Liver Oxygen Consumption Research Articles

104 Effects of providing vitamin and mineral supplementation throughout gestation on subsequent F1 replacement heifer liver and muscle oxygen consumption and mitochondrial function throughout pregnancy

Abstract The study aimed to assess the impact of maternal vitamin/mineral supplementation during gestation on the energy utilization in liver and muscle tissues of F1 heifer offspring. We hypothesized that VTM supplementation during gestation would mitigate mitochondrial energy loss and uncoupling of liver and muscle tissue in pregnant offspring. At breeding, crossbred Angus heifers [n = 31; initial body weight (BW) = 412.5 ± 53.68 kg] were randomly assigned to either a basal diet targeting BW gains of 0.45 kg•heifer-1•d-1(CON; n = 14) or the basal diet plus 113 g•heifer-1•d-1of a vitamin/mineral supplement (VTM; n = 17). Heifers remained on their respective dietary treatments until calving. After calving, all F1 heifer calves were raised in a single management group with their dams until weaning (BW = 209 ± 4.82 kg; VTM, n = 17; CON, n = 12) and fed a common diet throughout postnatal development. At breeding and throughout gestation, F1 heifers (n = 9, VTM; n = 7, CON) were allotted daily feed intakes of 1.5% of BW on a dry-matter basis as a total mixed ration and used to evaluate cellular energy metabolism throughout pregnancy. Liver and Longissimus dorsi muscle biopsies were collected on d 179 and d 247 +/- 3 of gestation and mitochondrial function was assessed via high resolution respirometry (Oroboros Instruments, Innsbruck, Austria). The following respiratory states were evaluated: LEAK respiration (L), OXPHOS capacity (P), NADH-linked OXPHOS (PI), and electron transfer capacity (E). Data were analyzed using the MIXED procedure of SAS with repeated measures where appropriate. No treatment × trimester interaction (P ≥ 0.13) was observed for any of the respiratory states in liver and muscle. Hepatic oxygen consumption was not influenced by maternal gestational VTM supplementation (P ≥ 0.68), or stage of gestation (P ≥ 0.27). In muscle, L respiration was the only state affected by maternal dietary treatments, being greater (P = 0.05) in CON than VTM heifers. Additionally, L respiration was greater (P = 0.02) during the third trimester compared with the second trimester. This observation in CON heifers is important because increased L respiration (proton leak) results in decreased ATP production. Protons that escape the intermembrane space, are dissipated as heat rather than be used for ATP synthesis; thus, oxygen consumption increases to compensate for the reduction of protons entering ATP turnover. Results show increased muscle mitochondria inefficiency in offspring with dams lacking supplementation during pregnancy. The increased incidence of proton leak results in potentially less efficient cattle as protons escape the electron transport chain rather than be transformed for energy utilization need of the animal, thus VTM supplementation throughout pregnancy impacts fetal development with potential effects in live offspring 18 to 22 mo post-parturition.

Effect of Pravastatin and Simvastatin on the Reduction of Cytochrome C.

Statins are used to treat hypercholesterolemia, with several pleiotropic effects. Alongside their positive effects (for example, decreasing blood pressure), they can also bring about negative effects/symptoms (such as myopathy). Their main mechanism of action is inducing apoptosis, the key step being the release of cytochrome c from the mitochondria. This can be facilitated by oxidative stress, through which glutathione is oxidized. In this research, glutathione was used as a respiratory substrate to measure the mitochondrial oxygen consumption of rat liver with an O2 electrode. The reduction of cytochrome c was monitored photometrically. Hydrophilic (pravastatin) and lipophilic (simvastatin) statins were used for the measurements. Pravastatin reduces the reduction of cytochrome c and the oxygen consumption of the mitochondria, while simvastatin, on the other hand, increases the reduction of cytochrome c and the mitochondrial oxygen consumption. The results make it seem probable that statins influence the mitochondrial oxygen consumption through cytochrome c. Simvastatin could enhance the oxidizing capacity of free cytochrome c, thereby increasing oxidative stress and thus facilitating apoptosis. The observed effects could further the understanding of the mechanism of action of statins and thereby aid in constructing optimal statin therapy for every patient.

