Calf Liver Research Articles

142 13C NUCLEAR MAGNETIC RESONANCE (NMR) EVALUATION OF GLYCOGEN CONTENT IS USEFUL IN THE DIAGNOSIS OF GLYCOGEN STORAGE DISEASES IN CHILDREN

Case report: a 10 month-old infant was admitted to hospital for stunted growth (-4SD). The diagnosis of glycogen storage disease (GSD) was suspected. RBC phosphorylase kinase activity was found to be normal. Liver needle biopsy confirmed the diagnosis of GSD and excluded GSD type I. 13C NMR was then performed. GSD type VI was later proved by the enzymatic assay.Methods: Ten minutes 1H decoupled 13C spectra of calf muscles and liver were obtained at 2T. Spectra were acquired with a 160° pulse at the coil center repeated every 200ms and with a 40ms acquisition time. The glycogen content was evaluated from the area of the glycogen C1 peak at 100.5 ppm. It was normalized to the area of the creatine peak at 157 ppm. 7 normal subjects and 11 patients suffering from GSD (3 type la, 2 type III, 4 type V, 1 type VI and 1 type VII) were studied.Results: The muscular glycogen/creatine ratio, R, was 2.0±0.7 in normal subjects and 2.1 in GSD type I. It was 13±1.7 in type V, 13.5 and 15 in type III, and 11.2 in type VII. In our patient, R was found to be 2.0. Semi quantitative (no internal reference for liver) evaluation of hepatic glycogen indicated high levels in the liver of our patient, as observed in GSD affecting the liver (types I, II, and VI).Conclusions: NMR allowed to exclude type III and strongly argued for type VI. Therefore, in vivo 13C NMR is able to distinguish GSD affecting the muscle from those not affecting it and also appears to detect abnormal hepatic glycogen content.

Hematology and clinical pathology of experimental Fascioloides magna infection in cattle and guinea pigs

The hematologic and clinico-pathologic response to Fascioloides magna infection in cattle and guinea pigs was investigated. Twelve calves (six infected and six controls) were monitored for 26 weeks after inoculation with 1000 metacercariae. All calves remained healthy and there were no significant differences in weight gains between infected and control groups. Flukes (mean = 9.2, range 1–32) were recovered from the liver and abdominal cavity of all infected calves. The only significant response observed in the complete blood counts was an eosinophilia present in the infected calves extending from Weeks 2 to 26 post-infection. There were no significant differences in serum levels of aspartate aminotransferase and only minor increases in the levels of gamma-glutamyl transferase and sorbitol dehydrogenase. A total of 48 infected and 48 control guinea pigs from three separate experiments were monitored for 16 weeks after inoculation with 20 metacercariae of Fascioloides magna. Infected guinea pigs died between 7 and 114 days after infection, and flukes (ean = 2.5, range 0–13) were recovered from the liver, abdominal cavity, lungs, thoracic cavity, skeletal muscle and subcutaneous tissue. There were no differences in weight gains between infected and control guinea pigs. Complete blood counts showed increases in white blood cell, monocyte and neutrophil counts from between the third and fourteenth weeks post-infection; however, the differences were not consistently significant. Infected guinea pigs developed a significant eosinophilia and basophilia from 2 to 16 weeks post-infection. There were no significant changes in the serum levels of alanine aminotransferase or gamma-glutamyl transferase. There was an increase in the serum levels of aspartate aminotransferase beginning at 5 weeks post-infection. The response observed in the guinea pigs was similar to that reported in sheep, suggesting the suitability of the guinea pig as a model for Fascioloides magna infection in the sheep.

