Frequencies and effects of alternative DRB3.2 alleles of bovine lymphocyte antigen for Holsteins in milk selection and control lines

Frequencies of alleles at the DRB3.2 locus of the bovine major histocompatibility complex for two genetic lines of Holsteins were compared, and allelic effects on yield and udder health traits were examined. The DRB3.2 genotypes of 186 cows and heifers were determined using DNA that had been extracted from blood samples. The cattle were from a designed selection experiment for milk yield. After edits, 173 cows and heifers with known alleles remained (54 from a control line and 119 from a selection line for milk yield). A total of 19 alternative DRB3.2 alleles appeared across the two lines, and frequencies ranged from 0.3 to 21.4%. Allelic frequencies differed significantly between the selection and control lines. Alleles 3, 8, 9, 10, 26, and 28 were more frequent in the control line, but alleles 16, 24, and 27 were more frequent in the selection line. An animal model was used to estimate gene substitution effects of alternative alleles on milk yield, fat yield, and protein yield, SCS, chronically elevated SCS, acutely elevated SCS, and mastitis incidence. First, second, and third lactations were analyzed separately. Allele 7 was associated with a significant increase in protein yield during first and second lactations and a significant increase in chronically elevated SCS and acutely elevated SCS during second lactation. Because of conflicting results with other studies and a limited numbers of cows, additional research is needed before specific alleles are confirmed to be associated with measures of yield and udder health.     

An immunological approach to the evaluation of welfare in Holstein Frisian cattle

Clinical immunological and haematological parameters, along with clinical conditions and growth rate, were studied in 413 male Holstein Frisian calves introduced into a large centre for genetic selection in different seasons of the year. Abnormalities were revealed by the laboratory tests in the great majority of calves after transportation stress, a general tendency to the restoration of physiological values being evident thereafter. Laboratory parameters were highly correlated with disease conditions: with three exceptions only, animals showed altered laboratory parameters some days before the occurrence of clinical symptoms. Eighteen per cent of animals showed altered laboratory parameters with no obvious clinical signs of disease; yet they experienced a reduced weight gain. Results suggest that clinical immunological and haematological parameters could be the foundation of a new, large-scale, robust approach to the control of welfare in cattle, which should be integrated preferably by a further range of records and measures.     

Genetic evaluation of body weight of lactating Holstein heifers using body measurements and conformation traits

Genetic and phenotypic parameters of body weight (BW), hip height, heart girth, and linear conformation traits were estimated from field data for 7344 lactating Holstein heifers from 560 herds. Mean BW was around 540 kg at calving, decreased to a minimum of 514 kg by wk 6, and increased to > 590 kg by wk 45 of lactation. The statistical model for BW included month of recording, lactation stage, pregnancy stage, proportion of Holstein genes, parity of the dam, calving age, herd, and animal. Variation between herds accounted for 33% of the phenotypic variation of BW. Estimated heritability was 0.33 for BW and ranged from 0.32 to 0.54 for heart girth, hip height, and conformation traits. Genetic correlations of BW with heart girth, hip height, body depth, rump width, and muscularity ranged from 0.48 to 0.77. The accuracy of a selection index was 0.90 for direct selection and 0.83 for indirect selection using information on those conformation traits of 50 daughters. Genetic variation of BW is considerable, and genetic evaluation of BW can be based on data for conformation traits from a field recording system with only a limited loss of accuracy.     

Level of viral antigen in blood leucocytes from cattle acutely infected with bovine viral diarrhoea virus

Blood samples from 24 calves undergoing experimental acute infection with a non-cytopathogenic bovine viral diarrhoea virus (BVDV) were examined for viral antigen in peripheral leucocytes with an enzyme-linked immunosorbent assay (ELISA), and for presence of virus in blood plasma in a cell culture assay. With the antigen ELISA, low positive values were detected in leucocytes sampled on days 3-4 from two of eight animals inoculated intranasally, and on days 11-13 from three of 16 animals inoculated intramuscularly. From 22 of the animals, low titres of BVDV were detected in blood plasma obtained 2-9 days after inoculation. All other samples, drawn between 2 and 21 days after inoculation, were negative for viral antigen. All animals seroconverted 3-4 weeks after inoculation, some after having shown mild and transient signs of disease.     

