Relationship between estimated breeding value for somatic cell count and prevalence of intramammary infection in dairy goats.

UNLABELLED: Abstract AIM: The objective of the study was to determine if dairy goats selected as having either Low or High estimated breeding values for somatic cell count (EBV SCC) would differ in prevalence of intramammary infection (IMI). METHODS: The EBV SCC for all does in two dairy goat herds were calculated based on the three or four herd test results for somatic cell count (SCC) from the preceding lactations. Within each herd does were categorised into six age groups (i.e. 2, 3, 4, 5, 6 and >6 years old) and ranked on EBV SCC. Approximately the top (High group; n=149) and bottom (Low group; n=148) 12.5% of the does within each age group within each herd had milk samples collected from each gland on one calendar day for subsequent microbiology. The presence of any IMI or IMI due to a major pathogen at goat level was modelled using a GLM with a binomial link function. RESULTS: There was no difference between the High and Low groups in terms of age, days in milk at the time of sampling or in the proportion of the Saanen breed. Mean EBV SCC was -1.0 (SD 0.4) vs 0.8 (SD 0.4) for the Low and High groups, respectively, and there was no overlap in EBV SCC between groups. Goats in the Low group had lower prevalence of any IMI than those in the High group (0.67 (95% CI=0.58-0.76) vs 0.81 (95% CI=0.74-0.88); p=0.002). Goats in the High group were 8.4 (95% CI=1.9-38.0) times more likely to have IMI due to a major pathogen infection than goats in the Low group (p=0.006). CONCLUSIONS: Does with a high EBV SCC had a higher prevalence of any IMI and were more likely to have an IMI due to a major pathogen than does with a low EBV SCC. Thus selection for EBV SCC is likely to result in a lower SCC and also lower prevalence of IMI.

Composition and distribution of fatty acids in triglycerides from goat infant formulas with milk fat.

Most infant formulas use vegetable oils in place of milk fat to provide an overall fatty acid profile similar to that of breast milk. Vegetable oils have 5 to 20% saturated fatty acids in the sn-2 position of triglycerides unless they are modified by interesterification. Interesterification is increasingly used for the fat for infant formulas to raise the level of saturated fatty acids in the sn-2 position to 40 to 60%. The objective of this study was to verify an alternative approach to providing the appropriate fatty acid profile, including in the sn-2 position, for a goat infant formula. In this method, 55% of total fat was made from goat milk fat and 45% from a mixture of unmodified high oleic sunflower, canola, and sunflower oils in a ratio of 44:30:26. The fatty acid profile was measured by gas-liquid chromatography and the relative percentage of fatty acids in the sn-2 position of triglycerides was measured via partial deacylation with Grignard reagent using trimethylsilyl derivatives of monoacylglycerols. Mixing goat milk fat with vegetable oils produced a formula with a profile of essential fatty acids and a ratio of linoleic:alpha-linolenic fatty acids within the required interval of 5 to 15:1 recommended for infant formula. The proportion of palmitic acid in the sn-2 position was 31%.

Effects on adhesion molecule expression and lymphocytes in the bovine mammary gland following intra-mammary immunisation.

Changes to adhesion molecule expression and lymphocyte populations were evaluated in alveolar mammary tissue collected from cows following an immunisation protocol that involved intra-mammary inoculation to induce an IgA response in mammary secretions. The right quarters of the udder were immunised; the left side acted as a control. Antibody titres in secretions showed that at least two animals responded with antigen-specific IgA. Numbers of T-lymphocytes were 4-fold higher in immunised glands compared with controls (P<0.05). IgA-, IgM- and IgG-positive cell numbers were significantly higher (P<0.01) in immunised glands compared with controls in three of the four cows. No mucosal addressin molecule-1 (MAdCAM-1), vascular cell-adhesion molecule-1 (VCAM-1) or peripheral node addressin (PNAd) protein expression was detected on smaller venules that stained positively for von Willebrand factor in alveolar mammary tissues, from either immunised or control glands. Both VCAM-1 and PNAd were detected on smaller venules in supramammary lymph nodes, however, there was no significant difference between immunised and control glands. Quantification of MAdCAM-1 mRNA showed very low expression in both immunised and control alveolar tissue compared with Peyer's patch positive-control tissue. These findings suggest that the bovine mammary gland is capable of a mucosal antibody response; however, MAdCAM-1 is not involved with lymphocyte homing to the mammary gland in this species.

cDNA microarray analysis reveals that antioxidant and immune genes are upregulated during involution of the bovine mammary gland.