Role of hydrogen sulfide in the regulation of respiration, blood flow and bile secretory function of the liver

In acute experiments on laboratory rats, intra-portal administration of L-cysteine (20 mg/kg), the precursor of hydrogen sulfide synthesis, stimulated oxygen consumption of liver by 38.6% and reduced oxygen tension by 37.1%. Activation of tissue respiration occurred due to the strengthening of oxygen-dependent synthetic processes in liver, in particular those associated with mitochondrial enzyme-catalysed bile acid biosynthesis through the acidic pathway. The concentrations of taurocholic acid and mixtures of taurodeoxycholic and taurohenodeoxycholic acids increased by 10.3 and 17.9%, respectively, compared to the initial levels. In addition, the level of free cholesterol was decreased by 33.9% and esterification processes were intensified, as indicated by an increase in the concentration of esterified cholesterol by 22.6% in the bile of rats. The latter was to some extent confirmed by a decrease in the level of free bile acids (by 15.8%) involved in the biosynthesis of cholesterol esters and intensification of tissue respiration in the liver. L-cysteine dilated intrahepatic vessels, resulting in a significant decrease of the systemic blood pressure and blood pressure in the portal vein by 17.6 and 24.5%, respectively. L-cysteine increased the rate of local blood flow in the liver and blood supply by 28.2 and 24.4%, respectively. Blockade of cystathionine-γ-lyase by DL-propargylglycine (11 mg/kg) significantly inhibited the L-cysteine-induced tissue respiration and bile acid biosynthesis in the liver. Administration of DL-propargylglycine resulted in constriction of blood vessels of the liver and, as a consequence, to an increased blood pressure and a decreased blood flow rate in tissue. Our data point to an involvement of hydrogen sulfide in the regulation of liver tissue respiration and bile secretory function.

Berberine alleviates lipid metabolism disorders via inhibition of mitochondrial complex I in gut and liver.

This study is to investigate the relationship between berberine (BBR) and mitochondrial complex I in lipid metabolism. BBR reversed high-fat diet-induced obesity, hepatic steatosis, hyperlipidemia and insulin resistance in mice. Fatty acid consumption, β-oxidation and lipogenesis were attenuated in liver after BBR treatment which may be through reduction in SCD1, FABP1, CD36 and CPT1A. BBR promoted fecal lipid excretion, which may result from the reduction in intestinal CD36 and SCD1. Moreover, BBR inhibited mitochondrial complex I-dependent oxygen consumption and ATP synthesis of liver and gut, but no impact on activities of complex II, III and IV. BBR ameliorated mitochondrial swelling, facilitated mitochondrial fusion, and reduced mtDNA and citrate synthase activity. BBR decreased the abundance and diversity of gut microbiome. However, no change in metabolism of recipient mice was observed after fecal microbiota transplantation from BBR treated mice. In primary hepatocytes, BBR and AMPK activator A769662 normalized oleic acid-induced lipid deposition. Although both the agents activated AMPK, BBR decreased oxygen consumption whereas A769662 increased it. Collectively, these findings indicated that BBR repressed complex I in gut and liver and consequently inhibited lipid metabolism which led to alleviation of obesity and fatty liver. This process was independent of intestinal bacteria.

Chlorella sorokiniana Dietary Supplementation Increases Antioxidant Capacities and Reduces Ros Release in Mitochondria of Hyperthyroid Rat Liver.