Effect of Excess Dietary Manganese on Lipid Composition of Calf Blood Plasma, Heart, and Liver

Preruminant calves were fed milk replacer containing control (40ppm) or two high concentrations (200 and 1000ppm) of Mn to assess the effect of excessive Mn intakes on plasma, heart, and liver lipids.The two higher Mn intakes had no effect on lipid classes in liver and heart, except for elevated triglycerides in liver and lower sphingomyelin in heart (for 1000ppm of Mn). At 1000ppm of Mn intake, but not at 200ppm, marked increases occurred in plasma total lipids, phosphatidylcholine, cholesterol, cholesterol esters, sphingomyelin, and triglycerides. The highest intake altered the essential fatty acid composition of liver phosphatidylcholine. Linoleic and linolenic acids were increased, but arachidonic and eicosapentaenoic acids were decreased, suggesting that very high excess of Mn interfered with hepatic desaturation and elongation of the essential fatty acids.Thus, high Mn intake (200ppm) caused relatively few tissue lipid changes, whereas very high intake (1000ppm) markedly increased plasma lipid classes and apparently interfered with essential fatty acid metabolism in liver.

Hepatic vitamin A and carotene levels in the newborn foal

Summary Previous reports have indicated that at birth, the livers of calves, rats and rabbits contain very little vitamin A or its precursor carotene. A definitive report of the level of hepatic vitamin A and carotene in neonatal foals has not been found. To avoid having to sacrifice neonates, a cooperative collection program was initiated with local veterinarians to utilize livers from newborn foals that had not survived. The collection protocol was defmed as: a) the foal must have been born full term; b) the foal must have never nursed; and c) the foal must have appeared normal to a veterinarian or veterinary diagnos- tic laboratory. Ten whole livers were collected over a two year period. Liver samples were analyzed for total vitamin A and beta carotene. The mean (±SE) vitamin Alevelofthe ten livers was 20.7 ± 6.1 mcg/g of wet tissue. The beta carotene concentration in all livers was minimal:

Calcium-dependent autophosphorylation of the glucose-regulated protein, Grp78

The characteristics of phosphorylation of the 78-kDa glucose-regulated protein (Grp78), also known as the immunoglobulin heavy chain binding protein, were studied in vitro and in vivo. The purified protein from either calf liver or bovine kidney cells (MDBK) could be phosphorylated in vitro with [γ- 32P]ATP, in a reaction that is stimulated by Ca 2+ and inhibited by the Ca 2+-chelator ethylene glycol bis(β-aminoethyl ether) N,N′-tetraacetic acid (EGTA). In the presence of EGTA, excess Ca 2+ increased the rate of phosphorylation about 18-fold. Based on EGTA Ca 2+ titrations, the optimal Ca 2+ concentration for phosphorylation was estimated to be 10–50 m m. Other divalent cations such as Mg 2+, Mn 2+, and Zn 2+ were found to be inhibitory as was the Ca 2+ antagonist lanthanum (La 3+). The in vivo phosphorylation of Grp78 was studied in MDBK cells labeled with 32 P i. In the presence of inducers of Grp78 synthesis, such as ionomycin, tunicamycin, or 2-deoxyglucose, there was a large increase in the level of Grp78 in the cells but a decrease in the amount of phosphorylated protein. Two-dimensional gel analysis of Grp78 purified from bovine liver and MDBK cells identified at least four isoforms. After in vivo and in vitro phosphorylation of Grp78 all the acidic isoforms contained radioactivity but not the most basic isoform. Phosphoamino acid analysis of Grp78 showed that serine and threonine were phosphorylated in vivo and only threonine was phosphorylated in vitro.

