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Water-soluble terpolymers of acrylamide, acrylic acid and N -[(4-decyl)phenyl]acrylamide have been synthesized by a micellar polymerization technique. Neutralization of the acid groups after polymerization yields hydrophobically modified polyelectrolytes. The ratio of the surfactant to the hydrophobic comonomer (SMR) was varied in successive polymerizations to yield a series of terpolymers with varied amphiphilic microstructures. Although each terpolymer in the series was identical in overall composition and molecular dimensions, profound differences in associative behaviour were observed. Terpolymers synthesized at low SMR exhibit large enhancements in viscosity owing to hydrophobic associations in 0.5 M NaCl. Those synthesized at high SMR behave in a similar fashion to a control acrylamide/acrylic acid copolymer. While viscometric studies indicate associative thickening behaviour, pyrene probe fluorescence studies fail to indicate the presence of well-organized hydrophobic microdomains.

We screened a chicken liver cDNA expression library with a probe spanning the distal region of the chicken vitellogenin II (VTGII) gene promoter and isolated clones for a transcription factor that we have named VBP (for vitellogenin gene-binding protein). VBP binds to one of the most important positive elements in the VTGII promoter and appears to play a pivotal role in the estrogen-dependent regulation of this gene. The protein sequence of VBP was deduced from a nearly full length cDNA copy and was found to contain a basic/zipper (bZIP) motif. As expected for a bZIP factor, VBP binds to its target DNA site as a dimer. Moreover, VBP is a stable dimer free in solution. A data base search revealed that VBP is related to rat DBP. However, despite the fact that the basic/hinge regions of VBP and DBP differ at only three amino acid positions, the DBP binding site in the rat albumin promoter is a relatively poor binding site for VBP. Thus, the optimal binding sites for VBP and DBP may be distinct. Similarities between the VBP and DBP leucine zippers are largely confined to only four of the seven helical spokes. Nevertheless, these leucine zippers are functionally compatible and appear to define a novel subfamily. In contrast to the bZIP regions, other portions of VBP and DBP are markedly different, as are the expression profiles for these two genes. In particular, expression of the VBP gene commences early in liver ontogeny and is not subject to circadian control.

The endogenous chicken vitellogenin II (VTGII) gene is transcribed exclusively in hepatocytes in response to estrogen. We previously identified two estrogen response elements (EREs) upstream of this gene. We now present an analysis of the VTGII promoter activated by these EREs in response to estrogen. Chimeric VTGII-CAT genes were cotransfected into LMH chicken hepatoma cells along with an estrogen receptor expression vector, and transient CAT expression was assayed after culturing the cells in the absence or presence of estrogen. An analysis of constructs bearing deletions downstream of the more proximal ERE indicated that promoter elements relevant to transcription in LMH cells extend to between -113 and -96. The relative importance of sequences within the VTGII promoter was examined by using 10 contiguous linker scanner mutations spanning the region from -117 to -24. Although most of these mutations compromised VTGII promoter function, one dramatically increased expression in LMH cells and also rendered the VTGII promoter capable of being activated by cis-linked EREs in fibroblasts cotransfected with an estrogen receptor expression vector. Gel retardation and DNase I footprinting assays revealed four factor-binding sites within this promoter. We demonstrate that three of these sites bind C/EBP, SP1, and USF (or related factors), respectively; the fourth site binds a factor that we denote TF-V beta. The biological relevance of these findings is suggested by the fact that three of these binding sites map to sites previously shown to be occupied in vivo in response to estrogen.