We have used cDNA microarray analysis to identify genes that play a role in bovine mammary involution. Involution was induced by termination of milking, and alveolar tissue was collected from 48 nonpregnant Friesian cows in mid lactation sacrificed at 0, 6, 12, 18, 24, 36, 72, and 192 h (n = 6/group) postmilking. The most highly upregulated genes were those associated with oxidative stress. Quantitative real-time reverse-transcription PCR analysis confirmed that mRNA expression of spermidine/spermine N(1)-acetyltransferase was increased by 24 h, superoxide dismutase 2 and metallothionein 1A by 36 h, and glutathione peroxidase by 72 h postmilking. The mRNA expression of the host defense proteins lactoferrin and lingual antimicrobial peptide were increased by 192 h postmilking. A dramatic increase in the protein expression of lactoferrin by 192 h postmilking was also detected by Western analysis. Decreased mRNA expression of the milk protein genes alpha(S1)-, beta-, and kappa-casein, and alpha-lactalbumin were early events in the process of involution occurring within 24 to 36 h postmilking, whereas beta-lactoglobulin mRNA was decreased by 192 h postmilking. Decreases in alpha-lactalbumin and beta-lactoglobulin protein levels in alveolar tissue occurred by 24 and 192 h postmilking, respectively, and the cell survival factors beta1-integrin and focal adhesion kinase were decreased by 72 and 192 h postmilking, respectively. The results demonstrate that in the bovine mammary gland, decreased milk protein gene expression and cell survival signaling are associated with multiple protective responses to oxidative stress that occur before the induction of immune responses and mammary epithelial cell apoptosis during involution.

Adhesion molecule expression in the bovine mammary gland.

The bovine mammary gland requires lymphocytes for immune protection of the gland from foreign pathogens and, in addition, to transfer immune protection to the neonate via colostrum and milk. The process of homing primed lymphocytes to tissues is mediated by the interaction of cell-adhesion molecules displayed on the surface of lymphocytes and counter receptors displayed on the vascular endothelium. This study was conducted to identify the cell-adhesion molecules involved in homing lymphocytes to the bovine mammary gland at four different physiological stages; pregnant, colostral, lactation and involution. The expression and distribution of adhesion molecules in alveolar tissues and supramammary lymph nodes from the mammary glands of healthy cows was determined in situ by immunohistochemical analysis and compared with bovine Peyer's patch, used as a typical mucosal-associated lymphoid tissue and positive control. The mucosal addressin molecule, MAdCAM-1, was not detected in bovine mammary tissues at any of the four different physiological stages. Absence of MAdCAM-1 expression was verified by quantitative real-time RT-PCR analysis. Transcription levels of MAdCAM-1 mRNA were found to be more then 5 x 10(3)-fold lower in mammary alveolar tissues compared with bovine Peyer's patch tissues. In contrast to MAdCAM-1, phase-dependent protein expression of VCAM-1 was detected in both mammary alveolar tissues and the supramammary lymph nodes, with the highest expression observed in colostral phase cows. The protein expression in mammary alveolar tissues was limited to larger venules, although in colostral phase cows, VCAM-1 was also detected around the alveoli perimeter. In the supramammary lymph node, VCAM-1 protein was observed on both small and large venules. PNAd was detected in supramammary lymph nodes at all physiological stages of the mammary gland; however, it was not found in mammary alveolar tissues. Lymphocytes expressing beta7 were not detected in mammary tissues and lymphocytes expressing CD62L were only observed in the supramammary lymph nodes. Overall the data suggest that MAdCAM-1 and VCAM-1 are not involved in homing lymphocytes to the bovine mammary gland; whereas, VCAM-1 and PNAd may have this role in the supramammary lymph node.

Mineral retention in three-week-old piglets fed goat and cow milk infant formulas.

