The acute-phase protein serum amyloid A3 is expressed in the bovine mammary gland and plays a role in host defence.

The serum amyloid A protein is one of the major reactants in the acute-phase response. Using representational difference analysis comparing RNA from normal and involuting quarters of a dairy cow mammary gland, we found an mRNA encoding the SAA3 protein (M-SAA3). The M-SAA3 mRNA was localized to restricted populations of bovine mammary epithelial cells (MECs).

Genetically hypertensive Brown Norway congenic rat strains suggest intermediate traits underlying genetic hypertension.

Aim: To determine the independent and combined effects of three quantitative trait loci (QTL) for blood pressure in the Genetically Hypertensive (GH/Omr) rat by generating and characterizing single and combined congenic strains that have QTL on rat chromosomes (RNO) 2, 6, and 18 from the GH rat introduced into a hypertension resistant Brown Norway (BN) background.

Methods: Linkage analysis and QTL identification (genome wide QTL scan) were performed with MapMaker/EXP to build the genetic maps and MapMaker/QTL for linking the phenotypes to the genetic map. The congenic strains were derived using marker-assisted selection strategy from a single male F1 offspring of an intercross between the male GH/Omr and female BN/Elh, followed by 10 generations of selective backcrossing to the female BN progenitor strain.

The BSP30 salivary proteins from cattle, LUNX/PLUNC and von Ebner's minor salivary gland protein are members of the PSP/LBP superfamily of proteins.

Saliva influences rumen function in cattle, yet the biochemical role for most of the bovine salivary proteins (BSPs) has yet to be established. Two cDNAs (BSP30a and BSP30b) from bovine parotid salivary gland were cloned and sequenced, each coding for alternate forms of a prominent protein in bovine saliva. The BSP30 cDNAs share 96% sequence identity with each other at the DNA level and 83% at the amino acid level, and appear to arise from separate genes.

Differential expression of the whey acidic protein gene during lactation in the brushtail possum (Trichosurus vulpecula).

The whey acidic protein (WAP) is a whey protein found in the milk of a number of species. We have isolated and characterised a WAP cDNA clone from the brushtail possum (Trichosurus vulpecula) and examined its expression in the mammary gland. The amino acid sequences of WAP from the possum and another marsupial, the tammar wallaby, share 69% identity, however, less sequence identity exists between the marsupial and eutherian WAP sequences (30-37%).

Somatostatin enhances cAMP-dependent short-circuit current in human colon via somatostatin receptor subtype-2.

Somatostatin inhibits colonic ion secretion in animal models and cultured intestinal cell lines via somatostatin receptor subtype 2 and subtype 1, respectively. In a recent in vitro ion transport study of the human colon, somatostatin was shown to stimulate short-circuit current, a measure of electrogenic ion transport. In this study we have used the reverse-transcription polymerase chain reaction (RT-PCR) and measurements of changes in short-circuit current (Isc) in response to receptor subtype-specific analogs of somatostatin, to define the somatostatin receptor subtype responsible for the stimulation of short-circuit current in human colon.

Somatostatin and octreotide stimulate short-circuit current in human colonic epithelium.

In vitro somatostatin is a potent inhibitor of intestinal ion secretion in animal models and cultured human cell lines, providing a rationale for its use in secretory diarrheas. However, the effects of somatostatin on ion transport in native human colonic epithelium have not been reported. In this study the effects of somatostatin and octreotide on the basal short-circuit current and the cAMP- and Ca2+-stimulated short-circuit current were studied in isolated human colonic mucosa mounted in Ussing chambers.

Nuclear localisation of the transcription factor Stat5b is associated with ovine milk protein gene expression during lactation but not during late pregnancy or forced weaning.

Localisation patterns of the transcription factor Stat5b in the udders from pregnant, lactating and involuting ewes were compared with the expression patterns of two major milk protein genes alpha-lactalbumin and alphaS1 casein. Stat5b was detected in the cytoplasm and nuclei of epithelial cells at all stages of mammary gland development. A consistent positive relationship between the nuclear localisation of Stat5b in lactating mammary alveolar epithelial cells, and the presence of milk protein gene mRNA was apparent during lactation and early involution.

