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Biomedical subjects

G L Bennett

Publications and source records attributed to G L Bennett.

At least 55 records · Page 3Linked to original sources

Initial results of genomic scans for ovulation rate in a cattle population selected for increased twinning rate.

Genomic scans were conducted with 273 markers on 181 sires from a cattle population selected for increased twinning rate to identify chromosomal regions containing genes that influence ovulation rate. Criteria used for selecting markers were number of alleles, ease of scoring, and relative position within linkage group. Markers were multiplexed or multiple-loaded on the gels to reduce the costs and labor required to obtain genotypic data. This approach reduced the number of gels by 45% when compared with running each marker independently. Male animals selected for the genomic scan sired the majority of the population. A modified interval analysis was used in a granddaughter design to compare effects of each allele within sire for 10 different sire families. The midparent deviation of the son's estimated breeding value for ovulation rate was used as the phenotype. Forty-one potential peaks were identified with a nominal significance level < or = 0.05. The 10 peaks with the highest significance levels (P < 0.02) were selected for further analysis. Markers were genotyped across daughters of the sire where nominal significance was found for each of the 10 peaks. One peak (BTA5, relative position 40 cM) was found to be nominally significant in the daughters. The nominal significance levels were P = 0.01 for the sons (n = 32) and P = 0.02 for the daughters (n = 94) of sire 784403. A combined genomewide significance value (P = 0.07) was calculated that accounted for the 10 analyses with sons and the 10 analyses with daughters. These results strongly suggest that this region contains a gene(s) that is involved in the follicular recruitment and development process.

Animals↗

Effects of sire growth potential, growing-finishing strategy, and time on feed on performance, composition, and efficiency of steers.

Beef production systems that increase use of unharvested forages and use animals with greater potential for gain affect age and size of animals placed on a finishing regimen. This experiment was conducted to evaluate effects of genetic potential for gain, age at the start of a finishing period, and time on feed on composition, quantity, and quality of beef produced and efficiency of production during finishing. Crossbred cows were bred by AI to Charolais or Line 1 Hereford bulls that represented potentially high (HG) or moderate growth (MG) rates, respectively, to produce spring- or fall-born calves. Steer calves from these matings were placed on an individually fed finishing diet at three ages (A). Spring-born steers were started at 6 or 18 mo of age (A6 and A18), and fall-born steers were started at 12 mo of age (A12). Slaughter times (T) were at 0, 90, 180, and 270 d for A6; 68, 136, and 204 d for A12; and 0, 45, 90, and 135 d for A18. Data collected on each animal included feed intake, growth, chemical composition of the complete body and carcass, and quantitative and qualitative assessment of the meat produced. Four steers of each sire group were slaughtered in each of the 11 A-T treatment groups, and the experiment was repeated for 2 yr in the A12 groups and 3 yr in the A6 and A18 groups (n = 237). Steers sired by HG bulls were larger and produced larger carcasses and more carcass protein than MG-sired steers (S, P < .05 or .01). Steers sired by MG bulls were fatter, had higher quality grades, and accumulated fat at a faster rate than HG-sired steers, and this effect was greater in older steers (G and GA, P < .05 or .01). Sire growth potential did not affect gain, intake, live weight efficiency, tenderness, or taste panel scores (P > .2). Steers sired by HG bulls were more efficient at producing carcass weight and carcass protein at A12 and A18 than were MG-sired steers. At the end of the finishing period, older (A18), HG-sired steers were too large with insufficient fat by current industry standards, and younger (A6), MG-sired steers were too small. Our conclusions are that both HG- and MG-sired steers can produce acceptable carcasses for current market standards with comparable efficiencies of live-weight gain, but the growing and finishing strategy must be adapted to the genotype.

Animal Feed↗

GDNF is abundant in the adult rat gut.