Goat milk and cow milk are commonly used in infant formula preparations and, as such, understanding the nutritional characteristics of infant formulas made from these milks is important. In this study, a goat milk infant formula was compared with an adapted (whey-enhanced) cow milk infant formula with respect to mineral absorption and deposition using the 3-wk-old piglet as a model for the 3-mo-old infant. Equal numbers of piglets (n = 8) were fed either the goat milk formula or the cow milk formula. The mineral composition of the prepared goat milk formula was higher than that of the prepared cow milk formula for most minerals, including calcium (75.1 vs. 56.7 mg/100 mL) but excluding iron, which was higher in the prepared cow milk formula (0.92 vs. 0.74 mg/100 mL). The amounts of calcium, phosphorus, and manganese absorbed by the piglets were significantly higher for the goat milk formula, whereas the amounts of zinc, iron, and magnesium absorbed were significantly higher for the cow milk formula. Apparent mineral absorption, relative to intake, was statistically higher in the cow milk formula for calcium and phosphorus, although the actual differences were very small (less than 1.3%). For copper, zinc, iron, and magnesium there was no significant difference between treatments in apparent mineral absorption, whereas for manganese, absorption was higher for the goat milk infant formula. The absolute mineral deposition was higher in piglets fed the goat milk formula for calcium, phosphorus, and manganese, whereas iron deposition was higher in the piglets fed cow milk formula. For all other minerals tested, there were no significant differences between treatments. The goat milk infant formula provided a pattern of mineral retention in the 3-wk-old piglet very similar to that of the adapted cow milk infant formula. The minor differences observed between the 2 appeared to be due to the different mineral contents of the 2 formulas.

True ileal amino acid digestibility of goat and cow milk infant formulas.

Goat milk is used as an alternative to cow milk for the production of infant formulas. However, little is known about the protein quality and, specifically, about the digestible AA pattern of goat milk formulas compared with their cow milk counterparts. In this study, the true ileal AA digestibility of a goat milk infant formula was compared with a premium cow milk infant formula. The 3-wk-old piglet was used as a model for the 3-mo-old infant. Both milk formulas were prepared as described by the manufacturer, with titanium dioxide added as an indigestible marker. The formulas were fed to the piglets over a 2-wk trial period. Digesta from the terminal ileum were collected post euthanasia and analyzed for AA content, along with samples of the formulas. True AA digestibility was determined after correcting for endogenous AA loss at the terminal ileum of pigs fed an enzyme-hydrolyzed casein-based diet, followed by ultrafiltration (5,000 Da) of the digesta. Total urine and feces collection was also undertaken to determine the nitrogen retention from the diets. The true ileal AA digestibility was similar between the goat and cow milk infant formulas for all AA except Gly and Trp. There was no significant difference in the nitrogen retention of piglets fed the two different formulas. The goat milk infant formula and the premium cow milk infant formula were similar in terms of protein quality.

Mammary blood flow does not limit milk yield in lactating goats.

There is a close relationship between mammary blood flow (MBF) and milk production, but whether MBF is limiting milk yield has not been determined. Five lactating goats received close arterial (external pudic) infusion of PBS or the nitric oxide donor diethylamine NONOate (0.5 mg/h; NONate) for 6 h, according to a crossover design. Goats were hand milked (with oxytocin) every 2 h starting 2 h before and ending 6 h after the end of the infusion. In one goat, a transit time flow probe was implanted around the infused and noninfused artery, whilst in another goat a flow probe was implanted around the infused artery only. Infusion of PBS did not affect MBF or milk production. As with previous results (Lacasse et al., 1996), NONate induced a rapid increase (up to 250% of preinfusion level) in MBF in the infused gland only. Mammary blood flow was still above the preinfusion level at the end of the infusion period. Despite this increase in MBF, NONate did not affect milk production. Milk yield ratio (infused/noninfused gland) averaged 1.20, 1.12, and 1.17 for the preinfusion, infusion and post infusion periods, respectively. Similarly, protein, fat and lactose yields were not affected by PBS or NONate infusion. These results provide no support to the contention that increasing MBF can enhance milk production.

Stat5 phosphorylation status and DNA-binding activity in the bovine and murine mammary glands.

