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Isolation of micro- and macro-droplet fractions from needle biopsy specimens of human liver and determination of the subcellular distribution of the accumulating liver lipids in alcoholic fatty liver.

Needle biopsy specimens of liver were obtained from six control subjects with histologically normal liver and 11 chronic alcoholics with fatty liver. Micro- and macro-lipid droplet fractions were isolated by differential flotation. These fractions, together with the sedimenting membranes, were assayed for cholesterol, cholesteryl ester, phospholipid, free fatty acids and triglyceride. Electron microscopy demonstrated marked differences in the range of lipid droplet sizes in the two fractions and biochemical analysis suggested that the microdroplet lipid corresponded to pre-very low density lipoprotein (VLDL) particles. Studies on biopsies from patients with alcoholic fatty liver showed a 2-3-fold increase in triglyceride in both lipid droplet fractions but most of the accumulating triglyceride was sedimentable and membrane-bound. Needle biopsy specimens from two patients with alcoholic fatty liver were fractionated with a vertical pocket re-orientating rotor. The principal organelles were separated and the subcellular distribution of triglyceride, phospholipid and free cholesterol determined. Triglyceride showed a bimodal distribution to a particulate fraction tentatively located to Golgi particles and to droplet-lipid remaining in the sample layer.

Biopsy, Needle↗

Regulation of the liver fatty acid-binding protein gene by hepatocyte nuclear factor 1alpha (HNF1alpha). Alterations in fatty acid homeostasis in HNF1alpha-deficient mice.

Hepatocyte nuclear factor 1alpha (HNF1alpha)-null mice have enlarged fatty livers and alterations in the expression of genes encoding enzymes involved in the synthesis, catabolism, and transport of fatty acids. Elevations in the expression of genes encoding fatty acid synthetic enzymes (fatty acid synthase and acyl-CoA carboxylase) and peroxisomal beta-oxidation enzymes (CYP4A3, bifunctional enzyme, and thiolase) were observed in the livers of HNF1alpha-null mice, whereas hepatic mitochondrial beta-oxidation gene (medium and short chain acyl-CoA dehydrogenase) expression levels remain unchanged relative to HNF1alpha-heterozygous controls. An elevation in the levels of fatty acid transporter gene expression was also observed. In contrast, there was a marked reduction of liver fatty acid-binding protein (l-FABP) gene expression in the livers of HNF1alpha-null mice. Isolation and sequence analysis of the 5'-flanking region of the mouse l-FABP gene revealed the presence of two HNF1alpha regulatory elements. The results of transient transfection studies indicate that HNF1alpha is required to trans-activate the expression of the l-FABP promoter. Taken together, these data define a critical role for HNF1alpha in the pathogenesis of a phenotype marked by fatty infiltration of the liver and in the regulation of the l-FABP gene, the expression of which may have a direct impact on the maintenance of fatty acid homeostasis.

Animals↗

Plasma lipids in normal cows around partus and in cows with metabolic disorders with and without fatty liver.

Free fatty acids, cholesterol and phospholipids in plasma were studied from 6 weeks before to 6 weeks after calving in 16 normal multiparous cows. The same plasma lipids were studied the day after calving in 20 normal primiparous cows. Ten of these were fed according to standards and the other 10 were overfed the last 3 weeks prepartum. The plasma lipids were also analysed in 16 cows with left displacement of the abomasum and fatty liver, and in 16 cows with ketosis with no or only slight fat infiltration of the liver. In the normal cows there was a rise in FFA-level and a reduction in cholesterol and phospholipids from 6 weeks before to the day after calving. Thereafter there was a reduction of FFA-level and rise in cholesterol and phospholipids. Increased feed intensity had no effect on plasma lipids at calving. The level of the plasma lipids in cows with fatty liver differed very much from the amounts in normal cows at corresponding time from calving. Cows with ketosis had high FFA-level but the amount of cholesterol and phospholipids differed very little from normal cows.

Acidosis↗

Inhibitory effect of a new alpha-glucosidase inhibitor on fatty liver in Zucker fatty rats.

