Source of dietary protein influences kinetics of plasma gut regulatory peptide concentration in response to feeding in preruminant calves

Anaemia in pregnancy in developing countries continues to be a public health problem of significant proportion. At least 50% of the anaemia has been blamed on iron deficiency. In populations where chronic inflammation and iron deficiency anaemia coexist, the criteria to accurately define iron status are not always clear. Similarly, in pregnancy, with marked physiological changes, cut-off points for biochemical parameters need to be re-examined. In this study we examined the diagnostic accuracy of iron parameters including mean cellular volume (MCV), serum iron, transferrin, total iron binding capacity (TIBC) and its saturation, zinc protoporphyrin (ZPP), ferritin and serum transferrin receptor (TfR) for the assessment of iron status in a population of anaemic pregnant women in Malawi. Stained bone marrow aspirates were used as the standard for comparison. Results show that for the purpose of screening, serum ferritin is the best single indicator of storage iron provided a cut-off point of 30 microg/l is used. A number of other commonly used parameters of iron status were shown to have limited diagnostic accuracy. Logistic regression was used to obtain mathematical models for the prediction of bone marrow iron status using a combination of available parameters.     

Kinetics of pancreatic juice secretion in relation to duodenal migrating myoelectric complex in preruminant and ruminant calves fed twice daily

Daily secretion of pancreatic juice, including postprandial responses to food, was investigated in two groups of calves: preruminant (fed with liquid food) and ruminant (fed with solid food). Male Friesian calves (1 week old and 6 weeks old) were surgically fitted with a pancreatic duct catheter, duodenal cannula and two duodenal electrodes. Continuous 24 h collections of pancreatic juice and myoelectrical recordings were performed with minimal restraint and disturbance of animals. In both groups of calves clear periodic fluctuations in pancreatic juice secretion (volume, protein output and trypsin activity) coinciding with duodenal migrating myoelectric complexes (MMC) were recorded. Secretion of juice per cycle and per day was greater in ruminant calves, but the frequency and amplitude of cycles were lower in this group. There were no differences between day and night-time preprandial pancreatic cycles and duodenal MMC in preruminant calves, whilst in ruminant calves, evening MMC were longer than morning MMC. The pancreatic cephalic phase (increase of volume flow, protein output and trypsin activity during and just after food intake) was significant only in preruminant calves following morning feeding. Postprandial pancreatic cycles did not differ from preprandial cycles, except the pancreatic cycle (juice volume and trypsin activity) in which food was offered in preruminant calves. No gastric or intestinal phase was observed in either group of calves. In conclusion, biological cycles of the gastrointestinal tract are present in both preruminant and ruminant calves, and these cycles evolve along with the change from liquid to solid food.     

The effects of dietary soybean versus skim milk protein on plasma and hepatic concentrations of zinc in veal calves

We assessed the zinc status of veal calves that were fed milk replacers containing either skim milk protein as the sole source of protein or a mixture of skim milk protein and soybean protein. After the milk replacers had been fed for 26 wk, mean body weight gain was 3 kg lower for calves fed the skim milk plus soybean proteins; this decrease was not significant. Inclusion of dietary protein from soybeans versus milk protein alone reduced plasma concentrations of zinc by 43% and reduced hepatic concentrations of zinc by 81%. The impairment of zinc status that was induced by the inclusion of soybean protein was probably caused by its phytate component. The effect of soybean protein on zinc status was rather specific because plasma and hepatic concentrations of copper were unaffected. Despite the high concentration of zinc (142 mg/kg of dry matter) in the milk replacer that contained milk plus soybean proteins, calves displayed a shortage of zinc because their plasma and hepatic concentrations of zinc were significantly reduced.     

