Peripartum factors associated with variation in voluntary postpartum hay intake in dairy cows.

The objective of this research was to assess variation in postpartum hay intake when offered alongside total mixed ration (TMR) as free choice, and identify factors related to the hay intake. Twenty multiparous cows were fed a closeup TMR (21.5% starch, 39.1% neutral detergent fiber [NDF] on a dry matter [DM] basis). After calving, cows were offered free choice timothy hay (61.6% NDF, 9.6% crude protein) in addition to a fresh cow TMR (26.8% starch, 33.0% NDF) for the first 5 d postpartum. Cows were fed individually with separate mangers for TMR and hay, each offered ad libitum. Prepartum DM intake (DMI) was recorded, and baseline blood samples were collected after calving, but before the first postpartum feeding. Free choice hay intake ranged from 0 to 4.7 kg/d (DM basis) or 0 to 55.2% (% of total DMI). Cows that consumed more hay (% of total DMI) from d 1 through 5 postpartum had lower DMI 2 d before calving (r = -0.63), and greater baseline concentrations of plasma ß-hydroxybutyrate (r = 0.60) and serum haptoglobin (r = 0.68). Additionally, hay intake (% of total DMI) from d 1 through 5 postpartum tended to be positively related to baseline plasma fatty acid concentration (r = 0.41). These findings suggest that cows with lower intake before calving and cows with greater ketone production and inflammation at calving may consume more hay, when offered separate from TMR.

Effects of offering free-choice hay for the first 5 days postpartum on productivity, serum inflammatory markers, gut permeability, and colon gene expression in fresh dairy cows.

The objective of the present study was to evaluate the effects of offering free-choice hay to cows during the first 5 d immediately after calving on feed intake, milk yield, plasma metabolites, serum inflammatory markers, rumination, gut permeability, and colon gene expression. It was hypothesized that cows offered free-choice hay would have lower gut permeability, lower inflammation, and higher milk production, compared with cows not offered hay. Thirty-two multiparous cows were fed a closeup total mixed ration (TMR; 21.5% starch, 32.1% forage neutral detergent fiber [NDF] on a dry matter basis) until calving. In the postpartum period, all cows were fed a fresh cow TMR (26.8% starch and 23.4% forage NDF) from calving until 21 DIM, and were assigned randomly to receive 1 of 2 treatments as follows: (1) free-choice timothy hay (61.6% NDF; 9.6% crude protein), offered outside of the TMR in a separate manger, for the first 5 d postpartum (FCH; n = 20), or 2) no free-choice hay (NH; n = 12). The FCH cows tended to have lower serum haptoglobin concentration on d 3, compared with NH (0.95 vs. 1.52 mg/mL). Within the FCH group, cows with greater hay intake had a smaller increase in serum amyloid A from d 1 to 3 after calving (r = 0.37), and tended to have a smaller increase in serum haptoglobin concentration (r = 0.36). Cows in the FCH group had a lower ratio of starch intake (kg) to forage NDF intake (kg) on d 1 and 2, compared with NH (0.91 vs. 1.14 ± 0.03), and cows that had a lower starch:forage NDF ratio tended to have a smaller increase in serum haptoglobin concentration from d 1 to 3 after calving (r = 0.32). Cows in the FCH group had lower TMR dry matter intake (DMI; 15.0 vs. 17.1 ± 0.93 kg/d) and lower total DMI (TMR + hay DMI; 15.9 vs. 17.1 ± 0.87 kg/d), from d 1 to 5 when free-choice hay was offered, compared with NH. However, the hay treatment did not affect plasma energy metabolite concentration, gut permeability, colon gene expression, milk yield, rumination time, or change in body weight or body condition score. Overall, these findings suggest that offering free-choice hay for the first 5 d after calving may reduce serum inflammatory marker concentration, but milk yield may not increase, due to lower intake.

Effects of dietary butyrate supplementation and oral nonsteroidal antiinflammatory drug administration on uterine inflammation and interval to first ovulation in postpartum dairy cows.