Seroprevalence to bovine immunodeficiency virus and lack of association with leukocyte counts in Italian dairy cattle

We report herein on the first serological detection of antibodies to bovine immunodeficiency virus (BIV) in Italy. According to criteria of a stratified-random sampling of dairy cattle reared in the Parma area (a province in the Po Valley, Northern Italy), sera from 3166 cows belonging to 272 herds were collected. In addition, sera of 138 bulls from eight artificial-insemination (AI) centres were sampled. Seventy-eight cows (2.5%) from 16 herds (5.8%) and seven bulls (5.1%) from two AI centres were positive for BIV-R29 antibodies in the IFA-test. IFA-positive sera assayed by Western blot had reaction to different viral proteins: 81 out of 85 sera showed antibody to p26 (considered the BIV major internal core protein); four sera reacted to other viral proteins but not to p26. Peripheral blood leukocytes of 60 seropositive and 60 seronegative animals, belonging to eight BIV-infected herds, were enumerated to assess any effect of BIV infection on white-blood cells. No significant differences were detected between the two groups. These data indicate that BIV infection is present in Italian dairy cattle--but the role of BIV in inducing disease remains unclear.     

Lack of association of genetic traits with alcoholism; C3, Ss and ABO systems

A study of association between markers of the Ss system, the C3 system and the ABO system showed no evidence of any association between alcoholism and specific phenotypes.     

Serine 257 phosphorylation regulates association of polyomavirus middle T antigen with 14-3-3 proteins

Polyomavirus middle T antigen (MT) is phosphorylated on serine residues. Partial proteolytic mapping and Edman degradation identified serine 257 as a major site of phosphorylation. This was confirmed by site-directed mutagenesis. Isoelectric focusing of immunoprecipitated MT from transfected 293T cells showed that phosphorylation on wild-type MT occurred at near molar stoichiometry at S257. MT was previously shown to be associated with 14-3-3 proteins, which have been connected to cell cycle regulation and signaling. The association of 14-3-3 proteins with MT depended on the serine 257 phosphorylation site. This has been demonstrated by comparing wild-type and S257A mutant MTs expressed with transfected 293T cells or with Sf9 cells infected with recombinant baculoviruses. The 257 site is not critical for transformation of fibroblasts in vitro, since S257A and S257C mutant MTs retained the ability to form foci or colonies in agar. The tumor profile of a virus expressing S257C MT showed a striking deficiency in the induction of salivary gland tumors. The basis for this defect is uncertain. However, differences in activity for the wild type and mutant MT lacking the 14-3-3 binding site have been observed in transient reporter assays.     

Molecular genetic characterization of new bovine kappa-casein alleles CSN3F and CSN3G and genotyping by PCR-RFLP

In order to characterize the two new kappa-casein variants F and G (CSN3F and CSN3G) recently detected in Ayrshire and Pinzgauer cattle, exon IV of CSN3 from heterozygous animals was amplified by polymerase chain reaction (PCR), cloned and sequenced. The sequencing data revealed single point mutations at nucleotide positions 10530 (G-->A) for CSN3F and 10790 (C-->T) for CSN3G, corresponding to amino acid exchanges in positions 10 (Arg-->His) and 97 (Arg-->Cys) respectively. These mutations alter recognition sites for the restriction enzymes HhaI and MaeII, which were subsequently used to confirm these polymorphisms in cattle carrying CSN3F or CSN3G. A PCR-restriction fragment length polymorphism (RFLP) genotyping procedure for all currently known CSN3 alleles (CSN3A, CSN3B, CSN3C, CSN3E, CSN3F, CSN3G) was developed.     