Fatty acid metabolism and very low density lipoprotein secretion in liver slices from rats and preruminant calves

Diethyldithiocarbamate methyl ester (DDTC-Me) is a precursorto the formation of S-methyl-N,N-diethylthiolcarbamate sulfoxide, the active metabolite proposed to be responsible for the alcohol deterrent effects of disulfiram. The present study investigated the role of human cytochrome P-450 (CYP) enzymes in sulfoxidation and thiono-oxidation of DDTC-Me, intermediary steps in the activation of disulfiram. Several approaches were used in an attempt to delineate the particular P-450 enzyme(s) involved in the sulfoxidation and thiono-oxidation of DDTC-Me. These approaches included the use of cDNA-expressed human P-450 enzymes, correlation analysis with sample-to-sample variation in human P-450 enzymes in a bank of human liver microsomes, and chemical and antibody inhibition studies. Multiple human P-450 enzymes (CYP3A4, CYP1A2, CYP2A6, and CYP2D6) catalyzed the sulfoxidation of DDTC-Me, as determined with cDNA-expressed enzymes. Several lines of evidence suggest that the sulfoxidation of DDTC-Me by human liver microsomes is primarily catalyzed by CYP3A4/5, including (1) a high correlation between DDTC-Me sulfoxidation and testosterone 6beta-hydroxylation; (2) increased DDTC-Me sulfoxidation in the presence of alpha-naphthoflavone, an activator of CYP3A enzymes; (3) inhibition of this reaction by inhibitors of CYP3A4/5 enzymes, such as troleandomycin and ketoconazole; and (4) inhibition of DDTC-Me sulfoxidation by antibodies against CYP3A enzymes. On the other hand, several lines of evidence suggested that the thiono-oxidation of DDTC-Me by human liver microsomes is catalyzed in part by CYP1A2, CYP2B6, CYP2E1, and CYP3A4/5, including (1) these human P450 enzymes among others have the capacity to catalyze this reaction, as determined with cDNA-expressed enzymes; (2) a high correlation between DDTC-Me thiono-oxidation and testosterone 6beta-hydroxylation, weak inhibition by ketoconazole, troleandomycin, and anti-CYP3A antibodies suggested a minor role for CYP3A4; (3) a high correlation with immunoreactive CYP2B6 suggested involvement of this enzyme; (4) weak inhibition of DDTC-Me thiono-oxidation by furafylline and anti-CYP1A antibody suggested involvement of CYP1A2; and (5) inhibition of DDTC-Me thiono-oxidation by DDTC and anti-CYP2E antibodies suggested a role for CYP2E1. Collectively, these data suggested CYP3A4/5 enzymes are the major contributors to the sulfoxidation of DDTC-Me by human liver microsomes, and CYP1A2, CYP2B6, CYP2E1, and CYP3A4/5 contribute toward DDTC-Me thiono-oxidation by human liver microsomes. This study, in conjunction with others (Madan et al., Drug Metab. Dispos. 23:1153-1162, 1995), may help explain the variability in disulfiram's effectiveness as an alcohol deterrent.     

Nitrogen and fat balances in very low birth weight infants fed human milk fortified with human milk or bovine milk protein

The study was designed to compare two different human milk fortifiers in a group of very low birth weight (VLBW) infants by analysing nitrogen and fat balances, serum concentrations of alpha-amino-nitrogen, urea, and prealbumin as well as growth rates when human milk enriched with one of the two studied fortifiers was fed to the infants. Fortifier A contained different bovine proteins, peptides and amino acids and had an amino acid composition comparable to that of the nutritional available proteins in human milk, with carbohydrates, and minerals. Fortifier B was composed of freeze-dried skimmed human milk and minerals to achieve a similar macronutrient composition in both fortifiers. Eleven infants were fed with human milk enriched with fortifier A and 13 with fortifier B. After a 10-day equilibration period, a 3-day metabolic balance was performed. On the 14th day of the study blood was obtained preprandially for serum analysis and growth rates were estimated. The nitrogen absorption rate (93.8% vs 93.5%) as well as the retention rate (80.8% vs 78.5%) were no different between the groups. The fat absorption rate (92.3% vs 91.5%) as well as the weight gain (32.1 vs 31.1 g/day) were similar and there were no differences in the serum parameters studied. The results indicate that feeding VLBW infants with human milk enriched with a well-balanced bovine fortifier fulfil their nutritional requirements as well as diets composed exclusively of human milk protein.     

