Multicenter study of acetaminophen hepatotoxicity reveals the importance of biological endpoints in genomic analyses.

Gene expression profiling is a widely used technique with data from the majority of published microarray studies being publicly available. These data are being used for meta-analyses and in silico discovery; however, the comparability of toxicogenomic data generated in multiple laboratories has not been critically evaluated. Using the power of prospective multilaboratory investigations, seven centers individually conducted a common toxicogenomics experiment designed to advance understanding of molecular pathways perturbed in liver by an acute toxic dose of N-acetyl-p-aminophenol (APAP) and to uncover reproducible genomic signatures of APAP-induced toxicity.

Rates of ethanol metabolism decrease in sons of alcoholics following a priming dose of ethanol.

Rapid changes in rates of ethanol metabolism in response to acute ethanol administration have been observed in animals and humans. To examine whether this phenomenon might vary by risk for alcoholism, 23 young men with a positive family history of alcoholism (family history positive [FHP]) were compared to 15 young men without a family history of alcoholism (family history negative [FHN]). Rates of ethanol metabolism were measured in all subjects first after an initial ethanol dose (0.

WY-14,643 induced cell proliferation and oxidative stress in mouse liver are independent of NADPH oxidase.

Long-term exposure of rodents to peroxisome proliferators leads to increases in peroxisomes, hepatocellular proliferation, oxidative damage, suppressed apoptosis, and ultimately results in the development of hepatic adenomas and carcinomas. Peroxisome proliferators-activated receptor (PPAR)alpha was shown to be required for these pleiotropic responses; however, Kupffer cells, resident liver macrophages, were also identified as playing a role in peroxisome proliferators-induced effects, independently of PPARalpha. Previous studies showed that oxidants from NADPH (nicotinamide adenine dinucleotide phosphate, reduced) oxidase mediate acute effects of peroxisome proliferators in rodent liver.

Negative energy balance increases periprandial ghrelin and growth hormone concentrations in lactating dairy cows.

The reported effects of feeding on growth hormone (GH) secretion in ruminants have been inconsistent, and are likely influenced by energy status of animals. High-producing dairy cows in early lactation and late lactation were used to assess the effects of energy balance on temporal variation of plasma metabolites and hormones. Cows were fed a single diet once daily, and feed was withdrawn for 90 min prior to feeding.

Short communication: Rate of propionate infusion within meals does not influence feeding behavior.

Propionate has been shown to depress the feed intake of ruminants, but whether the rate of propionate infusion influences this response is unknown. To test this possibility, the rate of propionate infused within meals was altered while the total amount of propionate infused was held constant. Eight multiparous Holstein cows (51 +/- 19 d in milk, 44.

Phlorizin induces lipolysis and alters meal patterns in both early- and late-lactation dairy cows.

Phlorizin is known to increase whole-body glucose demand, but it has also stimulated lipolysis in past studies in ruminants. Increased lipolysis complicates studies of dry matter intake (DMI) regulation by hepatic oxidation by providing the liver with additional oxidative substrate. Therefore, to assess whether increased glucose demand selectively increases DMI for cows in negative energy balance, phlorizin was administered to early- and late-lactation cows.

Phlorizin administration does not attenuate hypophagia induced by intraruminal propionate infusion in lactating dairy cattle.

Infusion data from ruminants has shown that propionate stimulates satiety and decreases meal size, possibly because of increased propionate oxidation in the liver. In this experiment, phlorizin was used to increase glucose demand, which was expected to decrease propionate oxidation and attenuate the decrease in dry matter intake (DMI) caused by propionate infusion. Twelve multiparous, ruminally-cannulated Holstein cows (49+/-33 d in milk, 40+/-7 kg/d milk; mean+/-SD) were randomly assigned to square and treatment sequence in a replicated 4x4 Latin square experiment with a 2x2 factorial arrangement of treatments.

Sustained formation of alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone radical adducts in mouse liver by peroxisome proliferators is dependent upon peroxisome proliferator-activated receptor-alpha, but not NADPH oxidase.

