Association of polymorphisms in solute carrier family 27, isoform A6 (SLC27A6) and fatty acid-binding protein-3 and fatty acid-binding protein-4 (FABP3 and FABP4) with fatty acid composition of bovine milk.

The main goal of this study was to develop tools for genetic selection of animals producing milk with a lower concentration of saturated fatty acids (SFA) and a higher concentration of unsaturated fatty acids (UFA). The reasons for changing milk fatty acid (FA) composition were to improve milk technological properties, such as for production of more spreadable butter, and milk nutritional value with respect to the potentially adverse effects of SFA on human health. We hypothesized that genetic polymorphisms in solute carrier family 27, isoform A6 (SLC27A6) fatty acid transport protein gene and fatty acid binding protein (FABP)-3 and FABP-4 (FABP3 and FABP4) would affect the selectivity of FA uptake into, and FA redistribution inside, mammary epithelial cells, resulting in altered FA composition of bovine milk. The objectives of our study were to discover genetic polymorphisms in SLC27A6, FABP3, and FABP4, and to test those polymorphisms for associations with milk FA composition. The results showed that after pairwise comparisons between SLC27A6 haplotypes for significantly associated traits, haplotype H3 was significantly associated with 1.37 weight percentage (wt%) lower SFA concentration, 0.091 lower SFA:UFA ratio, and 0.17 wt% lower lauric acid (12:0) concentration, but 1.37 wt% higher UFA and 1.24 wt% higher monounsaturated fatty acid (MUFA) concentrations compared with haplotype H1 during the first 3 mo of lactation. Pairwise comparisons between FABP4 haplotypes for significantly associated traits showed that haplotype H3 was significantly associated with 1.04 wt% lower SFA concentration, 0.079 lower SFA:UFA ratio, 0.15 wt% lower lauric acid (12:0), and 0.27 wt% lower myristic acid (14:0) concentrations, but 1.04 wt% higher UFA and 0.91 wt% higher MUFA concentrations compared with haplotype H1 during the first 3 mo of lactation. Percentages of genetic variance explained by H3 versus H1 haplotype substitutions for SLC27A6 and FABP4 ranged from 2.50 to 4.86% and from 4.91 to 7.22%, respectively. Tag single nucleotide polymorphisms were identified to distinguish haplotypes H3 of SLC27A6 and FABP4 from others encompassing each gene. We found no significant associations between FABP3 haplotypes and milk FA composition. In conclusion, polymorphisms in FABP4 and SLC27A6 can be used to select for cattle producing milk with lower concentrations of SFA and higher concentrations of UFA.

Sterol regulatory element binding transcription factor 1 (SREBF1) polymorphism and milk fatty acid composition.

Milk is known to contain high concentrations of saturated fatty acids-such as palmitic (16:0), myristic (14:0), and lauric (12:0) acids-that can raise plasma cholesterol in humans, making their presence in milk undesirable. The main objective of our candidate gene study was to develop genetic markers that can be used to improve the healthfulness of bovine milk. The sterol regulatory element binding transcription factor 1 (SREBF1) known to regulate the transcription of lipogenic genes together with SREBF chaperone and insulin induced gene 1 were the candidate genes. The results showed significant association of the overall SREBF1 haplotypes with milk production and variations in lauric (12:0) and myristic (14:0) acid concentrations in milk. Haplotype H1 of SREBF1 was the most desirable to improve milk healthfulness because it was significantly associated with lower lauric (12:0) and myristic (14:0) acid concentrations compared with haplotype H3 of SREBF1, and lower lauric acid (12:0) concentration compared with haplotype H2 of SREBF1. Haplotype H1 of SREBF1, however, was significantly associated with lower milk production compared with haplotype H3 of SREBF1. We did not detect any significant associations between genetic polymorphisms in insulin induced gene 1 (INSIG1) and SREBF chaperone and milk fatty acid composition. In conclusion, genetic polymorphisms in SREBF1 can be used to develop genetic tools for the selection of animals producing milk with healthier fatty acid composition.

Identification of genetic markers for productive life in commercial sows.