The livers of Zucker fatty (fa/fa) and lean (Fa/-) rats treated with standard rat chow diet containing 0, 10 or 50 ppm alpha-glucosidase inhibitor (AO-128) for 10 weeks were studied morphologically and biochemically. Light microscopic examination of livers from untreated Zucker fatty rats showed severe steatosis. The triglyceride content in the livers from Zucker fatty rats was significantly higher than that from lean rats (73 +/- 9 micrograms/mg protein for Zucker fatty rats vs. 30 +/- 10 for lean rats, p less than 0.01). Administration of the inhibitor caused a marked decrease in the number and size of lipid droplets in the hepatocytes from Zucker fatty rats and a decrease in the triglyceride content in the liver (73 +/- 9 micrograms/mg protein for untreated, 54 +/- 16 for 10-ppm-treated and 48 +/- 23 for 50-ppm-treated rats, p less than 0.05). This is the first report showing an inhibitory effect of an alpha-glucosidase inhibitor on steatosis in Zucker fatty rats.

Animals↗

[The behavior of lipid metabolism parameters in liver cirrhosis and fatty liver during glucose load].

In altogether 31 patients with liver cirrhosis, fatty degeneration of the liver or a morphologically normal liver the free fatty acids, glycerin, ketone bodies and triglycerides were examined as parameters of the fat metabolism after nocturnal alimentary abstinence and under 2-hour glucose infusion. Parallel to this estimations of the immune-reactive insulin and of the blood glucose were performed. In liver cirrhoses increased levels of free fatty acids, low ketone body levels, the absent correlation between ketone bodies and immune-reactive insulin under basis conditions as well as the relatively smaller reduction of the TG-values in hyperinsulinaemia were to be established. They may be explained as an expression of restricted metabolic functions of the cirrhotic liver. On the other hand from the prompt decrease of free fatty acids and glycerin under glucose-induced hyperinsulinaemia was concluded to an unrestricted efficacy of the insulin in the fatty tissue in the sense of the furthering of lipogenesis and inhibition of the lipolysis.

Fatty Acids, Nonesterified↗

Early effects of feeding excess vitamin A: mechanism of fatty liver production in rats.

Oral administration of vitamin A (30,000 IU daily for 2 days) to young rats caused a marked increase in hepatic glycogen, cholesterol, and glycerides, while hepatic phospholipid content remained almost unaltered. In an examination of the pathogenesis of the lipid accumulation, it was found that more glucose-(14)C was incorporated into liver lipids in vitamin A-fed rats, whereas incorporation of glucose-(14)C and dl-glycine-(14)C into liver protein remained unaltered. The increase in glucose-(14)C incorporation was confined to the glyceride-glycerol portion of the lipids; incorporation into liver fatty acids was inhibited. Plasma free fatty acid concentrations were elevated. It is postulated that in the vitamin A-fed rats, increased accumulation of lipids in the liver is caused by a stimulation of fatty acid mobilization from adipose tissue and enhanced formation of glycerophosphate through glycolysis, with consequent increase in the glyceride synthesis in the liver. The weight of the adrenals was increased, whereas cholesterol concentration in the gland was decreased, after administration of vitamin A to rats. This indicates adrenocortical stimulation. Interestingly enough, vitamin A feeding did not affect either the level of liver lipids or of plasma FFA in adrenalectomized rats.

Adrenal Glands↗

[Liver fibrosis indexes in nonalcoholic fatty liver disease].

OBJECTIVE: To assess the clinical utility of liver fibrosis indexes determination in patients with nonalcoholic fatty liver disease. METHODS: Liver fibrosis indexes of 79 patients with fatty liver, diagnosed by B ultrasonic echography, 37 patients with posthepatitic cirrhosis patients and 32 healthy subjects were determined. RESULTS: The level of procollagen III(PC III), hyaluronic acid (HA), collagen IV(C IV), laminin (LN) in the fatty liver cases were significantly higher than those of healthy subjects (P<0.05 - 0.01), but lower than those in the liver cirrhosis cases (P<0.01). The difference of PC III, C IV, LN between the severe and early fatty liver cases was significant (P<0.05 - 0.01). Positive rate of these indexes was higher in severe cases. CONCLUSION: There is a trend of liver fibrosis in severe fatty liver patients. Combined determination of liver fibrosis indexes may help the diagnosis of liver fibrosis in the fatty liver patients.

