HIF-1α associated logistic regression model serves for predicting decompensation of hepatitis B cirrhosis

Background HIF-1α is relevant to inflammation and fibrosis in hepatitis B virus (HBV)-related liver diseases. Thus, we designed a predictive model for decompensated cirrhosis. Methods Peripheral plasma HIF-1α levels were measured in 52 subjects, including 20 patients with HBV-related-compensated-cirrhosis (HBV-CC), 20 patients with HBV-related-decompensated-cirrhosis (HBV-DC) that underwent transjugular intrahepatic portosystemic shunt (TIPS), and 12 healthy controls (HC). Portal plasma HIF-1α levels were detected in HBV-DC patients. The correlation between clinical data and HIF-1α levels was assessed, logistic regression and nomogram were used to develop prediction model. Results Plasma HIF-1α levels were significantly higher in HBV-DC patients than that in HBV-CC patients and healthy controls (DC: 656.34±417.96, CC: 294.23±138.03, HC: 194.63±54.14, pg/ml; P = 0.0004). Plasma HIF-1α levels were positively correlated with total bile acid, total bilirubin, APRI, FIB-4, and MELD scores, and negatively correlated with albumin and platelets. Multivariate logistic regression manifested that total bilirubin (OR = 19.439; 95% CI: 1.486–254.320, P = 0.024), spleen thickness (OR = 75.144; 95% CI: 4.157–1358.440, P = 0.003) and HIF-1α concentrations above 341.78 pg/ml (OR = 23.580; 95% CI: 1.842–301.781, P = 0.015) were markedly associated with HBV-DC and thus included in the nomogram. The terrific cut-off value for the probability of HBV-DC was > 45%, and area under the curve was 0.954 (P < 0.001), with 95% sensitivity and specificity. Conclusions HIF-1α is related to biochemical liver parameters, cirrhosis grade, and progression to HBV-DC. Our model has preferable predictive value for HBV-DC.

Iberian pig adaptation to acorn consumption: II. Net portal appearance of amino acids

In Iberian pig outdoor production, pigs are fed equilibrated diets until the final fattening period when grazing pigs consume mainly acorns from oak trees. Acorns are rich in energy but poor in crude protein where lysine is the first limiting amino acid (AA). Net portal appearance (NPA) is very useful to ascertain AA available for liver and peripheral tissues. The aim of this study was to determine NPA of AA in Iberian gilts fed with acorns and to ascertain if there was an effect of acorn feeding over time. Two sampling periods were carried out (after one day and after one week of acorn feeding) with six gilts (34 kg average BW) set up with three catheters: in carotid artery and portal vein for blood sampling, and ileal vein for a marker infusion to measure portal plasma flow (PPF). Pigs were fed at 2.5 × ME for maintenance a standard diet in two meals, at 09:00 (0.25) and 15:00 h (the remaining 0.75). The day previous to first sampling, pig diet was replaced by 2.4 kg of acorn. A serial blood collection was done at −5 min, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5 and 6 h after feeding 0.25 of total daily acorn ration. Following identical protocol, one week later the second sampling was done. NPA of sum of essential AA (EAA) was poor. Although increased NPA of histidine (P < 0.001), leucine, phenylalanine and valine (0.05 < P < 0.08) was found after one week of acorn consumption, the sum of EAA did not change. Furthermore, fractional absorption (NPA/AA intake) of EAA, non-essential AA (NEAA) and total AA was 97, 44 and 49% lower, respectively, at the beginning of eating acorn than a week later. Supplementation, with some of the EAA and NEAA to Iberian pigs during the grazing period would be beneficial to overcome the increased portal-drained viscera (PDV) utilization of AA observed in the present study.

Iberian pig adaptation to acorn consumption: I. Net portal appearance of metabolites

Most valuable cured products from Iberian pigs come from pure bred animals raised for a final grazing-fattening period where pigs eat mainly acorns, a low protein energy rich fruit. This is a nutritional challenge for animals fed equilibrated diets from weaning. The aim of the study was to determine net portal appearance (NPA) of metabolites in gilts fed acorns and evaluate adaptational changes after one week of feeding. Two sampling periods were carried out (after one day and after one week of acorn feeding) with six gilts (34 kg average BW) set up with three catheters: in carotid artery and portal vein for blood sampling, and ileal vein for para-aminohippuric acid (PAH) infusion to measure portal plasma flow (PPF). Pigs were fed at 2.5 × ME for maintenance a standard diet in two portions, at 09:00 (0.25) and 15:00 h (the remaining 0.75). On the day prior to the first sampling period, pigs were fed 2.4 kg of oak acorns. After feeding 0.25 of ration a 6 h serial blood collection was initiated. Following an identical protocol, a second sampling session was performed 1 week later. Adaptation to acorn consumption decreased NPA of ammonia (47%,P < 0.001). Although there was a transfer of urea from the gastrointestinal tract to the circulation in both sampling periods, no differences in NPA of urea was found (P > 0.05). NPA of glucose was not influenced by sampling period (P > 0.05), but NPA of lactate was greatly increased (231%,P < 0.001). There was a negative NPA of albumin although adaptation to acorn feeding did not alter it. Although NPA of triglycerides and cholesterol were unchanged, a subtle increase in arterial and portal cholesterol was noticed (9.6%,P < 0.01). Pigs fed a protein deficient diet for one week adapted decreasing NPA of ammonia for saving metabolic energy as less ammonia would become available for conversion to urea.