Intestinal barrier function and cow's milk sensitization in guinea pigs fed milk or fermented milk

BACKGROUND: The respective effect of milk and fermented milks on intestinal barrier capacity and on sensitization to beta-lactoglobulin was studied using a guinea pig model of cow's milk allergy. METHODS: Guinea pigs were fed a control diet or the same diet supplemented with milk, fermented milk (Streptococcus thermophilus and Bifidobacterium breve), or dehydrated fermented milk. Intestinal barrier capacity to macromolecules was assessed in an Ussing chamber, and sensitization to cow's milk proteins was measured by systemic anti-beta-lactoglobulin immunoglobulin G1 titers and by intestinal anaphylaxis, the latter assessed by the beta-lactoglobulin-induced increase in short-circuit current of jejunal fragments (deltaIsc(beta-LG)). RESULTS: The electrical resistance of jejunum was similar in the four groups (approximately 80 omega/cm2) suggesting the same paracellular permeability. The transport of 14C-beta-lactoglobulin from mucosa to serosa was significantly decreased in the animals fed dehydrated fermented milk (403+/-131 ng / hr x cm2) compared with that in control animals or animals fed milk (767+/-250 ng / hr x cm2 and 749+/-475 ng / hr x cm2, respectively; p < 0.05). Milk fermentation did not modify native beta-lactoglobulin concentration but anti-beta-lactoglobulin immunoglobulin G1 titers were higher in fermented milk and dehydrated fermented milk (log10 titer = 2.86 and 2.79, respectively) than in guinea pigs fed milk (log10 titer = 2.5; p < 0.007). However, beta-lactoglobulin-induced intestinal anaphylaxis remained the same in the three groups (deltaIsc(beta-LG), 9.6+/-4.1 microA/cm2, 8.5+/-4.3 microA/cm2, and 8.5+/-3.4 microA/cm2 in milk-fed, fermented milk-fed, and dehydrated fermented milk-fed guinea pigs, respectively). CONCLUSIONS: The intestinal barrier capacity to milk proteins seems to be reinforced by dehydrated fermented milk, but milk and fermented milks are equally efficient in inducing cow's milk allergy in guinea pigs.     

Blocking of protein pancreatic secretion by maximal secretion infusion in the dog

In conscious chronic gastric and pancreatic fistula dogs (Thomas cannula), secretin was perfused for three hours with a submaximal (GIH, 1.0 C.U./kg.) and a maximal dose (GIH, 8.0 c.u./kg.), according to the following schedule: 1. First hour submaximal stimulus; 2. second hour maximal stimulus; 3. third hour submaximal stimulus. The alkaline and protein components of pancreatic secretion were analyzed in 20-minute sample collections thoughout the three hours. The same protocol was followed in anesthetized dogs subjected to a mind line laparotomy. A biopsy of the pancreatic gland was taken before (control) and at the end of each perfused dose. The secretion showed a significant increase of protein concentration and output when passing from the maximal to the last submaximal secretin perfusion dose. These findings correlated well with the piling up of zymogen and prozymogen granules in the apical zone of the acinar cells during maximal secretin perfusion, with their subsequent discharge into the acinar lumen upon abrupt reversal to the initial secretin submaximal dose. The study confirms that secretin influences pancreatic protein secretion and indicates in addition, that pharmacologic doses of the hormone, have the capacity to block acinar cell zymogen granule release.     

Exocrine pancreatic secretion in suckling goats. Adaptative effects of maternal milk and a milk substitute

McNtcp.24 cells are rat hepatoma cells that were made competent to take up conjugated bile acids actively from the culture medium. Treatment of McNtcp.24 cells with certain species of bile acids caused significant changes in cell structure. Incubation of McNtcp.24 cells in medium containing 100 microM taurocholic acid induced a profound alteration of cellular morphology. Very larger vesicles, visible by phase contrast microscopy, were the most prominent feature of bile acid-treated McNtcp.24 cells. Staining of cells with Oil red O and filipin indicated that the vesicles did not contain neutral lipids or free cholesterol. The vesicles remained in the cells after efflux of radiolabeled taurocholic acid from bile acid loaded cells, indicating that these structures are not intracellular stores of bile acids. Electron microscopic analysis of bile acid-treated McNtcp.24 cells confirmed that the vesicles were localized within the cells. Taurine-conjugated bile acid species were generally potent inducers of the morphological changes, although tauroursodeoxycholic acid did not have a significant effect. Unconjugated bile acid species were ineffective or only mildly effective. Bile acid treatment also caused profound alteration of mitochondrial structure. Surprisingly, there was no significant effect on the ability of treated cells to oxidize fatty acids. The bile acid-treated cells remained viable and upon withdrawal of bile acids from the culture medium, the cells returned to normal morphology by 24 h. The morphological changes observed after treatment of McNtcp.24 with bile acids are reminiscent of the morphological changes observed in hepatocytes following induction of cholestasis.     

