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Biomedical subjects

W A Phillips

Publications and source records attributed to W A Phillips.

At least 55 records · Page 3Linked to original sources
PubMed1995

Silage or limit-fed grain growing diets for steers: II. Empty body and carcass composition.

The influence of energy source (silage- or grain-based) on empty body and carcass composition and adipocyte cellularity independent of rate of gain was tested. Sixty-four Angus steers were allotted to either a forage (ad libitum) or grain (limit-fed) diet for a growing phase (145 d) followed by 45, 75, or 105 d of ad libitum access to a grain-based diet. Eight steers were slaughtered initially and eight from each treatment were slaughtered at the end of the growing phase, and at each of the termination dates. The silage growing diet consisted (DM basis) of 55% sorghum silage (approximately 24% dry matter), 22% alfalfa hay, 11% ground shelled corn, and 11% soybean meal. The grain-based growing diet was composed of 77% ground shelled corn, 5% soybean meal, 14% cottonseed hulls, 3% molasses, and 1% salt and mineral; it was limit-fed to produce the same rate of gain as the silage diet. No implants or ionophores were used. At the end of the growing phase, the steers fed grain were heavier and had a higher percentage of fat in the empty body (24 vs 19% fat) and the carcass (26 vs 21% fat) than did steers fed forage. Rate of gain during the growth phase was related positively to percentage of carcass fat; when corrected for fill, data for both diets fit one regression line for fat vs rate of gain. When adjusted for gain during the growing phase, fat content was not different in empty body or carcass, but internal fat was higher (P < .10) for steers fed grain. After 45 d on the finishing diet, carcass fat remained low (23%), but after 75 and 105 d, fat content reached 27%. Source of energy did not detectably affect carcass composition independent of rate of gain. Cell size of adipocytes from four adipose depots increased with time on feed but were not affected by diet during the growing phase. Lean Choice beef can be produced in only 45 d in the feedlot with medium-framed Angus cattle.

Adipose Tissue↗
PubMed1995

Effect of protein supplementation on forage intake and nitrogen balance of lambs fed freshly harvested wheat forage.

Wether lambs (mean BW = 32 kg) fed wheat forage freshly harvested daily (Exp. 1, n = 28; Exp. 2, n = 25) were used to evaluate the effects of supplemental energy and protein on forage intake, diet digestibility, and N retention. Experiment 1 was conducted in the winter of 1988 and the four treatments were 1) CONTROL, no supplement; 2) ENERGY, low-protein corn-based supplement; 3) CSM, 22% CP supplement with cottonseed meal; and 4) CGM, 22% CP supplement with corn gluten meal. Experiment 2 was conducted in the spring of 1989, and the five treatments were 1) CONTROL; 2) ENERGY; 3) CSM, 23% CP with cottonseed meal; 4) FTM, 25% CP supplement with feather meal and corn gluten meal; and 5) BM, 25% CP supplement with blood meal and corn gluten meal. All supplements were isoenergetic and composed 22 and 24% of total DMI in Exp. 1 and 2, respectively. In Exp. 1, forage intake was not affected but total and digestible DMI were increased (P < .01) by energy and protein supplementation. Protein supplements did not (P > .10) increase total and digestible DMI compared to ENERGY supplementation but tended (P < .10) to increase N retention (g/d). In Exp. 2, digestible DMI and N retention did not differ among the five treatments. Results suggest that lambs consuming wheat forage in the vegetative stage (Exp. 1) will have greater digestible DMI if supplemental energy or protein is provided. When more mature wheat forage was fed (Exp. 2), energy seemed to limit animal performance.

Animal Feed↗
PubMed1994

Activation of the macrophage respiratory burst by phorbol myristate acetate: evidence for both tyrosine-kinase-dependent and -independent pathways.

We have investigated the relationship between tyrosine phosphorylation and respiratory-burst activity in murine bone-marrow-derived macrophages (BMM) stimulated with phorbol myristate acetate (PMA). In unprimed BMM, a good correlation was observed between the net level of tyrosine phosphorylation and the activity of the respiratory burst. The phosphotyrosine phosphatase inhibitor, vanadate, enhanced both tyrosine phosphorylation and respiratory-burst activity triggered by PMA. Furthermore, the tyrosine kinase inhibitor, ST638, abolished both tyrosine phosphorylation and respiratory-burst activity stimulated by PMA. However, in BMM primed by preexposure to TNF alpha, the correlation between net tyrosine phosphorylation and respiratory-burst activity triggered by PMA was not maintained. ST638 was found to only partially inhibit the PMA-triggered respiratory burst under conditions where PMA-stimulated tyrosine phosphorylation was abolished. We conclude that PMA can activate the macrophage respiratory burst by both tyrosine-kinase-dependent and -independent pathways.

Animals↗
PubMed1994

Expression of p47-phox and p67-phox proteins in murine bone marrow-derived macrophages: enhancement by lipopolysaccharide and tumor necrosis factor alpha but not colony stimulating factor 1.

