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At least 271 records · Page 15Linked to original sources
PubMed1999

A quantitative trait locus for live weight maps to bovine chromosome 23.

A multiple-marker mapping approach was used to search for quantitative trait loci (QTLs) affecting production, health, and fertility traits in Finnish Ayrshire dairy cattle. As part of a whole-genome scan, altogether 469 bulls were genotyped for six microsatellite loci in 12 families on Chromosome (Chr) 23. Both multiple-marker interval mapping with regression and maximum-likelihood methods were applied with a granddaughter design. Eighteen traits, belonging to 11 trait groups, were included in the analysis. One QTL exceeded experiment level and one QTL genome level significance thresholds. Across-families analysis provided strong evidence (P(experiment) = 0.0314) for a QTL affecting live weight. The QTL for live weight maps between markers BM1258 and BoLA DRBP1. A QTL significant at genome level (P(genome) = 0.0087) was mapped for veterinary treatment, and the putative QTL probably affects susceptibility to milk fever or ketosis. In addition, three traits exceeded the chromosome 5% significance threshold: protein percentage of milk, calf mortality (sire), and milking speed. In within-family analyses, protein percentage was associated with markers in one family (LOD score = 4.5).

Alleles↗
PubMed1994

Cloning and sequencing of four structural genes for the Na(+)-translocating NADH-ubiquinone oxidoreductase of Vibrio alginolyticus.

Oligonucleotide probes based on the N-terminal amino acid sequences of the NqrA and NqrC subunits were used to clone genes for the Na(+)-dependent NADH-ubiquinone oxidoreductase complex from Vibrio alginolyticus. Four consecutive ORFs were identified encoding subunit proteins of 48.6, 46.8, 27.7 and 22.6 kDa, respectively (NqrA-D). A further ORF, showing 71% homology to the BolA protein of Escherichia coli, was located upstream. From sequence comparisons, we conclude that the Na(+)-dependent NADH-ubiquinone oxidoreductase complex of V. alginolyticus is clearly distinct from the corresponding H(+)-dependent enzymes of both prokaryotes and eukaryotes.

Amino Acid Sequence↗
PubMed1988

Qualitative variation in the immune response to ovarian follicular fluid proteins in cattle.

In two experiments 44 heifers were immunised with inhibin preparations of partially purified ovarian follicular fluid (PPFF) of ovine, porcine and equine origin in non-ulcerative Freund's adjuvant. In the first experiment 20 cattle were immunised with ovine PPFF1. In the second, a further 24 were immunised with either a different sheep PPFF (ovine PPFF2) or PPFF from pigs or horses. Large molecules of over 30 kDa initiated a strong immune response in some animals, but none in others, although there was no obvious relationship with the MHC Class I (BoLA) phenotype. Immunisation with ovine partly purified follicular fluid affected ovulation rate in some cattle, and only these cows produced detectable antibodies to an approximately 40 kDa protein. These data show that the immunological response of cattle to a mixture of antigens is highly variable between individuals and may explain the variation in the ovarian response to immunological manipulations of ovulation rate seen in this species.

Animals↗
PubMed1987

The development and analysis of species specific and cross reactive monoclonal antibodies to leukocyte differentiation antigens and antigens of the major histocompatibility complex for use in the study of the immune system in cattle and other species.