Adult↗

Prevalence and risk factors of nonalcoholic fatty liver disease in an adult population of taiwan: metabolic significance of nonalcoholic fatty liver disease in nonobese adults.

BACKGROUND: The prevalence of nonalcoholic fatty liver disease (NAFLD) is rarely reported in Taiwan. GOALS: To determine the prevalence and risk factors of NAFLD in an adult population of Taiwan. STUDY: The cross-sectional community study examined 3245 adults in a rural village of Taiwan. The diagnostic criteria for NAFLD included no excessive alcohol intake, no chronic viral hepatitis, no known etiologies of liver disease, and ultrasonography consistent with fatty liver. RESULTS: The prevalence of NAFLD was 11.5% (372/3245). The risk factors for NAFLD in the general population were male sex [odds ratio (OR), 1.44; 95% confidence interval (CI), 1.09-1.90], elevated alanine aminotransferase (ALT) (OR, 5.66; 95% CI, 3.99-8.01), obesity (OR, 7.21; 95% CI, 5.29-9.84), fasting plasma glucose > or =126 mg/dL (OR, 2.08; 95% CI, 1.41-3.05), total cholesterol > or =240 mg/dL (OR, 1.50; 95% CI, 1.06-2.13), triglyceride > or =150 mg/dL (OR, 1.76; 95% CI, 1.32-2.35), and hyperuricemia (OR, 1.53; 95% CI, 1.16-2.01). Age > or =65 years was inversely related to NAFLD (OR, 0.53; 95% CI, 0.36-0.77). The only NAFLD risk factors among nonobese subjects were age between 40 and 64 years (OR, 2.35; 95% CI, 1.34-4.11, P=0.003), elevated ALT (OR, 15.45; 95% CI, 8.21-29.09, P<0.001), and triglyceride > or =150 mg/dL (OR, 2.48; 95% CI, 1.42-4.32, P=0.001). In subjects with NAFLD, the prevalence of elevated ALT in the presence of each metabolic risk factor, such as obesity, fasting plasma glucose > or =126 mg/dL, total cholesterol > or =240 mg/dL, triglyceride > or =150 mg/dL, and hyperuricemia, did not differ from that of subjects with normal ALT levels. CONCLUSIONS: NAFLD is closely associated with elevated ALT, obesity, diabetes mellitus, hypercholesterolemia, hypertriglyceridemia, and hyperuricemia. Among the metabolic disorders, only hypertriglyceridemia was related to NAFLD in nonobese subjects. Serum ALT level was not a good predictor of metabolic significance in subjects with NAFLD.

Adult↗

In situ binding of fatty acids to the liver fatty acid binding protein: analysis using 3-[125I]iodo-4-azido-N-hexadecylsalicylamide.

A photoactivatable radioiodinated fatty acid analogue, 3-[125I]iodo-4-azido-N-hexadecylsalicylamide (125I-AHS) has been synthesized and used to investigate the involvement of cellular lipid carriers in hepatic fatty acid utilization. Photoactivation of Hep G2 internalized 125I-AHS revealed that several cellular proteins were crosslinked with the radiolabeled fatty acid analogue. Three predominant proteins in the membrane fraction of the cell with molecular masses 17, 50 and 127 kDa were crosslinked with the lipid analogue, as determined using autoradiography after SDS-PAGE. Three other proteins in the soluble fraction of the cell, with molecular masses 14, 24 and 35 kDa, were also labeled in situ. In contrast to the other labeled proteins, the fatty acid analogue accumulated on the cytoplasmic 14 kDa protein in a time and temperature dependent fashion. The in situ-labeled 14 kDa protein was identified from primary rat hepatocytes as the liver fatty acid binding protein by partial purification and its ability to be immunoprecipitated with immunospecific L-FABP antiserum. Collectively the results indicate that fatty acids traverse the plasma membrane and are bound cytoplasmically by the liver fatty acid binding protein, as well as other proteins in the cell. This represents the first demonstration in intact hepatocytes that the liver fatty acid binding protein participates in the process of intracellular fatty acid trafficking, and supports a model in which cytoplasmic lipid carriers solubilize fatty acids as a step in their metabolic utilization.