Two experiments evaluated the ability of maternal fatty acid supplementation to alter conceptus and endometrial fatty acid composition. In Exp. 1, treatments were 1) the control, a corn-soybean meal diet; 2) flax, the control diet plus ground flax (3.75% of diet); and 3) protected fatty acids (PFA), the control plus a protected fish oil source rich in n-3 PUFA (Gromega, JBS United Inc., Sheridan, IN; 1.5% of diet). Supplements replaced equal parts of corn and soybean meal. When gilts reached 170 d of age, PG600 (PMSG and hCG, Intervet USA, Millsboro, DE) was injected to induce puberty, and dietary treatments (n = 8/treatment) were initiated. When detected in estrus, gilts were artificially inseminated. On d 40 to 43 of gestation, 7 gilts in the control treatment, 8 gilts in the PFA treatment, and 5 gilts in the flax treatment were pregnant and were slaughtered. Compared with the control treatment, the flax treatment tended to increase eicosapentaenoic acid (EPA: C20:5n-3) in fetuses (0.14 vs. 0.25 +/- 0.03 mg/g of dry tissue; P = 0.055), whereas gilts receiving PFA had more (P0.05) docosahexaenoic acid (DHA: C22:6n-3) in their fetuses (5.23 vs. 4.04 +/- 0.078 mg/g) compared with gilts fed the control diet. Both the flax and PFA diets increased (P0.05) DHA (0.60, 0.82, and 0.85 +/- 0.078 mg/g for the control, flax, and PFA diet, respectively) in the chorioallantois. In the endometrium, EPA and docosapentaenoic acid (C22:5n-3) were increased by the flax diet (P0.001; P0.05), whereas gilts receiving PFA had increased DHA (P0.001). The flax diet selectively increased EPA, and the PFA diet selectively increased DHA in the fetus and endometrium. In Exp. 2, gilts were fed diets containing PFA (1.5%) or a control diet beginning at approximately 170 of age (n = 13/treatment). A blood sample was collected after 30 d of treatment, and gilts were artificially inseminated when they were approximately 205 d old. Conceptus and endometrial samples were collected on d 11 to 19 of pregnancy. Plasma samples indicated that PFA increased (P0.005) circulating concentrations of EPA and DHA. Endometrial EPA was increased (P0.001) for gilts fed the PFA diet. In extraembryonic tissues, PFA more than doubled (P0.001) the EPA (0.13 vs. 0.32 +/- 0.013 mg/g) and DHA (0.39 vs. 0.85 +/- 0.05 mg/g). In embryonic tissue on d 19, DHA was increased (P0.05) by PFA (0.20 vs. 0.30 +/- 0.023 mg/g). Supplementing n-3 PUFA, beginning 30 d before breeding, affected endometrial, conceptus, and fetal fatty acid composition in early pregnancy. Dynamic day effects in fatty acid composition indicate this may be a critical period for maternal fatty acid resources to affect conceptus development and survival.

Maternal diet influences fetal growth and postnatal development. We hypothesized that conceptuses gestated in sows provided ad libitum vs. restricted feed intake would differ in the milieu of hormones, growth factors, nutrients, and metabolites associated with growth and metabolism. This hypothesis was tested in two experiments by providing fourth-parity sows (Pig Improvement Co. C15 bred to Line 326 boars) with either 1.81 kg/d (as-fed basis; control) or ad libitum access to gestation diet. In Exp. 1, control (n = 6) or ad libitum (6.4 +/- 0.11 kg/d; n = 9) treatments were provided from d 29 to 45 (onset of estrus is d 0), and sows were slaughtered on d 46. Ad libitum sows gained more weight from d 29 to 45 than controls (34.0 vs. 4.32 kg, respectively; P0.01). No differences were observed on d 46 for the number of fetuses, conceptus attachment length, allantoic + amniotic fluid volume, placental weight, fetal weight, and fetal crown-to-rump length. Variation in fetal crown-to-rump length was less (P0.03) in sows fed ad libitum. Sows fed ad libitum had greater (P0.01) IGF-I and insulin concentrations in plasma than controls on d 43. In Exp. 2, sows were fed 1.81 kg/d (n = 6) or ad libitum (7.0 +/- 0.11 kg/d; n = 4) from d 30 to 56 of gestation, when sows were anesthetized and samples were collected surgically from their gravid uteri. Sows fed ad libitum gained more weight (P0.01) than did controls and had more (P0.06) IGF-I in their plasma and the plasma collected from umbilical veins of their fetuses. No differences were found for concentrations of insulin or glucose in plasma of sows or fetuses, but urea N concentrations were greater (P0.05) in maternal plasma and in the plasma, and allantoic and amniotic fluids of conceptuses from sows fed ad libitum. Combined data from Exp. 1 and 2 revealed a treatment x fetal number interaction (P0.05) for average fetal weight. The expected negative relationship between within-litter average fetal weight and the number of fetuses per uterus was observed for control sows (y = 115.4 -1.75 x fetal number; P0.05), but litters of ad libitum sows did not show this effect. The hypothesis that providing feed in excess of established requirements in early gestation affects the in utero milieu is supported by these results. Data further reveal that, at least at mid-gestation, the restraint to fetal growth that is exhibited when fetal number increases in control sows is not exhibited when sows are fed ad libitum.