Antibiotic and sulfonamide agents in bob veal calf muscle, liver, and kidney

Summary During the fiscal year 1988, USDA-FSIS detected 3,095 antimicrobial violations in bob veal calves, using the calf antibiotic and sulfonamide test. Of the 3,095 carcass submissions involved, 945 were tested further to identify the causative agents. The results of tests on the available kidney, liver, and muscle specimens are reported. Kidney specimens yielded a specific agent most often (71.2%), with neomycin (42.6%) being cited most among agents found in kidneys. Neomycin was found less frequently in liver (4.5%) and muscle (0.2%). Among all tissues, unidentified microbial inhibitors were either the largest or second largest category found (kidney, 10.5%; liver, 27.1%; muscle, 7.8%), and no other agent exceeded 7.0% (streptomycin in kidney). The proportion of liver and muscle specimens that had unidentified microbial inhibitors is particularly important because the next most common classes were streptomycin in liver at 5.5% and sulfamethazine in muscle at 2%. The frequency of unidentified microbial inhibitors may justify the addition of tests to the FSIS battery for identification of agents. Not all tissues were tested for sulfonamides, hence these agents are likely to have been underreported. Less than 10% of the muscle specimens evaluated yielded an agent, suggesting most calf antibiotic and sulfonamide test-positive carcasses may have been safe with regard to residues in meat, although organs might have been adulterated. Specimens for verification were not selected completely randomly from the population of all calf antibiotic and sulfonamide test-positive animals and calves selected for testing were not chosen strictly by random sampling; therefore, extrapolation of the contents of this report to the bob veal calf industry must be done with caution.

Multielement Concentrations in Liver and Kidney Tissues from Five Species of Canadian Slaughter Animals

The present paper describes results of a national survey conducted between 1982 and 1989 to determine residues of arsenic, cadmium, copper, mercury, lead, selenium, and zinc in Canadian slaughter animals. Liver and kidney tissues from cattle, swine, poultry, horses, calves, and sheep were tested. Arsenic was found in most avian and porcine samples, and their respective means of 0.36 and 0.26 micrograms/g in liver were 7 to 12 times higher than mean concentrations found in the other species. Cadmium was found in the tissues of all species; however, levels were consistently highest in equine samples with mean values of 3.09 and 27.7 micrograms/g in liver and kidney, respectively. Copper levels greater than 150 micrograms/g were found predominantly in liver from calves and sheep, with values considerably lower in the remaining species. Mercury levels were low or not detected in all species except horses. Ninety % of equine kidneys and 54% of equine livers had mercury concentrations greater than 0.01 microgram/g, with mean values of 0.18 and 0.06 microgram/g, respectively. Lead was found in tissues of all species; however, values greater than 2 micrograms/g were found only in 2 kidneys from adult cattle and 1 kidney from a horse. Selenium, tested only in cattle, was found at mean concentrations of 0.28 microgram/g in liver, and 0.92 microgram/g in kidney. Relatively high zinc levels were found in livers of horses, pigs, and calves, with respective mean concentrations of 67.3, 65.6, and 70.2 micrograms/g.

Estimation of pulses in ultrasound B-scan images

It is shown, based on an expression for the received pressure field in pulsed medical ultrasound systems, that a common one-dimensional pulse can be estimated from individual A-lines. An autoregressive moving average (ARMA) model is suggested for the pulse, and an estimator based on the prediction error method is derived. The estimator is used on a segment of an A-line, assuming that the pulse does not change significantly inside the segment. Several examples of the use of the estimator on synthetic data measured from a tissue phantom and in vitro data measured from a calf's liver are given. They show that a pulse can be estimated even at moderate signal-to-noise ratios.

Differential effects of dietary fatty acids on fatty acid composition of phosphotidylinositol in calf tissues

Fatty acid compositions of phosphatidylinositol (PI) in calf liver, heart, muscle, and brain were measured, as affected by dietary treatments. High Cu intake (1000 ppm) decreased 18:0 in liver PI and increased 18:1 n-9 and 18:2 n-6 in liver and heart; the reversed changes were observed in skeletal muscle PI. Additional high Zn intake (1000 ppm) inhibited the high Cu effects. Feeding an essential fatty acid (EFA) deficient diet markedly decreased 18:2 n-6, 20:3 n-6, and 20:4 n-6, and increased 18:1 and 20:3 n-9 in liver and heart PI. A high intake of 18:2 n-6 (corn oil) increased 18:2 n-6, but decreased 20:4 n-6, in liver, heart and muscle PI. Dietary fish oil concentrate (high in eicosapentaenoic acid) increased 20:5 n-3 in liver, heart, and muscle PI. Fatty acid composition of brain PI was resistant to different dietary regimens. Dietary imposed changes in fatty acid composition of PI in liver, heart, and muscle may have important implications in transmembrane signalling, eicosanoid production and other functions attributed to PI in tissues.