We have investigated the relationship between the expression of the p47-phox and p67-phox cytosolic components of the NADPH oxidase and priming of the macrophage respiratory burst. Western blot analysis revealed that murine bone marrow-derived macrophages (BMM) contain immunoreactive proteins detected by antisera raised against recombinant human p47-phox and p67-phox. Priming BMM by exposure to tumor necrosis factor alpha (TNF-alpha) or lipopolysaccharide (LPS) increased the levels of p47-phox and p67-phox. Colony-stimulating factor 1 (CSF-1), which we previously found to have a negative effect on the priming of murine macrophages, had no effect on the level of p47-phox but down-regulated that of p67-phox. Our results suggest that the regulatory effects of LPS, TNF-alpha, and CSF-1 on the respiratory burst of BMM may be due to modulation of the expression of the p47-phox and p67-phox cytosolic components of the NADPH oxidase.

Animals↗
PubMed1994

Essential role of YopD in inhibition of the respiratory burst of macrophages by Yersinia enterocolitica.

The respiratory burst is a key element of the bactericidal armamentarium of phagocytes. In this study we have shown that a virulent strain of Yersinia enterocolitica serogroup O:9 completely inhibited the ability of murine bone marrow-derived macrophages to mount a respiratory burst in response to stimulation by zymosan. This property of the bacterium was abrogated by curing the strain of its 71.5-kb virulence plasmid and by transposon mutagenesis of the plasmid-borne yopD gene. Derivatives of the bacterium which were unable to inhibit the respiratory burst were also less able to disrupt cytoskeletal actin and to resist phagocytosis. yopD mutants also showed an impaired ability to dephosphorylate phosphotyrosine residues in macrophage proteins and were completely avirulent for mice. All of these defects were fully or partly restored by trans-complementation of a yopD mutant with a cloned yopD gene. The results of this study and those of previous work with YopD (R. Rosqvist, A. Forsberg, and H. Wolf-Watz, Infect. Immun. 59:4562-4569, 1991) suggest that YopD functions chiefly by facilitating the transport of virulence plasmid-encoded proteins, such as YopE, a cytotoxin, and YopH, a protein tyrosine phosphatase, across the cytoplasmic membrane to their targets within host cells. The combined action of these Yops on cytoplasmic proteins, especially actin, could explain the effects of virulent Y. enterocolitica on macrophage morphology, phagocytic capacity, and respiratory burst activity, all of which rely on cytoskeletal integrity to function normally.

Animals↗
PubMed1994

Haematopoietic colony stimulating factors CSF-1 and GM-CSF increase phosphatidylinositol 3-kinase activity in murine bone marrow-derived macrophages.

The activity of phosphatidylinositol (PI) 3-kinase was examined in murine bone marrow-derived macrophages (BMM) stimulated with the haematopoietic growth factors colony stimulating factor-1 (CSF-1) and granulocyte/macrophage-CSF (GM-CSF). PI 3-kinase was immunoprecipitated from cell lysates using anti-phosphotyrosine antibody or an antibody directed against the 85K subunit of PI 3-kinase, and the activity assayed by the phosphorylation of PI in the presence of [gamma 32P]-ATP. The results demonstrate that CSF-1 increases the activity of PI 3-kinase, as compared to the non-stimulated control, in murine macrophages. Maximum activity was seen after 10 min of stimulation with CSF-1 at 3000-5000 U/ml. The dose-response of CSF-1 is consistent with other biochemical effects of CSF-1 seen in the BMM. GM-CSF also stimulated PI 3-kinase activity although to a lesser extent than CSF-1, correlating well with their degree of mitogenic activity on the BMM. Non-mitogenic macrophage activating agents, such as the phorbol myristate acetate, lipopolysaccharide, concanavalin A and formyl-methionyl-leucyl-phenylalanine, did not significantly increase the PI 3-kinase activity. Furthermore, CSF-1 failed to stimulate PI 3-kinase activity in resident peritoneal macrophages, a population of macrophages with poor proliferative capacity. These results suggest that the PI 3-kinase activity may be involved in the haemopoietic growth factor signalling pathways regulating macrophage growth.

Animals↗
PubMed1992

Zymosan-triggered tyrosine phosphorylation in mouse bone-marrow-derived macrophages is enhanced by respiratory-burst priming agents.

We have investigated the relationship between tyrosine phosphorylation and respiratory-burst activity in mouse bone-marrow-derived macrophages (BMM). We demonstrate that zymosan, an agent known to trigger the macrophage respiratory burst, also triggers the activation of tyrosine kinase activity, resulting in rapid tyrosine phosphorylation on numerous proteins, and provide evidence for the role of tyrosine phosphorylation in the triggering of the BMM respiratory burst. Agents, such as tumour necrosis factor alpha (TNF alpha), interferon-gamma (IFN-gamma) or lipopolysaccharide (LPS), which prime the macrophage for an enhanced zymosan-triggered respiratory burst, increase tyrosine phosphorylation triggered by zymosan. The zymosan-triggered tyrosine phosphorylation and respiratory-burst activity were partially suppressed by the tyrosine kinase inhibitors alpha-cyano-3-ethoxy-4-hydroxy-5-phenylmethylcinnamide (ST638) and herbimycin A. In addition, pre-exposure of BMM to vanadate, a phosphotyrosine phosphatase inhibitor, greatly enhanced the ability of zymosan to induce tyrosine phosphorylation and trigger the respiratory burst. These data highlight the importance of the balance between tyrosine kinase and phosphotyrosine phosphatase activity in determining the ultimate level of tyrosine phosphorylation in BMM and suggest that zymosan-triggered tyrosine phosphorylation is an important biochemical signal for triggering of the respiratory burst.