The ability of aerobic organisms to cope with the attack of radicals and other reactive oxygen species improves by feeding on foods containing antioxidants. Microalgae contain many molecules showing in vitro antioxidant capacity, and their food consumption can protect cells from oxidative insults. We evaluated the capacity of dietary supplementation with 1% dried Chlorella sorokiniana strain 211/8k, an alga rich in glutathione, α-tocopherol, and carotenoids, to counteract an oxidative attack in vivo. We used the hyperthyroid rat as a model of oxidative stress, in which the increase in metabolic capacities is associated with an increase in the release of mitochondrial reactive oxygen species (ROS) and the susceptibility to oxidative insult. Chlorella sorokiniana supplementation prevents the increases in oxidative stress markers and basal oxygen consumption in hyperthyroid rat livers. It also mitigates the thyroid hormone-induced increase in maximal aerobic capacities, the mitochondrial ROS release, and the susceptibility to oxidative stress. Finally, alga influences the thyroid hormone-induced changes in the factors involved in mitochondrial biogenesis peroxisomal proliferator-activated receptor-γ coactivator (PGC1-1) and nuclear respiratory factor 2 (NRF-2). Our results suggest that Chlorella sorokiniana dietary supplementation has beneficial effects in counteracting oxidative stress and that it works primarily by preserving mitochondrial function. Thus, it can be useful in preventing dysfunctions in which mitochondrial oxidative damage and ROS production play a putative role.

Human Red Blood Cells as Oxygen Carriers to Improve Ex-Situ Liver Perfusion in a Rat Model.

Ex-situ machine perfusion (MP) has been increasingly used to enhance liver quality in different settings. Small animal models can help to implement this procedure. As most normothermic MP (NMP) models employ sub-physiological levels of oxygen delivery (DO2), the aim of this study was to investigate the effectiveness and safety of different DO2, using human red blood cells (RBCs) as oxygen carriers on metabolic recovery in a rat model of NMP. Four experimental groups (n = 5 each) consisted of (1) native (untreated/control), (2) liver static cold storage (SCS) 30 min without NMP, (3) SCS followed by 120 min of NMP with Dulbecco-Modified-Eagle-Medium as perfusate (DMEM), and (4) similar to group 3, but perfusion fluid was added with human RBCs (hematocrit 15%) (BLOOD). Compared to DMEM, the BLOOD group showed increased liver DO2 (p = 0.008) and oxygen consumption (2) (p < 0.001); lactate clearance (p < 0.001), potassium (p < 0.001), and glucose (p = 0.029) uptake were enhanced. ATP levels were likewise higher in BLOOD relative to DMEM (p = 0.031). 2 and DO2 were highly correlated (p < 0.001). Consistently, the main metabolic parameters were directly correlated with DO2 and 2. No human RBC related damage was detected. In conclusion, an optimized DO2 significantly reduces hypoxic damage-related effects occurring during NMP. Human RBCs can be safely used as oxygen carriers.

Noninvasive quantification of oxygen saturation in the portal and hepatic veins in healthy mice and those with colorectal liver metastases using QSM MRI.

PurposeThis preclinical study investigated the use of QSM MRI to noninvasively measure venous oxygen saturation (SvO2) in the hepatic and portal veins.MethodsQSM data were acquired from a cohort of healthy mice (n = 10) on a 9.4 Tesla MRI scanner under normoxic and hyperoxic conditions. Susceptibility was measured in the portal and hepatic veins and used to calculate SvO2 in each vessel under each condition. Blood was extracted from the inferior vena cava of 3 of the mice under each condition, and SvO2 was measured with a blood gas analyzer for comparison. QSM data were also acquired from a cohort of mice bearing liver tumors under normoxic conditions. Susceptibility was measured, and SvO2 calculated in the portal and hepatic veins and compared to the healthy mice. Statistical significance was assessed using a Wilcoxon matched‐pairs signed rank test (normoxic vs. hyperoxic) or a Mann‐Whitney test (healthy vs. tumor bearing).ResultsSvO2 calculated from QSM measurements in healthy mice under hyperoxia showed significant increases of 15% in the portal vein (P < 0.05) and 21% in the hepatic vein (P < 0.01) versus normoxia. These values agreed with inferior vena cava measurements from the blood gas analyzer (26% increase). SvO2 in the hepatic vein was significantly lower in the colorectal liver metastases cohort (30% ± 11%) than the healthy mice (53% ± 17%) (P < 0.05); differences in the portal vein were not significant.ConclusionQSM is a feasible tool for noninvasively measuring SvO2 in the liver and can detect differences due to increased oxygen consumption in livers bearing colorectal metastases.

