Prevalence,risk factors,and effects on fertility of cytological endometritis at the time of insemination in Norwegian Red cows

The present study aimed to assess the occurrence of cytological endometritis (CYTO), a nonsymptomatic inflammation of the endometrium, at first artificial insemination (AI) postpartum in Norwegian Red cows. Further, risk factors for CYTO manifestation and its effect on reproductive success and late embryo loss were evaluated. In total 1,648 cows located in 116 herds were included in the study. On mainly spontaneous estrus, endometrial cytology samples were collected using a cytotape technique, and a total of 300 representative epithelial cells and polymorphonuclear neutrophils (PMN) were counted at 400× magnification. Vaginal mucus obtained by Metricheck (Simcro) and body condition score were recorded. Milk samples for progesterone analysis were collected at AI and 21 d later. Pregnancy was diagnosed by rectal palpation or analysis of pregnancy-associated glycoproteins. Based on the constructions of a receiver operator characteristics curve, the cut-off level for PMN defined as CYTO was set to 3.0%, representing the level at which the PMN occurrence affected pregnancy outcome, with the highest summation of sensitivity (32.4%) and specificity (74.9%). Three logistic models with herd included as random factor were constructed. The outcome for the first model was the likelihood for CYTO based on the endometrial samples, in the second model pregnancy to first AI, and in the third model embryo loss. The proportion of CYTO was 28.0% (461/1,648). The average interval in days to first AI was 71.7 d (standard error ± 0.7) and the overall pregnancy incidence to first AI was 59.8% (866/1,449). The likelihood for CYTO at first AI was associated with AI personnel, calving to first AI interval, vaginal mucus characteristics, amount of red blood cells in sample, season, and barn type. Pregnancy to first AI was lower in CYTO-positive cows (odds ratio = 1.51, confidence interval = 1.17–1.94). Other factors affecting pregnancy to first AI were AI personnel, test day milk yield, barn type, and obstetrical conditions or fertility treatments before first AI. The proportion of late embryo loss and abortion was 8.6% (82/948) and 2.8% (24/866), respectively. Late embryo loss was associated with treatment against fertility disorders before first AI, but not associated with CYTO. Overall, our results suggest that even if Norwegian Red cows show a fairly high prevalence of CYTO in the endometrium at first AI, it does not seem to have a major effect on the reproductive performance. The Norwegian Red breeding program has emphasized fertility and health for decades, and a genetically advantageous uterine immunology might be one of the preserved mechanisms.     

Dynamische Zustandsbewertung einer Verbundbrücke: Beobachtung äußerer und nichtlinearer Einflüsse auf die modalen Eigenschaften

Bei der Anwendung dynamischer Untersuchungsmethoden zur Zustandsbewertung von Ingenieurbauwerken aus Stahl‐ und Spannbeton sowie Verbundbau spielen Veränderungen von Rand‐ und Umweltbedingungen auf die modalen Parameter eine wesentliche Rolle. Während man diese im Laborexperiment weitest gehend überwachen bzw. ausschließen kann, sind sie bei in‐situ Versuchen nur schwer zu kontrollieren. Am Beispiel einer Verbundbrücke soll demonstriert werden, welche Größenordnung die Veränderung von Randbedingungen (z. B. Veränderung von anregenden Kräften) und Umwelteinflüsse (z. B. Temperatur) auf die dynamischen Parameter einer in‐situ Struktur haben können. Die Größenordnung dieser Veränderungen wird mit Veränderungen, die aus wirklichen Strukturschäden resultieren können, verglichen und bewertet. Dynamic Condition Assessment of a Composite Bridge: Investigation of External and Nonlinear Influences on the Modal Properties Using dynamic investigation methods to assess the state of civil constructions like reinforced and prestressed concrete structures as well as composite structures, changes in boundary conditions and environmental influences play a decisive role. Under laboratory conditions it is possible to control respectively to exclude these influences but it is not always possible to control and to exclude them when testing in‐situ. By means of dynamic measurements which are conducted on a composite bridge, possible changes in dynamic properties resulting from changes in the amplitude of the excitation force and resulting from changes in the temperature conditions are demonstrated. The dimension of these changes is compared with changes in modal properties resulting from real structural defects and structural damages.     