Polyspermic fertilization and embryo quality are important issues for the in vitro production of pig embryos. We hypothesized that oocyte donor (prepubertal gilt vs. sow) affects polyspermy and blastocyst development in vitro and that the sexual maturity of the oocyte donor affects the response to sperm concentration in the fertilization medium. In Exp. 1, oocytes of sows and gilts were mounted and stained 12 h after insemination to provide fertilization data. In Exp. 2, putative embryos were developed in vitro to 144 h post-insemination before mounting. In both experiments, cumulus-oocyte complexes (COC) were collected from ovaries of prepubertal gilts and adult sows. Sperm were added after maturation of COC for 40 to 44 h. Sperm from two boars at 0.5 to 5.0 x 10(6) sperm/mL was used for insemination. More (P0.01) monospermic fertilizations were observed in oocytes derived from gilts than for oocytes from sows. There were fewer (P0.02) penetrated sperm per fertilized oocyte in oocytes from gilts compared with sows. There were effects of semen donor (boar) on the percentage of monospermic (P0.01) and polyspermic (P0.002) fertilizations, and on the number of penetrated sperm/fertilized oocyte (P0.02). In Exp. 2, cleavage and blastocyst formation was evaluated at 2 and 6 d postinsemination, respectively. More (P0.001) blastocysts developed from sow-derived oocytes than from gilt-derived oocytes. More (P0.05) total cells per blastocyst were observed in embryos from sow-derived oocytes than from gilt-derived oocytes. Semen donor affected the percentage of oocytes cleaving (P0.02), and a boar x sperm concentration interaction affected (P0.05) the incidence of blastocyt formation. Results indicate that sexual maturity of the donor is not responsible for the high incidence of polyspermy in porcine in vitro fertilization. However, blastocyst development is improved by the use of oocytes from sows rather than from prepubertal gilts.

A series of three experiments was conducted to test the functional status of the uterus and embryos in prepubertal gilts. In Exp. 1, gilts were induced to ovulate by treating with gonadotropins followed by hCG 72 or 96 h later, and were artificially inseminated 24 h after hCG. Five of the 10 gilts treated at 120 d of age, but none of the gilts treated at 100 of age, maintained pregnancies. We next tested the function of the uterine environment by transferring embryos from postpubertal females into gilts of various ages that had been induced to ovulate but not inseminated (Exp. 2). Pregnancy rate at d 50 of gestation was 44% (4/9) for 100-d-old recipients, 67% (2/3) for 140-d-old recipients, and 60% (3/5) for postpubertal recipients (P0.20). Therefore, uteri of 100-d-old gilts are able to maintain pregnancies with conceptuses from postpubertal gilts. In Exp. 3, embryos from 100-d-old and postpubertal gilts were transferred into postpubertal recipients. Uterine horns of recipients were surgically separated before transfer, and embryos from 100-d-old and post-pubertal females were transferred to opposite horns of some recipients (experimental). Other recipients received embryos from postpubertal females in both uterine horns (control). When examined on d 50 to 60 of gestation, three of five control gilts were pregnant and three of seven experimental gilts were pregnant (P0.50). In experimental recipients, the survival of embryos from 100-d-old gilts was 38% (8/21) compared to 57% (15/26) for embryos from postpubertal gilts (P0.30). Because all uterine horns of pregnant recipients contained fetuses, these results support the hypothesis that embryos from 100-d-old gilts are able to initiate and maintain pregnancies in the uteri of postpubertal gilts. Therefore, the uterine environment of 100-d-old gilts provides an environment that supports development of embryos produced by postpubertal gilts, and the embryos produced by 100-d-old gilts can survive and develop in the uteri of postpubertal gilts. It was only the combination of embryos and uteri of 100-d-old gilts that did not permit pregnancy to be maintained.

A genetic mapping strategy was employed to identify chromosomal regions harboring genes that influence the absorption of intestinal cholesterol in the mouse. Analysis of seven inbred strains of male mice (129P3, AKR, BALB/c, C3H/He, C57BL/6, DBA/2, and SJL, all from Jackson Laboratories) revealed substantial differences in their abilities to absorb a bolus of cholesterol delivered by gavage. Crosses between high (AKR, 129) and low (DBA/2, SJL) absorbing strains revealed evidence for the presence of dominant genes that increase and decrease cholesterol absorption. Backcrosses between F1 offspring and parental strains (DBA/2xAKD2F1 and 129xSJL129F1) followed by linkage analyses revealed four quantitative trait loci that influenced cholesterol absorption. Analyses of recombinant inbred strains identified an additional three loci affecting this phenotype. These seven quantitative trait loci, which map to different chromosomes and are termed Cholesterol absorption 1-7 (Chab1-7) loci, together influence the absorption of intestinal cholesterol in mice and are likely to be involved in different steps of this complex pathway.