Escalating replacement rates and production costs warrant attention on sow productive life (SPL). Increasing average SPL by one-tenth of 1 parity would result in an annual revenue increase of over $15 million in the United States. Research in model organisms has revealed conserved genes and gene pathways that lead to longer lifespan. The most prominent gene pathways are those involved in growth, most notably genes in the IGF pathway that serve to mimic the response of caloric restriction. The objective of this research was to test the hypothesis that these well conserved genes and gene pathways could also play a role in SPL, even though the productive life of sows is both a measure of longevity and their reproductive performance. Preliminary research on 3 distinct populations of over 2,000 animals suggested that several genes were associated with components of SPL. Genetic markers were then analyzed against the corresponding records of the sows for reproductive and longevity traits using a validation population of 2,000 commercial females. Right censored data were used to test associations of genetic markers with survival to defined time points. Three distinct models of survival analysis were implemented using nonparametric estimates of the survival distribution in a sequential order, using a parametric accelerated failure time model with a Weibull distribution of the error term, and a Cox proportional hazards model, which is a semiparametric model that uses an unspecified baseline hazard function. The genetic markers CCR7 and CPT1A were significantly associated (P < 0.05) with survival using the nonparametric model and tended (P < 0.1) toward significance using the parametric and semiparametric models with significantly different effects (P < 0.05) between some genotype classes. Genetic markers for MBL2, IGFBP3, and WARS2 also tended (P < 0.1) toward significance for survival traits, but were not consistent. Mixed model analyses were used to determine the associations of these genetic markers with reproductive traits. The genetic markers for IGFBP1, MBL2, CPT1A, CCR7, SLC22A5, and ACE were significantly (P < 0.05) associated with at least 1 reproductive trait. These results show that molecular markers should be considered for use in marker-assisted selection to improve SPL.

Acute metabolic responses of postpartal dairy cows to subcutaneous glucagon injections, oral glycerol, or both.

This study examined the effects of multiple subcutaneous glucagon injections with or without co-administration of oral glycerol on energy status-related blood metabolites and hormones of Holstein dairy cows in the first 2 wk postpartum. Twenty multiparous cows were fed a dry cow ration supplemented with 6 kg of cracked corn during the dry period to increase the likelihood of developing postpartal fatty liver syndrome. Cows with a body condition score of >or=3.5 points (1- to 5-point scale) were assigned randomly to 1 of 4 treatment groups: saline, glucagon, glycerol, or glucagon plus glycerol. Following treatment, serial blood samples were collected over an 8-h period to determine the effects of glucagon and glycerol on blood metabolites and hormones. Treatment effects were determined by comparing the concentrations of metabolites and hormones during the first 4-h period and the entire 8-h period after treatment administration (time 0) with the concentration of the same compounds at time 0 on d 1, 7, and 13 postpartum. Administration of glucagon alone increased concentrations of plasma glucagon and insulin on d 1, 7, and 13 and increased plasma glucose and decreased plasma nonesterified fatty acids (NEFA) on d 7 and 13 postpartum relative to the saline group. Administration of glycerol alone increased plasma glucose on d 7 and plasma triacylglycerols on d 1 postpartum. Glycerol administration also decreased plasma glucagon and NEFA on d 1, 7, and 13 and plasma beta-hydroxybutyrate (BHBA) on d 1 postpartum relative to the saline group. Administration of glucagon plus glycerol increased and sustained concentrations of plasma glucagon, glucose, and insulin on d 1, 7, and 13 and decreased plasma NEFA on d 1, 7, and 13 and BHBA on d 1 and 7. Early postpartal treatment of dairy cows with glucagon plus glycerol increased plasma glucose and insulin, decreased plasma NEFA and BHBA, and increased secretion of liver NEFA as plasma triacylglycerols. This suggests that glucagon and glycerol, when co-administered, act to decrease the likelihood of metabolism-related syndrome development in dairy cows.

Spatial Heterogeneity of Leaf Wetness Duration in Apple Trees and Its Influence on Performance of a Warning System for Sooty Blotch and Flyspeck.