Genetic and phenotypic (co)variances for production traits of intact male populations of purebred and composite beef cattle

Least squares means, genetic (sigma g) and phenotypic (sigma p) standard deviations, and phenotypic coefficients of variation (CV) were estimated for growth traits of intact males from 12 breed groups combined, for nine purebreds combined, and for the F1, F2, F3, and F4 generations of three composite populations to which the nine purebreds contributed. Heritabilities (h2) and genetic (rg) and phenotypic (rp) correlations were estimated for growth traits, calving difficulty of calves with dams of different ages, and gestation length. Coefficients of variation and sigma g generally were similar for composites and contributing purebreds. Generally, estimates of h2 were similar for all breed groups combined, contributing purebreds combined, and composites combined. Estimates of h2 for calving difficulty were higher for calves with 2-yr-old dams than for calves with dams > or = 3 yr old and were sufficiently high (.27 and .31) to be a useful selection criterion for reducing calving difficulty. Mean h2 pooled within all breed groups ranged from .35 for 200-d weight and 368-d weight to .48 for 368-d height. Estimates of h2 for subjective scores of anatomical traits were only slightly lower than those for growth and size traits. The h2 of scrotal circumference (.43) was similar to those for growth and size traits. Genetic correlations between birth weight and calving difficulty were similar for 1) calves with dams of all ages, 2) calves with 2-yr-old dams, and 3) calves with dams > or = 3 yr old.(ABSTRACT TRUNCATED AT 250 WORDS)     

Genetic association of apolipoprotein E with age-related macular degeneration

Several investigators have studied the deficit in maximal voluntary force that is said to occur when bilateral muscle groups contract simultaneously. A true bilateral deficit (BLD) would suggest a significant limitation of neuromuscular control; however, some of the data from studies in the literature are equivocal. Our purpose was to determine whether there is a BLD in the knee extensors of untrained young male subjects during isometric contractions and whether this deficit is associated with a decreased activation of the quadriceps, increased activation of the antagonist muscle, or an alteration in motor unit firing rates. Twenty subjects performed unilateral (UL) and bilateral (BL) isometric knee extensions at 25, 50, 75, and 100% maximal voluntary contraction. Total UL and BL force (delta 3%) and maximal rate of force generation (delta 2.5%) were not significantly different. Total UL and BL maximal vastus lateralis electromyographic activity (EMG; 2.7 +/- 0.28 vs. 2.6 +/- 0.24 mV) and coactivation (0.17 +/- 0.02 vs. 0.20 +/- 0.02 mV) were also not different. Similarly, the ratio of force to EMG during submaximal UL and BL contractions was not different. Analysis of force production by each leg in UL and BL conditions showed no differences in force, rate of force generation, EMG, motor unit firing rates, and coactivation. Finally, assessment of quadriceps activity with the twitch interpolation technique indicated no differences in the degree of voluntary muscle activation (UL: 93.6 +/- 2.51 Hz, BL: 90.1 +/- 2.43 Hz). These results provide no evidence of a significant limitation in neuromuscular control between BL and UL isometric contractions of the knee extensor muscles in young male subjects.     

IL-3 enhances both presentation of exogenous particulate antigen in association with class I major histocompatibility antigen and generation of primary tumor-specific cytolytic T lymphocytes

Recent studies have reported that APC can present particulate exogenous Ag in the context of class I MHC to CD8+ CTL, and our laboratory demonstrated that IL-3 could enhance CTL generation to exogenous Ag. In this paper, we wished to determine whether presentation of particulate Ag could be enhanced by IL-3. A T cell hybridoma, B3Z86/90.14 (B3Z) restricted to Ova/Kb, was used as an indicator for presentation of particulate Ag with class I MHC. When activated, this hybridoma expresses lacZ, allowing a simple colorimetric measurement of Ag-specific T cell stimulation. We demonstrated that bone marrow cells stimulated by IL-3 in vivo and in vitro exhibited significantly increased presentation of exogenous OVA linked to beads. Lysate from OVA-transfected line 1 murine lung adenocarcinoma cells (line 1/OVA) was also presented by IL-3-stimulated bone marrow cells, suggesting that these APC can process tumor fragments or debris. Studies using TAP1/2-deficient mice and Ag presentation inhibitors indicate that this exogenous Ag presentation is mediated via the conventional class I MHC pathway. Adoptive transfer of IL-3-stimulated bone marrow cells pulsed with lysate from line 1/OVA tumor cells into naive recipient mice led to the generation of a potent CTL response. These observations indicate that use of such cells may provide a new avenue for development of tumor vaccines.     