We examined the potential of developing a set of species specific and cross reactive monoclonal antibodies (MoAbs) for use in the study of the phylogenetic and functional relation of class I and class II antigens of the major histocompatibility complex (MHC) and leukocyte differentiation antigens in cattle and other species. Comparing immunization strategies demonstrated the number of hybrids producing cross reactive antibodies can be increased by hyperimmunization of mice with lymphoid cells from multiple species. Comparing various methods of assay (antibody-complement mediated cytotoxicity [CT], enzyme linked immunosorbent assay [ELISA] and flow microfluorimetry [FMF]), revealed FMF is the most useful technique for the primary assay of hybridomas producing MoAbs of potential interest. By using dual parameter and dual fluorescence analysis, we could determine whether a given MoAb reacted with mononuclear cells (lymphocytes and monocytes) and/or granulocytes, and also whether any two MoAbs of different isotype and specificity recognized antigens present on identical or separate populations of leukocytes. Comparing the patterns of MoAb reactivity with leukocytes obtained from cows, goats, sheep, pigs, horses and humans, as well as comparing the patterns of reactivity with a panel of lymphoid cell lines derived from cattle (with enzootic bovine leukemia) and humans (with various forms of leukemia), revealed sets of MoAbs reactive with unique antigenic determinants present on BoLA class I (15 MoAbs) and class II (9 MoAbs) antigens, and also MoAbs reactive with determinants present on leukocyte differentiation antigens (36 MoAbs). Dual fluorescence analysis demonstrated the antigens detected by some MoAbs are predominantly expressed on one lineage of leukocytes while others are expressed on two or more lineages of leukocytes. Dual and single fluorescence analysis also demonstrated the PNA receptor(s) is: expressed on T cells, granulocytes and class II antigen monocytes and absent or expressed in low amount on sIgM+ B cells and a newly defined Non T/Non B population of cells. The strategies described for identifying and analyzing the specificity of MoAbs demonstrate the feasibility of developing a set of cross reactive MoAbs for identifying homologous molecules in multiple species and delineating their functional and phylogenetic relation.

Animals↗
PubMed1989

Selected phenotypic and cloning properties of a bovine lymphoblastoid cell line, BL20.

A bovine lymphoblastoid cell line, BL20, was shown to express a BoLA antigen and surface IgM, implying that it was probably of B-cell origin. At low density (less than 10(5)/ml), the cells failed to grow. Inclusion of growth factor-containing supernatants from concanavalin A or pokeweed mitogen (PWM)-activated bovine lymphocytes or from thymus fibroblast-like cells did not improve cloning of the cells. Feeder cells also did not enhance cloning of the cells except for bovine thymus fibroblast-like cell monolayer.

Animals↗
PubMed1989

Recombinant bovine interferon-gamma enhances expression of class I and class II bovine lymphocyte antigens.

Recombinant bovine interferon-gamma augments expression of class I and class II histocompatibility antigens on the surface membrane of bovine lymphocytes. Immunofluorescence techniques using a series of monoclonal anti-HLA antibodies demonstrate that this enhancement is detectable as early as 24 h after incubation with rBoIFN, while maximum surface expression is obtained within 3-5 days. A concentration as low as 10 units of rBoIFN is effective. Such results may be useful for characterizing the BoLA gene products.

Animals↗
PubMed1996

Altered expression of class I major histocompatibility antigens on bovine cells infected with the protozoan parasite Theileria annulata.

Theileria annulata infection is controlled by MHC class I restricted cytotoxic T cell killing of infected cells. The long term persistence of parasite suggests that it employs a mechanism to avoid immune surveillance. In this paper we describe the modification of BoLA class I antigens on the surface of Theileria annulata transformed lymphoblastoid cells. The extent of modification is different between individuals varying from extensive to undetectable.

Animals↗
PubMed2005

Mitochondrial leader sequence--plasmid DNA conjugates delivered into mammalian cells by DQAsomes co-localize with mitochondria.

In the last decade the increase in therapeutic strategies aimed at mitochondrial targets has resulted in the need for novel delivery systems for the selective delivery of drugs and DNA into mitochondria. In this study, we have continued our efforts towards the development of the first mitochondriotropic drug and DNA delivery system (DQAsomes). Prepared from derivatives of the self-assembling mitochondriotropic bola-amphiphile dequalinium chloride, these vesicles bind and transport DNA to mitochondria in living mammalian cells where upon they have been shown to release the DNA on contact with mitochondrial membranes. We present data to demonstrate that oligonucleotides as well as plasmid DNA conjugated to a mitochondrial leader sequence (MLS) co-localize with mitochondria when delivered into mammalian cells by DQAsomes. In contrast to a commercially available DNA delivery vector, our vesicles appear to have a pronounced specificity for mitochondria. Further, the data strongly suggest that linear conjugates might be better suited to delivery into mitochondria and that in the absence of a mitochondria specific vector, the presence of a MLS-peptide conjugated to the DNA is alone not sufficient to direct the accumulation of DNA at mitochondria.