To determine the effect of sensor placement on the performance of a disease-warning system for sooty blotch and flyspeck (SBFS), we measured leaf wetness duration (LWD) at 12 canopy positions in apple trees, then simulated operation of the disease-warning system using LWD measurements from different parts of the canopy. LWD sensors were placed in four trees within one Iowa orchard during two growing seasons, and in one tree in each of four orchards during a single growing season. The LWD measurements revealed substantial heterogeneity among sensor locations. In all data sets, the upper, eastern portion of the canopy had the longest mean daily LWD, and was the first site to form dew and the last to dry. The lower, western portion of the canopy averaged about 3 h less LWD per day than the top of the canopy, and was the last zone where dew formed and the first to dry off. On about 25% of nights when dew occurred in the top of the canopy, no dew formed in the lower, western canopy. Intracanopy variability of LWD was more pronounced when dew was the sole source of wetness than on days when rainfall occurred. Daily LWD in the upper, eastern portion of the canopy was slightly less than reference measurements made at a 0.7-m height over turfgrass located near the orchard. When LWD measurements from several canopy positions were input to the SBFS warning system, timing of occurrence of a fungicide-spray threshold varied by as much as 30 days among canopy positions. Under Iowa conditions, placement of an LWD sensor at an unobstructed site over turfgrass was a fairly accurate surrogate for the wettest part of the canopy. Therefore, such an extra-canopy LWD sensor might be substituted for a within-canopy sensor to enhance operational reliability of the SBFS warning system.

Acid-base balance of the domestic turkey during thermal panting.

The objectives of this research were to evaluate the effects of thermal panting in domestic turkeys on arterial blood values for the acid-base variables, pH(a), bicarbonate concentration ([HCO(-) (3)](a)), partial pressure of carbon dioxide (P(a)CO(2)), and hemoglobin concentration [Hb]. In addition, body temperature and partial pressure of oxygen (P(a)O(2)) were measured to determine the effectiveness of panting in their control. Nine adult (23 wk) broad-breasted white turkey toms, all from the same hatch and reared contemporaneously in the same facility, were acclimated to room conditions of 19 degrees C and 65% RH. After a 1-wk control period, a 3-wk heat-stress period (32 degrees C, 65% RH) was induced, for a heat-stress group of 9 turkeys. Thermal panting began at this time and continued to its end. A 1-wk recovery period followed (19 degrees C, 65% RH) during which panting ceased. An age-matched group of 8 turkeys was similarly acclimated (19 degrees C, 65% RH) but was continued at this level to the end of the experiment. During the heat-stress period, the bicarbonate concentration increased, whereas pH(a) and P(a)CO(2) did not change significantly. Body temperature changes were not significant. Parabronchial ventilation was not compromised by panting, as noted by a significant increase in P(a)O(2). Hemoglobin concentration decreases were significant. The only significant change that occurred for the age-matched group was an increase in [Hb]. Domestic turkeys, reared in confinement, have the ability to resist changes in blood pH and prevent the development of respiratory alkalosis while panting in response to thermal stress. Normal body temperature and oxygenation of the blood are also maintained.

Effects of storage time and temperature after blood sampling from turkeys on plasma concentrations of potassium, sodium, and chloride.

The objectives of this research were to study the effect of several time intervals of turkey blood storage from sampling to centrifugation on plasma K, Na, and Cl concentrations and to study the effect of ambient temperature of turkey blood storage on these same variables. In the first study, 6 consecutive blood samples were obtained from each turkey. The first sample was centrifuged immediately (control), and each of the others was respectively stored for 20, 40, 60, 120, or 360 min at 22 degrees C before centrifugation and plasma harvest. In the second study, 4 consecutive blood samples were obtained from each turkey. The first sample was centrifuged immediately (control), and each of the others was respectively stored at ambient temperatures of 9, 22, and 30 degrees C for 120 min before centrifugation and plasma harvest. Plasma K concentration declines and is significant (P < 0.01) for each of the 20, 40, 60, 120, and 360 min values. Plasma Na concentration increases and Cl decreases were not significant (P < 0.01) until 360 and 120 min, respectively. Significant (P < 0.01) reductions occurred for plasma K concentration for each of the ambient temperature values but were the least at 9 degrees C and greatest when stored at 30 degrees C. Plasma Na concentration decreased (P < 0.01) when stored at 9 degrees C. Though increased (P < 0.01) when stored at 22 and 30 degrees C, the difference was not significant (P < 0.01) between samples stored at either 22 or 30 degrees C. Plasma Cl concentration was decreased (P < 0.01) from control values, but there was no significant difference (P < 0.01) among samples stored at 9, 22, and 30 degrees C. Clot formation and its retraction from serum are unduly delayed after blood is withdrawn from turkeys because birds lack the intrinsic mechanism for blood coagulation. Accordingly, serum is not appropriate for the determination of K concentration in turkeys, and plasma harvested immediately after blood sampling is the fluid of choice for K analysis.