Effects of intraduodenal administration of tarazepide on pancreatic secretion and duodenal EMG in neonatal calves

The influence of CCK-A receptor antagonism on pancreatic exocrine secretion and duodenal EMG, and the mechanism(s) involved in CCK-induced pancreatic secretion were studied in conscious calves. Seven 1-week-old calves were fitted with a pancreatic duct catheter, duodenal cannula and duodenal electrodes. Pancreatic exocrine secretion and duodenal EMG were studied following intraduodenal CCK-A receptor antagonist (Tarazepide), intravenous atropine, and intravenous or intraduodenal CCK-8 administrations. Tarazepide decreased duodenal electric activity, reduced interdigestive pancreatic secretion, especially protein; reduced cephalic and early postprandial (milk) induced secretion of bicarbonate and protein. Pancreatic protein secretion to intravenous CCK-8 was little affected by atropine, but was significantly reduced by Tarazepide+/-atropine; in contrast, protein secretion to intraduodenal CCK-8 was abolished by Tarazepide or atropine. We conclude that pre- and especially early postprandial pancreatic secretion are partly controlled via CCK-A (mainly mucosal) mediated mechanisms.     

Plasma lipids, ketone bodies, and glucose concentrations in calves fed high- and low-fat milk replacers

Twenty-four 5-day-old male calves were fed twice daily milk replacers containing either 5% (low-fat) or 25% (high-fat) lard. Plasma lipids, blood glucose, and ketone bodies were determined in jugular blood before feeding and every hour during 8 h after feeding. The high-fat diet caused in the 1st h after feeding a sharp increase of triglycerides and phospholipids followed by a sharp decrease; these two increased slowly during the following 5 h. Within the first 2 h after feeding, there was an increase of cholesterol esters, free cholesterol, and nonesterified fatty acids. With the low-fat diet, triglycerides and cholesterol esters showed a small increase during the 4 h following meal whereas phospholipids, free cholesterol, and nonesterified fatty acids were not affected significantly. With both diets, blood glucose reached a maximum of 110 mg/100ml 1 h after feeding; ketone bodies were not altered significantly. With the high-fat diet, lipid digestion would occur in two phases; firstly, part of the fat would be lipolyzed quickly by pregastric esterase before clot formation in the abomasum; secondly, the rest of the lipids, slowly released by progressive lysis of the coagulum would be digested under the action of gastric and pancreatic lipases. The first phase did not occur with the low-fat diet.     

Milk production and plasma gossypol of cows fed cottonseed and oilseed meals with or without rumen-undegradable protein

Twenty-four multiparous Holstein cows were randomly assigned at calving to treatment diets using a modified split-plot design to determine the effects of protein source on milk production and composition. The treatment diets consisted of an 80:20 combination of corn and alfalfa silages and whole cottonseed at 12% of the dietary dry matter (DM). The treatment diets were formulated to contain 17% crude protein (CP) and 20% acid detergent fiber on a DM basis. One of the following sources of supplemental CP was included in each treatment diet: 1) cottonseed meal, 2) cottonseed meal plus a rumen-undegradable protein (RUP) supplement, 3) soybean meal, and 4) soybean meal plus an RUP supplement. Cows were fed the initial treatment diet for 6 wk and then were switched to the other oilseed meal source but continued to receive the same amount of RUP during the second period of the study. Milk production and composition were not affected by treatment diet. Cows fed treatment diets without RUP supplementation consumed more DM and thus more CP. Supplementation with RUP resulted in greater milk production efficiency per unit of DM consumed. Cows fed treatment diets containing cottonseed meal had higher plasma gossypol concentrations than did cows fed treatment diets containing soybean meal. Plasma gossypol concentrations for all cows in each group were below the recommended upper limit that is considered to be safe. Data suggest that cottonseed meal in the diet can be substituted for soybean meal, resulting in similar milk production and composition.     