Glial derived neurotrophic factor (GDNF) is essential for the development of the enteric nervous system (ENS). Although previous work has measured GDNF mRNA levels, little is known about the concentration of GDNF protein produced in developing or adult tissues. The aim of this study was to quantitate the concentration of GDNF protein in various tissues of the developing and adult rat and in adult human gut. A two site antibody immunoassay was used to quantitate GDNF using recombinant rat GDNF as a standard. In the adult rat gastrointestinal tract the intestine contained the highest concentration of GDNF while the stomach and esophagus have the lowest concentrations. The isolated muscular wall of the intestine has approximately four times the GDNF concentration of the intact intestine. Other tissues with smooth muscle such as the aorta and urinary bladder contain moderate GDNF concentrations. In contrast, GDNF is barely detectable in the adult kidney and liver. High concentrations of GDNF were also detected in human colon and jejunum. As development proceeds in the rat, there is a tendency for the concentration of GDNF to increase in the intestine but decrease in other tissues. Treatment of the jejunum with the cationic surfactant benzyldimethyltetradecylammonium chloride (BAC) results in an increase in the number of smooth muscle cells, a decrease in myenteric neurons, and an increase in the concentration of GDNF in homogenates of intestine. The observations that GDNF concentrations are high in the adult intestine suggest that this growth factor may be important for the maintenance of the adult ENS.

Adult↗

Anatomic investigation of the deep posterior compartment of the leg.

The purpose of this study is to critically investigate the anatomy of the deep posterior compartment of the leg. Specifically, the relationship of the deep posterior compartment to the superficial posterior compartment and their insertion onto the posteromedial border of the tibia are assessed. Cross-sectioning of 10 fresh-frozen cadaver legs was performed at 2-cm increments. The inferior surface of each section was photographed. The photographs were visually analyzed, and the fascial separation between the posterior compartments along with their relationship to the posteromedial border of the tibia were recorded for each specimen. Magnetic resonance images in the axial plane of 10 healthy, normal volunteers' lower extremities at 2-cm increments were obtained and analyzed. All specimens and images demonstrated that the medial fascial attachment of the deep posterior compartment was along the posteromedial aspect of the tibia in the proximal third of the leg and was not superficially accessible. In the proximal third of the leg, the superficial posterior compartment fascial attachment overlapped the deep posterior compartment by inserting medial and anterior to the deep posterior compartment fascial attachment. In the middle and distal thirds of the leg, the medial fascial attachment of the deep posterior compartment shifted medially and anteriorly, making the deep posterior compartment superficially accessible. The surgeon must appreciate the change in the anatomic relationships along the medial side of the leg while performing double-incision four-compartment fasciotomy release to obtain a complete release of the muscular portion of the deep posterior compartment.

Cadaver↗

A model of litter size distribution in cattle.

Genetic increases in twinning of cattle could result in increased frequency of triplet or higher-order births. There are no estimates of the incidence of triplets in populations with genetic levels of twinning over 40% because these populations either have not existed or have not been documented. A model of the distribution of litter size in cattle is proposed. Empirical estimates of ovulation rate distribution in sheep were combined with biological hypotheses about the fate of embryos in cattle. Two phases of embryo loss were hypothesized. The first phase is considered to be preimplantation. Losses in this phase occur independently (i.e., the loss of one embryo does not affect the loss of the remaining embryos). The second phase occurs after implantation. The loss of one embryo in this stage results in the loss of all embryos. Fewer than 5% triplet births are predicted when 50% of births are twins and triplets. Above 60% multiple births, increased triplets accounted for most of the increase in litter size. Predictions were compared with data from 5,142 calvings by 14 groups of heifers and cows with average litter sizes ranging from 1.14 to 1.36 calves. The predicted number of triplets was not significantly different (chi2 = 16.85, df = 14) from the observed number. The model also predicted differences in conception rates. A cow ovulating two ova was predicted to have the highest conception rate in a single breeding cycle. As mean ovulation rate increased, predicted conception to one breeding cycle increased. Conception to two or three breeding cycles decreased as mean ovulation increased because late-pregnancy failures increased. An alternative model of the fate of ova in cattle based on embryo and uterine competency predicts very similar proportions of singles, twins, and triplets but different conception rates. The proposed model of litter size distribution in cattle accurately predicts the proportion of triplets found in cattle with genetically high twinning rates. This model can be used in projecting efficiency changes resulting from genetically increasing the twinning rate in cattle.

Animals↗

The incidence of Escherichia coli on beef carcasses and its association with aerobic mesophilic plate count categories during the slaughter process.