Longitudinal studies of immune function in cattle experimentally infected with bovine immunodeficiency-like virus and/or bovine leukemia virus

The effects of single or dual infection with bovine immunodeficiency-like virus (BIV) and/or, bovine leukemia virus (BLV) on bovine immune function were examined over a 4 year period. Holstein calves were infected with BIV (four calves), BLV (five calves), BIV and BLV (five calves), or sham inoculated (three calves). Lymphocyte blastogenesis to mitogens, seven tests of neutrophil function, and mononuclear cell subset analysis by flow cytometry (BoCD4, BoCD8, BoCD2, BoWC1, sIgM+, and monocytes) were performed at regular intervals to 49 months post-infection. These data were analyzed for main effects of each virus and interaction as a 2 x 2 factorial. BIV infected cattle had lower neutrophil antibody-dependent cell-mediated cytotoxicity and iodination responses during 2 of the 4 years post-infection (P < 0.05). BIV infection was not associated with any long-term significant changes in lymphocyte blastogenesis to mitogens or changes in mononuclear cell subset numbers in blood. There was a tendency for animals infected with BIV alone to have decreased lymphocyte blastogenic responses to mitogens, but this was not statistically significant. BLV infection caused an increase in total mononuclear cells with no dramatic shift in the relative proportions of the various subsets. Co-infection with BIV and BLV did not consistently cause a different response than either virus did individually. One BIV infected animal died of non-BLV lymphosarcoma 7 months after infection. All other animals had no unusual clinical signs. In summary, infection with BIV caused a significant, temporary decrease in neutrophil function with no consistent statistically significant alteration in lymphocyte blastogenesis or mononuclear cell numbers during the first 4 years after infection. BLV infection caused an increase in lymphocyte numbers, and there appeared to be no synergism between the viruses.     

Detection of virus infection-associated antigen and 3D antibodies in cattle vaccinated against foot and mouth disease

The analysis of sera obtained from animals vaccinated or revaccinated with inactivated vaccines against foot and mouth disease (FMD) virus showed that these vaccines induced antibodies against the virus infection-associated (VIA) antigen, detectable by agar gel immunodiffusion (AGID). The present study evaluates the antibody response to protein 3D and the VIA antigen (VIAA) of FMD virus induced by different vaccines in a group of 51 calves. This response was detected using AGID and a liquid-phase blocking sandwich enzyme-linked immunosorbent assay (ELISA) for anti-3D antibodies (ELISA-3D). No anti-VIAA or anti-3D antibodies were detected after the initial vaccination. Following revaccination, animals giving positive results were detected by both methods. This immune response disappeared 60-120 days post-revaccination (dprv) according to the AGID method, and 90-180 dprv when ELISA-3D was used. Samples of oesophageal-pharyngeal fluid obtained from animals that remained positive for anti-VIAA antibodies at 90-120 dprv gave negative results for viral isolation, indicating that the transitional antibody response induced by the vaccine was due to the presence of non-structural antigens in the vaccine and not to viral infection. These results indicate that the ELISA-3D method could be used as a complementary method for sero-epidemiological studies as an indirect indicator of viral activity, as long as the age and vaccination status of the animals being sampled are taken into consideration.     

Detection of viral genome in non-neural tissues of cattle experimentally infected with bovine herpesvirus 1

