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Biomedical subjects

R J Collier

Publications and source records attributed to R J Collier.

At least 217 records · Page 12Linked to original sources
PubMed1985

Adipose tissue fatty acid metabolism during pregnancy in swine.

In vitro adipose tissue fatty acid pool size (POOL), fatty acid release (FAR) and esterification (EST) were measured in peritoneal (PFP) and subcutaneous mammary (MFP) fat pads of swine at d 15, 30, 45, 60, 75, 90, 105 and 112 of pregnancy. Plasma free fatty acids (FFA) and triglycerides (TG) were not altered by stage of pregnancy. Basal EST in PFP was generally constant across pregnancy with a peak at d 75. Basal EST in MFP was elevated at d 30, 75 and 112. Esterification in response to norepinephrine stimulus (NE) was lower than basal rates in both fat depots. Basal FAR was constant throughout pregnancy in PFP, but elevated at d 75 and 90 in MFP. Fatty acid release in response to NE was biphasic with peaks at d 30 and in late pregnancy (in MFP, micromolar FAR in response to NE was 69.3% greater on d 75 to 112 than on d 45 to 60). Basal POOL was constant throughout pregnancy in both depots and lower than NE-stimulated POOL. All responses to NE were greater in MFP than in PFP, indicating that adipose tissue surrounding the developing mammary gland had higher metabolic activity and a greater response to NE than peritoneal adipose. Changes in fatty acid metabolism during pregnancy in swine are temporally related to published values for plasma steroids, fetal growth and mammary development. Metabolic adaptations in adipose and mannary epithelial tissue occur in synchrony with changing plasma estrogen concentrations, redirecting energy flow from maternal adipose tissue toward developing mammary and fetal tissue.

Adipose Tissue↗
PubMed1985

Periparturient endocrine changes of conceptus and maternal units in Holstein heifers bearing genetically different conceptuses.

Holstein heifers (n = 21) were balanced across sires and assigned to three service-sire-breed groups in which heifers were inseminated artificially to either purebred Angus (n = 7), Holstein (n = 7) or Brahman (n = 7) bulls. Semen from four bulls was used for each service sire-breed group. Blood samples were collected from a jugular vein thrice weekly from d 160 to 265 of pregnancy, daily thereafter until 15 d postpartum, and then thrice weekly until d 60 postpartum. Concentrations of progesterone, estrone, estradiol, and estrone sulfate from 23 d prepartum to parturition, and of 15-keto-13,14-dihydro-prostaglandin F2 alpha (PGFM) from 2 d prepartum to d 15 postpartum were measured by radioimmunoassay. Heifers within the Brahman-service-sire group had longer gestations (P less than .05) than those of Holstein- or Angus-service-sire groups (285.0 vs 278.7), 279.0 d). Calf birth weight was lower (P less than .05) in Angus- than Holstein- and Brahman-service-sire groups (30.6 vs 36.1, 43.4 kg). Daily trends of prepartum maternal progesterone concentrations were approximately 1 ng/ml lower (P less than .01) in Angus- than Holstein- or Brahman-service-sire groups until luteolysis occurred. Heifers bearing crossbred Angus conceptuses had lower daily trends of prepartum estrogens concentrations (P less than .01), whereas heifers of the Holstein- and, even more dramatically, of the Brahman-service-sire groups had a higher magnitude and greater rise of plasma estrogens concentrations between d -10 and -1 prepartum (less than .01). Postpartum mean concentrations (P less than .05) and response curves of PGFM were lower (P less than .01) in the Angus- than in the Holstein- or Brahman-service-sire groups. Calf birth weights were correlated with least-squares means for maternal concentrations of prepartum estrone (r = .57), estradiol (r = .59) and estrone sulfate (r = .64) and postpartum maternal concentrations of PGFM (r = .56). Functional responses of the conceptus (e.g., estrogens) and maternal units (e.g., progesterone and PGFM) were influenced by conceptus genotype during the periparturient period.

Animals↗
PubMed1985

Carry-over effects of periparturient endocrine changes on postpartum reproductive function of Holstein heifers bred to genetically different service sires.