The transcription factor LF-A1 interacts with a bipartite recognition sequence in the promoter regions of several liver-specific genes

The transcription factors LF-A1 and LF-B1 are required for the cell-specific expression of the human alpha 1-antitrypsin gene in hepatocytes. We report here the purification and preliminary characterization of LF-A1. This protein, purified to homogeneity from calf liver nuclei by site-specific DNA affinity chromatography and reverse-phase HPLC, has a molecular mass of 40 kDa. Binding sites of LF-A1 are present in the promoter regions of several genes expressed in the liver (alpha 1-antitrypsin, apolipoproteins A1, B1, A4 and pyruvate kinase). Interestingly, the binding site of LF-A1 is bipartite and consists of two short sequence motifs (consensus: TGGACT/CT/C and TGGCCC) separated by a variable 'spacer' region. Insertion or deletion of 1-4 nucleotides in the 'spacer' region of the site in the alpha 1-antitrypsin promoter does not abolish DNA binding.

Large-scale purification of prosomes from calf's liver

Prosomes, cytoplasmatic ribonucleoprotein complexes containing small ribonucleic acid (19S small cytoplasmic RNPs), are ubiquitous in eukaryotic organisms. A new method for the preparation of prosomes in large amounts, starting with ca. 2 kg of calf's liver, is described. A combination of centrifugation and low- and high-pressure chromatography was used to purify intact particles. An alternative purification of prosomes with Solanum tuberosum agglutinin bound to divinyl sulphone-activated agarose is discussed. Calf's liver prosomes have a similar protein composition and RNA content to prosomes isolated from other tissues.

Effects of Dietary Corn Oil and Fish Oil Concentrate on Lipid Composition of Calf Tissues

Lipid composition of calf blood plasma, liver platelets, muscle, heart, and brain was measured, as affected by high dietary intake of linoleic acid from corn oil or of polyunsaturated fatty acids from fish oil concentrate. Plasma total lipids, phosphatidylcholine, and cholesteryl esters were reduced by corn oil and fish oil concentrate. Dietary fatty acid composition had no influence on percentage distribution of the major phospholipid components of liver, heart, muscle, and brain, but did alter the polyunsaturated fatty acid composition of major phospholipids in plasma, liver, platelets, muscle, and heart. In general, high linoleic acid intake increased linoleic acid and decreased oleic, arachidonic, and linolenic acids in tissue phospholipids, and fish oil concentrate high in eicosapentaenoic and docosahexaenoic acids increased phoslipid concentrations of these fatty acids. The fatty acid composition of brain phosphatidylcholine and phosphatidyl-ethanolamine was relatively resistant to dietary lipid alterations. The fatty acid changes in tissue phospholipids that resulted from dietary lipid alterations may have important implications in eicosanoid metabolism and in the structure and function of cell membranes.

Characterization of particulate cyclic nucleotide phosphodiesterases from bovine brain: purification of a distinct cGMP-stimulated isoenzyme