The difference between NOAEL and BMD approach

Oxygenation is necessary for aerobic metabolism, which maintains adenosine triphosphate within the graft organ. In recent years, some studies have demonstrated that subnormothermic machine perfusion (SNMP) with hemoglobin-based oxygen carriers has the potential to improve oxygen metabolism.The aim of this study was to evaluate the effectiveness of perfusate with human-derived hemoglobin vesicles (HbV) under SNMP in a pig model of donation after cardiac death.In this study, pig livers were procured with a warm ischemic time of 60 minutes and were preserved in 3 groups for 240 minutes. The preservation conditions were as follows: 4°C cold storage (Group 1); SNMP with University of Wisconsin perfusate alone (Group 2); and SNMP (21°C) with University of Wisconsin solution and HbV (hemoglobin, 0.6 mg/dL) perfusate (Group 3). All livers were perfused for 120 minutes using pig autologous blood machine perfusion (reperfusion phase). We investigated the aspartate transaminase level and hemodynamics (portal vein resistance and oxygen consumption) in the preservation and reperfusion phases. A histologic study (hematoxylin-eosin staining) was performed after 240 minutes of preservation.The portal vein resistance of Group 3 was not increased in comparison with Group 2. During preservation, the oxygen consumption of Group 3 was higher than that of Group 2. However, the level of aspartate transaminase did not differ between Groups 2 and 3.The present study revealed that perfusate with HbV increased the oxygen consumption of the donor liver during SNMP.

Body and organ metabolic rates of a cave fish, Triplophysa rosa: influence of light and ontogenetic variation

Triplophysa rosa is a typical species of cave-dwelling fish distributed throughout Wulong County, Chongqing, China. This study aimed to test whether T. rosa has a low metabolic level as a cave species and how the metabolic rate of this fish responds to light stimulation. The whole body and organ (including brain, heart, and liver) oxygen consumption rates ([Formula: see text]) and several blood parameters related to oxygen carrying were compared between T. rosa acclimated in constant dark and those in regular photoperiod conditions. No significant changes in any variables were observed between the regular photoperiod fish and the dark fish, suggesting that the metabolic consumption of T. rosa is not light sensitive, which may be attributed to the highly degraded eyes of this cave species. The average mass-specific resting [Formula: see text] of T. rosa was 38.3 mgO2 kg- 1 h- 1 and was lower than many other fish species. One possible explanation for the low metabolic level of T. rosa can be due to its highly degraded eyes and small brain size. Whole-organ [Formula: see text] of the brain, heart, and liver were on average responsible for 8.18%, 3.55%, and 8.61% of the body resting [Formula: see text], respectively. Both heart mass and liver mass increased with increasing body mass; however, brain mass did not correlate with body mass. Maintaining a small brain size throughout ontogeny suggests energy-saving advantages for this cave species.

Determination of Minimal Hemoglobin Level Necessary for Normothermic Porcine Ex Situ Liver Perfusion.