Genetic linkage analysis in the laboratory mouse identified chromosomal regions containing genes that contribute to cholesterol accumulation in the liver and plasma. Comparisons between five inbred strains of mice obtained from the Jackson Laboratory (DBA/2, AKR, C57BL/6, SJL, and 129P3) revealed a direct correlation between intestinal cholesterol absorption and susceptibility to diet-induced hypercholesterolemia. This correlation was lost in the F1 generation arising from crosses between high- and low-absorbing strains. Linkage analyses in AKxD recombinant inbred strains and 129xSJL129F1 N2 backcross mice identified four quantitative trait loci (QTL) that influenced Liver cholesterol accumulation (Lcho1-4) and one locus that affected Plasma cholesterol accumulation (Pcho1). These loci map to five chromosomes and, with one exception, are different from the seven QTL identified previously that influence intestinal cholesterol absorption. We conclude that a large number of genes affects the amount of cholesterol absorbed in the small intestine and its accumulation in the liver and plasma of inbred mice.

Recent changes in management and medical nutrition therapy for diabetes mellitus have produced a need to retrain many practicing dietitians. To meet this need, a multidisciplinary group experienced in medical nutrition therapy and educational methods used a formal needs-assessment process to design a new training program. Sugar is Not a Poison (SNAP): The Dietitian's New Role in Diabetes Management is a 2 1/2-day program that uses written text, didactic presentation, and exercises that simulate patient encounters to accomplish 12 learning objectives. Program evaluations show high levels of participant satisfaction. Mean (+/- standard deviation) scores on pre- and postests of knowledge and problem solving were 69 +/- 13% and 86 +/- 9%, respectively (P0.01). The SNAP program needs assessment, training methods, and knowledge problem-solving test are relevant to all types of education programs in clinical dietetics.

The effectiveness of different habituation criteria was examined by means of computer simulations. A criterion based on fitting a second-order polynomial regression function to the looking time data was described. This criterion produced more accurate estimation of looking times as well as higher experimental power for detecting novelty effects, compared to the traditional windowed running average criterion or to a criterion based on linear regression. The polynomial regression approach probably has this advantage because it utilizes all of the available looking time data, rather than just the data in the current windowed average, and because it is sensitive to nonlinear trends in looking time. This new habituation criterion is easy to implement on a laboratory computer, and it should increase session lengths by no more than one trial or so, compared to windowed average criteria. With regard to test-retest reliability, all of the habituation criteria that were evaluated appear to have low reliability on average, with high sample-to-sample variability. These undesirable reliability characteristics are attributable to the high variability of infants' attentional behavior.

The aim of this investigation was to establish a homologous culture system for study of the transcriptional mechanisms underlying endometrial expression of the pregnancy-associated genes encoding antileukoproteinase (ALP), an elastase/cathepsin G protease inhibitor, and uteroferrin (Uf), a transplacental iron transport protein. Glandular epithelial (GE), Luminal epithelial (LE), and stromal (ST) cells were isolated from pig endometrium at Day 12 of pregnancy by differential enzymatic digestion and sieve filtration. The three cell populations differed with respect to their morphology in culture and with respect to their expression of ALP and Uf. Expression of the ALP gene was much higher in GE than in LE cells and was undetectable in ST cells. Similarly, GE had the highest expression of the Uf gene, and expression in ST was lower but distinct. Western blot analysis of conditioned media (72 h) from GE, LE, and ST, using antiporcine Uf antiserum, detected significant levels of secreted Uf only in GE. The steroid hormone responsiveness of GE cells was monitored by changes in steady-state levels of ALP mRNA after 24-h exposure to estradiol 17 beta (E2; 10 nM) and/or progesterone (P; 10 nM). Glandular epithelial cells treated with E2, P, and E2 + P had increased (p0.05) ALP mRNA levels relative to those in control cultures. Glandular epithelial cells were transiently transfected with reporter constructs containing the 5'-flanking genomic regions of each gene. For ALP, the 1266-nucleotide (nt) region of the ALP 5'-flanking genomic DNA, and progressive 5' deletions within this region, were coupled to a luciferase reporter gene (LUCE). The most proximal 119-bp fragment (-119ALP LUCE), which contains the TATAA box (-21 to -26 nt) and a GC-rich sequence (-66 to -74 nt), was sufficient to confer transcriptional activity to the reporter vector. Progressively longer 5'-genomic fragments had promoter activities higher than or similar to those of the 119-nt fragment. Estrogen had no effect on the transcriptional activities of any of the ALP constructs. Uteroferrin 5'-flanking and promoter DNA constructs containing the chloramphenicol acetyl transferase (CAT) reporter gene also exhibited transcriptional activity in GE cells. The presence of multiple interacting cis-regulatory sequences within this region was demonstrated by increased promoter activity, relative to that of the smallest construct (-182 UFCAT-E; basal activity), with the inclusion of sequences between -182 and -484 nf, and drastic reduction to basal activity with the inclusion of sequences between -484 and -831 nt. In summary, primary cultures of GE from early-pregnant porcine endometrium express ALP and Uf, are steroid hormone-responsive, and support the transcriptional activity of endometrial-associated gene promoter and regulatory sequences. The use of primary GE cells thus provides a convenient in vitro system for further study of the endocrine, paracrine, and autocrine factors regulating endometrial gene expression during pregnancy.