Nitrogen retention in men fed isolated soybean protein supplemented with L-methionine, D-methionine, N-acetyl-L-methionine, or inorganic sulfate

The ability of various sulfur-containing compounds to replace L-methionine (L-Met) was investigated by metabolic balance studies in man. N-acetyl-L-Methionine (AcMet), D-methionine (D-Met), and sodium sulfate (Na2SO4) were used to supplement a diet deficient in sulfur amino acids. The daily diet contained 4.5 g nitrogen (N) from isolated soybean protein (SB) and 4.5 g from glycine and alanine (9 g total N). SB diet was given alone or supplemented to six adult men for periods of 9 days after a standardization period with an equal N eggwhite diet, preceded by a 2-day zero N adaptation period. Supplements provided equivalent amounts of sulfur to that present in 420 ml L-Met, the amount added to SB to bring the total sulfur amino acid content to 900 mg/day. AcMet was as benfeficial as L-Met in improving N balance but D-Met was not as effective as L-Met. Difference between balances obtained with L-Met and Na2SO4 was not significant due to large variation in response to Na2SO4. While addition of D-Met to SB did not result in significantly greater N retention than unsupplemented SB, NA2SO4 addition did cause increased N retention.     

Growth effects of regulatory peptides and intracellular signaling routes in human pancreatic cancer cell lines

Spore germination is a defined developmental process that marks a critical point in the life cycle of Dictyostelium discoideum. Upon germination the environmental conditions must be conducive to cell growth to ensure survival of emerged amoebae. However, the signal transduction pathways controlling the various aspects of spore germination in large part remain to be elucidated. We have used degenerate PCR to identify dhkB, a two-component histidine kinase, from D. discoideum. DhkB is predicted to be a transmembrane hybrid sensor kinase. The dhkB-null cells develop with normal timing to give what seem to be mature fruiting bodies by 22 to 24 h. However, over the next several hours, the ellipsoidal and encapsulated spores proceed to swell and germinate in situ within the sorus and thus do not respond to the normal inhibitors of germination present within the sorus. The emerged amoebae dehydrate due to the high osmolarity within the sorus, and by 72 h 4% or less of the amoebae remain as spores, while most cells are now nonviable. Precocious germination is suppressed by ectopic activation of or expression of cAMP-dependent protein kinase A. Additionally, at 24 h the intracellular concentration of cAMP of dhkB- spores is 40% that of dhkB+ spores. The results indicate that DHKB regulates spore germination, and a functional DHKB sensor kinase is required for the maintenance of spore dormancy. DHKB probably acts by maintaining an active PKA that in turn is inhibitory to germination.     

Presence of bifidobacteria in the rumen of calves fed different rations

The silent period in the agonist prior to rapid voluntary movement (premotion silent period) was observed in the lower limb muscles. The subject was asked to respond to the flashing light by performing a vertical jump as quickly as possible. The most consistent results were obtained in the extensors, particularly the quadriceps. The frequency of the silence of six subjects was 53% in the rectus femoris, 61% in the vastus medialis, and 72% in the vastus lateralis. A neural switching mechanism, directing excitation from preparation phase of jumping into rapid contraction phase, seemed to be attributable to the occurrence of the premotion silent period.     