Cell Line, Tumor↗
PubMed2003

DQAsome-mediated delivery of plasmid DNA toward mitochondria in living cells.

DQAsomes are mitochondriotropic cationic 'bola-lipid'-based vesicles, which have been developed by us for the transport of drugs and DNA to mitochondria in living cells. This has made direct mitochondrial gene therapy feasible for the very first time. Our strategy for the delivery of DNA into the matrix of mitochondria is based upon the DQAsomal transport of a DNA-signal peptide conjugate to mitochondria, the selective liberation of this conjugate from DQAsomes at the mitochondrial membrane followed by DNA uptake via the mitochondrial protein import machinery. Using membrane-mimicking liposomes and isolated rat liver mitochondria we have shown earlier that DQAsome-DNA complexes (DQAplexes) selectively release pDNA when in contact with mitochondria-like membranes. Employing a newly developed protocol for selectively staining free pDNA in the cytosol of living cells and based on confocal fluorescence microscopic imaging we demonstrate here that DQAplexes appear to be able to escape from endosomes without loosing their pDNA load and transport the pDNA to the site of mitochondria at which at least a portion of the pDNA is released from its DQAsomal carrier. Free pDNA could not be detected anywhere else inside the cytosol of transfected cells demonstrating the target-selectivity of DQAsome-mediated DNA delivery to mitochondria.

Animals↗
PubMed2001

Identification of CD4+ T cell epitopes on the fusion (F) and attachment (G) proteins of bovine respiratory syncytial virus (BRSV).

To gain insight into the antigenic structure of the F and G proteins of BRSV, we have mapped CD4+ T cell epitopes on these proteins using synthetic peptides and lymphocytes from vaccinated, naturally infected or experimentally infected calves, in proliferation assays. Bovine CD4+ T cells recognised epitopes that were distributed predominantly within the F1 subunit of the F protein, some of which were adjacent to previously identified B cell epitopes. Bovine CD4+ T cell epitopes within the G protein were mainly located within the cytoplasmic tail. Several immunodominant bovine T cell epitopes within the F protein, that were recognised by calves with different haplotypes, are also recognised by human T cells. Thus, cattle and humans appear to recognise similar T cell epitopes on the F protein. Studies using antibodies to bovine MHC class II and BoLA DR-transfected CHO cells as antigen-presenting cells indicated that immunodominant regions of the F and G proteins contained both DR- and DQ-restricted epitopes. The finding that there was little recognition of the extracellular domain of the G protein by T cells has important implications for vaccine design based on the soluble form of this protein.

Animals↗
PubMed2001

Recombinant hemagglutinin protein of rinderpest virus expressed in insect cells induces humoral and cell mediated immune responses in cattle.

Rinderpest virus causes a highly contagious and often fatal disease in domestic and wild ruminants. The surface glycoproteins, hemagglutinin (H) and fusion (F) proteins of this enveloped virus are known to confer protective immunity in cattle. We have reported the generation of a recombinant baculovirus expressing H protein and studied its protective properties in cattle. In this report, we demonstrate that the recombinant baculovirus encoded H protein expressed in insect cells gets incorporated into extracellular baculovirus. Single administration of low doses of purified recombinant extracellular virus with or without adjuvant induces virus neutralizing antibody responses and bovine leukocyte antigen (BoLA) class II restricted helper T cell responses in cattle.

Animals↗
PubMed1992

Somatic cell mapping, polymorphism, and linkage analysis of bovine prolactin-related proteins and placental lactogen.