In current studies of ex situ liver perfusion there exists considerable variability in perfusate composition, including the type of oxygen carrier. Herein, we aim to clarify the minimal hemoglobin level necessary during normothermic porcine ex situ liver perfusion. Livers procured from 35 to 45 kg domestic pigs were connected to our experimental ex situ circuit (n = 10). In the treatment group, perfusate was sequentially diluted hourly to predetermined hemoglobin levels. At the end of each hemoglobin dilution, perfusate samples were analyzed for liver transaminases, lactate dehydrogenase (LD), total bilirubin, and lactate levels. Liver oxygen consumption was measured. In the control group, livers were perfused continually for a duration of 24 hours at target hemoglobin levels of 30 and 20 g/L. Rising liver transaminases, significantly higher lactate (P < 0.001), and LD levels (P < 0.001) were noted at lower perfusate hemoglobin levels in the treatment group. Liver oxygen utilization (P < 0.001) and hepatic artery oxygen delivery (P < 0.001) were significantly lower at lower hemoglobin levels, whereas liver vessel resistance remained relatively constant. Histology demonstrated increasing parenchymal damage at lower hemoglobin levels. In control livers, higher perfusate transaminases, higher lactate, and LD levels were noted at a perfusion hemoglobin level of 20 g/L. Ex situ liver function decompensated during perfusion between a mean hemoglobin level of 30 to 20 g/L, as evidenced by notably rising lactate and LD levels. This study demonstrates optimal hemoglobin concentration during normothermic ex situ liver perfusion to ensure a fully metabolically functioning graft.

Application of Perfusate With Human-Derived Oxygen Carrier Solution Under Subnormothermic Machine Perfusion for Donation After Cardiac Death Liver Grafts in Pigs

Oxygenation is necessary for aerobic metabolism, which maintains adenosine triphosphate within the graft organ. In recent years, some studies have demonstrated that subnormothermic machine perfusion (SNMP) with hemoglobin-based oxygen carriers has the potential to improve oxygen metabolism. ObjectiveThe aim of this study was to evaluate the effectiveness of perfusate with human-derived hemoglobin vesicles (HbV) under SNMP in a pig model of donation after cardiac death. Materials and MethodsIn this study, pig livers were procured with a warm ischemic time of 60 minutes and were preserved in 3 groups for 240 minutes. The preservation conditions were as follows: 4°C cold storage (Group 1); SNMP with University of Wisconsin perfusate alone (Group 2); and SNMP (21°C) with University of Wisconsin solution and HbV (hemoglobin, 0.6 mg/dL) perfusate (Group 3). All livers were perfused for 120 minutes using pig autologous blood machine perfusion (reperfusion phase). We investigated the aspartate transaminase level and hemodynamics (portal vein resistance and oxygen consumption) in the preservation and reperfusion phases. A histologic study (hematoxylin-eosin staining) was performed after 240 minutes of preservation. ResultsThe portal vein resistance of Group 3 was not increased in comparison with Group 2. During preservation, the oxygen consumption of Group 3 was higher than that of Group 2. However, the level of aspartate transaminase did not differ between Groups 2 and 3. ConclusionThe present study revealed that perfusate with HbV increased the oxygen consumption of the donor liver during SNMP.

The Relationship between Liver Dysfunction, Electrocardiographic Abnormalities and Metabolism in Rat

Introduction: Liver disorders may lead to many critical complications such as electrical heart abnormalities. The aim of this study was to investigate the relationship between the severity of liver dysfunctions, electrocardiogram variables and oxygen consumption. Methods: Female Sprague Dawley rats were divided into 4 groups of sham, partial portal vein ligation (PPVL), common bile duct ligation (CBDL), and combination of them (CBDL+ PPVL). 28 days after the first surgery, animals were anesthetized; ECG recorded and right femoral artery and vein cannulated. Blood gas parameters and complete blood count were measured and the arterial-venous oxygen content difference (A-VO2) calculated. Results: QT interval and QTc in the CBDL+PPVL and CBDL groups were longer than those of the sham and PPVL groups. Furthermore, there were abnormal and bizarre T waves in the CBDL and CBDL+PPVL groups. In the CBDL+PPVL group, arterial O2 pressure was lower while arterial CO2 pressure and plasma concentration of HCO3- were higher than those of the other groups. A-VO2 in the CBDL+PPVL group was also lower than those of the other groups, and in the CBDL group was less than that of the PPVL group. Conclusions: In this study, we detected bizarre T waves and increased QT interval and QTc in the ECG of cirrhotic rats, which could be partly linked to the severity of liver dysfunction and reduction of metabolism.