Reactive oxygen species are thought to be crucial for peroxisome proliferator-induced liver carcinogenesis. Free radicals have been shown to mediate the production of mitogenic cytokines by Kupffer cells and cause DNA damage in rodent liver. Previous in vivo experiments demonstrated that acute administration of the peroxisome proliferator di(2-ethylhexyl) phthalate (DEHP) led to an increase in production of alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN) radical adducts in liver, an event that was dependent on Kupffer cell NADPH oxidase, but not peroxisome proliferator-activated receptor (PPAR)alpha.

Propionate challenge tests have limited value for investigating bovine metabolism.

Two experiments were designed to assess the potential utility of the propionate challenge test (PCT) as an index of gluconeogenic capacity. In Expt. 1, the dose-response to jugular propionate infusion was assessed in a duplicated 4 x 4 Latin square experiment with 8 lactating dairy cows.

Lipopolysaccharide-binding protein modulates hepatic damage and the inflammatory response after hemorrhagic shock and resuscitation.

Hemorrhagic shock and resuscitation cause endotoxemia and hepatocellular damage. Because lipopolysaccharide-binding protein (LBP) enhances cellular responses to endotoxin, our aim was to determine whether LBP contributes to hemorrhage/resuscitation-induced injury by comparing LBP knockout and wild-type mice. Under pentobarbital anaesthesia, wild-type and LBP-deficient mice were hemorrhaged to 30 mmHg for 3 h and then resuscitated with shed blood plus half the volume of lactated Ringer solution.

Temporal correlation of pathology and DNA damage with gene expression in a choline-deficient model of rat liver injury.

Hepatocellular carcinoma (HCC) is the terminal event in chronic liver diseases with repeated cycles of cellular injury and regeneration. Although much is known about the cellular pathogenesis and etiological agents leading to HCC, the molecular events are not well understood. The choline-deficient (CD) model of rodent HCC involves the consecutive emergence of a fatty liver, apoptosis, compensatory proliferation, fibrosis, and cirrhosis that is markedly similar to the sequence of events typified by human HCC.

Phlorizin administration increases hepatic gluconeogenic enzyme mRNA abundance but not feed intake in late-lactation dairy cows.

Gluconeogenic capacity may be an important factor regulating dry matter intake (DMI) in lactating dairy cows. To determine whether increased glucose demand affects feed intake and hepatic gene expression, lactating Holstein cows were treated with phlorizin or vehicle (propylene glycol) for 7 d. Multiparous cows (n = 12; 269 +/- 65 d in milk, mean +/- SD) were randomly assigned to treatment sequence in a crossover design and were adapted to a common diet for 7 d before the beginning of the experiment.

The cow as a model to study food intake regulation.

Animal models have been invaluable for studying aspects of food intake regulation that for various reasons cannot be observed in humans. The dairy cow is a unique animal model because of an unrivaled energy requirement; its great drive to eat results in feeding behavior responses to treatments within the physiological range. Cows' docile nature and large size make them ideal for measuring temporal treatment effects because digestion and absorption kinetics and responses in endocrine systems, gene expression, metabolite pools and fluxes, and feeding behavior can be measured simultaneously.

Propionate is not an important regulator of plasma leptin concentration in dairy cattle.

Propionate was recently shown to increase leptin synthesis in rodents. To determine if a similar effect occurs in ruminants, propionate was administered to lactating dairy cows. In experiment 1, 31 cows were given an intrajugular Na propionate bolus (1,040 micromol/kg body weight), increasing plasma propionate from 160 to 5,680 microM and plasma insulin from 6.

Swift increase in alcohol metabolism (SIAM): understanding the phenomenon of hypermetabolism in liver.

The swift increase in alcohol metabolism (SIAM) is a phenomenon defined as a rapid increase in hepatic respiration and alcohol metabolism after administration of a bolus dose of alcohol. Continuous exposure to alcohol is known to produce adaptive changes in liver alcohol and oxygen metabolism. A considerable burst of hepatic respiration can also occur after administration of a single large dose of alcohol and results in a near doubling of alcohol metabolism, a high demand for oxygen, and downstream or pericentral hypoxia.

Standardizing global gene expression analysis between laboratories and across platforms.

To facilitate collaborative research efforts between multi-investigator teams using DNA microarrays, we identified sources of error and data variability between laboratories and across microarray platforms, and methods to accommodate this variability. RNA expression data were generated in seven laboratories, which compared two standard RNA samples using 12 microarray platforms. At least two standard microarray types (one spotted, one commercial) were used by all laboratories.