Various bone disorders become manifest as cystic lesions. The differential diagnosis must include benign and malignant tumors and also non-tumorous lesions, such as osteomyelitis. The most important and most frequent types of genuine bone cyst are juvenile bone cyst and aneurysmal bone cyst. When juvenile bone cysts occur in adults they are called solitary bone cysts. Despite intensive research the pathogenesis of bone cysts is still unknown to this day, so that successful causal therapy is impossible. The main problem in the treatment of bone cysts is their high rate of recurrence, rates ranging between 20% and 50% having been cited in the international literature. A critical review of the literature reveals few publications with helpful follow-up results. Most of the publications are case reports, and they frequently merely describe various forms of treatment. More recent reports are mainly concerned with such methods as curettage, steroid injections, and continuous decompression with perforated screws. Until the early 1980s, segmental bone resection was the treatment of choice. Because of its high complication rate it has since been abandoned. In the last analysis, the only well-established method for which long-term results obtained in studies of any size have been published, is curettage of the cyst and grafting with cancellous bone from the iliac crest. In our series, 41 patients were treated with this method, and we recorded a recurrence rate of 17.1%. Complications were rare. The risk of recurrence depended on the age of the patient. A higher recurrence rate must be expected in children under the age of 10 years. For this reason, operative treatment should not be performed until after that age if possible. Newer methods, such as steorid injections and continuous decompression by means of perforated screws, had better results in some studies, but only according to a few authors. Further research is needed to show whether our method will yield good results in the long term when applied in larger patient collectives.     

Differential association of traits of fear and anxiety with norepinephrine- and dark-induced pupil reactivity.

The relation between fear and anxiety remains unclear, though psychometric data strongly suggest they are independent emotional systems. Because central norepinephrine (NE) projection systems are at the core of models of both fear and anxiety, the present experiment explored whether this independence extends to NE functioning. Two different aspects of NE functioning were assessed in a healthy young adult sample (N?=?18): pupillary reactivity to (a) a specific NE alpha-1 agonist challenge to assess receptor reactivity and (b) a darkness challenge to assess contributions of central NE. Pupillary reactivity to the former was strongly and specifically related to A. Tellegen's (1982) Multidimensional Personality Questionnaire (MPQ) Harm Avoidance scale (i.e., trait fear), whereas the latter was strongly and specifically related to MPQ Negative Emotionality (i.e., trait anxiety). Implications for conceptualizing fear and anxiety as emotional systems are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Apical sorting of bovine enteropeptidase does not involve detergent-resistant association with sphingolipid-cholesterol rafts

Enteropeptidase is a heterodimeric type II membrane protein of the brush border of duodenal enterocytes. In this location, enteropeptidase cleaves and activates trypsinogen, thereby initiating the activation of other intestinal digestive enzymes. Recombinant bovine enteropeptidase was sorted directly to the apical surface of polarized Madin-Darby canine kidney cells. Replacement of the cytoplasmic and signal anchor domains with a cleavable signal peptide (mutant proenteropeptidase lacking the amino-terminal signal anchor domain (dSA-BEK)) caused apical secretion. The additional amino-terminal deletion of a mucin-like domain (HL-BEK) resulted in secretion both apically and basolaterally. Further deletion of the noncatalytic heavy chain (L-BEK) resulted in apical secretion. Thus enteropeptidase appears to have at least three distinct sorting signals as follows: the light chain (L-BEK) directs apical sorting, addition of most of the heavy chain (HL-BEK) inhibits apical sorting, and addition of the mucin-like domain (dSA-BEK) restores apical sorting. Inhibition of N-linked glycosylation with tunicamycin or disruption of microtubules with colchicine caused L-BEK to be secreted equally into apical and basolateral compartments, whereas brefeldin A caused basolateral secretion of L-BEK. Full-length BEK was not found in detergent-resistant raft domains of Madin-Darby canine kidney cells or baby hamster kidney cells. These results suggest apical sorting of enteropeptidase depends on N-linked glycosylation of the serine protease domain and an amino-terminal segment that includes an O-glycosylated mucin-like domain and three potential N-glycosylation sites. In contrast to many apically targeted proteins, enteropeptidase does not form detergent-resistant associations with sphingolipid-cholesterol rafts.     