This study evaluated the effects of dietary butyrate supplementation and oral nonsteroidal antiinflammatory drug (NSAID) administration on uterine inflammation and the interval from calving to first ovulation (ICFO; in days). We hypothesized that a combination of dietary butyrate and oral NSAID would reduce uterine inflammation and decrease ICFO. Sixty-five cows were enrolled in a 2 × 2 factorial design and assigned to receive an iso-energetic diet containing a supplement of either butyrate (fatty acid-coated calcium butyrate) or control (commercial fat and calcium carbonate mixture) at 1.42% of diet dry matter, during the calving transition period from -28 (±3) to +24 (±3) days in milk (DIM; calving = d 0). At 12 to 24 h postcalving, cows received an oral NSAID (1 mg of meloxicam/kg of BW) or a placebo (food dye). Ovarian ultrasonography was performed weekly from 14 DIM until first ovulation or up to 56 DIM. Endometrial cytology was performed at 28 DIM to assess uterine inflammation based on polymorphonuclear leukocytes (PMN). No interactions were detected between treatments. The proportions of cows with high (>18%) endometrial PMN did not differ between butyrate and control diets or between NSAID and placebo. Mean (± standard error of mean) ICFO did not differ between butyrate (28 ± 2 d) and control (25 ± 2 d) or between NSAID (29 ± 2 d) and placebo (24 ± 2 d). However, the ovulation rate up to 56 DIM (hazard ratio: 0.61; 95% confidence interval: 0.35 to 1.04) established by survival analysis tended to be lower in NSAID than in placebo. In conclusion, dietary butyrate supplementation and oral NSAID administration did not reduce endometrial inflammation or the mean ICFO, but NSAID-treated cows tended to have a lower rate of ovulation up to 56 DIM.

Effects of dietary butyrate supplementation and oral nonsteroidal anti-inflammatory drug administration on serum inflammatory markers and productivity of dairy cows during the calving transition.

Dairy cattle experience inflammation during the calving transition period, and butyrate and nonsteroidal anti-inflammatory drugs (NSAID) are expected to reduce the inflammation. Our objective was to evaluate the effects of dietary butyrate supplementation and oral NSAID administration on feed intake, serum inflammatory markers, plasma metabolites, and milk production of dairy cows during the calving transition period. Eighty-three Holstein cows were used in the experiment with a 2 × 2 factorial arrangement of treatments. The cows were blocked by parity and calving date, and randomly assigned to a dietary butyrate or control supplement, and NSAID or a placebo oral administration. Experimental diets were iso-energetic containing calcium butyrate at 1.42% of diet dry matter (DM) or the control supplement (1.04% commercial fat supplement and 0.38% calcium carbonate of diet DM). The close-up diets contained 13.3% starch and 42.4% neutral detergent fiber on a DM basis, and were fed from 28 d before expected calving date until calving. The postpartum diets contained 22.1% starch and 34.1% neutral detergent fiber on a DM basis and were fed from calving to 24 d after calving. Oral NSAID (1 mg of meloxicam/kg of body weight) or placebo (food dye) was administered 12 to 24 h after calving. Dietary butyrate supplementation and oral NSAID administration did not affect milk yield or postpartum serum concentrations of amyloid A and haptoglobin. However, butyrate-fed cows increased plasma fatty acid concentration on d -4 relative to calving (501 vs. 340 µEq/L) and tended to increase serum haptoglobin concentration (0.23 vs. 0.10 mg/mL). There was a supplement by drug interaction effect on plasma glucose concentration on d 4; in cows administered the placebo drug, butyrate supplementation decreased plasma glucose concentration compared with control-fed cows (62.8 vs. 70.1 mg/dL). Butyrate-fed cows tended to have lower milk crude protein yield compared with cows fed the control diet (1.21 vs. 1.27 kg/d). Dietary butyrate supplementation and oral NSAID administration did not have overall positive effects on production performance of dairy cows during the calving transition period.

Effects of feeding hay and calf starter as a mixture or as separate components to Holstein calves on intake, growth, and blood metabolite and hormone concentrations.