Adipose tissue insulin receptor and glucose transporter 4 expression,and blood glucose and insulin responses during glucose tolerance tests in transition Holstein cows with different body condition

Glucose uptake in tissues is mediated by insulin receptor (INSR) and glucose transporter 4 (GLUT4). The aim of this study was to examine the effect of body condition during the dry period on adipose tissue mRNA and protein expression of INSR and GLUT4, and on the dynamics of glucose and insulin following the i.v. glucose tolerance test in Holstein cows 21 d before (d ?21) and after (d 21) calving. Cows were grouped as body condition score (BCS) ≤3.0 (thin, T; n = 14), BCS = 3.25 to 3.5 (optimal, O; n = 14), and BCS ≥3.75 (overconditioned, OC; n = 14). Blood was analyzed for glucose, insulin, fatty acids, and β-hydroxybutyrate concentrations. Adipose tissue was analyzed for INSR and GLUT4 mRNA and protein concentrations. During the glucose tolerance test 0.15 g/kg of body weight glucose was infused; blood was collected at ?5, 5, 10, 20, 30, 40, 50, and 60 min, and analyzed for glucose and insulin. On d ?21 the area under the curve (AUC) of glucose was smallest in group T (1,512 ± 33.9 mg/dL × min) and largest in group OC (1,783 ± 33.9 mg/dL × min), and different between all groups. Basal insulin on d ?21 was lowest in group T (13.9 ± 2.32 µU/mL), which was different from group OC (24.9 ± 2.32 µU/mL. On d ?21 the smallest AUC 5–60 of insulin in group T (5,308 ± 1,214 µU/mL × min) differed from the largest AUC in group OC (10,867 ± 1,215 µU/mL × min). Time to reach basal concentration of insulin in group OC (113 ± 14.1 min) was longer compared with group T (45 ± 14.1). The INSR mRNA abundance on d 21 was higher compared with d ?21 in groups T (d ?21: 3.3 ± 0.44; d 21: 5.9 ± 0.44) and O (d ?21: 3.7 ± 0.45; d 21: 4.7 ± 0.45). The extent of INSR protein expression on d ?21 was highest in group T (7.3 ± 0.74 ng/mL), differing from group O (4.6 ± 0.73 ng/mL), which had the lowest expression. The amount of GLUT4 protein on d ?21 was lowest in group OC (1.2 ± 0.14 ng/mL), different from group O (1.8 ± 0.14 ng/mL), which had the highest amount, and from group T (1.5 ± 0.14 ng/mL). From d ?21 to 21, a decrease occurred in the GLUT4 protein levels in both groups T (d ?21: 1.5 ± 0.14 ng/mL; d 21: 0.8 ± 0.14 ng/mL) and O (d ?21: 1.8 ± 0.14 ng/mL; d 21: 0.8 ± 0.14 ng/mL). These results demonstrate that in obese cows adipose tissue insulin resistance develops prepartum and is related to reduced GLUT4 protein synthesis. Regarding glucose metabolism, body condition did not affect adipose tissue insulin resistance postpartum.     

Pregnancy incidence in Norwegian red cows using nonreturn to estrus, rectal palpation, pregnancy-associated glycoproteins, and progesterone

The objectives of the study were to estimate pregnancy incidence and calving rate after first artificial insemination (AI) in Norwegian Red cows undergoing spontaneous estrus, to assess the relationship between pregnancy and management factors at herd or cow level, to evaluate differences between 60-d nonreturn rate (NRR60d) and pregnancy incidence, and to compare the accuracy of pregnancy diagnosis by rectal palpation and plasma pregnancy-associated glycoproteins (PAG) analysis supported by progesterone measurements. In total, 829 animals (n = 229 heifers, 234 first-lactation, 173 second-lactation, and 193 >second-lactation cows) were included. Milk samples for progesterone analysis were collected both at AI and 3 wk later. Cows with progesterone concentrations <3 ng/mL at AI were considered in estrus or having nonactive ovaries, whereas cows with progesterone concentrations >7 ng/ mL 3 wk later were considered pregnant. Blood sampling for PAG analysis and pregnancy diagnosis by rectal palpation were conducted 57.6 ± 0.92 d after AI. Pregnancy-associated glycoprotein concentrations equal to 2.5 ng/mL gave the greatest sensitivity (94.3%) and specificity (94.6%) in the assessment of pregnancy. The number of days from calving to first AI was 85.3 ± 1.71. Overall NRR60d after first AI was 72.5%. The corresponding values for heifers, first-lactation, second-lactation, and >second-lactation cows were 76.9, 67.1, 69.9, and 76.2%. Overall pregnancy incidence after first AI was 63.7%. The corresponding values for heifers, first-lactation, second-lactation, and >second-lactation cows were 70.0, 58.2, 61.6, and 64.9%. Overall calving rate to first AI was 57.2%. The corresponding values for heifers, first-lactation, second-lactation, and >second-lactation cows were 64.9, 54.3, 54.7, and 53.9%. The overall difference between NRR60d and pregnancy incidence was 8.8%, whereas the parity-specific differences were 6.9, 8.9, 8.3, and 11.3% for heifers, first-lactation, second-lactation, and >second-lactation cows, respectively. Eight animals with PAG <2.5ng/mL and classified as pregnant by rectal palpation calved, whereas 5 animals with PAG ≥2.5 ng/mL and classified as non-pregnant by rectal palpation also calved. The study showed that Norwegian Red cows have relatively high reproductive performance. Breeding for fertility traits over 35 yr is probably an important reason for such high fertility.     