High and low activity alleles of catechol-O-methyltransferase gene: ethnic difference and possible association with Parkinson's disease

Catechol-O-methyltransferase (COMT) is an enzyme that inactivates catecholamines such as adrenaline, noradrenaline, dopamine, and levodopa. Recently an amino acid change (Val-108-Met) of the COMT protein was found to determine high and low activity alleles of the COMT gene. We genotyped 109 Japanese patients with Parkinson's disease (PD) and 153 controls by using polymerase chain reaction (PCR) amplification and digestion by the restriction enzyme NlaIII. The frequency of low activity allele in the controls was 0.29, which was significantly different from that reported in Caucasians (0.50). When comparison was made between patients with PD and controls, homozygosity for the low activity allele was significantly more common among the patients than among the controls (P = 0.017; odds ratio, 2.8, 95% CI 1.2-6.5), suggesting that homozygosity for the low activity allele may increase susceptibility to PD.     

Severe deficiency of 1,25-dihydroxyvitamin D3 in human immunodeficiency virus infection: association with immunological hyperactivity and only minor changes in calcium homeostasis

The serum level of 1,25-dihydroxyvitamin D3 [1,25-(OH)2D], the biologically most potent metabolite of vitamin D, is tightly regulated within narrow limits in human healthy adults. 1,25-(OH)2D deficiency is rare and is associated with disturbances in calcium and bone metabolism. We have previously reported a marked decrease in serum levels of 1,25-(OH)2D in human immunodeficiency virus (HIV)-infected patients. The present study was designed to further examine the causes and consequences of severe 1,25-(OH)2D deficiency in these patients. The design was a prospective cohort study. Fifty-four HIV-infected patients clinically classified according to the revised criteria from Centers for Disease Control and Prevention and healthy controls were studied. Parameters related to vitamin D and calcium metabolism as well as immunological and nutritional status were determined. Twenty-nine of the patients (54%) had serum levels of 1,25-(OH)2D below the lower reference limit, and 18 of these had undetectable levels. In contrast, HIV-infected patients had normal serum levels of 25-hydroxyvitamin D and vitamin D-binding protein. HIV-infected patients as a group had modestly depressed serum calcium and PTH levels. There were, however, no correlations between these parameters and serum levels of 1,25-(OH)2D. There were no differences in serum calcium or PTH levels or nutritional status when patients with severe 1,25-(OH)2D deficiency were compared to other patients, but patients with undetectable 1,25-(OH)2D had significantly elevated serum phosphate levels. Furthermore, patients with undetectable 1,25-(OH)2D levels were characterized by advanced clinical HIV infection, low CD4+ lymphocyte counts, and high serum levels of tumor necrosis factor-alpha (TNFalpha). We conclude that inadequate 1alpha-hydroxylation of 25-hydroxyvitamin D seems to be the most likely cause of 1,25-(OH)2D deficiency in HIV-infected patients, possibly induced by an inhibitory effect of TNFalpha. The low 1,25-(OH)2D and high TNFalpha levels observed may impair the immune response in HIV-infected patients both independently and in combination and may represent an important feature of the pathogenesis of HIV-related immunodeficiency. Markedly depressed 1,25-(OH)2D serum levels are also present in certain other disorders characterized by immunological hyperactivity. Thus, the findings in the present study may not only represent a previously unrecognized immune-mediated mechanism for induction of 1,25-(OH)2D deficiency in human disease, but may also reflect the importance of adequate serum levels of 1,25-(OH)2D for satisfactory performance of the immune system in man.     

Increased ratio of bcl-2/bax expression is associated with bovine leukemia virus-induced leukemogenesis in cattle

To further investigate the molecular basis underlying the dysregulation of B cell homeostasis associated with bovine leukemia virus disease progression in cattle, bovine bax was cDNA cloned and sequenced. The predicted amino acid sequence of bovine Bax revealed a 192-amino-acid protein having extensive identity with the human (97%), murine (93%), and rat (94%) homologues. Because the ratio of Bcl-2 to Bax is believed to predetermine the susceptibility to a given apoptotic stimulus, the relative expression of the genes encoding these oncoproteins was evaluated in cattle naturally infected with BLV. In BLV-infected cattle an increase in the ratios of bcl-2/bax mRNA and protein expression correlated with advancing stages of disease. These findings suggest that in addition to the maintenance of BLV-associated hematopoietic malignancies, the reciprocal expression of Bcl-2/Bax may modulate the induction of B cell expansion typical of BLV disease progression.