An analysis of 535 prefabricated beef carcass samples taken in three processing plants demonstrated an association between the mesophilic aerobic plate count (APC) class and the incidence of obtaining an Escherichia coli-positive sample. Beef carcasses were sampled from three separate plants; one was a fed-beef processing plant and the other two were cow/bull plants. Samples were obtained by sponging and were analyzed for APC and E. coli. When samples were classified into four APC levels or classes (class 1: < 2, class 2: > or = 2 and < 3, class 3: > or = 3 and < 4, and class 4: > or = 4 log CFU/cm2), a trend indicating that samples from higher APC classes were more likely to be positive for E. coli biotype 1 was observed. Of the APC class 4 samples (> or = 4 log CFU/cm2), 88% were positive for the presence of E. coli, as opposed to 21% in APC class 1 (< 2 log CFU/cm2). Univariate chi-square analysis of the resulting contingency tables from reclassified data (class 1: < 2, class 2: > or = 2 and < 3, and class 3: > or = 3 log CFU/cm2) indicated a strong association between APC class and the incidence (presence or absence) of an E. coli-positive sample. Using multivariate analysis to account for influences of plant and within plant processing site, the data indicated a strong positive linear trend between the presence of E. coli and the APC class.

Abattoirs↗

A GPI-linked protein that interacts with Ret to form a candidate neurturin receptor.

Glial-cell-line-derived neurotrophic factor (GDNF) and neurturin (NTN) are two structurally related, potent survival factors for sympathetic, sensory and central nervous system neurons. GDNF mediates its actions through a multicomponent receptor system composed of a ligand-binding glycosyl-phosphatidylinositol (GPI)-linked protein (designated GDNFR-alpha) and the transmembrane protein tyrosine kinase Ret. In contrast, the mechanism by which the NTN signal is transmitted is not well understood. Here we describe the identification and tissue distribution of a GPI-linked protein (designated NTNR-alpha) that is structurally related to GDNFR-alpha. We further demonstrate that NTNR-alpha binds NTN (K[d] approximately 10 pM) but not GDNF with high affinity; that GDNFR-alpha binds to GDNF but not NTN with high affinity; and that cellular responses to NTN require the presence of NTNR-alpha. Finally, we show that NTN, in the presence of NTNR-alpha, induces tyrosine-phosphorylation of Ret, and that NTN, NTNR-alpha and Ret form a physical complex on the cell surface. These findings identify Ret and NTNR-alpha as signalling and ligand-binding components, respectively, of a receptor for NTN and define a novel family of receptors for neurotrophic and differentiation factors composed of a shared transmembrane protein tyrosine kinase and a ligand-specific GPI-linked protein.

Amino Acid Sequence↗

Human platelets as a model for the binding and degradation of thrombopoietin.

Recent studies have shown that plasma thrombopoietin (TPO) levels appear to be directly regulated by platelet mass and that removal of plasma TPO by platelets via binding to the c-Mpl receptor is involved in the clearance of TPO in rodents. To help elucidate the role of platelets in the clearance of TPO in humans, we studied the in vitro specific binding of recombinant human TPO (rhTPO) to human platelet-rich plasma (PRP), washed platelets (WP), and cloned c-Mpl. Using a four-parameter fit and/or Scatchard analysis, the approximate affinity of rhTPO for its receptor, which was calculated from multiple experiments using different PRP preparations, was between 128 and 846 pmol/L, with approximately 25 to 224 receptors per platelet. WP preparations gave an affinity of 260 to 540 pmol/L, with approximately 25 to 35 receptors per platelet, and erythropoietin failed to compete with 125I-rhTPO for binding to WP. Binding and dissociation studies conducted with a BiaCore apparatus yielded an affinity of 350 pmol/L for rhTPO binding to cloned c-Mpl receptors. The ability of PRP to bind and degrade 125I-rhTPO was both time- and temperature-dependent and was blocked by the addition of excess cold rhTPO. Analysis of platelet pellets by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that 125I-rhTPO was degraded into a major fragment of approximately 45 to 50 kD. When 125I-rhTPO was incubated with a platelet homogenate at pH = 7.4, a degradation pattern similar to intact platelets was observed. Together, these data show that human platelets specifically bind rhTPO with high affinity, internalize, and then degrade the rhTPO.

Adult↗

Tear hepatocyte growth factor (HGF) availability increases markedly after excimer laser surface ablation.

