease inhibitor involved in several biological activities. It is progressively expressed in chronic liver disease, but not in normal liver. The role in vascular reactivity of this serpin, belonging to the same family of Angiotensin II, is still unknown. Our aim was to evaluate the in vivo and in vitro effects of SerpinB3 on systemic and splanchnic hemodynamics.Material and methodsDifferent hemodynamic parameters were evaluated by ultrasonography in two colonies of mice (transgenic for human SerpinB3 and C57BL/6J controls) at baseline and after chronic carbon tetrachloride (CCl₄) treatment. In vitro SerpinB3 effect on mesenteric microvessels of 5 Wistar-Kyoto rats was analyzed measuring its direct action on: (a) preconstricted arteries, (b) dose–response curves to phenylephrine, before and after inhibition of angiotensin II type 1 receptors with irbesartan. Hearts of SerpinB3 transgenic mice and of the corresponding controls were also analyzed by morphometric assessment.ResultsIn SerpinB3 transgenic mice, cardiac output (51.6 ± 21.5 vs 30.1 ± 10.8 ml/min, p = 0.003), hepatic artery pulsatility index (0.85 ± 0.13 vs 0.65 ± 0.11, p < 0.001) and portal vein blood flow (5.3 ± 3.2 vs 3.1 ± 1.8 ml/min, p = 0.03) were significantly increased, compared to controls. In vitro, recombinant SerpinB3 had no direct hemodynamic effect on mesenteric arteries, but it increased their sensitivity to phenylephrine-mediated vasoconstriction (p < 0.01). This effect was suppressed by inhibiting angiotensin II type-1 receptors.ConclusionsIn transgenic mice, SerpinB3 is associated with a hyperdynamic circulatory syndrome-like pattern, possibly mediated by angiotensin receptors.     

Intestinal permeability is increased in children with non-alcoholic fatty liver disease,and correlates with liver disease severity

BackgroundIncreased intestinal permeability seems to play a major role in non-alcoholic liver disease development and progression.AimTo investigate the prevalence of altered intestinal permeability in children with non-alcoholic fatty liver disease, and to study its potential association with the stage of liver disease.MethodsWe performed a case–control study examining intestinal permeability in children using the lactulose–mannitol bowel permeability test.ResultsOverall, 39 consecutive patients (30 males, median age 12 years) and 21 controls (14 males, median age 11.8 years) were included. The lactulose/mannitol ratio resulted impaired in 12/39 patients (31%) and none of the controls. Intestinal permeability was higher in children with non-alcoholic fatty liver disease (lactulose/mannitol ratios: 0.038 ± 0.037 vs. 0.008 ± 0.007, p < 0.05). Within the non-alcoholic fatty liver disease group, intestinal permeability was increased in children with steatohepatitis compared to those with steatosis only (0.05 ± 0.04 vs. 0.03 vs. 0.03, p < 0.05). Pathological lactulose/mannitol ratio correlated with portal inflammation (p = 0.02), fibrosis (p = 0.0002), and ballooning of hepatocytes (p = 0.003). Blood lipopolysaccharides levels were higher in children with steatohepatitis (2.27 ± 0.68 vs. 2.80 ± 0.35, p < 0.05).ConclusionsIntestinal permeability is increased in children with non-alcoholic fatty liver disease, and correlates with the severity of the disease.