Increased inflammation of the peripheral airways has been implicated as a cause of pulmonary function impairment. However, little information is available on the correlation between subclinical decrements of pul monary function and inflammation in asymptomatic individuals. A relationship between markers of inflam mation and lung function may be useful in predicting the early onset of lung function impairment. The purpose of this study was to investigate the correlation of hematolo gic markers of inflammation and spirometry in asympto matic smokers and nonsmokers. The specific objectives of this study were twofold. The first objective was to quantify and compare the spirometric measures of lung function in smokers and nonsmokers having similar demographic and lifestyle characteristics. The second objective was to define the correlation between these spirometric measure ments and hematologic markers of inflammation (white blood cells, monocytes, basophils, PGE1, IgG, and IgE). Systemic blood samples and spirometric measurements were obtained from 61 age-matched (33 ± 9 years) healthy, asymptomatic smokers and nonsmokers, with similar self- reported lifestyles (i.e., food, alcohol, vitamin consump tion and exercise). Both male and female smokers self- reported a higher coffee consumption (P < 0.05) compared to nonsmokers. Male smokers self-reported a trend toward current blue-collar versus white-collar occupation when compared with the nonsmokers. Body weight (77.6± 16.6 kg) did not differ between the smokers and non smokers. The male nonsmokers were taller than the male smokers (P < 0.05). All subjects were asymptomatic and had clinically normal spirometry. Compared to male nonsmokers, the male smokers had lower FEF25-75%and FEF75-85%values (P1/FVC, FEV1and FVC were significantly different. The female smokers did not differ from the female nonsmokers (P1) or immunologic endpoints (IgE) and spirometric measure ments were observed in female smokers, female non smokers and male nonsmokers. No statistically significant correlations involving immunologic or inflammatory endpoints were observed in the male smokers. A better mechanistic understanding of the observed relationship between elevated hematologic inflammatory endpoints and reduced lung function may provide valuable insight into the clinical significance of these correlations.

Autosomal recessive mutations in the 17 beta-hydroxysteroid dehydrogenase 3 gene impair the formation of testosterone in the fetal testis and give rise to genetic males with female external genitalia. Such individuals are usually raised as females, but virilize at the time of expected puberty as the result of increases in serum testosterone. Here we describe mutations in 12 additional subjects/families with this disorder. The 14 mutations characterized to date include 10 missense mutations, 3 splice junction abnormalities, and 1 small deletion that results in a frame shift. Three of these mutations have occurred in more than 1 family. Complementary DNAs incorporating 9 of the 10 missense mutations have been constructed and expressed in reporter cells; 8 of the 9 missense mutations cause almost complete loss of enzymatic activity. In 2 subjects with loss of function, missense mutations testosterone levels in testicular venous blood were very low. Considered together, these findings strongly suggest that the common mechanism for testosterone formation in postpubertal subjects with this disorder is the conversion of circulating androstenedione to testosterone by one or more of the unaffected 17 beta-hydroxysteroid dehydrogenase isoenzymes.

Of the several sources of acoustic information for distance perception, those arising from motion of the listener or sound source have received little attention. This motion-related information (recently called acoustic tau) is described, and experiments evaluating its utilization are presented. Accuracy and consistency at walking to the locations of briefly presented sounds were better when people listened while walking than while standing still. Manipulations of the sound to simulate shorter or longer target distances produced appropriate undershooting but not overshooting. The results indicate that people use motion-related acoustic information about distance to guide their locomotor actions, although they do not take full advantage of this information.

In the DE-52 fraction 19 of the crude cytosolic extract of Saccharomyces cerevisiae, the 31-kDa endogenous substrate(s) of protein kinase C were detected by SDS-polyacrylamide gel electrophoresis and autoradiography. Phosphorylation of the substrate(s) depended on Ca2+, phosphatidylserine and diacylglycerol. It was also enhanced by phosphatidylinositol, phosphatidylethanolamine and phosphatidylglycerol (dioleoyl), but inhibited by arachidonic acid and sphingosine.