This study investigated how providing hay mixed with calf starter to dairy calves affected their solid feed intake, feed sorting, growth, and plasma metabolite and hormone concentrations. Forty Holstein heifer calves were fed a texturized calf starter (23.4% crude protein, 32.3% starch on a dry matter basis) and chopped Klein grass hay as separate components (CONT) or the same starter and hay mixed at a 90:10 ratio on an as-fed basis (MIX) ad libitum from the date transported to the research farm (4-7 d of life) to 90 d of life. Calves were provided milk replacer (28% crude protein, 15% fat) at up to 557 g/d before the study, 737 g/d from d 14 to 20, 1,105 g/d from d 21 to 41, 737 g/d from d 42 to 48, and 557 g/d from d 49 to 55 on a dry matter basis. calves were fully weaned on d 56. Feed sorting for the MIX calves was evaluated using the Penn State Particle Separator; the sorting index was calculated as the actual intake as a percentage of predicted intake, with values >100% indicating sorting for and values <100% indicating sorting against. Treatment did not affect solid feed intake, growth performance, or plasma metabolite or hormone concentration during the preweaning or weaning periods. However, calves in the MIX treatment had less neutral detergent fiber intake as a percentage of solid feed intake than CONT calves in the preweaning (23.3 vs. 37.0%) and weaning (23.5 vs. 25.8%) periods, although MIX calves sorted (107.2%) for long particles, which were primarily hay, during weaning. During the postweaning period, MIX calves had greater neutral detergent fiber intake as a percentage of solid feed intake compared with CONT calves (23.4 vs. 22.7%), although they sorted against long particles (84.4%), and decreased solid feed dry matter intake compared with CONT calves (3,292 vs. 3,536 g/d) and average daily gain (1.20 vs. 1.31 kg/d). Weaned calves in the MIX treatment also had lower plasma concentration of glucagon-like peptide 2 compared with CONT (0.46 vs. 0.77 ng/mg) but had higher plasma concentrations of ghrelin (0.05 vs. 0.03 ng/mg). These results suggest that feeding a mixture of texturized calf starter and chopped hay at the 90:10 ratio to postweaned calves may decrease solid feed intake and growth.

Identification of BV/ODV-C42, an Autographa californica nucleopolyhedrovirus orf101-encoded structural protein detected in infected-cell complexes with ODV-EC27 and p78/83.

orf101 is a late gene of Autographa californica nucleopolyhedrovirus (AcMNPV). It encodes a protein of 42 kDa which is a component of the nucleocapsid of budded virus (BV) and occlusion-derived virus (ODV). To reflect this viral localization, the product of orf101 was named BV/ODV-C42 (C42). C42 is predominantly detected within the infected-cell nucleus: at 24 h postinfection (p.i.), it is coincident with the virogenic stroma, but by 72 h p.i., the stroma is minimally labeled while C42 is more uniformly located throughout the nucleus. Yeast two-hybrid screens indicate that C42 is capable of directly interacting with the viral proteins p78/83 (1629K) and ODV-EC27 (orf144). These interactions were confirmed using blue native gels and Western blot analyses. At 28 h p.i., C42 and p78/83 are detected in two complexes: one at approximately 180 kDa and a high-molecular-mass complex (500 to 600 kDa) which also contains EC27.

Humic acid-induced echinocyte transformation in human erythrocytes: characterization of morphological changes and determination of the mechanism underlying damage.

Blackfoot disease (BFD) is a peripheral arterial occlusive disease found among human inhabitants along the southwest coast of Taiwan. Well water used for drinking and cooking contains humic acid (HA), which may be a possible etiological factor. In this study, HA toxicity was investigated in human erythrocytes and was found to induce echinocytic formation. Morphological changes occurred in both a concentration- and time-dependent fashion. The presence of HA was also observed to facilitate the loading of erythrocytes with excess Ca(2+) (1 mM), which may have occurred following permeability changes in cell membranes, leading to echinocytic transformations. Sodium dodecyl sulfate (SDS) gel electrophoresis indicated that echinocyte formation was due to the oxidation of normal membrane proteins that were replaced by high-molecular-weight proteins. Humic acid also induced hemoglobin oxidation in erythrocytes. Data show that oxidative stress generated by HA as well as direct effects were exerted on the cytoskeleton of erythrocytes, and these may be significant factors in the etiology of BFD.