Comparative aspects of milk caseins.

The caseins comprise the major protein component of milk of most mammals and are secreted as micelles that also carry high concentrations of calcium. They are phosphoproteins that represent the products of four genes, equivalent to those that encode the bovine alpha s1, alpha s2, beta, and kappa-caseins. There is considerable variation in the relative proportions of the particular caseins across species.

Cloning and expression of the transferrin and ferritin genes in a marsupial, the brushtail possum (Trichosurus vulpecula).

Transferrin and ferritin cDNAs have been isolated and characterised from the common brushtail possum (Trichosurus vulpecula), the first marsupial examples of these genes. The transferrin cDNA encodes a 711 amino acid pre-protein which shows high levels of amino acid identity with eutherian transferrins (58-60%) and lactoferrins (54-56%). Phylogenetic analysis suggests that the possum transferrin has evolved independently along a pathway distinct from that of the eutherian transferrins and lactoferrins.

Identification, characterisation and cDNA cloning of two caseins from the common brushtail possum (Trichosurus vulpecula)1.

Two major caseins have been isolated from the milk of the common brushtailed possum (Trichosurus vulpecula). These have been identified as alpha- and beta-casein on the basis of the similarity of their N-terminal sequences to those of the caseins of another marsupial (Macropus eugenii). Both proteins appear to exist in multiple forms.

Cosegregation of the Tnfalpha locus with cardiovascular phenotypes in the F2 generation of a New Zealand genetically hypertensive and Brown Norway cross.

1. The association of the Tnfalpha locus with several cardiovascular phenotypes and body mass has been studied in the F2 generation of a reciprocal cross between rats of the New Zealand genetically hypertensive (GH) and the normotensive Brown Norway (BN) strains. In the total F2 population the GH allele of Tnfalpha cosegregated with increased intra-arterial blood pressure (BP) in a recessive manner.

Differential expression of milk protein genes during lactation in the common brushtail possum (Trichosurus vulpecula).

In the common brushtail possum (Trichosurus vulpecula) lactation lasts for 200 days and consists of two distinct phases. Milk composition changes dramatically between phase 2 and 3, which correspond to early and late lactation respectively (phase 1 corresponds to pregnancy). RNA expression patterns have been established for eight major milk protein genes throughout lactation in possum mammary glands.

Phylogenetic analysis of three lipocalin-like proteins present in the milk of Trichosurus vulpecula (Phalangeridae, Marsupialia).

Three proteins have been identified in the milk of the common brush tail possum. Trichosurus vulpecula that from sequence analysis are members of the lipocalin family. They include beta-lactoglobulin, which appears to have two forms; a homologue to the late-lactation protein found in tammar, Macropus eugenii; milk; and a novel protein termed trichosurin.

Lysozyme and alpha-lactalbumin from the milk of a marsupial, the common brush-tailed possum (Trichosurus vulpecula).

Lysozyme and alpha-lactalbumin have been identified using N-terminal sequence analysis of whey proteins from the common brush-tailed possum, Trichosurus vulpecula after separation by two-dimensional denaturing electrophoresis. Both proteins were purified from pooled possum milk using ion exchange chromatography and gave mass values of 14,896 and 13,985 Da respectively by MALDI-TOF mass spectrometry. Clones containing the full coding sequences of the genes for both proteins were isolated from a possum mammary cDNA library and the DNA sequence of the coding region determined.

Glucose transport in a murine mammary epithelial cell line.

The glucose transport systems of the COMMA-D cell line (a murine mammary epithelial cell line) were examined using 2-deoxyglucose as substrate. The kinetics and inhibition studies with other sugars including xylose suggested that the transport system had properties of both GLUT-1 and Glut-3. Subsequent analysis of mRNA transcripts using cDNAs for GLUT-1 to 4 showed that only GLUT-1 was expressed in the COMMA-D cells.