In the absence of detergent, approximately 80-85% of the total cGMP-stimulated phosphodiesterase (PDE) activity in bovine brain was associated with washed particulate fractions; approximately 85-90% of the calmodulin-sensitive PDE was soluble. Particulate cGMP-stimulated PDE was higher in cerebral cortical gray matter than in other regions. Homogenization of the brain particulate fraction in 1% Lubrol increased cGMP-stimulated activity approximately 100% and calmodulin-stimulated approximately 400-500%. Although 1% Lubrol readily solubilized these PDE activities, approximately 75% of the cAMP PDE activity (0.5 microM [3H]cAMP) that was not affected by cGMP was not solubilized. This cAMP PDE activity was very sensitive to inhibition by Rolipram but not cilostamide. Thus, three different PDE types, i.e., cGMP stimulated, calmodulin sensitive, and Rolipram inhibited, are associated in different ways with crude bovine brain particulate fractions. After solubilization and purification by chromatography on cGMP-agarose, heparin-agarose, and Superose 6, the brain particulate cGMP-stimulated PDE cross-reacted with antibody raised against a cGMP-stimulated PDE purified from calf liver supernatant. The brain enzyme exhibited a slightly greater subunit Mr than did soluble forms from calf liver or bovine brain, as evidenced by protein staining or immunoblotting after polyacrylamide gel electrophoresis under denaturing conditions. Incubation of brain particulate and liver soluble cGMP-stimulated PDEs with V8 protease produced several peptides of similar size, as well as at least two distinct fragments of approximately 27 kDa from the brain and approximately 23 kDa from the liver enzyme.(ABSTRACT TRUNCATED AT 250 WORDS)

12. Growth performance, liver and thyroid functions in buffalo calves reared on milk replacers supplemented with hydrogenated oils

12. Growth performance, liver and thyroid functions in buffalo calves reared on milk replacers supplemented with hydrogenated oils

Influence of soybean and corn gluten proteins as substitutes for milk protein in milk replacers on growth, liver and thyroid functions in buffalo calves

Influence of soybean and corn gluten proteins as substitutes for milk protein in milk replacers on growth, liver and thyroid functions in buffalo calves

Protein disulfide-isomerase is a substrate for thioredoxin reductase and has thioredoxin-like activity.

We have demonstrated that calf liver protein disulfide-isomerase (Mr 57,000) is a substrate for calf thymus thioredoxin reductase and catalyzes NADPH-dependent insulin disulfide reduction. This reaction can be used as a simple assay for protein disulfide-isomerase during purification in place of the classical method of reactivation of incorrectly oxidized ribonuclease A. Protein disulfide-isomerase contains two redox-active disulfides/molecule which were reduced by NADPH and calf thioredoxin reductase (Km approximately 35 microM). The isomerase was a poor substrate for NADPH and Escherichia coli thioredoxin reductase, but the addition of E. coli thioredoxin resulted in rapid reduction of two disulfides/molecule. Tryptophan fluorescence spectra were shown to monitor the redox state of protein disulfide-isomerase. Fluorescence measurements demonstrated that thioredoxin--(SH)2 reduced the disulfides of the isomerase and allowed the kinetics of the reaction to be followed; the reaction was also catalyzed by calf thioredoxin reductase. Equilibrium measurements showed that the apparent redox potential of the active site disulfide/dithiols of the thioredoxin domains of protein disulfide-isomerase was about 30 mV higher than the disulfide/dithiol of E. coli thioredoxin. Consistent with this, experiments using dithiothreitol or NADPH and thioredoxin reductase-dependent reduction and precipitation of insulin demonstrated differences between protein disulfide-isomerase and thioredoxin, thioredoxin being a better disulfide reductase but less efficient isomerase. Protein disulfide-isomerase is thus a high molecular weight member of the thioredoxin system, able to interact with both mammalian NADPH-thioredoxin reductase and reduced thioredoxin. This may be important for nascent protein disulfide formation and other thiol-dependent redox reactions in cells.