Antigen-presenting function and B7 expression of murine sinusoidal endothelial cells and Kupffer cells

BACKGROUND & AIMS: Inflammatory liver disease as well as rejection of liver allografts are thought to be mediated by resident antigen-presenting cells in the liver. At the same time, in vivo antigen presentation in the liver appears to be a more tolerogenic than systemic antigen challenge. The aim of this study was to show and characterize the antigen-presenting capability of sinusoidal endothelial cells and Kupffer cells. METHODS: Purified murine sinusoidal endothelial cells and Kupffer cells were studied for their ability to serve as accessory cells and antigen-presenting cells by proliferation assays. They were also studied for their expression of interleukin 1 and the B7 costimulatory molecules by Northern blotting, polymerase chain reaction, and flow cytometry. RESULTS: Both cell types expressed interleukin 1 messenger RNA and could serve equally well as accessory and antigen-presenting cells. B7-2 messenger RNA and surface expression on sinusoidal endothelial cells and on Kupffer cells was shown. Antibodies to the B7 molecules inhibited antigen presentation. Addition of interleukin 10 as a regulatory cytokine secreted by Kupffer cells was suppressive. CONCLUSIONS: Sinusoidal endothelial cells carry functional B7-2 molecules and can serve as effective antigen-presenting cells. However, antigen presentation by sinusoidal endothelial cells may be locally down-regulated by interleukin 10.     

Human immunoglobulin D segments encoded in tandem multigenic families

A family of germ-line immunoglobulin D-region genes has been cloned and mapped at regular intervals along a 33-kilobase length of human chromosomal DNA. Each member of the family varies slightly in sequence, but precisely conserves the recombinational signals and spacing that flank each gene. This region seems to have been formed by the tandem amplification of large and still well conserved segments of genomic DNA. Further, structural comparisons of germ-line and rearranged D segments suggest that D segments may recombine with each other.     

Linking the fields--the interplay of organic synthesis, biophysical chemistry, and cell biology in the chemical biology of protein lipidation

Research in the biological sciences has undergone a fundamental and dramatic change during the last decades. Whereas biology was more phenomenologically oriented for a long time, today many biological processes are investigated and understood in molecular detail. It has become evident that all biological phenomena have a chemical basis: Biology is based on chemical principles. In the past, this insight had led to the development of biochemistry, molecular biology, and modern pharmacology. Today it increasingly determines the manner in which various biological phenomena are studied. The tools provided by classical biological techniques often are not sufficient to address the prevailing issues in precise molecular detail. Instead, the strengths of both chemical and biological methodology have to be used. Several recent research projects have proven that combining the power of organic synthesis with cell biology may open up entirely new and alternative opportunities for the study of biological problems. In this review we summarize the successful interplay between three disciplines-organic synthesis, biophysics, and cell biology-in the study of protein lipidation and its relevance to targeting of proteins to the plasma membrane of cells in precise molecular detail. This interplay is highlighted by using the Ras protein as a representative example. The development of methods for the synthesis of Ras-derived peptides and fully functional Ras proteins, the determination of their biophysical properties, in particular the ability to bind to model membranes, and finally the use of synthetic Ras peptides and Ras proteins in cell biological experiments are addressed. The successful combination of these three disciplines has led to a better understanding of the factors governing the selective targeting of Ras and related lipid-modified proteins to the plasma membrane.     

Exploiting the substrate tolerance of farnesyltransferase for site-selective protein derivatization

The site-selective modification of proteins with a functional group is an important biochemical technique, but covalent attachment of a desired group to a chosen site is complicated by the reactivity of other amino acid side chains, often resulting in undesired side reactions. One potential solution to this problem involves exploiting the activity of protein-modifying enzymes that recognize a defined protein sequence. Protein farnesyltransferase (FTase) covalently attaches an isoprenoid moiety to a cysteine unit in the context of a short C-terminal sequence that can be easily grafted onto recombinant proteins. Here we describe the synthesis of four phosphoisoprenoids functionalized with biotin, azide, or diene groups. These phosphoisoprenoids bound to FTase with affinities comparable to that of the native substrate. With the exception of the biotin-functionalized analogue, all the phosphoisoprenoids generated could be transferred to peptide and protein substrates by FTase. Unlike proteins modified with farnesyl moieties, Ypt7 prenylated with (2E,6E)-8-(azidoacetamido)-3,7-dimethylocta-2,6-dienyl groups did not oligomerize and showed no detectable increase in hydrophobicity. To assess the suitability of the functionalized isoprenoids for protein modifications they were further derivatized, both by Diels-Alder cycloaddition with 6-maleimidohexanoic acid and by Staudinger ligation with a phosphine. We demonstrate that the Staudinger ligation proceeds more rapidly and is more efficient than the Diels-Alder cycloaddition. Our data validate the use of FTase as a protein-modification tool for biochemical and biotechnological applications.