The bovine prolactin gene family includes novel members expressed in the fetal placenta that are distinct from placental lactogen. In this study, we investigated the genetic organization of four members of this gene family (PRP1, PRP3, PRP6, and PRP10) as well as placental lactogen (PL). Using a bovine-rodent hybrid somatic cell panel, all five genes were assigned to bovine chromosome 23, which contains prolactin and the major histocompatibility group (BOLA). Restriction fragment length polymorphisms were detected by all probes in breeding populations with the restriction enzyme MspI, whereas no polymorphisms were detected with BamHI. EcoRI, HindIII, TaqI, and PstI produced polymorphic fragments with some but not all of the probes tested. A PRP10 polymorphism, which is apparently the result of a insertion/deletion event, detected polymorphism frequency differences between Bos indicus and Bos taurus. No recombinational events were observed with these probes and prolactin using linkage analysis involving 91 American Holsteins. The bovine prolactin gene family was incorporated into a linkage group containing CYP21. Our studies demonstrate that members of the bovine prolactin gene family have a close physical association with each other, and all members demonstrate genetic variability in the breeding population.

Animals↗
PubMed2006

Detection of smoked paprika "Pimentón de La Vera" adulteration by free zone capillary electrophoresis (FZCE).

The purpose of this work was to develop a procedure based on protein analysis by free zone capillary electrophoresis (FZCE) that can be used in the determination of smoked paprika "Pimentón de La Vera" adulteration with paprika elaborated from varieties of pepper foreign to the "La Vera" region, in central western Spain. Two autochthonous varieties of pepper, Jaranda and Bola, and the variety Papri Queen, foreign to the "La Vera" region, were used in the study. Several aqueous solutions for solubilization of the methanol-soluble proteins were tested, and the FZCE conditions of capillary dimensions, FZCE buffer concentrations, and detection wavelengths were optimized. On the basis of the results, 30% (v/v) acetonitrile was adopted as the suspending solution for routine analysis, and the optimal FZCE parameters were 75 microm inner diameter and 57 cm total length capillaries, 8.75 mM phosphate/20.6 mM tetraborate as run buffer, and 256 nm as detection wavelength. This method was found to give excellent repeatability of the corrected migration time (CMT) with coefficients of variation (RSD %; n = 5) of <1% for most of the proteinaceous compounds analyzed and showed greater effectiveness in discriminating paprika varieties than the SDS-PAGE technique. Four peaks found in the FZCE electropherograms were investigated as a basis for detecting and estimating the adulteration of smoked paprika with paprika elaborated from the Papri Queen variety. The adulteration detection limits varied from 5 to 40% of the Papri Queen variety within a satisfactory working range of mixture (5-80%) sufficiently large to cover the adulteration levels of interest. The use of peak 6 as a marker for determining adulteration gave the best results, with an adulteration detection limit of 5-10% (w/w).

Buffers↗
PubMed1997

Shortcut Syntheses of Naturally Occurring 5-Alkylresorcinols with DNA-Cleaving Properties.

Resorcinols such as 1-5 bearing long alkyl- or alkenyl substituents at the C-5 position, including bola-formed bis-resorcinol derivatives, have recently been isolated from natural sources and were shown to exhibit exceptional DNA-cleaving properties under oxidative conditions. Previous synthetic approaches to such compounds seem inappropriately lengthy with regard to their structural simplicity. Disclosed is a very flexible synthesis which assembles these targets from triflate 7 and well accessible alkenes, dienes, enynes, or dienynes, respectively, by means of a boron-mediated reaction manifold. As a typical example, hexadeca-1,15-dien-8-yne 11 is hydroborated with 9-H-9-BBN at all possible sites, the alkenyl borane entity of the resulting tris-borane 12 is selectively cleaved off to afford the desired (Z)-alkene group in a stereoselective manner, the remaining two terminal alkylboranes are treated with NaOMe, and the bis-borate complex 13 thus formed is finally used as the nucleophile for a palladium-catalyzed Suzuki cross-coupling reaction with triflate 7. This sequence is carried out in one pot and provides product 14 in 62% overall yield. Demethylation of 14 (and analogues) can be conveniently achieved by means of 9-iodo-9-BBN to afford the natural product 5. The efficiency and flexibility of this unprecedented approach which combines different features of classical and modern boron chemistry is further demonstrated by the synthesis of anacardic and ginkgolic acid derivatives.