Establishing associations between environmental agents and disease presents challenges to both epidemiologists and toxicologists, particularly in cases of complex gene-environment interactions and when there is a long latency between exposure and disease. Biologic markers, physiological signals that reflect exposure, early cellular response, or inherent or acquired susceptibilities, provide a new strategy for resolving some of these problems. Biomarker research assumes that toxicant-induced diseases are progressive and that injury proceeds from entry of the toxicant into target cells, which induces subcellular biochemical events, to cell- and organ-level events that eventually induce irreversible or persistent organism dysfunction. The epidemiologic value of a biomarker lies in its ability to predict backward toward exposure and forward toward risk of clinical outcome, which is largely unknown. Research in mechanistic toxicology will advance the range of useful biomarkers in epidemiology and clinical medicine.

The objective of the present study was to evaluate the feasibility of delivering feed and detecting estrous behavior by computer-controlled equipment in a nonconfinement environment. In Exp. 1, gilts were assigned to treatment when detected pregnant by ultrasound at 30 to 35 d after artificial insemination. They were assigned to be fed individually in stalls once/day (0830) with a scoop (controls, n = 20) or with an electronic sow feeding station (ESF, n = 20). The ESF gilts received their feed in 98.6-g aliquots at 80-s intervals as they visited the feeding station. Control vs ESF gilts did not differ (P.8) for backfat (2.2 vs 2.1 cm) or weight (170 vs 172 kg) before farrowing, total and live pigs/litter (9.3 and 8.7 vs 9.1 and 8.8), or litter birth weight (12.7 vs 12.1). In Exp. 2, proceptive behavior, as measured by visits to a boar's pen, were recorded electronically, and observed estrus was evaluated in two groups of sows during their first (n = 11) and second and third (n = 19) estrous cycles and in one group of gilts (n = 14). A partition prevented visual and physical contact between the boar and the visiting females except where the electronic estrus detection (EED) station was installed. Feed delivery software was used to monitor boar visitation even though no feed delivery equipment was present at the boar pen.(ABSTRACT TRUNCATED AT 250 WORDS)

Protein kinases and their endogenous substrates from the crude cytosolic extract of Saccharomyces cerevisiae were coeluted in the fraction 13 on DE-52 column chromatography. Analyses of SDS-polyacrylamide gel electrophoresis and autoradiography revealed that the peptides between 14 and 34 kDa were the major phosphorylated substrates. In the presence of Ca2+ and Mg2+, the phosphorylation was suppressed strongly by the regulatory subunit of cAMP-dependent protein kinase and slightly by oleic acid, whereas it was augmented appreciably by phosphatidylglycerol (dioleoyl) and phosphatidylinositol.

Phosphorylation of endogenous yeast substrates in DE-52 fractions were analysed by SDS-polyacrylamide gel electrophoresis and autoradiography. In fractions 29 and 45, phosphatidylinositol inhibited the phosphorylation of multiple peptides with a wide range of molecular mass whereas it enhanced the phosphorylation of peptides smaller than 14 kD. Similar dual modulation of the phosphorylation by arachidonic acid was observed in fraction 37 which also contained potent phosphatidylserine-activating protein kinase C.

Primiparous and multiparous sows received a single dietary supplement of either L-tyrosine, L-phenylalanine, or L-glutamate in their feed on the day after weaning and effects on various reproductive traits were evaluated in three experiments. In Exp. 1 and 2, sows received either 0 (control; n = 22 and 64, respectively) or 100 mg of L-tyrosine/kg BW (n = 24 and 62, respectively) on the day after weaning. In Exp. 1, days from weaning to estrus (5.5 +/- .3 vs 5.3 +/- .3 d) and ovulation rate (15.6 +/- .9 vs 15.6 +/- 1) were similar in control and tyrosine-supplemented sows. In Exp. 2, interval from weaning to estrus was extended (P.01) in tyrosine-supplemented sows (6.4 +/- .5 d) compared with controls (4.5 +/- .5 d), but this was due to long return intervals in 7 of 62 tyrosine-supplemented sows. Total number of pigs born (10.2 +/- .4 vs 10.0 +/- .4) was similar in control and tyrosine-supplemented sows. In Exp. 3, sows received either no supplemental amino acids (n = 31) or their diet was supplemented with 100 mg/kg BW of either L-tyrosine (n = 31), L-phenylalanine (n = 33), or L-glutamate (n = 32). Neither days from weaning to estrus nor subsequent farrowing traits were altered in sows that received supplemental amino acids on the day after weaning. In conclusion, a single dietary supplementation of either tyrosine, phenylalanine, or glutamate to sows on the day after weaning failed to improve interval from weaning to estrus, ovulation rate, or litter traits at subsequent farrowing.