Isolation from fetal bovine serum of an apolipoprotein-H-like protein which inhibits thymidine incorporation in fetal calf erythroid cells

A 46 kDa heparin-binding protein which inhibits thymidine incorporation in cultures of fetal calf liver erythroid cells was isolated from fetal bovine serum by affinity chromatography on heparin-Sepharose, ion-exchange chromatography, gel filtration and reversed-phase h.p.l.c. The N-terminal sequence of the first 22 amino acids showed 81% identity with the published sequence of human apolipoprotein H. The isolated protein inhibited thymidine incorporation with an ED50 (concn. producing 50% of maximal effect) of 36 nM. A 100% inhibition of thymidine incorporation and a 40% decrease in cell numbers in cultures of fetal calf erythroid cells were observed at a protein concentration of 840 nM. No effects could be seen in cultures of 3T3 cells used as controls. Human apolipoprotein H had no inhibitory activity in any of the cell cultures tested, suggesting a species-specificity or a different structure or function for the bovine heparin-binding protein.

Structural studies of cytochrome b5: complete sequence-specific resonance assignments for the trypsin-solubilized microsomal ferrocytochrome b5 obtained from pig and calf

We report complete sequence-specific proton resonance assignments for the trypsin-solubilized microsomal ferrocytochrome b5 obtained from calf liver. In addition, sequence-specific resonance assignments for the main-chain amino acid protons (i.e., C alpha, C beta, and amide protons) are also reported for the porcine cytochrome b5. Assignment of the majority of the main-chain resonances was rapidly accomplished by automated procedures that used COSY and HOHAHA peak coordinates as input. Long side chain amino acid spin system identification was facilitated by long-range coherence-transfer experiments (HOHAHA). Problems with resonance overlap were resolved by examining differences between the two-dimensional 500-MHz NMR spectra of rabbit, pig, and calf proteins and by examining the temperature-dependent variation of amide proton resonances. Calculations of the aromatic ring-current shifts for protons that the X-ray crystal structure indicated were proximal to aromatic residues were found to be useful in corroborating assignments, especially those due to the large shifts induced by the heme. Assignment of NOESY cross peaks was greatly facilitated by a prediction of intensities using a complete relaxation matrix analysis based on the crystal structure. These results suggest that the single-crystal X-ray structure closely resembles that of the solution structure although there is evidence that the solution structure has a more dynamic character.

Postmortal degradation of furazolidone and furaltadone in edible tissues of calves

Rapid and complete postmortal degradation of furazolidone and furaltadone occurred in liver, kidney and muscle tissues of veal calves. Different degradation half-lives were observed between these tissues, the mean ranging from less than 7 minutes to 63 minutes. At 24 h after slaughter the parent nitrofurans were no longer detectable in edible tissues. For residue monitoring purposes plasma and/or urine can be used, if these matrices are treated in a specific way immediately after slaughter; muscle tissues and organs are unsuitable for residue monitoring of parent nitrofurans.

Evidence for the absence of cysteineS-conjugateN-acetyltransferase activity in the metabolism of propachlor, naphthalene, and dichlobanil in calves

1. The glutathione conjugate of 2-chloro-N-isopropyl[1-14C]acetanilide (14C-propachlor) was perfused through a calf kidney in situ; 23% of the dose was excreted in the perfused kidney urine as the cysteine conjugate, no mercapturic acid was detected. 2. A 5-day-old calf dosed orally with 14C-propachlor excreted 70% dose in the urine as the cysteine conjugate; no mercapturic acid was detected. Rumen microflora were established in the calf (5 weeks older) and the experiment was repeated with the same results. 3. When the same calf was dosed 1 week later with 14C-naphthalene, 99% dose was excreted in the urine, mostly as the dihydrodiol-glucuronide (34%) and the dihydrohydroxy-cysteine conjugate (47%); no mercapturate was detected. 4. A 9-day-old calf dosed orally with 2,6,-dichlorobenzo[14C]nitrile (14C-dichlobanil) excreted 67% dose in the urine as cysteine conjugates (34%), and products of cysteine conjugate beta-lyase cleavage of cysteine conjugates (30%); no mercapturates were detected. 5. Cysteine S-conjugate N-acetyltransferase activity in calf kidney and liver was about 10% of that in the corresponding rat tissues.