Effects of halothane and methoxyflurane on the hypothalamic-pituitary-adrenal axis in rat.

Effects of acute exposure (2 h) to either 1.5% halothane or 0.5% methoxyflurane on chemical mediators of the hypothalamic-pituitary-adrenal (HPA) axis were evaluated in male Sprague-Dawley rats immediately after exposure, after the righting reflex (4 h), or 24 h postexposure. Effects of these anesthetics on hippocampal corticotropin releasing factor (CRF) were also evaluated. Methoxyflurane caused significant elevations in pituitary adrenocorticotropin hormone (ACTH)-like immunoreactivities in all three of the experiment's time groups, yet halothane failed to cause the same response immediately after exposure. Serum ACTH-like immunoreactivities were significantly elevated immediately after exposure to both anesthetics, but were not elevated at 4 and 24 h postexposure. Corticosterone (CORT)-like immunoreactivities were significantly elevated by halothane in all experimental groups, and in the 2- and 24-h groups following methoxyflurane exposure. Hippocampal CRF-like immunoreactivities remained unaffected by either anesthetic. Results indicate that a 2-h exposure to either halothane or methoxyflurane results in significant activation of the rat hypothalamic-pituitary-adrenal axis, and that the activation appears to be sustained over a 24-h period.

Alterations in catecholamine turnover in specific regions of the rat brain following acute exposure to nitrous oxide.

The effects of nitrous oxide (N2O) on steady-state concentrations and turnover rates of catecholamines in the olfactory bulb, hypothalamus, brain stem, hippocampus, striatum, thalamus, cerebral cortex, and spinal cord were determined in rats. Animals were exposed for 2 h to either 60% N2O or air. Immediately following exposure, all animals were injected intraperitoneally with alpha-methylparatyrosine (alphaMPT), a competitive inhibitor of tyrosine hydroxylase, and sacrificed at 0, 30, or 90 min postinjection. Brain catecholamine concentrations were determined using high-performance liquid chromatography coupled with electrochemical detection (HPLC-EC). Results indicate that N2O exposure significantly elevates steady-state concentrations of norepinephrine (NE) in the hypothalamus and striatum yet decreases amine levels in the brain stem region. Steady-state levels of dopamine (DA) were not significantly altered in any region of the CNS by N2O exposure. Acute exposure to N2O also resulted in significant decreases in the turnover rate of NE in the brain stem, yet it increased turnover of this amine in the olfactory bulb, hypothalamus, and striatum. Acute exposure to N2O resulted in a decreased turnover rate of DA in the hippocampus and striatum. In contrast, N2O appears to increase DA turnover in the olfactory bulb. These results indicate that acute exposure to N2O in rats causes region-specific alterations in steady-state levels and turnover rates of DA and NE within the central nervous system.

Effects of halothane and methoxyflurane on regional brain and spinal cord substance P-like and beta-endorphin-like immunoreactivities in the rat.

Effects of acute exposure (2 hr) to either 1.5% halothane or 0.5% methoxyflurane were investigated in the Sprague Dawley rat. Pituitary (PIT) and central nervous system (CNS) substance P (SP)-like and beta-endorphin (beta-end)-like immunoreactivities were evaluated immediately after anesthetic exposure (2 h), after righting reflex (4 h) or 24 hr postexposure (24 h). Only halothane significantly reduced SP-like immunoreactivity in olfactory bulbs in both the 2-h and 4-h groups. Halothane elevated SP-like immunoreactivity of hippocampus at all three time periods, and in the hypothalamus at 2 h. Both anesthetics significantly depleted thalamic concentrations of SP-like immunoreactivity. Methoxyflurane anesthesia resulted in a drastic decrease in SP-like immunoreactivity in PIT at all three time periods periods, while halothane elevated PIT concentrations of this peptide at 4 h. Both anesthetics significantly decreased beta-end-like immunoreactivity in the olfactory bulbs and thalami at 2, 4, and 24 h. However, halothane alone significantly elevated beta-end-like immunoreactivity in the spinal cord at 24 h. Halothane significantly elevated PIT beta-end-like immunoreactivity at 2 and 24 h, while methoxyflurane significantly lowered it in the 4-h group, but elevated the levels of the same in the 24-h group. Brain stem beta-end immunoreactivity were significantly reduced at 2 h by both anesthetics, and at 4 h by methoxyflurane. Results indicate that halothane and methoxyflurane may differ significantly in their actions on SP and beta-end secreting neurons in the CNS.