Seasonal and ontogenetic changes modulate oxygen consumption and antioxidant defenses in the cutlassfish Trichiurus lepturus (Pisces, Trichiuridae)

Several oxidative stress markers and liver oxygen consumption were measured in different tissues of the marine fish Trichiurus lepturus in late summer and late winter, as well as in juveniles and adult females. Oxygen consumption in liver, superoxide dismutase (SOD) and catalase (CAT) activity in liver, red cells, lens and roe, vitamin E, ubiquinol10, β-carotene in liver, red cells, and roe, as well as contents of reduced glutathione (GSH) and lipoperoxidation (TBARS) in red cells were evaluated. Regarding ontogeny, compared to adult fish, juveniles showed significant higher SOD activity in liver and lens, as well as higher liver contents of vitamin E. In contrast, adult females showed higher contents of vitamin E in roe, ubiquinol10 in liver and roe, and higher GSH levels in red cells, while the other markers remained unchanged. Regarding seasonal changes, no differences were detected in adult females for liver CAT and ubiquinol10, CAT in roe, vitamin E in roe and in red cells, liver and red cell ubiquinol10, and in GSH in red cells. However, and coinciding with the spawning period of late summer, liver oxygen consumption, SOD and CAT activity and ubiquinol10 contents in roe and SOD activity in red cells, and red cell TBARS contents were higher compared to late winter. These temporal antioxidant adjustments of Trichiurus lepturus seem to be parallel to the higher oxygen consumption typical of juvenile forms and also to the intense spawning and foraging activities of adult females in late summer.

Respiratory Surveillance and Ca&lt;sup&gt;2+&lt;/sup&gt;-ATPase Enzyme Activity Studies of &lt;i&gt;Clarias gariepinus &lt;/i&gt;Exposed to Acute Toxicity of Cyanide

Potassium cyanide, a highly contaminating and toxic aquatic ecosystems pollutant was investigated for acute toxicity on the freshwater fish Clariasgariepinus. Its effect on the Ca2+ -ATPase activities in the liver, gills, muscle and intestinal tissues and oxygen consumption index was studied. Short-term toxicity test was carried out by static renewal bioassay test over a 96 h period using a lethal concentration (LC50) value of 0.361mg/mL. Potassium cyanide was highly toxic to the animal tested. Results reveal that normal respiratory activity (O2 consumption) of the fish was significantly affected and there was significant decreased in the Ca2+ - ATPase activities at the end of exposure periods (24, 48, 72 and 96 h). Correlation analysis reveals a strong relationship between oxygen consumption index and ATPase enzyme activity of Clariasgariepinus exposed to the toxicant. This study reflects the toxic effect of potassium cyanide to the freshwater fish, Clariasgariepinus and suggestion on the possible application of Ca2+ -ATPase activities and oxygen consumption index as possible biomarkers for early detection of cyanide poisoning in aquatic bodies.

Respiratory Surveillance and Ca&lt;sup&gt;2+&lt;/sup&gt;-ATPase Enzyme Activity Studies of &lt;i&gt;Clarias gariepinus &lt;/i&gt;Exposed to Acute Toxicity of Cyanide

Potassium cyanide, a highly contaminating and toxic aquatic ecosystems pollutant was investigated for acute toxicity on the freshwater fish Clariasgariepinus. Its effect on the Ca2+ -ATPase activities in the liver, gills, muscle and intestinal tissues and oxygen consumption index was studied. Short-term toxicity test was carried out by static renewal bioassay test over a 96 h period using a lethal concentration (LC50) value of 0.361mg/mL. Potassium cyanide was highly toxic to the animal tested. Results reveal that normal respiratory activity (O2 consumption) of the fish was significantly affected and there was significant decreased in the Ca2+ - ATPase activities at the end of exposure periods (24, 48, 72 and 96 h). Correlation analysis reveals a strong relationship between oxygen consumption index and ATPase enzyme activity of Clariasgariepinus exposed to the toxicant. This study reflects the toxic effect of potassium cyanide to the freshwater fish, Clariasgariepinus and suggestion on the possible application of Ca2+ -ATPase activities and oxygen consumption index as possible biomarkers for early detection of cyanide poisoning in aquatic bodies.