The transcription factors Stat5a and Stat5b are mediators of prolactin signalling in mammary epithelial cells, and are thought to play a role in lactogenesis. In cultured cells, activation of Stat5 activity through phosphorylation results in Stat5 binding to the promoters of at least some of the milk protein genes, thereby stimulating their transcription. However, the mammary biology of Stat5 differs between species, and the role of Stat5 in the bovine mammary gland is not fully understood. We have generated an antibody that specifically recognises the phosphorylated forms of Stat5a and Stat5b and used it to compare the levels of phosphorylated Stat5 with Stat5 DNA-binding activity in bovine and murine mammary tissue. Both Stat5 DNA-binding activity and phosphorylation status in the bovine mammary gland were at near-maximal levels at late pregnancy (27-35 days prior to calving), when at least three of the major milk proteins are not highly expressed. In addition, these studies revealed significant animal-to-animal variation in the level of Stat5 activity in both species. The results are consistent with a role in terminal differentiation of mammary epithelial cells. They also suggest that the stimulation of high-level expression of milk protein genes in the bovine mammary gland is not through activation of the prolactin receptor-Jak2-Stat5 pathway.

Effects of pasture allowance on the yield and composition of milk from cows of different beta-lactoglobulin phenotypes.

The objective of this study was to determine whether the differences in the composition of milk from cows of different beta-lactoglobulin beta-LG) phenotypes are affected by the amount of pasture available and, hence, pasture dry matter intake. Twenty-two Friesian cows of each of the AA and BB variants of the beta-LG phenotype were subjected to ad libitum grazing or restricted grazing in crossover experiments during spring (early lactation, approximately 60 d in milk) and summer (mid to late lactation, approximately 180 d in milk). Milk samples were collected from each cow at the end of each 8-d treatment period and analyzed for composition. Cows of the AA variant of the beta-LG phenotype had higher concentrations of whey protein and beta-LG, but lower concentrations of casein (CN), alpha-CN, kappa-CN (summer only), and BSA, than cows of the BB variant. Compared with cows with a restricted allowance, cows grazing ad libitum had higher milk yields and concentrations of protein, casein, whey protein, and all individual proteins except BSA and immunoglobulin. There were no interactions between effects of pasture allowance and phenotype on milk yield or composition. The data show that having adequate pasture for grazing cows is important not only to maximize milk yield, but also to optimize concentrations of protein and casein, and hence the manufacturing potential of milk. Further, the differences in composition of milk from cows of differing beta-LG phenotypes persisted during short-term restrictions in pasture allowance, and between spring and summer.

Effects of mammary engorgement and feed withdrawal on microvascular function in lactating goat mammary glands.

The responses of the mammary microvasculature in lactating goats (n = 8) during feed withdrawal (18-20 h) and mammary engorgement (26-28 h of milk accumulation) were compared using an indicator-dilution technique with FITC-albumin and [(14)C]sucrose as the intravascular and diffusible indicators, respectively. Feed withdrawal and mammary engorgement caused a 50-60% decrease in mammary arterial flow and in the permeability-surface area product (PS) values for sucrose. Only feed withdrawal increased the mean transit time [from 17.3 to 30.0 s, SE of the difference (SED) = 2.16, P < 0.01] of FITC-albumin, whereas only mammary engorgement reduced sucrose extraction (0.63 to 0.51, SED = 0.04, P < 0.05). Mammary engorgement also caused a substantial reduction in the sucrose-accessible extravascular space from 92 to 44 ml (SED = 15.2, P < 0.01). In a separate experiment using five goats, milking after mammary engorgement did not immediately restore arterial flow or sucrose extraction, indicating that the effect of milk accumulation was not mediated simply via increased intramammary pressure. In conclusion, feed withdrawal resulted in slower flow in the capillary bed but apparently no change in capillary recruitment, whereas mammary engorgement caused capillary derecruitment.

Effect of milking frequency and pasture intake on milk yield and composition of late lactation cows.

Twenty-four monozygous twinsets in late lactation (> 210 d in milk) were used to examine the effects of feed restriction and milking frequency prior to drying off on milk yield and composition in a pastoral dairying system. Cows were assigned to one of four treatment groups for 26 d and were milked either twice or once daily and given either unrestricted or restricted access to feed. Dry matter intakes averaged 16 or 8 kg per cow per day, and diets comprised ryegrass and white clover pasture supplemented with 15% pasture silage. Feed restriction and once daily milking reduced milk yield and increased concentrations of milk fat and protein. Somatic cell count was increased by feed restriction only. Production losses caused by feed restriction were nearly threefold higher than were those for once daily milking. Yields of components that were mammary synthesized and serum derived were reduced by feed restriction, in accordance with milk volume reduction. Plasma lactose concentration increased with once daily milking only and indicated enhanced permeability of mammary tight junctions. Both feed restriction and once daily milking compromised milk quality, but increased leakage of serum components into milk via mammary tight junctions was deemed to occur only for once daily milking.