Cytochrome P450 CYP2E1, but not nicotinamide adenine dinucleotide phosphate oxidase, is required for ethanol-induced oxidative DNA damage in rodent liver.

The occurrence of malignant tumors of the upper gastrointestinal tract and liver is, based largely on epidemiological evidence, causally related to the consumption of ethanol. It is widely recognized that oxidants play a key role in alcohol-induced liver injury; however, it is unclear how oxidants may be involved in DNA damage. We asked whether nicotinamide adenine dinucleotide phosphate oxidase, cytochrome P450 CYP2E1, or both are responsible for the production of DNA damage.

Milk fat responses to a change in diet fermentability vary by production level in dairy cattle.

The effects of dietary starch fermentability on plasma metabolites and hormones, milk production, and milk fatty acid profile were evaluated in a crossover study. Thirty-two multiparous Holstein cows (121 +/- 48 d in milk, 41 +/- 9 kg/d 3.5% fat-corrected milk [FCM]; mean +/- SD) were randomly assigned to treatment sequence and were fed a diet intermediate to the treatments during an initial 21-d period.

Antifibrotic effects of a tissue inhibitor of metalloproteinase-1 antibody on established liver fibrosis in rats.

Liver fibrosis is characterized by increased synthesis, and decreased degradation, of extracellular matrix (ECM) within the injured tissue. Decreased ECM degradation results, in part, from increased expression of tissue inhibitor of metalloproteinase-1 (TIMP-1), which blocks matrix metalloproteinase (MMP) activity. TIMP-1 is also involved in promoting survival of activated hepatic stellate cells (HSCs), a major source of ECM.

Cre-loxP mediated control of PrP to study transmissible spongiform encephalopathy diseases.

Expression of the PrP glycoprotein is essential for the development of the transmissible spongiform encephalopathy (TSE) or prion diseases. Although PrP is widely expressed in the mouse, the precise relevance of different PrP-expressing cell types to disease remains unclear. To address this, we generated two lines of floxed PrP gene-targeted transgenic mice using the Cre recombinase-loxP system.

Contribution of angiotensin II to alcohol-induced pancreatic fibrosis in rats.

The mechanisms by which alcohol causes pancreatic fibrosis remain unknown. Recent studies have demonstrated that angiotensin II contributes to the development of fibrosis in liver, kidney, and heart injury. Here, the effects of angiotensin-converting enzyme inhibitor (captopril) and angiotensin II receptor antagonist (losartan) on alcohol-induced pancreatic fibrosis were examined in an intragastric ethanol-feeding model.

CD14 mediates the innate immune responses to arthritopathogenic peptidoglycan-polysaccharide complexes of Gram-positive bacterial cell walls.

Bacterial infections play an important role in the multifactorial etiology of rheumatoid arthritis. The arthropathic properties of Gram-positive bacteria have been associated with peptidoglycan-polysaccharide complexes (PG-PS), which are major structural components of bacterial cell walls. There is little agreement as to the identity of cellular receptors that mediate innate immune responses to PG-PS.

Expression of base excision DNA repair genes is a sensitive biomarker for in vivo detection of chemical-induced chronic oxidative stress: identification of the molecular source of radicals responsible for DNA damage by peroxisome proliferators.

Oxidative stress to DNA is recognized as one of the mechanisms for the carcinogenic effects of some environmental agents. Numerous studies have been conducted in an attempt to document the fact that chemical carcinogens that are thought to induce production of oxidants also cause the formation of oxidative DNA lesions. Although many DNA adducts continue to be useful biomarkers of dose/effect, changes in gene expression have been proposed to be a practical novel tool for studying the role of chemically induced oxidative DNA damage.

The CYP inhibitor 1-aminobenzotriazole does not prevent oxidative stress associated with alcohol-induced liver injury in rats and mice.

Cytochrome P450 (CYP) 2E1 is induced by ethanol and is postulated to be a source of reactive oxygen species during alcoholic liver disease. However, there was no difference in liver pathology and radical formation between wild-type and CYP2E1 knockout mice fed ethanol. Other CYP isoforms may contribute these effects if CYP2E1 is inhibited or absent.