Effect of the Combination of Ezetimibe and Simvastatin on Gluconeogenesis and Oxygen Consumption in the Rat Liver

The aim of this work was to investigate the effects of chronic treatment with the combination of ezetimibe and simvastatin on gluconeogenesis in rat liver. Rats were treated daily for 28 days with the combination of ezetimibe and simvastatin (10/40 mg/kg) by oral gavage. To measure gluconeogenesis and the associated pathways, isolated perfused rat liver was used. In addition, subcellular fractions, such as microsomes and mitochondria, were used for complementary measures of enzymatic activities. Treatment with the combination of simvastatin and ezetimibe resulted in a decrease in gluconeogenesis from pyruvate (-62%). Basal oxygen consumption of the treated animals was higher (+22%) than that of the control rats, but the resulting oxygen consumption that occurred after pyruvate infusion was 43% lower in animals treated with the combination of simvastatin and ezetimibe. Oxygen consumption in the livers from treated animals was completely inhibited by cyanide (electron transport chain inhibitor), but not by proadifen (cytochrome P450 inhibitor). Chronic treatment with ezetimibe/simvastatin decreased the activity of the key enzymes glucose-6-phosphatase and fructose-1,6-bisphosphatase by 59% and 45%, respectively, which is probably the major reason for the decreased gluconeogenesis seen in ezetimibe-/simvastatin-treated rats. It is also possible that part of the effect of this combination on gluconeogenesis and on the oxygen consumption is related to the impairment of mitochondrial energy transduction.

Influence of diet on serum glucose and insulin concentrations, insulin receptor concentration in adipose and muscle tissues, and oxygen consumption in liver and muscle in gilts

Raw samples of 14 muscles: Mm. biceps femoris (BF), quadriceps femoris (CF), diaphragm (DI), flexor digitorum (FD), gluteus medius (GM), infraspinatus (IE), longissimus lumborum (LL), longissimus thoracis (LT), psoas major (PM), pectoralis profundus (PP), semimembranosus (SM), semitendinosus (ST), sternomandibularis (STER) and triceps brachii (TB) from four Swiss Brown (485±15 days old) young bull carcasses and weighing approximately 300 kg were evaluated for some chemical and physical properties. PM (2.11 kg) and DI (2.24 kg) were the muscles which had the lowest Warner–Bratzler shear force values, while PP (6.66 kg) had the greatest shear force (P<0.05). FD and IE muscles had the highest concentration of total collagen content while PM and DI had the lowest (P<0.05) contents, TB and IE muscles presented the highest insoluble collagen concentration while PM and LT had the lowest (P<0.05) contents. High positive correlation between total collagen content and Warner–Bratzler shear force of raw samples was found (r=0.723; P<0.01) and between insoluble collagen content and Warner–Bratzler shear force was (r=0.661; P<0.01). Significant differences (P<0.05) were observed among muscles for differential scanning calorimetry, sarcomere length, pH and colour parameters.

Changes in liver gluconeogenesis during the development of Walker‐256 tumour in rats

Few studies have investigated liver gluconeogenesis in cancer and there is no agreement as to whether the activity of this pathway is increased or decreased in this disease. The aim of this study was to evaluate gluconeogenesis from alanine, pyruvate and glycerol, and related metabolic parameters in perfused liver from Walker-256 tumour-bearing rats on days 5 (WK5 group), 8 (WK8 group) and 12 (WK12 group) of tumour development. There was reduction (P < 0.05) of liver glucose production from alanine and pyruvate in WK5, WK8 and WK12 groups, which was accompanied by a decrease (P < 0.05) in oxygen consumption. Moreover, there was higher (P < 0.05) pyruvate and lactate production from alanine in the WK5 group and a marked reduction (P < 0.05) of pyruvate and urea production from alanine in the WK12 group. In addition, liver glucose production and oxygen consumption from glycerol were not reduced in WK5, WK8 and WK12 groups. Thus the, the results show inhibition of hepatic gluconeogenesis from alanine and pyruvate, but not from glycerol, on days 5, 8 and 12 of Walker-256 tumour development, which can be attributed to the metabolic step in which the substrate enters the gluconeogenic pathway.