Metabolic effects of amylin in lactating goats.

The objective of this study was to investigate the role of amylin (a pancreatic hormone) in regulating metabolism in support of lactation. Rat amylin was infused (320 pmol.kg LW(-1).h(-1)) for 6 h via an external pudic (mammary) artery into six lactating goats. This dose of amylin led to a sixfold increase in plasma concentrations of amylin relative to baseline. Amylin infusion increased plasma concentrations (jugular) of glucose and NEFA up to 16 and 168%, respectively, relative to saline infusion. In contrast, plasma concentrations of Ca and PO4 during amylin infusion were reduced by 18 and 30%, respectively, relative to saline infusion. Plasma concentrations of IGF-I, insulin, and Mg were not different between the two treatments, although IGF-I concentrations in the amylin-infused group, 1 and 6 h postinfusion, were significantly higher than those in the saline-infused group. Similarly, amylin infusion failed to affect milk yield and major constituents of milk except protein; milk protein content decreased progressively until the end of amylin infusion and remained low thereafter. Amylin also had no effect on minerals in milk (Ca, PO4, Mg, Fe, Sr, S, K, or Na) except Zn, which was significantly decreased from 56.8+/-5.8 micromol/L at 0 h to 44.5+/-2.4 micromol/L at 6 h postinfusion. Mammary blood flow (measured with a transit-time blood flow probe) increased up to 26% during amylin infusion, although this effect lasted only for the first 3 h. In conclusion, amylin increased plasma concentrations of glucose and NEFA, and mammary blood flow, while decreasing plasma concentrations of Ca and PO4. Despite these metabolic changes, amylin infusion did not increase milk yield of lactating goats.

No evidence for basolateral secretion of milk protein in the mammary gland of lactating goats.

Recent research suggests that a small percentage of milk proteins may be secreted basolaterally, which would have implications for our work on the permeability of tight junctions in the mammary epithelium. In our work, the presence of alpha-lactalbumin (LA) or lactose in plasma is used as an indicator of permeability. The aim of this study was to examine basolateral secretion by determining the presence of milk proteins in efferent mammary lymph. Five Saanen goats were fitted with mammary lymph catheters and were administered intramammary isosmotic bolus infusions of sucrose control solutions or ethylene glycolbis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid to induce leaky tight junctions. Lymph samples were collected before and approximately 5 h after infusion. Lymph was analyzed by Western blotting for the presence of alpha-casein (CN), beta-CN, and alpha-LA No alpha-CN or beta-CN was detected in lymph, but alpha-LA was detected in all lymph samples. Moreover, the signal was much stronger in samples from goats that were treated with ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, and concentrations of alpha-LA in lymph were significantly increased with this treatment. These changes and the absence of casein in lymph suggest increased permeability of tight junctions rather than basolateral secretion. In summary, these data do not support basolateral secretion.

Time course of milk accumulation-induced opening of mammary tight junctions, and blood clearance of milk components.

Eight cows in early lactation were used to study the effect of milk accumulation on the state of mammary tight junctions and to examine alpha-lactalbumin as an indicator of tight junction permeability in vivo. During three successive periods, the cows were milked twice (4 days), once (6 days), and twice daily (4 days). Plasma lactose, alpha-lactalbumin, and milk sodium concentrations were used as indicators of tight junction permeability. Furthermore, four cows were used to study the clearance of lactose and alpha-lactalbumin from the blood. Milk yield during once-daily milking decreased by 15.4% (P < 0.001). All indicators of mammary tight junction patency increased (P < 0.05) transiently during once-daily milking and indicated that tight junctions opened after approximately 18 h. Plasma alpha-lactalbumin and lactose were highly correlated (r = 0.82, P < 0.001), indicating the suitability of plasma alpha-lactalbumin as an indicator of tight junction status in vivo. Clearance of alpha-lactalbumin and lactose from the blood was best described by a biexponential model. Elimination half-lives for lactose and alpha-lactalbumin were 44 and 40 min, respectively. This study showed that milk stasis during early established lactation induces tight junctions to switch to a leaky state after approximately 18 h and to revert to the closed state shortly after milking.