Tear cytokines and growth factors are likely to modulate the wound healing process following corneal epithelial injury. Hepatocyte growth factor (HGF) is a paracrine mediator of epithelial proliferation, motility, and differentiation that is produced by keratocytes and the lacrimal gland. Tear samples were collected preoperatively and one, two, and seven days postoperatively in eyes undergoing excimer laser surface ablation [photorefractive keratoplasty (PRK) or phototherapeutic keratoplasty (PTK)]. Tear HGF concentration was measured with a sensitive ELISA assay. Tear HGF production was calculated using the tear flow rate in the collection capillary and HGF concentration. Although the instantaneous concentration of HGF in tears decreased significantly in the days following PRK, a large increase in tear flow resulted in a marked increase in HGF bioavailability. The heparin-binding characteristics of HGF would result in increased binding to glycosaminoglycans and other heparin-like matrix components and, therefore, increased growth factor availability to the cognate recptor. This is the first report documenting changes in tear film HGF production. HGF may have an important function in maintenance and wound healing of the ocular surface epithelium since HGF is present in the normal tear film and the HGF secretion rate increases markedly in parallel with aqueous tear production following corneal surgical injury.

Enzyme-Linked Immunosorbent Assay↗

Cloning and characterization of a human leptin receptor using a biologically active leptin immunoadhesin.

Leptin, the product of the ob gene, is a hormone secreted by fat cells which is primarily involved in the regulation of body weight. We have generated a leptin immunoadhesin (leptin-IgG) which was more potent than leptin alone at reducing body weight and food intake when injected into ob/ob mice. This molecule was used to identify high affinity binding sites on human embryonic 293 kidney cells and subsequently to isolate a cDNA encoding the leptin receptor from this cell line by expression cloning. This receptor corresponds to the short form of the recently isolated leptin receptor. Analysis of the expression pattern of the two forms of receptor by Northern blot, in situ hybridization and quantitative PCR showed that the receptor is expressed in most tissues but that the long form is prevalent in the hypothalamus.

Animals↗

A simulation model including ovulation rate, potential embryonic viability, and uterine capacity to explain litter size in mice: I. Model development and implementation.

Litter size in mice was studied using a model including ovulation rate, potential embryonic viability, and uterine capacity. Simulated results were compared with experimental results from a selection experiment with mice. The four criteria of selection were selection on number born (LS), selection on an index of ovulation rate and ova success (IX), selection on number born to unilaterally ovariectomized females (UT), and unselected control (LC). Comparisons were made to statistics of the base generation and to responses after 13 generations of selection. Phenotypic and genetic statistics for uterine capacity were generated so that simulations produced the experimental means, standard deviations, and correlations between left and right litter size, as well as responses in number born using the LS, IX, and UT criteria. Statistics for the simulated data generally agreed with observed values. Simulated heritability in the base generation for uterine capacity was .065. Experimental and simulated responses per generation in litter size through 13 generations of selection were .15 and .16, .17 and .18, and .10 and .11 for LS, IX, and UT, respectively. Simulated responses in uterine capacity after 13 generations were 2.19, 1.60, and 3.40 for LS, IX, and UT, respectively. Simulated means for the base generation were 13.22 and 16.30 for ovulation rate and uterine capacity, respectively. Uterine capacity was an important component of the variability in litter size; however, ovulation rate was the more limiting component.

Algorithms↗

A simulation model including ovulation rate, potential embryonic viability, and uterine capacity to explain litter size in mice: II. Responses to alternative criteria of selection.

Direct selection for litter size was compared with selection for ovulation rate, ova success, or uterine capacity and for indexes of ovulation rate with ova success or uterine capacity. Selection was simulated for 10 generations in a mouse population based on a model integrating ovulation rate, potential embryonic viability, and uterine capacity. Two indexes including ovulation rate (OR) and ova success (OS) were I = .291 x OR + 2.19 x OS and I = .165 x OR + .736 x OS. Heritabilities for ovulation rate and ova success, assumed in the simulation and to derive the indexes, were .25 and .06, respectively. Both indexes resulted in the same response in litter size, 12.9% greater than response to direct selection for litter size. Two indexes including OR and uterine capacity (TUC = true total uterine capacity; UC = uterine capacity measured as number born for a female with right ovary excised) were I = .881 x OR + .223 x TUC and I = .876 x OR + .568 x UC. Heritabilities assumed for uterine capacity were .09 (TUC) and .065 (UC). The first index assumed true parameters for uterine capacity (TUC) and resulted in a response in litter size that was 23.9% greater than direct selection. The second index was calculated using parameters estimated under a unilateral-ovariectomy model and resulted in response that was 14.7% greater than direct selection. Selection for OR, TUC, UC, or OS resulted in responses that were 4.5, 48.5, 38.7, or 74.8%, respectively, less than that from direct selection for litter size.