Effects of fetal sire on postpartum reproductive changes of the dam were studied in 21 Holstein heifers whose pregnancy had been initiated by either Angus (n = 7), Holstein (n = 7) or Brahman (n = 7) bulls. After parturition, all heifers were managed uniformly. Heifers in each service-sire-breed group were bled via jugular venipuncture thrice weekly from d 160 to 265 of pregnancy, daily thereafter until 15 d postpartum, and three times per week until d 60 postpartum. Ability of heifers to release prolactin (PRL) and luteinizing hormone (LH) was evaluated on d 10 postpartum after a simultaneous injection of thyrotropin releasing hormone (TRH; 100 micrograms) and gonadotropin releasing hormone (GnRH; 100 micrograms). Between d 5 and 60 postpartum, the reproductive tract of each heifer was examined rectally thrice weekly after collection of blood samples. Basal concentrations of LH from d 1 to 10 postpartum, as well as ability of the pituitary gland to release LH and PRL after the GnRH-TRH challenge, did not differ between service-sire-breed groups (P greater than .1). Means and profiles of progesterone concentrations during the first 60 d postpartum did not differ between service-sire-breed groups (P greater than .1). However, increases in progesterone concentrations following the GnRH-TRH challenge were synchronized more precisely in Angus (P less than .02) than in Holstein- and Brahman-service-sire groups. Daily rates of reduction in cervical and uterine horn diameters were higher (P less than .01) in Holstein- and Brahman- than in Angus-service-sire groups and were associated with higher profiles of postpartum 15-keto-13,14-dihydro-prostaglandin F2 alpha (PGFM) concentrations. Within-cow PGFM concentrations were correlated positively with cervical (r = .36) and uterine horn (r = .32) diameters. Postpartum ovarian responsiveness and uterine involution in Holstein heifers may be affected by genotypes of the conceptus they bore during pregnancy.

Animals↗
PubMed1985

Effects of climatic and management factors on conception rate of dairy cattle in subtropical environment.

Breeding records, representing 12,038 inseminations at Bassett's Dairy Farm (Monticello, FL), were analyzed to document effects of environmental and management factors on fertility of dairy cattle from January 1, 1975 to December 31, 1977. Conception rates of lactating cows decreased sharply when maximum air temperature on day after insemination exceeded 30 degrees C. In contrast, conception rates for heifers did not decline until 35 degrees C. Virgin heifers had higher conception rates for all services (50%) than lactating cows (34%) and suffered only slight depression of fertility during summer months. Heifers required 1.5 services per conception compared with 2.3 for lactating cows. Relationship between conception rate and rainfall on day after insemination was negative and curvilinear. Jerseys had higher conception rates (45%) than Holstein (39%) and Brown Swiss (41%). Services per conception were 1.7, 2.0, and 1.9. Substantial decreases of fertility were associated with advancing service number. Estrous status (standing; positive heat detection patch; mounting activity), inseminator, and year of service were related to variation of conception rate. Seasonal effects on fertility of lactating cows were marked. Thus, environmental management of the postpartum cow during hot summer months is warranted to maximize fertility.

Animal Husbandry↗
PubMed1985

Mammary blood flow and regulation of substrate supply for milk synthesis.

In ruminants, mammary supply of substrate varies with rate of mammary blood flow and concentrations of blood substrates. Blood concentrations of most mammary substrates, except acetate and tryptophan, do not vary greatly with feed intake, short term. Fasting reduces mammary blood flow, whereas milking and injection of growth hormone or thyroxine increase flow. It is proposed that the fraction of cardiac output that perfuses the udder of lactating ruminants plays a role in regulation of nutrient partitioning between milk and body tissues. In fed animals this fraction is 15 to 16% of cardiac output, which declines on fasting to 8 to 9% and increases slightly following growth hormone treatment to 17.6%. Following realimentation of fasted cows or goats, mammary blood flow takes several hours to return to normal. Investigation of the mechanism of this response, in terms of the ability of the animal to recognize its nutritional status and partition nutrients accordingly, should prove fruitful to understanding causes of variations of milk production in response to feed quantity and quality. Several substrates show increased mammary arteriovenous difference with increasing blood concentrations. This may reflect differing ratios of blood flow:milk yield. The steep gradient of concentration of substrates across the mammary epithelial cell membrane suggests that a major impediment to substrate supply for milk synthesis is the rate of substrate transport across the membrane.

3-Hydroxybutyric Acid↗
PubMed1985

Production and physiological responses of dairy cows to varying dietary potassium during heat stress.