Journal Article↗
PubMed2005

Organized assemblies in bolaamphiphile/oppositely charged conventional surfactant mixed systems.

Five ionic bolaamphiphiles were synthesized and the aggregation behavior of bola single systems and bola/oppositely charged conventional surfactant mixed systems was studied. Small spherical vesicles were formed in all these mixed systems revealed by transmission electron microscopy (TEM). Variation of the structure of the hydrophobic chain of bolaamphiphiles has great influences on the vesicle formation ability. Vesicles were also found in the single system of a carboxylate bolaamphiphile, which was attributed to the hydrolysis of the bolaamphiphile. The results of FT-IR and X-ray diffraction (XRD) showed that bolaamphiphiles spanned through the vesicle membranes in these mixed systems. Super thermostability of the vesicles in this kind of mixed system was also investigated.

Cations↗
PubMed1996

The PCR typing of MHC-DRB genes in the sheep using primers for an intronic microsatellite: application to nematode parasite resistance.

The strong association between polymorphisms in an intronic microsatellite and the coding sequences for (BoLA)-DRB3 genes, previously described for demonstrating alleles of class II major histocompatibility complex (MHC) in the cow, was examined in sheep to see if similar polymorphisms could be demonstrated in the DRB region of the MHC. The bovine primes LA53 and LA54, previously used to amplify the bovine DRB3 microsatellites, were used with DNA from Australian sheep, eight DRB alleles were identified by length polymorphisms of polymerase chain reaction (PCR) products amplified from the DRB microsatellite region. Incomplete amplification of both alleles was sometimes found for sheep DNA samples using bovine primers, so a modified primer (LA53b) was used, and found to amplify the microsatellite next to intron 2 of the MHC more reliably than the LA53 primer. Two additional primers (LA31 and LA32), used in amplification of the exon 2 region of bovine DRB3, were used in the sheep, and the PCR products were analysed by single-stranded conformation polymorphism (SSCP). These primers successfully amplified the variable region of the ovine DRB region coded by exon 2, and the SSCP technique demonstrated polymorphisms with sheep DNA. Family studies demonstrated the segregation of alleles, by amplification both of intronic microsatellites and of the exon 2 variable region. Close correspondence was found between the two regions for several alleles, suggesting that the intronic microsatellites were closely linked to DRB-variable region alleles. Three families of Merino sheep with different antibody responses to intestinal nematode parasites were examined. The sire group with the highest antibody levels possessed two microsatellite alleles of closely similar length (alleles 3 and 4) inherited from the sire and present in high frequency in the lambs. In contrast, the other two sires did not possess these two alleles and the alleles were in low frequency in their progeny. Further studies are required in unrelated sheep to confirm whether these two alleles are associated with resistance to nematode parasites.

Alleles↗
PubMed1998

Mapping the T cell epitopes of the Babesia bovis antigen 12D3: implications for vaccine design.

The Babesia bovis antigen 12D3 was analysed to identify potential T-cell epitopes. Two predictive algorithms identified 13 possible sites but there was minimal agreement between the different predictive methods. Experimental determination of the T-cell epitopes recognized by nine cattle was achieved using a panel of overlapping peptides which identified seven different epitopes, five of which were clustered together around residues 210-320 of the molecule. No T cell epitopes were located within the tightly disulphide bonded core of 12D3. Using a series of truncated peptides, the location of two of the epitopes was mapped to residues 35-43 and 266-275. The sequences of these two epitopes was compared with a database of previously described binding motifs for MHC II alleles and each epitope was found to contain three sequence motifs recognized by HLA-DR alleles. The BoLA-DRB3 alleles occurring in these cattle were determined by a sequence specific oligonucleotide hybridization assay. Within those cattle whose T cells proliferated in response to 12D3, there was a consistent pattern of epitope recognition and presence of particular DRB3 alleles. The implications for effective vaccine design are discussed.

Alleles↗