Absorption, Metabolism, and Excretion by Freely Moving Rats of 3,4-DHPEA-EDA and Related Polyphenols from Olive Fruits (Olea europaea)

Absorption, metabolism, and excretion of 3,4-DHPEA-EDA, oleuropein, and hydroxytyrosol isolated from olive fruits were newly evaluated after oral and intravenous administration in freely moving rats cannulated in the portal vein, jugular vein, and bile duct. Orally administered 3,4-DHPEA-EDA, an important bioactive compound in olive pomace, was readily absorbed and metabolized to hydroxytyrosol, homovanillic acid, and homovanillyl alcohol, as shown by dose-normalized 4 h area under the curve (AUC0→4 h/Dose) values of 27.7, 4.5, and 4.2 μM·min·kg/μmol, respectively, in portal plasma after oral administration. The parent compound 3,4-DHPEA-EDA was not observed in the portal plasma, urine, and bile after oral and intravenous administration. Additionally, hydroxytyrosol, homovanillic acid, and homovanillyl alcohol in the portal plasma after oral administration of hydroxytyrosol showed 51.1, 22.8, and 7.1 μM·min·kg/μmolAUC0→4 h/Dose, respectively. When oleuropein, a polar glucoside, was injected orally, oleuropein in the portal plasma showed 0.9 μM·min·kg/μmolAUC0→4 h/Dose. However, homovanillic acid was detected from oleuropein in only a small amount in the portal plasma. Moreover, the bioavailability of hydroxytyrosol and oleuropein for 4 hours was 13.1% and 0.5%, respectively. Because the amount of 3,4-DHPEA-EDA in olive fruits is about 2-3 times greater than that of hydroxytyrosol, the metabolites of 3,4-DHPEA-EDA will influence biological activities.

Effect of tryptophan supplementation on diet-induced non-alcoholic fatty liver disease in mice

Intestinal serotonin (5-hydroxytrypamine, 5-HT) metabolism is thought to play a role in gut functions by regulating motility, permeability and other functions of the intestine. In the present study, we investigated the effect of tryptophan (TRP), the precursor of 5-HT, supplementation on intestinal barrier functions and non-alcoholic fatty liver disease (NAFLD). An established mouse model of NAFLD induced by feeding a fructose-rich diet (N group) was used in the present study. TRP was administered orally for 8 weeks to C57BL/6J control or NAFLD mice. NAFLD-related liver parameters (hepatic TAG and Oil Red O staining), intestinal barrier parameters (tight-junction protein occludin and portal plasma lipopolysaccharides (LPS)) and 5-HT-related parameters (5-HT, 5-HT transporter (SERT) and motility) were measured. We observed reduced duodenal occludin protein concentrations (P= 0·0007), high portal plasma LPS concentrations (P= 0·005) and an elevated liver weight:body weight ratio (P= 0·01) in the N group compared with the parameters in the control group. TRP supplementation led to an increase in occludin concentrations (P= 0·0009) and consecutively reduced liver weight:body weight ratio (P= 0·009) as well as overall hepatic fat accumulation in the N group (P= 0·05). In addition, the N group exhibited reduced SERT protein expression (P= 0·002), which was normalised by TRP supplementation (P= 0·02). For the first time, our data indicate that oral TRP supplementation attenuates experimental NAFLD in mice. The underlying mechanisms are not clear, but probably involve stabilisation of the intestinal barrier in the upper small intestine and amelioration of the dysregulated intestinal serotonergic system.