Effect of atropine on milk protein yield by dairy cows with different beta-lactoglobulin phenotypes.

The objective of this study was to determine the response of individual milk proteins to a reduction in amino acid (AA) availability induced by atropine and to determine whether the response was different in cows with different beta-lactoglobulin (LG) phenotypes. Six cows that were homozygous for the A variant of beta-LG and six cows that were homozygous for the B variant of beta-LG were each given a single subcutaneous injection of saline or 20 mg of atropine. In both groups of cows, atropine decreased milk yield by 30% and reduced the concentration of alpha-lactalbumin (LA) by 25 to 30% at 8 h following injection. Eight hours after atropine injection, yield of beta-LG was 41% lower than it was following saline injection, and yield of beta-casein (CN) after atropine injection declined 16% relative to saline. Concentrations of BSA and the ratio of gamma-CN to beta-CN, which reflects plasmin activity in milk, were significantly increased after administration of atropine. Although the response to atropine tended to be more pronounced in cows that were homozygous for beta-LG B, they were not significantly different from the response of cows that were homozygous for beta-LG B, they were not significantly different from the response of cows that were homozygous for beta-LG A. The differential response of individual proteins to a reduction in AA concentrations in whole blood suggested that susceptibility to restriction in substrate availability differed for individual proteins. The concentration of lactose in plasma did not change, which implied that the integrity of the mammary epithelial barrier was not compromised when AA derived from blood were diminished. The consistent concentration of lactose combined with the minimal increase in total yield of BSA in milk following atropine treatment indicated that the increased concentration in milk of proteins derived from serum was due to the concentrating effect of lower milk volume.

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. The results highlight the fact that kinetic and substrate specificity are not sufficient, by themselves, for the identification and characterisation of GLUT isoforms in cultured cells.

The effect of bovine somatotropin and diet on somatotropin binding sites in hepatic tissue of lactating dairy cows.

In the lactating cow, galactopoiesis is stimulated by treatment with recombinant bovine somatotropin (bST) and by an improved plane of nutrition. The present study determined the interaction between these variables and examined whether a positive galactopoietic effect was accompanied by a change in hepatic binding sites for bST. Lactating dairy cows received one of three diets with increasing nutrient density; diet 1, 150 g/kg of dry matter (DM) of crude protein (CP) and 10.5 MJ/kg of DM of metabolizable energy; diet 2, 170 g/kg of DM of CP and 11.3 MJ/kg of DM of metabolizable energy; and diet 3, 190 g/kg of DM of CP and 12.1 MJ/kg of DM of metabolizable energy. At 90 d after calving, half of the cows in each dietary group were treated with bST every 14 d for the rest of the lactation. Both nutrient density and administration of bST increased milk yield significantly in mid and late lactation; there was no significant treatment by diet interaction. Treatment with bST significantly increased plasma concentrations of insulin-like growth factor (IGF)-I compared with IGF-I concentrations in controls in both mid and late lactation. Comparisons within diet revealed that concentrations of IGF-I were significantly higher in cows fed diet 3 than in cows fed diets 1 and 2 at both stages of lactation. Increases in plasma insulin were confined to cows in late lactation, and no changes were observed for nonesterified fatty acids. Liver biopsies showed that concentrations of hepatic binding sites for bST were not affected significantly by bST treatment but were increased in midlactation for cows fed diet 3. Concentration of hepatic binding sites per unit weight of tissue were greater for cows in midlactation than for cows in late lactation. In summary, exogenous bST treatment and increased nutrient density were associated with elevated plasma IGF-I concentrations and increased milk yield; however, only nutrient density in midlactation increased the number of hepatic binding sites for bST. Exogenous bST treatment had relatively little effect on the concentration of hepatic bST receptors compared with nutrient density.

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. Messenger RNA levels for the inducible NOS isoform were increased by IFN-gamma and TNF-alpha in a manner consistent with the elevation of NOS activity. EGF also stimulated thymidine incorporation into DNA which was attenuated by coexposure with IFN-gamma in a manner that appeared to be largely NO-independent.