Prevention of fatty liver in transition dairy cows by subcutaneous injections of glucagon.

The main objective of this study was to test the extent to which injecting glucagon subcutaneously for 14 d beginning at d 2 postpartum would prevent fatty liver development in transition dairy cows. Twenty-four multiparous Holstein cows were fed 6 kg of cracked corn in addition to their standard diet during the last 30 d of a dry period to induce postpartum development of fatty liver. Glucagon at either 7.5 or 15 mg/d or saline (control) was injected subcutaneously 3 times daily for 14 d beginning at d 2 postpartum. Glucagon at 15 mg/ d prevented liver triacylglycerol accumulation in postpartum dairy cows. Glucagon at 7.5 mg/d showed potential for fatty liver prevention. Glucagon increased concentration of plasma glucose and insulin and decreased plasma nonesterified fatty acid concentrations. No effects of glucagon were detected on plasma beta-hydroxybutyrate concentrations. Glucagon affected neither feed intake nor milk production. Moreover, milk composition was not altered by glucagon. Milk urea N concentrations decreased, and plasma urea N concentrations tended to decrease during glucagon administration, indicating that glucagon may improve protein use. Liver glycogen concentrations were not affected by glucagon. No significant differences in body condition scores were detected among treatments throughout the study. These results indicate that subcutaneous glucagon injections can prevent fatty liver in transition dairy cows without causing major production and metabolite disturbances.

Microsatellite markers associated with resistance to Marek's disease in commercial layer chickens.

The objective of the current study was to identify QTL conferring resistance to Marek's disease (MD) in commercial layer chickens. To generate the resource population, 2 partially inbred lines that differed in MD-caused mortality were intermated to produce 5 backcross families. Vaccinated chicks were challenged with very virulent plus (vv+) MD virus strain 648A at 6 d and monitored for MD symptoms. A recent field isolate of the MD virus was used because the lines were resistant to commonly used older laboratory strains. Selective genotyping was employed using 81 microsatellites selected based on prior results with selective DNA pooling. Linear regression and Cox proportional hazard models were used to detect associations between marker genotypes and survival. Significance thresholds were validated by simulation. Seven and 6 markers were significant based on proportion of false positive and false discovery rate thresholds less than 0.2, respectively. Seventeen markers were associated with MD survival considering a comparison-wise error rate of 0.10, which is about twice the number expected by chance, indicating that at least some of the associations represent true effects. Thus, the present study shows that loci affecting MD resistance can be mapped in commercial layer lines. More comprehensive studies are under way to confirm and extend these results.

Genetic correlations between lean growth and litter traits in U.S. Yorkshire, Duroc, Hampshire, and Landrace pigs.

The objective of this study was to estimate breed-specific genetic correlations between lean growth and litter traits for four U.S. swine breeds. Records for lean growth and litter traits on Yorkshire, Duroc, Hampshire, and Landrace pigs collected between 1990 and April 2000 in herds on the National Swine Registry Swine Testing and Genetic Evaluation System were analyzed. A bivariate animal model and restricted maximum likelihood procedures were used to estimate genetic and environmental correlations between lean growth rate, days to 113.5 kg, backfat, and loin muscle area with litter traits of number born alive, litter weight at 21 d, and number weaned. Most genetic correlation estimates between lean growth and litter traits were small in magnitude and consistent across breeds. Backfat had the largest within-breed genetic correlations with number born alive (0.18 to 0.20) and litter weight at 21 d (-0.27 to -0.30). Estimates of genetic correlations between lean growth traits and number weaned were very small. Estimates of the environmental correlations between lean growth and litter traits also were very small for all traits and for all four breeds. Results indicate that selection for lean growth traits could have a long-term effect on litter traits. Including lean growth traits in a maternal-line evaluation using a multiple-trait model could increase the accuracy of the genetic evaluation for litter traits.

Genetic parameters and trends for litter traits in U.S. Yorkshire, Duroc, Hampshire, and Landrace pigs.