The phosphorylation of histones and glycogen synthase by protein kinases was analysed by SDS-polyacrylamide gel electrophoresis and autoradiography. The phosphorylation of histone III-S by the catalytic subunit of cyclic AMP-dependent protein kinase (A-PK) or cGMP-dependent protein kinase (G-PK) was inhibited by archidonic acid, sphingosine and staurosporine. Using the catalytic subunit of A-PK, the phosphorylation of histone VIII-S was inhibited by Ca2+, arachidonic acid and staurosporine; the phosphorylation of histone II-S was inhibited by phosphatidyl ethanolamine, phosphatidyl inositol, arachidonic acid and staurosporine; and the phosphorylation of glycogen synthase was inhibited by arachidonic acid and staurosporine. After being phosphorylated by the catalytic subunit of A-PK, calpain II with 4 microM Ca2+ was less effective in degrading histone III-S, which had been prephosphorylated by PK-C.

The uterus plays a central role in the reproductive biology of mammals. Adaptation of the uterus from an oviparous to a viviparous nature required changes that involved production of a uterine environment that could support the development of the embryo and fetus. Production of a suitable environment includes the synthesis and secretion of products by the uterine endometrium. However, the uterine endometrium is not a single homogeneous unit, but rather consists of several cell populations. Recent accomplishments in cell culture techniques provide a means for examining the contributions and secretory control of different endometrial cell populations. Furthermore, it is possible to recombine specific cell types to study their interaction. It is clear that the luminal epithelium, glandular epithelium and endometrial stroma produce different secretory products. Some secretions (for example uteroferrin) are secreted by only one cell type; others (for example prostaglandins, PGs) are secreted by all types of cell. There is much to be learned about the functions and regulations of endometrial secretions and there are important aspects of the role of the endometrium in pregnancy that present concepts do not address. For example, there is no explanation for the required synchrony between the embryo and uterus before day 10 and the implications of control of the uterine environment by progesterone from day 4 to day 10 are not understood. Almost all of the uterine secretory proteins are produced after day 10. In this review, we consider the protein and prostaglandin products from the different cell populations of the pig endometrium and propose a model to explain the integration of multiple sources of PGs and multiple regulators of PG secretion. Our purpose is to facilitate a more complete understanding of the individual uterine cell populations and a better understanding of how these cell types interact to function as a complete unit.

The proteolysis of the 32P-labeled holoenzyme of cyclic AMP-dependent protein kinase (A-PKII:DEAE, peak II fraction) was analysed by SDS-polyacrylamide gel electrophoresis and autoradiography. The contaminants of the A-PKII and calpain II apparently did not interfere with the accuracy of this highly sensitive analysis. Phosphorylation of calpain II by the catalytic subunit of cyclic AMP-dependent protein kinase (A-PK) greatly enhanced the proteolysis of A-PKII, whereas phosphorylation by protein kinase C (PK-C) or cyclic GMP-dependent protein kinase (G-PK) slightly altered the proteolysis.

Research into the nature of aggressive behavior in youths has demonstrated that these youths are often the victims of abuse, exhibit aggressive behavior in early childhood, and remain aggressive into young adulthood. The treatment approach described in this article is a modification of Monahan's [1981] model of the prediction of violent behavior and the anger-management approach of Novaco [1985], and integrates the developmental models of Piaget [1963] and Erikson [1959]. The program is a combination of cognitive, behavioral, and expressive therapies and is targeted to the reduction of dysfunctional cognitive, affective, behavioral, and problem-solving patterns of aggressive youths. As referrals of such aggressive clients are often involuntary, interventions with unwilling and resistant clients are also presented.