Physiology of the endocrine pancreas.

The endocrine pancreas is composed of nests of cells called the islets of Langerhans, which comprise only about 20% of pancreatic cell mass and secrete insulin, glucagon, somatostatin, and pancreatic polypeptide. Insulin is anabolic, increasing storage of glucose, fatty acids and amino acids, while glucagon namely stimulates hepatic glycogenolysis, gluconeogenesis, and ketogenesis. Somatostatin acts as a paracrine agent to inhibit both insulin and glucagon release, and, therefore, to modulate their output. This article explores factors controlling release of these hormones, as well as the way in which they affect fuel metabolism in the whole animal.

Biochemical and physiological manifestations of acute insulin withdrawal.

When insulin is withdrawn acutely from a severely diabetic patient, a sequence of intricately interconnected events begins which, without intervention, can result in coma and death. These events involve not only carbohydrate metabolism, but protein and lipid metabolism, electrolyte and fluid balance as well. Acute insulin withdrawal permits the unopposed action of the counter-regulatory hormones (which are attempting to elevate the plasma glucose concentration), namely glucagon, cortisol, growth hormone, and epinephrine. This article details interrelationships existing between the many biochemical and physiological events that are occurring more or less simultaneously in the animal following acute insulin withdrawal.

Influence of halothane and methoxyflurane on regional brain and spinal cord concentrations of methionine-enkephalin in the rat.

Rats were exposed to either oxygen (controls), 1.5% halothane in oxygen, or methoxyflurane (0.5%) in oxygen over a period of 2 h, then sacrificed at the end of exposure (2-h group), 4 h after removal from environmental chamber (4-h group), or at 24 h following anesthetic exposure (24-h group). Pituitary (excluding the neural lobe, Pit), brain, and spinal cord areas were isolated and processed with Met-enkephalin tissue concentrations determined. In halothane-exposed animals, Met-enkephalin concentrations in pit and across CNS areas studied were significantly lower at 2 h following anesthetic exposure than in control animals. Concentrations of Met-enkephalin in many areas of CNS and Pit of 4-h group approached control levels. Concentrations of Met-enkephalin in all areas studied except spinal cord returned to basal levels by 24 h following halothane exposure. Exposure to methoxyflurane resulted in less dramatic changes in Met-enkephalin concentrations across CNS regions examined. Exposure to methoxyflurane resulted in significant decreases in Met-enkephalin levels in olfactory bulb, thalamus, and hippocampus only. Met-Enkephalin levels did not change significantly in other areas of the central nervous system following methoxyflurane exposure. These results indicate that halothane and methoxyflurane may have differential effects on the endogenous opioid system.

Effect of chronic DDC treatment on LHRH and substance P amidation processes in the rat.