Objectives were to study influences of heat stress and dietary potassium content on production and physiological responses of 8 Jersey and 10 Holstein cows blocked by breed and assigned randomly to no shade or shade environments. Each cow received a different dietary potassium treatment (.66, 1.08, and 1.64% of dry matter) in each of three 30-day periods. Rates of potassium loss from skin were almost five times greater for no shade as for shade cows during the hottest part of the day (1300 to 1500 h). Overall, cows with no shade ate 56% less during the daytime (0800 to 1600 h), 19% more during nighttime (1600 to 0800 h), and 13% less total feed than cows with shade. Interactions of environment and breed with dietary potassium treatment suggest differences in feed intake and milk yield responses to increasing dietary potassium content. Total daily feed intake and milk yield of cows with no shade responded in curvilinear fashion to increasing dietary potassium, whereas responses in shade were small. Largest responses in no shade were as dietary potassium increased from .66 to 1.08%. Milk yield of Holsteins increased with increasing dietary potassium, but yield of Jerseys did not. Combined effects of elevated potassium loss from skin and reduced potassium and dry matter intake during heat stress suggested that lactating dairy cows may benefit from supplemental potassium.

Analysis of Variance↗
PubMed1985

Interrelationships of milk yield, body weight, and reproductive performance.

Records (2263) from a single north Florida herd for 3 yr were evaluated in a series of analyses. Environment was subtropical. Data set included only cows that had normal milk records and became pregnant. Holsteins and Jerseys averaged 6799 and 4504 kg milk, 587 and 418 kg postpartum body weight, 164 and 141 days open, and 2.3 and 2.1 services per subsequent pregnancy. Pooled within breed, repeatabilities and heritabilities of these performance measures were .37, .15; .53, .25; .20, .06; and .30, 0. Estimable genetic correlations were milk yield and body weight -.09, milk yield and services per conception .38, and services per conception and body weight .37; range of standard errors was .22 to .39. Substantial genetic antagonism may exist between milk yield and body weight, and efficient reproduction. If results of this research are verified elsewhere, breeders should be aware that selection for increased weight could lead to decreased reproductive efficiency.

Analysis of Variance↗
PubMed1984

Interaction of diphtheria toxin with adenylyl-(3',5')-uridine 3'-monophosphate. I. Equilibrium and kinetic measurements.

Purified diphtheria toxin from various sources contains tightly, but noncovalently, bound nucleotides, the major component of which is adenylyl-(3',5')-uridine 3'-monophosphate (ApUp). We used ApUp radiolabeled with 32P to measure equilibrium dissociation constants (KD), and association and dissociation rate constants (k+1 and k-1, respectively) under various conditions. Diphtheria toxin bound 1 molar equivalent of ApUp, regardless of the temperature. Values of KD were 0.2 nM (25 degrees C) and 1.8 nM (37 degrees C) as determined by flow dialysis. No difference in KD was observed between the nicked and intact forms of toxin. The dissociation rate constant showed marked variation with temperature, ranging from 1.8 X 10(-4) s-1 at 5.5 degrees C (t 1/2 of the complex = 64 min) to 2.5 X 10(-2) s-1 at 25 degrees C (t 1/2 of the complex = 28 s). By contrast, k+1 varied only by a factor of 5 over the same temperature range (2.0 X 10(7) M-1 s-1 at 5.5 degrees C; 9.6 X 10(7) M-1 s-1 at 25 degrees C). The KD at 5.5 degrees C, calculated from the ratio of k-1/k+1, is 9 pM, which represents the strongest affinity of a dinucleotide for a protein ever reported. Affinity was maximal in the range of pH 6.5 to 7.1 and was sensitive to ionic strength. Thermodynamic parameters of the system were calculated.

Chemical Phenomena↗
PubMed1984

Interaction of diphtheria toxin with adenylyl-(3',5')-uridine 3'-monophosphate. II. The NAD-binding site and determinants of dinucleotide affinity.