Affinity Labels↗

Efficacy and effectiveness of liver screening program to detect fatty liver in the periodic health check-ups.

To determine whether the current liver screening program for fatty liver has sufficient scientific evidence to justify its continued implementation. The liver screening program to detect fatty liver was performed on 411 Japanese workers utilizing serum aspartate aminotransferase (ALT), alanine aminotransferase (AST), and gammaglutamyl transpeptidase (gamma-GTP). Based on the preceding studies, subjects with viral and alcohol hepatitis were excluded from the evaluation. The diagnosis of fatty liver was based on ultrasound findings. The program was evaluated by efficacy and effectiveness; efficacy was measured according to the receiver operating characteristic (ROC) curves in comparison with the Body Mass Index (BMI). Effectiveness, based on the efficacy determinations, was assessed by means of the positive predictive value (PPV) test performance, the disease characteristics, and the program price. The diagnostic performances of ALT and BMI were nearly acceptable but far from excellent. The areas under the curves of the two indices were 0.69 and 0.63, respectively and these were statistically equivalent. The PPV ranged from 15 to 28% where the prevalence of fatty liver was 12.3%. The price of the program was estimated at US 4 dollars per person based on the medical reimbursement fee rate. The efficacy of the liver screening program was found to be insufficient and BMI monitoring may provide a more suitable and inexpensive alternative. Furthermore, the effectiveness of the program is open to question, considering the generally benign prognosis of the disease in the absence of any accompanying morbid conditions and the high price of the program.

Adult↗

Technique for expanding the donor liver pool: heat shock preconditioning in a rat fatty liver model.

Fatty liver is a common predisposing risk factor for postoperative liver failure and accounts for most discarded livers during triage of donors. We investigated the effect of heat shock preconditioning (HPc) on recipient survival in a rat fatty liver transplantation model. Fatty liver donor rats were exposed to brief whole-body hyperthermia (10 minutes at 42.5 degrees C) and allowed to recover. HPc induced heat shock proteins (HSPs) (HSP72, HSP90, and heme oxygenase [HO]-1) in donor livers, with levels peaking 12 to 48 hours after HPc. Subsequently, donor livers were harvested 24 hours after HPc, placed in cold storage for 10 hours, and transplanted into normal rats. At 3 hours posttransplantation, HPc reduced serum liver enzymes in the recipients and almost completely suppressed the release of tumor necrosis factor (TNF)-alpha and interleukin (IL)-10. Histologic evaluation 3 and 24 hours after transplantation showed that HPc significantly reduced hepatic inflammation and hepatocellular necrosis without affecting the steatotic appearance of hepatocytes. One week after transplantation, control non-heat-shocked and heat-shocked fatty liver recipients exhibited survival rates of less than 10% and more than 80%, respectively. The evaluation of the survival of recipients receiving fatty livers at different times after HPc showed that the protective effect of HPc was significant when donor livers were transplanted 3 to 48 hours after HPc, with the maximum effect seen 6 to 48 hours after HPc. In conclusion, HPc is a promising avenue to salvage rejected donor fatty livers and enhance the survival rate of fatty liver recipients. We estimate that this technique could increase the annual donor pool by 600 livers.

Animals↗

Impaired blood rheology by remnant-like lipoprotein particles: studies in patients with fatty liver disease.

Fatty liver disease (FLD) characterised by a high plasma levels of lipoproteins and remnant-like lipoproteins (RLP) is a risk factor for impaired microvascular blood flow, endothelial cell dysfunction and atherosclerosis. Using an immunoseparation technique with a gel mixture containing human monoclonal antibodies to apo A-I and apo B-100, we separated and measured RLP cholesterol (RLP-C) levels which reflect RLP in patients with FLD (n=20). Whole blood transit time (TT) was determined by a microchannel method (MC-FAN) which allows blood flow to be viewed via a microscope connected to an image display unit. RLP-C levels were higher (P<0.01) in FLD, 15.6 +/- 1.0 mg/dl compared with 4.8 +/- 0.5 mg/dl for controls (n=20). Similarly, TT was longer (P<0.01) in FLD, 284.5 +/- 26.1 sec/100 microl compared with 82.8 +/- 1.0 sec/100 microl for controls. Since the liver is a major site for RLP formation and degradation, it is affected to a greater extent in patients with FLD. It is likely that high levels of RLP can impair microvascular perfusion in the liver tissue and contribute to the development and progression of FLD.