Supplementation of milk replacers containing soy protein with threonine, methionine, and lysine in the diets of calves

An attempt was made to improve the protein quality supplied in milk replacers containing soy protein by supplementing Thr, Met, and Lys to the milk replacers fed to calves. Six Holstein x indigenous male calves were fitted with single cannulas at the end of the ileum. Calves were fed milk replacers containing skim milk (86%) and whey (14%) proteins or skim (43%), whey (14%), and soy (43%) proteins either with or without amino acid (AA) supplementation according to a double 3 x 3 Latin square design. Average daily gain, N retention, and ileal digestibilities of dry matter, N, and AA were significantly higher for calves fed the milk replacer containing skim milk protein than for calves fed the milk replacer containing soy protein. Average daily gain, N retention, and ileal digestibilities of dry matter, N, and AA were significantly higher for calves fed the milk replacer containing soy protein plus AA supplementation than for calves fed the milk replacer containing the soy protein without AA supplementation. Therefore, supplementation of a milk replacer containing the soy protein without AA supplementation. Therefore, supplementation of a milk replacer containing soy protein with limiting AA that correspond to the AA found in milk protein can considerably improve the protein quality of that milk replacer for the preruminant calf.     

Brefeldin A inhibits the constitutive-like secretion of a sulfated protein in pancreatic acinar cells

Sulfation is a common posttranslational modification of secretory proteins and serves as a valuable marker of constitutive and regulated secretory pathways. We investigated the cellular localization and the secretory behavior of sulfated macromolecules in the mouse pancreatic acinar cell. The major sulfated proteins of the cell were present in isolated zymogen granules, as determined by metabolic labeling with [35S]sulfate and subcellular fractionation. The sulfated proteins fell into three groups: gp300 is not secreted and is a component of the zymogen granule membrane; pancreatic lipase (56 kDa) and a 40 kDa protein are soluble and exhibit regulated secretion kinetics; and p82 is initially granule membrane associated, but is released from the cell with constitutive-like kinetics as a 75 kDa protein (p75). Secretion of p75 could be stimulated for up to 4 h after pulse labeling, presumably from immature secretory granules, but not after 6 h of chase. Treatment of cells with brefeldin A (BFA) at the start of the [35S]sulfate pulse resulted in almost total inhibition of sulfation. Addition of BFA during the chase (0-2 h) allowed normal basal and stimulated secretion of regulated secretory proteins, but reversibly inhibited the constitutive-like secretion of p75. In this case, the behavior of p75 was maintained as that of a regulated secretory protein for up to 6 h of chase. In untreated cells, immunofluorescence of p82/p75 was along the acinar lumen, and in small punctate structures in the apical cytoplasm. In BFA-treated cells, immunolabeling of p82/p75 was lost from the acinar lumen, and cytoplasmic labeling was finer and appeared to be associated with the secretory granule membranes. These data suggest a role for brefeldin A-sensitive coat formation in maturation of secretory granules after they bud from the TGN.     

Effects of consumption of oat milk, soya milk, or cow''s milk on plasma lipids and antioxidative capacity in healthy subjects

The effect of hydration and benzene adsorption on 23Na resonance and the quadrupolar interaction in NaY zeolites is studied by triple-quantum MAS 23Na NMR spectroscopy. In the case of a C6D6/NaY system, the results show that with an increase in benzene loading, there is an up-field trend in isotropic chemical shift (delta CS) and a decreasing second order quadrupolar effect (chi s) for the site II sodium ions. It was found that adsorbed benzene molecules have a slight effect on the environment of sodium ions on site I. All the sodium sites in NaY are influenced upon hydration. The up-field shift of the sodium delta CS reflects the effect of coordination of oxygen atoms on sodium cations due to hydration. The magnitude of chi s for hydrated sodium sites increases and then falls off with water loading. The increase in chi s is due to the initial hydration among SI-, SI'- and SII-sodium ions, while the decrease is the result of approaching the final stage of saturated hydration.     

The effect of VIP/PACAP family of peptides on pancreatic blood flow and secretion in conscious dogs

Coherent oscillatory activity of a population of neurons is thought to be a vital feature of temporal coding in the brain. We focus on the question of whether a single neuron can transform a spike code into a rate code. More precisely, how does a neuron vary its mean output firing rate, if its input changes from random to coherent? We investigate the coincidence detection properties of an integrate-and-fire neuron in dependence upon internal parameters and input statistics. In particular, we show how coincidence detection depends on the membrane time constant and the threshold. Furthermore, we demonstrate that there is an optimal threshold for coincidence detection and that there is a broad range of near-optimal threshold values. Fine-tuning is not necessary.