We examined the effects of chronic diethyldithiocarbamate (DDC) treatment on the concentrations of methionine-enkephalin, mature and unamidated forms (-Gly) of luteinizing hormone releasing hormone (LHRH) and substance P (SP) in various regions of the central nervous system (CNS). Chronic DDC treatment resulted in elevations of LHRH-Gly like immunoreactivity in the preoptic area (POA) and the medial basal hypothalamus (MBH), as well as elevations in SP-Gly like immunoreactivity in all areas of the CNS examined. Castration altered the ratios of SP-G-like/SP-like immunoreactivity in the pons, and LHRH-Gly like immunoreactivity in the MBH. Met-enkephalin concentrations were significantly elevated in the pons and medulla of intact DDC-treated animals, and in the POA of both intact- and castrated DDC-treated animals. Results demonstrate that it is possible to detect basal levels of unamidated LHRH and SP in many areas of the CNS, with ratios of unamidated/amidated peptides representing a unique and sensitive method for determining altered posttranslational processing of these transmitters, especially under altered endocrine states such as castration. Pharmacological blockade of terminal enzymatic processing of these peptides may be useful in studying upstream regulatory events in peptidergic neurons.

Role of intracellular calcium and protein kinases in the activation of hepatic Na+/taurocholate cotransport by cyclic AMP.

Glucagon and dibutyryl cyclic AMP (Bt2cAMP) stimulate Na+/taurocholate (TC) cotransport and increase the intracellular Ca2+ concentration ([Ca2+]i) of hepatocytes. Whether the effect of cAMP is mediated via increases in [Ca2+]i, cAMP-dependent protein kinase (PKA), and/or protein kinase C (PKC) was investigated in this study. TC uptake and [Ca2+]i were determined in isolated rat hepatocytes using [14C]TC and the fluorescent dye quin-2, respectively. Bt2cAMP, forskolin, and 8-bromo-cAMP stimulated Na(+)-dependent, but not Na(+)-independent TC uptake. Bt2cAMP increased the maximal rate of Na+/TC cotransport without affecting the apparent Km. Increases in TC uptake and [Ca2+]i by Bt2cAMP were inhibited in hepatocytes preloaded with bis-(2-amino-5-methylphenoxy)-ethane-N,N,N',N'-tetraacetic acid (MAPTA) or preincubated with 8-diethylaminooctyl 3,4,5-trimethoxybenzoate (TMB8). Calmodulin antagonists inhibited Bt2cAMP-induced increases in TC uptake, but not [Ca2+]i. Other Ca(2+)-mobilizing agents (thapsigargin, vasopressin, phenylephrine, and ionomycin) increased [Ca2+]i but failed to stimulate TC uptake, indicating that an increase in [Ca2+]i alone is not a sufficient stimulus for TC uptake. However, increases in TC uptake by 1 and 10 microM Bt2cAMP were further increased by thapsigargin, indicating a permissive role for Ca2+/calmodulin. Bt2cAMP-induced increases in TC uptake and [Ca2+]i were inhibited by known inhibitors of PKA and by an activator of PKC, but they remained unaffected by a specific inhibitor of PKC. Unlike thapsigargin, vasopressin inhibited Bt2cAMP-induced increases in TC uptake. Taken together these results indicate that stimulation of hepatic Na+/TC cotransport by cAMP 1) is mediated via PKA; 2) is potentiated, but not mediated, by Ca2+/calmodulin-dependent processes; and 3) may be down-regulated by PKC.

Distribution of catecholamines in the central nervous system of the pig.

The objective of this study was to document, through comprehensive means, normal distribution and concentration of catecholamines in various regions of the CNS of pigs, an increasingly popular animal model used for transgenic manipulation of neural genes. The effects of gonadal steroidal status on this distribution were also assessed by comparing CNS catecholamine concentrations among mature male pigs (boars), immature (gilts) and mature female pigs (sows), and adult male pigs castrated prepuberally (barrows). Dissected tissue samples from the CNS were extracted in 2 N acetic acid, filtered through a 0.2 micron filter, then quantitated by reverse-phase high performance liquid chromatography using a C-18 reverse phase column with electrochemical detection. In both boars and sows the highest concentrations of norepinephrine (NE) were found in the diencephalic areas and brain stem. Gilts exhibited elevated concentrations of NE in the olfactory bulbs (OB), hypothalamus, pons, and corpus trapezoideum-locus ceruleus (LC) compared to lower concentrations in corresponding areas of sows. Prepuberal castration of the male was associated with significantly lower NE concentrations in the striatum, periaqueductal area (PAG), pons, LC, and spinal cord. The sow exhibited significantly lower NE concentrations in the mammillary area (Mam), PAG, pons, and spinal cord than those in corresponding areas of the boar. Dopamine concentrations appeared to be similar in all areas of the brain and spinal cord studied in the sow and boar. Results demonstrated that prepuberal castration of the male appears to significantly alter the DA content of the Mam and dorsal spinal cord, in contrast to gilts who possess significantly higher concentrations of DA. It is concluded from our studies that in general, catecholamine concentrations in various regions of the brain and spinal cord of sexually mature pigs parallel distributions of neuropeptides, substance P, and methionine enkephalin, as previously reported. In addition, significant association was found between gonadal activity and catecholamine concentrations in discrete areas of the pig brain.