Interaction of interferon-gamma and epidermal growth factor in the regulation of nitric oxide production and cellular proliferation in a cultured murine mammary cell line, COMMA-D.

The regulation of nitric oxide synthase (NOS) activity and gene expression by cytokines and growth factors has been studied using the murine mammary epithelial cell line, COMMA-D. NOS activity was stimulated by exposure to interferon-gamma (IFN-gamma) and could be further stimulated by tumour necrosis factor-alpha (TNF-alpha) and epidermal growth factor (EGF) although neither was affective alone. The maximal activity observed in the presence of IFN-gamma and EGF was not affected by the order in which cells were exposed.

A novel marsupial protein expressed by the mammary gland only during the early lactation and related to the Kunitz proteinase inhibitors.

A novel whey protein has been found in marsupial milk, the early lactation protein (ELP). The whey of Trichosurus vulpecula, the Australian common brush-tailed possum, contains two forms of ELP, estimated by protein electrophoresis at 8 and 16 kDa. The 16-kDa form contains approximately 60% N- linked carbohydrate.

Interleukin-1 beta and tumor necrosis factor-alpha stimulate the cat-2 gene of the L-arginine transporter in cultured vascular smooth muscle cells.

The production of nitric oxide (NO) from L-arginine by nitric oxide synthase (NOS) in cytokine-stimulated vascular smooth muscle cells (VSMC) is thought to play an important role in the pathophysiology of several vascular disease states including septic shock. This study examines the relationship between cytokine-stimulated NO production and L-arginine transport in cultured VSMC. Cultured VSMC from rat aorta were stimulated with interleukin-1 beta, tumor necrosis factor-alpha, and/or angiotensin II (Ang II); and the accumulation of nitrite, a stable product of NO metabolism, in the culture media and the rates of net L-arginine uptake were measured.

Angiotensin II stimulates system y+ and cationic amino acid transporter gene expression in cultured vascular smooth muscle cells.

The effect of angiotensin II (Ang II) on the transport of cationic amino acids has been examined in vascular smooth muscle cells (VSMC) isolated from rat aortae. Ang II stimulated the uptake rates of radiolabeled arginine and lysine in a time- and concentration-dependent manner. The stimulated arginine uptake could be blocked by pretreatments with cycloheximide and actinomycin D or co-treatment with valsartan, an antagonist specific for Ang II receptor subtype-1.

Expression of somatostatin receptor genes and their role in inhibiting Cl- secretion in HT-29cl.19A colonocytes.

Recent studies in a cultured model of the intestinal epithelium (HT-29cl.19A) have shown that somatostatin-14 (SS-14) inhibits the Cl- secretory process by acting at multiple G protein-dependent sites. These actions may underlie the antidiarrheal properties of SS peptides.

Isolation of cDNAs and tissue specific expression of ovine glucose transporters.

The facilitative glucose transporters are a family of proteins responsible for the transmembrane transport of glucose and other hexose sugars (1,2). In mammals, the seven glucose transporter isoforms display a characteristic tissue distribution reflecting the physiological requirement and metabolism of glucose. This report describes the isolation and sequencing of the full length ovine GLUT-3 cDNA and the tissue distribution of ovine GLUT-1 and GLUT-3 mRNA.

Heart mass and blood pressure have separate genetic determinants in the New Zealand genetically hypertensive (GH) rat.

Objective: To determine associations between cardiovascular parameters and genotype in 205 F2 rats of both sexes and lineages from reciprocal crosses made between rats of the New Zealand genetically hypertensive (GH) and Brown Norway (BN) rat strains.

Methods: Systolic tail blood pressure, mean arterial blood pressure, pulse rate, heart mass, body mass and relative heart mass were determined for each rat in the age range 17-19 weeks, and DNA polymorphisms were examined for the guanylyl cyclase A (GCA), angiotensin converting enzyme (ACE) and renin (REN) genes.

Results: The phenotypic data indicated the presence of genes on the X and Y chromosomes that affected blood pressure.