Records on 251,296 Yorkshire, 75,262 Duroc, 83,338 Hampshire, and 53,234 Landrace litters born between 1984 and April of 1999 in herds on the National Swine Registry Swine Testing and Genetic Evaluation System were analyzed. Animal model and restricted maximum likelihood procedures were used to estimate variances of animal genetic (a), maternal genetic (m), permanent environmental, and service sire, and the covariances between a and m for number born alive (NBA), litter weight at 21 d (L21WT), and number weaned (NW). Fixed effects of contemporary groups were included in the analysis. Based on a single-trait model, estimates of heritabilities were 0.10, 0.09, 0.08, and 0.08 for NBA; 0.08, 0.07, 0.08, and 0.09 for L21WT; and 0.05, 0.07, 0.05, and 0.05 for NW in the Yorkshire, Duroc, Hampshire, and Landrace breeds, respectively. Estimates of maternal genetic effects were low and ranged from 0.00 to 0.02 for all traits and all breeds. Estimates of permanent environmental effects ranged from 0.03 to 0.08. Estimates of service sire effects ranged from 0.02 to 0.05. A bivariate analysis was used to estimate the genetic correlations among traits. Average genetic correlations over the four breeds were 0.13, 0.15, and 0.71 for NBA with L21WT, NBA with NW, and L21WT with NW, respectively. Average genetic trends were 0.018 pigs/yr, 0.114 kg/yr, and 0.004 pigs/yr for NBA, L21WT, and NW, respectively. Although estimates of heritabilities for litter traits were low and similar across breeds, genetic variances for litter traits were sufficiently large to indicate that litter traits could be improved through selection. This study presents the first set of breed-specific estimates of genetic parameters available from large numbers of field records. It provides information for use in national genetic evaluations.

Genetic parameters and trends for lean growth rate and its comvonents in U.S. Yorkshire, Duroc, Hampshire, and Landrace pigs.

Records on 361,300 Yorkshire, 154,833 Duroc, 99,311 Hampshire, and 71,097 Landrace pigs collected between 1985 and April of 2000 in herds on the National Swine Registry Swine Testing and Genetic Evaluation System were analyzed. Animal model and REML procedures were used to estimate random effects of animal genetic, common litter, maternal genetic, and the covariances between animal and maternal for lean growth rate (LGR), days to 113.5 kg (DAYS), backfat adjusted to 113.5 kg (BF), and loin eye area adjusted to 113.5 kg (LEA). Fixed effects of contemporary group and sex were also in the statistical model. Based on the single-trait model, estimates of heritabilities were 0.44, 0.44, 0.46, and 0.39 for LGR; 0.35, 0.40, 0.44, and 0.40 for DAYS; 0.48, 0.48, 0.49, and 0.48 for BF; and 0.33, 0.32, 0.35, and 0.31 for LEA in the Yorkshire, Duroc, Hampshire, and Landrace breeds, respectively. Estimates of maternal genetic effects were low and ranged from 0.01 to 0.05 for all traits across breeds. Estimates of common litter effects ranged from 0.07 to 0.16. A bivariate analysis was used to estimate the genetic correlations between lean growth traits. Average genetic correlations over four breeds were -0.83, -0.37, 0.44, -0.07, 0.08, and -0.37 for LGR with DAYS, BF, and LEA, DAYS with BF and LEA, and BF with LEA, respectively. Average genetic trends were 2.35 g/yr, -0.40 d/yr, -0.39 mm/yr, and 0.37 cm2/yr for LGR, DAYS, BF, and LEA, respectively. Results indicate that selection based on LGR can improve leanness and growth rate simultaneously and can be a useful biological selection criterion.

Model to Enhance Site-Specific Estimation of Leaf Wetness Duration.

The ability of empirical models to enhance accuracy of site-specific estimates of leaf wetness duration (LWD) was assessed for 15 sites in Iowa, Nebraska, and Illinois during May to September of 1997, 1998, and 1999. Enhanced estimation of LWD was obtained by applying a 0.3-m height correction to SkyBit wind-speed estimates for input to the classification and regression tree/stepwise linear discriminant (CART/SLD) model (CART/SLD/Wind model), compared to either a proprietary model (SkyBit wetness) or to the CART/SLD model using wind speed estimates for a 10-m height. The CART/SLD/Wind model estimated LWD more accurately than the other models during dew-eligible (20:00 to 9:00) as well as dew-ineligible (10:00 to 19:00) periods, and for the period 20:00 to 9:00 regardless of rain events. Improvement of LWD estimation accuracy was ascribed to both the hierarchical structure of decision-making in the CART procedure and wind speed correction. Accuracy of the CART/SLD/Wind model identifying hours as wet or dry varied little among the 15 sites, suggesting that this model may be desirable for estimating LWD from site-specific data throughout the midwestern United States.