Equine fasting hyperbilirubinemia.

It appears that different mechanisms responsible for fasting hyperbilirubinemia may be operative in different mammalian species (and subspecies). An increase in bilirubin production does not seem to occur in the horse, but a decrease in the hepatic uptake of bilirubin has been supported by a number of studies. Even though the delay in plasma elimination could also result from a decrease in hepatic blood flow, this possibility does not seem to play a major role since the hepatic uptake of compounds with low intrinsic hepatic clearance (e.g., ICG and bilirubin) appear to be affected more during fasting than those with higher clearances (e.g., BSP, bile acid, antipyrine, acetaminophen, and lidocaine) (Table I). Other possibilities such as a decrease in the affinity of hepatocellular membrane carriers involved in the uptake of these compounds or altered content of intracellular proteins involved in cellular transport or storage of bilirubin have not been investigated in horses. Competition with free fatty acids for these carrier-mediated events seems likely, particularly because horses and ponies experience high degrees of hyperlipidemia during fasting. However, studies that have explored the competition hypothesis, while not entirely negative, do not fully support it as being the sole mechanism responsible for this phenomenon. Hepatocellular UDPGT activities have not been adequately investigated in horses, but it is apparent that intraduodenal infusion of glucose is effective in reducing fasting hyperbilirubinemia and also in increasing biliary bilirubin excretion. It therefore seems possible that UDP-glucose and UDPGA levels in the livers of horses could be reduced during fasting, thus resulting in substrate depletion for the conjugating enzymes. As pointed out by Freedland et al. (1991), it is also possible that the horse, like the Bolivian squirrel monkey, might also have a relatively high apparent Km and low Vmax for UDPGT, thus resulting in high steady-state levels of plasma bilirubin, particularly during fasting. Although little is known about the cause of equine fasting hyperbilirubinemia and the subtle factors that may be modulating slight changes in the production, hepatocellular uptake, binding, conjugation, and/or biliary excretion of this pigment, it is known that it can be rapidly reversed by refeeding native hay. Perhaps one direction for future research could point toward more fully exploring what aspects of feeding are responsible for reversing this intriguing physiological phenomenon.

Cocaine and lidocaine interfere with epinephrine-induced changes in intracellular calcium concentration and glucose efflux from rat hepatocytes.

The mechanism of cocaine-induced hepatotoxicity is not clearly understood. Recent studies show that fluctuations in intracellular Ca2+ ([Ca2+]i) and/or cyclic-AMP ([cAMP]) concentration play a major role in hormone action, and sustained elevations in [Ca2+]i may be involved in the initiation of hepatocellular damage. To evaluate the possible involvement of intracellular Ca2+ and/or cAMP, we investigated effects of cocaine and lidocaine on basal, epinephrine and dibutyryl cyclic-AMP (DBcAMP)-induced changes in [Ca2+]i and glucose efflux from isolated rat hepatocytes. [Ca2+]i was monitored continuously using a Ca2(+)-selective fluorescence indicator, Quin-2, and was calculated after correcting for autofluorescence. Neither cocaine nor lidocaine (0.1-5 mmol/l) affected basal [Ca2+]i, yet both agents decreased epinephrine (10 mumol/l) and DBcAMP (100 mumol/l)-induced increases in [Ca2+]i in a dose-dependent fashion. Half-maximal inhibition occurred at 0.75 mmol/l cocaine and 1.7 mmol/l lidocaine. Cocaine and lidocaine also decreased epinephrine and DBcAMP-induced glucose efflux. The dose-dependent effect on epinephrine-induced glucose efflux was similar to that of both anesthetics on epinephrine-induced increases in [Ca2+]i. However, 5 mmol/l cocaine or lidocaine decreased DBcAMP-induced glucose efflux by less than 50%, and [Ca2+]i by more than 80%. Taken together, these results indicate that cocaine and lidocaine decrease the ability of epinephrine to stimulate glucose efflux by interfering with the Ca2(+)-mediated, and not the cAMP-mediated intracellular pathway. It is therefore speculated that alterations in metabolic endocrine regulation may contribute to cocaine's hepatotoxic effect.