Variations in the Response of Pituitary Lactotrophs to Oxytocin during the Rat Estrous Cycle

Although removal of dopamine inhibition is established as a major factor in prolactin (PRL) release, a large body of evidence suggests that hypothalamic oxytocin (OT) may serve as a PRL-releasing hormone in the rat. PRL release is modulated by estradiol (E2), which rises between diestrus and proestrus of the estrous cycle, causing a PRL surge in the afternoon of proestrus. Given that E2 strongly modulates OT actions in both central and peripheral tissues, OT action on lactotrophs might also be modulated by the stage of the estrous cycle. To test this hypothesis, we have monitored PRL release and intracellular calcium levels ([Ca2+]i) induced by OT in pituitary lactotrophs obtained from female rats in either diestrus 1 or proestrus. We found that both secretory and [Ca2+]i responses to OT are significantly increased in lactotrophs obtained on proestrus. Moreover, we show that these differences are due to an increase in both the number of OT-responding lactotrophs and the magnitude of their individual [Ca2+]i responses. Both secretory and [Ca2+]i responses were abolished by a specific OT antagonist. Finally, dose-dependent studies show that the increased PRL-releasing effect of OT on proestrus is significant over a wide range of concentrations, particularly those observed in hypophyseal portal plasma. These results suggest that the rising E2 titers that culminate on proestrus facilitate the stimulatory action of OT on lactotrophs and support the notion that OT is a PRL-releasing hormone with an important role in the production of the proestrous surge of PRL.

Effects ofdl-2-hydroxy-4-methylthiobutyrate on the first-pass intestinal metabolism of dietary methionine and its extra-intestinal availability

The present study was conducted in a one-factorial arrangement to determine the effects ofdl-2-hydroxy-4-methylthiobutyrate (dl-HMTB) on the first-pass intestinal metabolism of dietary methionine and its extra-intestinal availability. Barrows (n6; aged 35 d; weight 8·6 kg), implanted with arterial, portal, mesenteric and gastric catheters, were fed a diet containingdl-methionine (dl-MET) ordl-HMTB once hourly and infused intramesenterically with 1 %p-aminohippurate and intragastrically with [1-13C]methionine at 7·0 μmol/kg body weight per h. Arterial and portal blood samples were taken at hourly intervals until 6 h of tracer infusion and pigs was then killed for collection of muscle, intestine, liver and kidney samples. The net portal appearance of methionine, expressed as the fraction of ingested directly availablel-methionine, was higher (P < 0·05) in thedl-HMTB than in thedl-MET diet, and there was no difference (P = 0·26) in the fractional portal balance of [1-13C]methionine between the diets. [1-13C]methionine enrichment (tracer:tracee ratio; mol/100 mol amino acid) in the jejunum, arterial and portal plasma, liver, kidney and muscle was also not different (P>0·05) between the groups. Over the 6 h period after the start of feeding, the average concentration of citrulline both in the arterial and portal plasma was higher (P < 0·05) in thedl-HMTB than in thedl-MET group, and arterial plasma ornithine and taurine concentration was also higher (P < 0·05) in thedl-HMTB than in thedl-MET group. However, plasma urea concentration both in the arterial and portal vein was lower (P < 0·05) in thedl-HMTB than in thedl-MET group. These results suggested that the potential difference in the first-pass use of methionine by the intestine between thedl-HMTB anddl-MET diets might affect intestinal and systemic metabolism of other amino acids, which may provide new important insights into nutritional efficiency of different methionine sources.

Comparative study of coagulation and thrombin generation in the portal and jugular plasma of patients with cirrhosis

SummaryPortal vein thromboses are frequent in cirrhotic patients and may be favoured by hypercoagulability in the splanchnic venous system. The coagulation balance and thrombin generation (TG) were evaluated in platelet-free plasma obtained from portal and systemic blood samples in 28 cirrhotic patients while undergoing transjugular intrahepatic porto-systemic shunt. TG assay (TGA) was performed with all samples from cirrhotic patients and with plasma samples from 14 healthy controls, with varying concentrations of tissue factor and phospholipids, with or without thrombomodulin. Screening tests and specific assays were also performed and activated partial thromboplastin time was shorter in portal plasma samples with higher FVIII and lower protein C levels, well correlated with Child-Pugh scores, and higher D-dimers and F1+2 levels However, all TGA parameters were similar in portal and jugular samples, possibly due in part to similar concentrations of factor II and antithrombin in these two sites of plasma sampling. TGA showed lower thrombin peaks and endogenous thrombin potential values in cirrhotic plasma compared to those of healthy controls. Importantly, a resistance to thrombomodulin that well correlated with factor VIII and PC levels, was evidenced in all samples from patients with cirrhosis, and was more significant in those from severely affected cases. This study therefore supports the existence of a relative hypercoagulability in the portal vein of cirrhotic patients that is likely due to protein C/S deficiency and to high FVIII levels.