Phenotypic trends in incidence of stillbirth for Holsteins in the United States.

The objectives for this study were to determine 1) if there was a trend in stillbirths for the U.S. Holstein population, 2) if stillbirths are the same trait in primiparous and multiparous cows, and 3) what was the role of dystocia in stillbirths. A sample of 666,341 births from the MidStates Dairy Records Processing Center and the National Association of Animal Breeders was used to examine the influence of sire, herd, year, season, sex of calf, parity of dam, calving ease, and gestation length on the survival of the calf. Parity was scored as an ordered variable (1, 2, 3+). Calving ease was scored on a scale of 1 (no assistance) to 3+ (needed assistance). An increasing trend in stillbirths was found in primiparous and multiparous cows. The percentage of stillborn calves in primiparous cows increased from 9.5 in 1985 to 13.2 in 1996. Stillbirths in multiparous cows increased from 5.0 to 6.6% from 1985 to 1996. Variation about the trend was greater in primiparous cows than in multiparous cows. Dystocia was a major determinant of stillbirth incidence, but the association was stronger in primiparous cows. Sex of calf had different associations with stillbirth incidence in primiparous and multiparous cows. Gestation length and season of birth also had significant associations with stillbirth incidence. Logistic regression models with fixed and random effects were fit to the data to preserve the binary nature of the stillbirth response. The expected probability of stillbirths for an average herd and sire was 10% for primiparous cows and 5% for multiparous cows. Replacement of stillborn calves is a substantial cost to the dairy industry at more than $125.3 million per year. Because of the increasing incidence of stillbirths, these costs have increased by $75.9 million from 1985 to 1996.

Interactions among factors affecting stillbirths in Holstein cattle in the United States.

Each year about 7% of the Holstein calves born in the United States die within 48 h of birth. The exact cause of death is unknown. The purpose of this article is to examine the complex interactions among factors (e.g., parity, season of birth, dystocia, year) contributing to stillbirth rates. A modified chi-squared automated interaction detection algorithm was used to develop classification trees explaining the most likely sequence of factors that result in a stillborn calf. The data were 666,341 births from the MidStates Dairy Records Processing Center and the National Association of Animal Breeders. Primiparous and multiparous cows clearly differ in the rate of stillbirths, 11.0 and 5.7%, respectively. Dystocia followed parity as the next most important factor within both primiparous and multiparous cows. In primiparous cows, season, year of birth, or gestation length ranked third as an important predictor for dystocia equal to 1, 2, or 3+, respectively. Gestation length ranked third in importance among the factors that affect stillbirth rates for all levels of dystocia in multiparous cows. Among multiparous cows needing assistance (dystocia 3+), stillbirth rates were greatest for shorter gestations less than the average of 280 d, 55.3% for -15 to -12 d, 45.5% for -11 to -9 d, 33.7% for -8 to -5 d, 23.8% for -4 to 13 d, and 35.4% for 14 to 15 d. Gestation length pinpointed the time when stillbirths occurred, as indicated by the increase from 23.8% stillbirth rate among calves born at or above the mean gestation length to 55.3% for those calves born -15 to -12 d below the mean gestation. Further investigation of the relationship between stillbirth rates and gestation length is needed to develop a more complete understanding of the biological processes resulting in the loss of calves at birth.

Cell-cycle arrest at G2/M and growth inhibition by apigenin in human colon carcinoma cell lines.

Apigenin, a common dietary flavonoid, has been shown to induce cell cycle arrest in both epidermal and fibroblast cells and inhibit skin tumorigenesis in murine models. The present study assessed the influence of apigenin on cell growth and the cell cycle in the human colon carcinoma cell lines SW480, HT-29, and Caco-2. Treatment of each cell line with apigenin (0-80 microM) resulted in a dose-dependent reduction in both cell number and cellular protein content, compared with untreated control cultures. DNA flow cytometric analysis indicated that treatment with apigenin resulted in G2/M arrest in all three cell lines in a time- and dose-dependent manner. Apigenin treatment (80 microM) for 48 h produced maximum G2/M arrest of 64%, 42%, and 26% in SW480 cells, HT-29 cells, and Caco-2 cells, respectively, in comparison with control cells (15%). The proportion of S-phase cells was not altered by apigenin treatment in each of the three cell lines. The G2/M arrest was reversible after 48 h of apigenin treatment in the most sensitive cell line SW480. The degree of G2/M arrest by apigenin was inversely correlated with the corresponding inhibition of cell growth measurements in all three cell lines (r = -0.626 to -0.917, P

Modification of the Greenwood formula for correlated response times.