2-Chloroadenosine↗

[Death of pregnant heifers at a dairy farm due to fatty liver disease].

Fatty liver disease usually occurs in periparturient dairy cows. However, this case study describes the occurrence of the disease in maiden heifers on a dairy farm during late gestation. It resulted in the death of 5 heifers (21-25 months of age) within 31 days after turning them out to grass. The diagnosis was confirmed by blood testing and gross pathology findings. Housing the animals and providing them with good quality grass silage prevented further clinical cases.

Animal Husbandry↗

Epidemiology of nonalcoholic fatty liver.

Nonalcoholic fatty liver (NAFL) is increasingly recognized as an important and common public health problem that can lead to cirrhosis and hepatic failure. Because it is often asymptomatic,many people may not know that they have it. NAFL is closely linked to obesity, which in the United States and other developed countries is becoming more common. Consequently, the proportion of the population affected by NAFL will likely increase. Despite the growing importance of this condition, knowledge of the epidemiology of NAFL is limited by the lack of an accurate,noninvasive measure for use in screening of the general population. This article reviews information available from studies with relatively unselected samples with regard to prevalence, demographics,and risk factors for NAFL.

Adult↗

Spleen enlargement in patients with nonalcoholic fatty liver: correlation between degree of fatty infiltration in liver and size of spleen.

Our purpose was to determine if there is an association between nonalcoholic fatty liver and spleen enlargement. Spleen volume was measured by computed tomography (CT) in 32 patients with nonalcoholic fatty liver (23 men and 9 women; age, 41.6+/-12.1, range, 22-69 years) and 34 patients with normal liver (19 men and 15 women; age, 51.1+/-16.2, range, 14-86 years). The values were compared with the patient's demographic data, the liver-to-spleen (L/S) ratio of CT Hounsefield unit measurements, and the results of liver function tests. Diagnosis of fatty liver was made if the L/S ratio was less than 1.0. The mean spleen volume was 73.0+/-24.4 cm3 (range, 21.1-106.1) in normal subjects and 141.2+/-54.1 cm3 (range, 44.1-267.3) in patients with fatty liver (P<0.0001). Multivariate linear regression analysis identified that only the L/S ratio (P<0.0001) and age (P<0.01) were significantly correlated with spleen volume. Using forward selection stepwise regression, the L/S ratio entered first (beta = -0.634) and age second (beta = -0.293). In conclusion, spleen enlargement was commonly seen in patients with nonalcoholic fatty liver, and the recognition of this association may halt further attempts to evaluate the cause of spleen enlargement.

Adult↗

[Serum immunoglobulin E levels in patient with fatty liver, whether associated with alcohol consumption or not].

It has been reported that total Immunoglobulin E levels (IgE) are elevated in patients with liver damage (fatty liver), associated with alcohol consumption, but the mechanism responsible for this increase is not completely understood. The objective of this investigation was to determine serum concentrations of IgE in patients with fatty liver, associated or not with alcohol consumption. During the period of February-August 2000, a total of 756 patients attended the outpatient Gastroenterology Service of the University Central Hospital "Antonio María Pineda" in Barquisimeto, Venezuela. Of these, 150 were diagnosed as suffering from fatty liver, but only 63 patients fulfilled the inclusion criteria. The IgE was determined by Photoemission Immunometric Enzyme Immunoassay (High Resolution Amplified Chemoluminescence). IgE serum levels were higher in patients that consumed alcohol (low risk consumer, mean 586.42 +/- 779.74 UI/mL; consumer at risk, mean 329.31 +/- 358.13 UI/mL) in comparison with abstainers (mean 77.51 +/- 56.95 UI/mL) (p < 0.05). There was no relationship between IgE levels and the severity of hepatic steatosis. IgE may be considered a biochemical marker for fatty liver associated with alcohol consumption.

Adult↗