Basal and bile salt-stimulated bile flow and biliary lipid excretion in ponies.

The role of bile salt in biliary lipid excretion was studied in 3 healthy ponies with chronic external biliary fistulas. After endogenous bile salt pool depletion, micelle-forming taurocholate or taurochenodeoxycholate was infused to replace excreted bile salt. Enterohepatic circulations were held open (total biliary diversion) throughout each study. Results indicated that biliary lipid excretion in ponies (113 +/- 21 nmol/min/kg of body weight) is approximately 10 times less than that reported in rodents. Although the lipid composition (4.4% cholesterol, 5.6% phospholipid, and 90% bile salt) was within the predicted range for a single phase of micellar (or vesicular) liquid in solution, it was supersaturated with cholesterol because of low absolute concentrations of bile salt and phospholipid. Ponies, like guinea pigs, were determined to have a high bile salt-independent secretion of biliary lipid with little (or no) coupling to endogenous bile salt output. However, bile salt excretion induced by higher taurocholate infusion rates (ie, those greater than the physiologic range of 61 to 125 nmol/min/kg) was positively correlated with an increase in biliary phospholipid excretion, but not cholesterol excretion, thus indicating that a threshold intracellular bile salt concentration may be associated with enhanced biliary phospholipid excretion in ponies. The apparent cholerectic effects of endogenous bile salts, taurocholate, and taurochenodeoxycholate (that is, the increment in bile flow per increment in bile salt recovered) were greater in ponies than reported for any other mammal.

Hepatotoxic bile acids increase cytosolic Ca++ activity of isolated rat hepatocytes.

Effects of bile acids on cystolic Ca++ activity and cell viability of isolated rat hepatocytes were studied to test the hypothesis that bile acids may produce hepatotoxicity by increasing cystolic Ca++ activity. Changes in cystolic Ca++ activity were calculated from time-dependent changes in fluorescence of quin-2 loaded hepatocytes. Release of lactate dehydrogenase and changes in propodium iodide fluorescence were used to assess cell viability. Bile acids studied were unconjugated and taurine-conjugated cholate, chenodeoxycholate (and taurochenodeoxycholate), deoxycholate (and taurodeoxycholate) and lithocholate (and taurolithocholate). With the exception of cholate and taurocholate, bile acids increased cystolic Ca++ activity within 10 to 30 sec in a concentration-dependent fashion (0.05 to 1.0 mM) and in the order lithocholate = taurolithocholate greater than chenodeoxycholate = taurochenodeoxycholate = deoxycholate = taurodeoxycholate. The initial increase in cystolic Ca++ activity by bile acids was not due to cell damage, since bile acid-induced decreases in cell viability were not significant until 2 to 3 min. At higher concentrations of unconjugated bile acid, there was a secondary increase in quin-2 fluorescence corresponding temporally to the increase in propodium iodide fluorescence, indicating cell damage after the initial increase in cystolic Ca++ activity. The ability of conjugated and unconjugated bile acids to increase cystolic Ca++ activity was abolished and decreased (60 to 90%), respectively, in the absence of extracellular Ca++, indicating that extracellular Ca++ is the major source of the bile acid-induced increase in cystolic Ca++ activity.(ABSTRACT TRUNCATED AT 250 WORDS)