Life-table methodology for interval-censored survival times is used to estimate marginal survival probabilities from data consisting of independent cohorts of correlated responses. We restrict our attention to situations where response times within cohorts are exchangeable and the marginal survival distributions are the same for all individuals within any cohort. A modification of the Greenwood formula is derived for estimating variances and covariances of the estimates of the marginal survival probabilities. Simulation results show that this formula provides accurate adjustments for within-cohort correlations in finite samples. Comparisons with a nonparametric maximum likelihood estimator indicate negligible loss in efficiency from using simple life-table estimators of marginal survival probabilities that ignore the within-cohort correlations. Data from an angioplasty study where more than one procedure is performed on some patients are used to illustrate this methodology.

Bootstrapping regression parameters in multivariate survival analysis.

Bootstrap methods are proposed for estimating sampling distributions and associated statistics for regression parameters in multivariate survival data. We use an Independence Working Model (IWM) approach, fitting margins independently, to obtain consistent estimates of the parameters in the marginal models. Resampling procedures, however, are applied to an appropriate joint distribution to estimate covariance matrices, make bias corrections, and construct confidence intervals. The proposed methods allow for fixed or random explanatory variables, the latter case using extensions of existing resampling schemes (Loughin, 1995), and they permit the possibility of random censoring. An application is shown for the viral positivity time data previously analyzed by Wei, Lin, and Weissfeld (1989). A simulation study of small-sample properties shows that the proposed bootstrap procedures provide substantial improvements in variance estimation over the robust variance estimator commonly used with the IWM.

Immunological parameters of periparturient Holstein cattle: genetic variation.

The genetic variability of blood neutrophil functions, lymphocyte blastogenic responses to mitogens, serum Ig concentrations, and serum complement and conglutinin activities was investigated from 35 d prepartum to 35 d postpartum for 137 Holstein cows. Periparturient cows experience an immunosuppression of various immunologic parameters at calving. Heritability estimates were obtained before, during, and after the episodes of immunosuppression. Significant genetic variability occurred in the periparturient changes for total number of neutrophils, neutrophil chemokinesis, assays of the neutrophil respiratory burst associated with phagocytosis (cytochrome c reduction, chemiluminescence, and iodination), serum concentrations of IgG1, IgG2, and IgM, and serum homolytic complement activity. This variability implies that immune profiles could be used for the selection of cattle with improved innate immune response without adverse effects on milk productivity. These results should be considered tentative, however, because the number of observations included in the data were limited.

Factors affecting dystocia and early calf mortality in Angus cows and heifers.

Calving performance records (965,417) from purebred American Angus herds throughout the United States were used to study dystocia and early calf mortality during the period from 1972 to 1985. A sample of 53 (n = 83,467) herds was used to establish reasonable limits on the expected frequency of dystocia and mortality within and among herds that have good reproductive management programs and to verify the frequencies of scores in all other herds (n = 4,130) reporting calving performance information. The data were analyzed by logistic regression models. Dystocia and perinatal mortality, to a lesser extent, were found to be more frequent in heifers than in cows. The odds of unassisted births vs births with major difficulty were 11.58 times greater in cows than in heifers. In heifers, the odds of an unassisted birth increased with age of dam and decreased with birth weight of calf. At a baseline age of 22 mo the odds of an unassisted birth for calves weighing 30 and 40 kg were .13 and .02 times lower than the odds for calves weighing 20 kg. Alternately, at 29 mo, the odds of an unassisted birth for heifers producing a 20-kg calf were 4.53 times greater than at 22 mo. Survival of calves to 24 h in heifers was primarily affected by birth weight. Heifers producing calves at intermediate weights of 29 kg had higher odds of producing live calves at all ages. Considering all heifers calving at 22 to 29 mo relative to 20-kg calves, the odds for survival to 24 h were 3.83 times greater for 29-kg calves and lower by a factor of .52 for 40-kg calves.