Diphtheria toxin (DT) binds NAD with a KD of about 10 microM and adenylyl-(3',5')-uridine 3'-monophosphate (ApUp) with KD values ranging from 9 pM to 1.8 nM, depending on temperature (Collins, C. M., Barbieri, J. T., and Collier, R. J. (1984) J. Biol. Chem. 259, 15154-15158). Here we report experiments to explore relationships between ApUp binding and NAD binding to DT and to identify structural features of ApUp that determine its high affinity for DT. NAD, adenine, and nicotinamide competitively inhibited ApUp binding to DT, and we confirmed that ApUp blocked the binding and hydrolysis of NAD. Binding of P-site ligands to the toxin blocked interactions with ApUp. CRM197, a mutant form of DT defective in NAD binding and hydrolysis, bound ApUp 5,000-fold less tightly than did DT. These results are consistent with models in which the ApUp- and NAD-binding sites on DT overlap or are identical. Various mono-, di-, and oligonucleotides were studied as competitors of ApUp binding or the NAD-glycohydrolase reaction. The results imply that the high affinity of ApUp for DT depends on the presence of the 3'-terminal phosphate and a 3'-5' internucleoside linkage. There was strong specificity for adenine as the 5' base, but only weak specificity for uracil as the 3' base. Oligoribonucleotides containing additional nucleotides at either or both ends of ApUp sequences bound to the toxin 1-3 orders of magnitude less avidly than ApUp. Oligodeoxyribonucleotides containing dApdT sequences bound with still lower affinities. In contrast to the case with whole toxin, ApUp bound to fragment A less avidly than did NAD, and elimination of the 3'-terminal phosphate of ApUp resulted in increased affinity for the protein. These differences may reflect the absence in free fragment A of interactions with the cationic P-site, located on the toxin's B moiety.

Chemical Phenomena↗
PubMed1984

Homologous somatotropin radioreceptor assay utilizing recombinant bovine growth hormone.

A homologous radioreceptor assay using recombinant bovine growth hormone and bovine liver membranes is described. The total specific binding of 125I-labeled recombinant bovine growth hormone to the 100 000 X g pellet was 48% in 24 h at 25 degrees C. Hormone binding was partially reversible, with 40% of the radiolabeled hormone being irreversibly bound. The amount of specific binding varied with assay pH, with the optimum occurring at pH 7.8. Specific binding was temperature-dependent, with greater specific binding occurring at 25 degrees C than at 5 degrees C or 37 degrees C during a 24 h period. Recombinant bovine growth hormone, human growth hormone, ovine growth hormone and recombinant porcine growth hormone competed effectively with 125I-labeled recombinant bovine growth hormone for binding sites, while bovine prolactin and ovine prolactin were needed in amounts 10(6)-fold the concentration of recombinant bovine growth hormone to displace the radiolabeled hormone. Surprisingly, human placental lactogen did not displace the radiolabeled hormone.

Animals↗
PubMed1984

Immunotoxins.

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Amino Acid Sequence↗
PubMed1984

NAD binding site of diphtheria toxin: identification of a residue within the nicotinamide subsite by photochemical modification with NAD.

We showed earlier that exposing mixtures of NAD and diphtheria toxin fragment A to ultraviolet radiation (253.7 nm) induced the formation of covalently linked protein-ligand photoproducts. Here we report that when [carbonyl-14C]NAD was employed in such procedures, the efficiency of labeling of the protein approached 1 mol/mol, and at least 94% of the incorporated label was associated with a single residue, glutamic acid at position 148. Fragment A photolabeled in this manner was enzymically inactive. The efficiency of photolabeling was much lower (less than 0.2 mol/mol) when NAD radiolabeled in either the adenine moiety or the adenylate phosphate was used, and the label was attached to different site(s) within fragment A. Efficient photochemical transfer of label from [carbonyl-14C]NAD occurred under identical conditions with the nucleotide-free form of whole diphtheria toxin, CRM-45, or activated exotoxin A from Pseudomonas aeruginosa, but not with nucleotide-bound diphtheria toxin, CRM-197, native exotoxin A, or any of several NAD-linked dehydrogenases. On the basis of these and other results we suggest that part or all of the nicotinamide moiety of NAD is efficiently transferred to glutamate-148 of fragment A under the influence of ultraviolet irradiation and that this residue is located within the nicotinamide subsite. This location implies that glutamate-148 is at or near the catalytic center of the toxin. Our data provide direct evidence for the location of the NAD site in an ADP-ribosylating toxin and demonstrate highly efficient and specific photolabeling by [carbonyl-14C]NAD.

Binding Sites↗
PubMed1984

Genetic analysis of the functional relationship between colicin E3 and its immunity protein.

Partial deletions in the immunity gene of the colicin E3 operon were used to study possible functions of the immunity protein besides protection against exogenous colicin. Nuclease BAL-31 was used to create a series of carboxyl-terminal deletions of the immunity gene. Mutants displaying lowered immunity against exogenous colicin were found, and six that had reduced but detectable levels of immunity were chosen for further analysis. DNA sequence analysis of the deletions showed that all six terminated within the last five codons of the immunity gene. The wild-type immunity gene was replaced by each of the six mutated immunity genes in a plasmid containing an otherwise functional colicin E3 operon. Transformants containing the resulting plasmids produced smaller colonies on solid medium and grew more slowly in liquid culture than transformants carrying the wild-type colicin and immunity genes. This result suggested that immunity protein was required to protect the cell against endogenous colicin E3. This idea was confirmed in experiments in which the colicin E3 and immunity genes were independently cloned on two compatible plasmid vectors.