Normothermic Acellular Ex Vivo Liver Perfusion (NEVLP): Development of a New Method for the Storage, Assessment, and Repair of Marginal Liver Grafts

Livers retrieved after cardiac death (DCD) are often declined for transplantation because of the increased risk for graft failure and bile duct injury. The current cold static preservation technique is associated with a high risk of biliary complications and does not offer the opportunity to assess graft injury and function. We compared the novel normothermic acellular ex vivo liver perfusion (NEVLP) with conventional cold static organ preservation for livers retrieved after cardiac death. We implemented spectrophotometric imaging as tool for online monitoring of blood flow and oxygen transport to tissue during NEVLP to predict organ function post reperfusion. Methods: First, pig livers were perfused (n=5) for 12 hours in a cell free, oxygenated perfusion solution (STEEN), followed by 12hr perfusion with whole blood as a model of transplantation. ALT and histology were evaluated as parameters of the liver injury, urea synthesis, bile production and oxygen consumption were determined as marker of the liver function. In a second approach, pig livers either subjected to 1hr warm ischemia plus 12hrs cold storage in UW (n=5) or 1hr warm ischemia plus 4hr cold storage plus 8hr NEVLP. After 12hr organ preservation the livers were perfused for 8hr in a pig liver transplant model. Spectrophotometric imaging was used to monitor blood flow and oxygen supply to liver tissue and bile duct tissue. Results: 12hr normothermic acellular ex vivo liver perfusion followed by 12hr whole blood reperfusion was not associated with liver injury. Serum ALT (mean 27U/L) remained normal, and histology did not show any evidence of necrosis (< 1%). Bile production (mean 3cc/hr), oxygen consumption (400mmHg), and BUN synthesis (1.8mmol/l) were within normal limits. In a second approach, NEVLP was compared to cold static storage in a DCD model using 1hr warm ischemia and 12hr preservation. NEVLP was associated with significantly decreased serum AST (26U/L vs 285U/L), decreased necrosis (10% vs 35%, p< 0.05), and increased oxygen consumption (400mmHg vs 230mmHg, p< 0.05). Cold static storage after DCD retrieval was associated with loss of peripheral arterial blood supply, while arterial blood flow was maintained in NEVLP preserved grafts. Accordingly, UW preserved grafts had massive biliary necrosis (90%), while bile ducts were normal in NEVLP preserved livers. Spectrophotometric imaging showed decreased oxygen supply especially in peripheral parts of liver and bile duct tissue. Conclusion: NEVLP allows prolonged organ storage with normal liver metabolism without inducing preservation injury. Bile duct injury after DCD liver retrieval develops in cold static, but not NEVLP preserved organs. NEVLP is a novel preservation technique for the assessment of marginal grafts. The implemented spectrophotometric imaging can monitor oxygen transport to tissue and seems to be a promising tool to predict organ function post reperfusion.

Selectivity of TMC207 towards Mycobacterial ATP Synthase Compared with That towards the Eukaryotic Homologue

The diarylquinoline TMC207 kills Mycobacterium tuberculosis by specifically inhibiting ATP synthase. We show here that human mitochondrial ATP synthase (50% inhibitory concentration [IC(50)] of >200 microM) displayed more than 20,000-fold lower sensitivity for TMC207 compared to that of mycobacterial ATP synthase (IC(50) of 10 nM). Also, oxygen consumption in mouse liver and bovine heart mitochondria showed very low sensitivity for TMC207. These results suggest that TMC207 may not elicit ATP synthesis-related toxicity in mammalian cells. ATP synthase, although highly conserved between prokaryotes and eukaryotes, may still qualify as an attractive antibiotic target.