Animals↗

Genetic and environmental parameters for ovulation rate, twinning rate, and weight traits in a cattle population selected for twinning.

A project was implemented in 1981 with the objective of increasing twinning rate in cattle. Daughters of foundation sires had twin calves at a frequency of from 8 to 13%, and foundation females had twin calves at an average frequency of 50%. Data were analyzed on twinning rate, ovulation rate, and weight traits. The h2 of ovulation rate increased from .11 to .38 for a single estrous cycle to the mean of eight estrous cycles. From all data, h2 for single observation of ovulation rate and twinning rate were .10 and .09, respectively. The r(g) between them was .75. The h2 of weight traits ranged from .42 to .54 when weight traits were analyzed pair-wise with ovulation rate and with twinning rate. The r(g) between weight traits with ovulation rate ranged from .15 to .30 and with twinning rate ranged from .24 to .39. Phenotypic mean twinning rate increased from 1.07 to 1.29 calves per parturition for females born 1981 through 1993, and adjusted mean predicted breeding value (PBV) increased from 1.07 to 1.33 calves per parturition. Phenotypic mean ovulation rate for fall of 1984 through fall 1994 birth groups increased from 1.11 to 1.26 per estrous cycle, and adjusted mean PBV for ovulation rate increased from 1.11 to 1.29 per estrous cycle. Because of the high r(g) (i.e., .75) between ovulation rate and twinning rate, and because of a h2 of .35 for ovulation rate for the mean of six estrous cycles, repeated records of ovulation rate in puberal heifers is an effective indirect selection criterion for twinning rate. The positive r(g) between growth traits and ovulation and twinning rate suggest the need for some compromise when the selection goal is increased twinning rate with no increase in growth and size.

Animals↗

Hepatocyte growth factor. A cytokine mediating endothelial migration in inflammatory arthritis.

OBJECTIVE: Angiogenesis is an integral component of the vasculoproliferative phase of rheumatoid arthritis (RA). Recently, a heparin-binding cytokine termed hepatocyte growth factor (HGF), or scatter factor (due to its ability to disperse cohesive epithelial colonies), was described. We conducted this study to investigate the hypothesis that this cytokine was present in the milieu of the inflamed joint, and that it contributed to the chemotaxis of endothelial cells in the synovial tissue. METHODS: We examined synovial fluid, synovial tissue, and peripheral blood from 91 patients with RA and other arthritides. We used 83 total samples in an enzyme-linked immunosorbent assay to quantitate the HGF in synovial fluids and peripheral blood. To determine whether the HGF was biologically active, an epithelial scatter factor assay was performed. Immunohistochemical analysis was used to determine localization in synovial tissues. To define a function for synovial HGF, we preincubated rheumatoid synovial fluids with neutralizing anti-HGF and measured the ability of these synovial fluids to induce endothelial chemotaxis. RESULTS: Synovial fluid from patients with RA contained a mean +/- SEM HGF concentration of 2.0 +/- 0.3 ng/ml, while synovial fluid from patients with other arthritides (including inflammatory arthritis) contained 2.4 +/- 0.7 ng/ml HGF. Osteoarthritis (OA) patient samples contained the smallest quantities of synovial fluid HGF at 0.9 +/- 0.1 ng/ml. RA synovial fluid contained significantly more HGF than did RA peripheral blood (1.1 +/- 0.2 ng/ml) (P < 0.05). Rheumatoid synovial fluids induced more scattering of cells than did OA synovial fluids, suggesting a role for this cytokine in rheumatoid joint destruction. Interleukin-1 beta induced expression of rheumatoid synovial tissue fibroblast antigenic HGF and scatter factor activity. Immunohistochemically, HGF, as well as the HGF receptor (the met gene product), localized to significantly more rheumatoid synovial tissue lining cells than normal lining cells (P < 0.05). Both HGF and its receptor immunolocalized to subsynovial macrophages as well. Levels of synovial tissue immunoreactive HGF correlated positively with the number of synovial tissue blood vessels. Anti-HGF neutralized a mean of 24% of the chemotactic activity for endothelial cells found in 10 rheumatoid synovial fluid samples. CONCLUSION: These results indicate that synovial HGF may contribute to the vasculoproliferative phase of inflammatory arthritides such as RA, by inducing HGF-mediated synovial neovascularization. These findings point to a newly described role for HGF in the fibroproliferative phase of RA-associated synovitis.

Arthritis, Rheumatoid↗