Epidermal growth factor (EGF) and its receptor have been implicated in the control of uterine cell growth and differentiation. The objectives of this study were to determine EGF binding characteristics and effects of EGF on prostaglandin (PG) production in vitro by glandular and stromal cells from porcine endometrium. Endometrial tissues were taken from 10 sows on Day 13 of pregnancy (first day of estrus = Day 0). Glandular and stromal cells were separated by enzymatic dispersion and sieve filtration and cultured for 3 days. EGF-binding assay was carried out at 20 degrees C in the presence of 0.2 nM 125I-EGF with increasing concentrations of unlabeled EGF (0-12 nM). Scatchard analyses revealed one class of high-affinity binding sites in each cell type with apparent equilibrium dissociation constants (n = 6) of 2.96 +/- 0.60 nM and 2.48 +/- 0.50 nM for stromal and glandular cells, respectively. The apparent binding capacities were 199.3 +/- 34.8 fmol/10(6) cells for stromal cells and 40.7 +/- 6.5 fmol/10(6) cells for glandular cells. Effects of EGF on PG production were determined by including 1, 5, 10, or 20 ng/ml EGF in the medium for the final 24 h of the 72-h culture. EGF increased PGE (p less than 0.01) and PGF2 alpha (p less than 0.05) secretion by stromal cells. The highest concentration (20 ng/ml) of EGF increased secretion of PGE and PGF2 alpha by 133% and 64%, respectively, over controls.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of various agents on the cleavage of serum albumin, interferon, immunoglobulin and complement component C1q by the extracellular protease from Staphylococcus aureus were analysed by SDS-polyacrylamide gel electrophoresis. Arachidonic acid moderately stimulated the proteolysis of serum albumin, interferon and complement component. Phosphatidic acid effectively enhanced the proteolysis of serum albumin and IgG, whereas it inhibited the cleavage of IgM. The proteolysis of IgG was appreciably enhanced by sphingosine. In contrast, phosphatidyl choline and phosphatidyl glycerol were shown to have an inhibitory effect on the proteolysis of IgG and IgM. Phosphatidyl serine, phosphatidyl inositol and phosphatidyl ethanolamine also inhibited the proteolysis of IgG. The failure of any of these agents to exert a persistent effect on the cleavage of all substrates, revealed the complexity of the interactions among the agent, the substrate and the protease.

The role of interaural time differences in infants' sound localization was investigated. One experiment on free-field sound localization corroborated previous findings that the minimum audible angle changes substantially toward the end of the first half year after birth, and 3 experiments explored interaural time discrimination in that age range. The first of these 3 experiments used an adaptive psychophysical procedure, showing that infants responded appropriately in a sound lateralization task that provides a direct measure of interaural time discrimination. The other 2 experiments improved on the psychophysical procedure by taking into account the ceiling level on performance in the task. Infants aged 16, 20, and 28 weeks had thresholds in the range of 50 to 75 microsec, with no apparent age difference. These thresholds were much lower than would be predicted from studies of free-field sound localization, indicating that sensitivity to interaural time differences is not a limiting factor for the precision of sound localization in this age range. Instead, age-related changes in free-field sound localization may reflect the need to integrate across different localization cues and to calibrate the changing values of cues due to head growth.

A case of fly larva infestation of the eye (ophthalmomyiasis) is reported in an American soldier serving in Operation Desert Shield. Background information, prognosis, and treatment are discussed.

Pig conceptuses display a surge in oestrogen and catecholoestrogen synthetic activity during the periimplantation period. However, the pathways of catecholoestrogen metabolism in pig conceptuses and endometrium are unknown. O-Methylation is an important route of catecholoestrogen metabolism. Therefore, the O-methylations of 2- and 4-hydroxy-oestradiols (2- and 4-OH-oestradiol) by cytosol of pig conceptuses and endometrium during the periimplantation period were studied. Kinetic studies performed in tissues obtained on day 13 of pregnancy (day 0 = first acceptance of the male) indicated that the O-methylation of 2-OH-oestradiol displayed simple Michaelis–Menten kinetics in both tissues. In blastocysts, the apparent Michaelis constant (Km) and maximum velocity (Vmax) for the O-methylation of 2-OH-oestradiol were 1·4 μmol/l and 11·27 pmol/mg protein per min respectively, and when 4-OH-oestradiol was used as substrate, the values were 2·53 μmol/l and 9·86 pmol/mg protein per min respectively. The apparent Km and Vmax values for the O-methylation of 2-OH-oestradiol in endometrium were 0·77 μmol/l and 19·6 pmol/mg protein per min respectively, and for the O-methylation of 4-OH-oestradiol were 2·44 μmol/l and 10·38 pmol/mg protein per min respectively. Ontogenesis of catechol-O-methyltransferase (COMT) in conceptuses and endometrium was studied from day 10 to day 19 of pregnancy. Conceptus COMT activity was lowest on day 10 and increased gradually to day 19 of pregnancy. Because a surge of oestradiol-2/4-hydroxylase activity occurs on days 11–13, less COMT activity on these days than on the later days of pregnancy is consistent with a role for catecholoestrogens in conceptus-maternal signalling during pregnancy establishment. Endometrial COMT activity on day 10 was higher than that on later days of pregnancy. Therefore, a role for COMT in modulation of catecholoestrogen and oestrogen function during the peri-implantation period is possible. Journal of Endocrinology (1990) 127, 77–84

A case of fly larva infestation of the eye (ophthalmomyiasis) is reported in an American soldier serving in Operation Desert Shield. Background information, prognosis, and treatment are discussed.