Amino Acid Sequence↗
PubMed1984

Effects of heat stress during pregnancy on postpartum reproductive changes in Holstein cows.

Effects of heat stress during the last third of gestation on reproductive changes postpartum were studied in Holstein cows. Cows and heifers 160 to 190 d of gestation were assigned in June 1978 to shade (n = seven cows and two heifers) or no shade (n = eight cows and two heifers) management treatments. After parturition, all cows (n = 19) were moved to the milking herd and managed uniformly. On the day of calving and on each Monday, Wednesday and Friday thereafter until d 50 postpartum, jugular blood samples were collected. Beginning approximately 7 d postpartum, the reproductive tract of each cow was examined rectally after collection of blood samples. Estrus was monitored twice daily and cows were inseminated after d 45 postpartum. Prepartum heat stress increased 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) concentrations postpartum and increased the rate of uterine involution. Regardless of prepartum treatment, progesterone concentrations indicated that luteal phases had begun by d 12.4 +/- 1.3 postpartum, which was about 3 d before PGFM was basal. The first luteal phase lasted only 10.7 +/- .9 d. First estrus was not detected until d 32.3 +/- 4.8 postpartum. The previously gravid uterine horn had a negative effect on ovarian volume, diameter of the largest follicle and percentage of ovaries with a corpus luteum. However, prepartum heat stress attenuated this effect. This study indicates that heat stress prepartum had residual effects on postpartum reproductive changes, and that the previously gravid uterine horn exerted some control, which was attenuated by heat stress, over ovarian recrudescense. Even though heat stress prepartum affected sensitive measures of postpartum reproductive function, it did not alter days to first estrus, days open (102.3 +/- 13.1) or services/conception (2.5 +/- .3).

Animals↗
PubMed1984

A review of endocrine regulation of metabolism during lactation.

Lactogenesis signals the shift from uterine nutrient transfer to the fetus to neonatal nourishment at the mammary gland. Metabolic adaptations involved in this process are under endocrine regulation. Key events include an increase in blood flow to mammary tissue, a decrease in nutrient utilization by peripheral tissues and an increase in nutrient utilization by mammary tissue for milk synthesis. Deficits of certain substrates during early lactation require mobilization of those substrates from depot stores. Changes in metabolism of various tissues are related to changes in hormone receptor populations of those tissues and hormone concentrations in blood. Hormone receptors are therefore the primary mechanism by which information from the endocrine systems is linked to cellular metabolism. Endocrine changes at parturition result in dramatic changes in receptor populations of key tissues such as adipose and mammary tissues. Knowledge in this area, however, is incomplete. Relationship between hormone receptors and specific cellular metabolic pathways remains unresolved.

Amino Acids↗
PubMed1984

Influence of dietary sodium and potassium bicarbonate and total potassium on heat-stressed lactating dairy cows.

Objectives were to study effects of heat stress, 0 or .85% sodium bicarbonate, 0 or 1.0% potassium bicarbonate, and 1.0 or 1.5% total dietary potassium on production and physiological responses of dairy cows. Eighteen lactating Holsteins were assigned to shade (control) or no shade (heat stress) lots continuously for three consecutive 35-day periods and to different dietary treatments each period. Basal diet was 25% cottonseed hulls and 75% concentrate. Daytime and nighttime feed intake and production were measured the last 2 wk of each period, and milk and blood were sampled the final day of each period. Black globe temperature, rectal temperature, respiration rate, and blood pH were higher in no shade. Daytime intake was 132% greater in shade, nighttime intake was not different between environments. Milk production was about 19% greater for evening and morning milkings in shade. Daytime intake, daytime and nighttime milk production were higher with sodium bicarbonate. Potassium bicarbonate reduced intake and production. Higher total dietary potassium increased total daily milk production. Lactating cows appear adept at withstanding environmental and dietary challenges to acid-base homeostasis. Supplementation of sodium bicarbonate and 1.5% dietary potassium, but not potassium bicarbonate, were beneficial to lactating dairy cows.

Animals↗