Animals↗
PubMed1992

Priming of the respiratory burst of bone marrow-derived macrophages is associated with an increase in protein kinase C content.

The biochemical mechanism(s) underlying the priming of the macrophage for an enhanced PMA-induced respiratory burst is not understood. Because the cellular receptor for PMA is thought to be protein kinase C (PKC), we have investigated the effects of priming agents on cellular PKC levels. Sonicates from unprimed bone marrow-derived macrophages (BMM) were found to contain PKC activity (309 +/- 51 pmol 32P-incorporated/mg/min; mean +/- SE, n = 17) as measured by the phospholipid-, diacylglycerol-, and calcium-dependent phosphorylation of histone. Exposure of BMM to priming agents such as TNF-alpha, LPS, and granulocyte/macrophage-CSF resulted in a significant increase in both histone-phosphorylating activity and levels of immunoreactive PKC protein in these cells. A minimum of 6-h exposure, with an increasing effect up to 48 h, was required for a detectable increase in PKC level. The activity from primed BMM, like that of the untreated cells, was predominantly cytosolic. The kinetics and concentration dependence of the priming agent-induced increase in the PKC content of BMM closely paralleled the enhancing effects of these agents on the PMA-stimulated respiratory burst. Furthermore, CSF-1, a cytokine that does not prime BMM, failed to increase PKC activity. We propose that the exposure of BMM to priming agents leads to an increase in the expression of a stimulatory isozyme(s) of PKC, resulting in an enhanced ability to mount a respiratory burst in response to stimulation with PMA.

Animals↗
PubMed1992

The effect of interleukin-4 on the macrophage respiratory burst is species dependent.

Preexposure of human monocytes to recombinant human interleukin-4 (IL-4) suppressed the respiratory burst response to a number of different stimuli, including phorbol myristate acetate, zymosan, platelet-activating factor and the chemotactic peptide, f-met-leu-phe. Under similar conditions, the respiratory burst of murine macrophages was enhanced by preexposure to recombinant murine IL-4. By conducting our studies on cells from different species under similar conditions we have demonstrated that there is a significant disparity in the effects of IL-4 on human and murine macrophages thus providing an explanation for some apparent inconsistencies in the literature and highlighting the need for caution when extrapolating data across species barriers.

Animals↗
PubMed1992

Protein kinase C has both stimulatory and suppressive effects on macrophage superoxide production.

Unlike resident peritoneal macrophages (RPM) or tumor necrosis factor alpha (TNF alpha)-primed bone marrow-derived macrophages (BMM), unprimed BMM do not generate superoxide in response to the protein kinase C (PKC) activator, phorbol myristate acetate (PMA). However, these cells do contain significant levels of PKC activity. In contrast to PMA, zymosan induces the generation of superoxide in unprimed BMM, as well as in TNF alpha-primed BMM and RPM. Staurosporine, a potent PKC inhibitor, failed to affect the zymosan-induced production of superoxide by unprimed and TNF alpha-primed BMM and RPM, in spite of substantial inhibition of PMA-induced superoxide production by the primed BMM and RPM. However, when PKC was depleted from unprimed BMM by prolonged (24 h) treatment with phorbol dibutyrate (PdBt) (10(-7) M) the ability of zymosan to induce the production of superoxide was greatly diminished. Such a result could be interpreted as suggesting a role for PKC in the zymosan-induced response, a conclusion which contrasts with the inhibitor data. However, PKC depletion, in this case, is achieved via the PdBt-induced activation of PKC. It is thus possible that it is the initial activation of PKC, rather than its depletion, that suppresses superoxide production. Consistent with this interpretation, the co-stimulation of unprimed BMM with both zymosan and PMA resulted in a reduced superoxide release compared to zymosan alone. The activation of PKC therefore appears to have a suppressive effect on the generation of superoxide by unprimed cells. We thus conclude that PKC is not required for zymosan-induced superoxide production by either primed or unprimed macrophages and suggest that PKC may be involved in regulatory mechanisms restricting superoxide production by macrophages. However, since PMA alone can initiate the release of superoxide from primed BMM and RPM, it would appear that PKC can mediate both stimulatory and suppressive signals for macrophage superoxide production.

Alkaloids↗
PubMed1992

Effect of initial restraint, weaning, and transport stress on baseline and ACTH-stimulated cortisol responses in beef calves of different genotypes.

The productivity and well-being of animals can be substantially affected by stress. This is particularly true in the case of beef calves that are subjected to a multitude of stressors over a short period during the first year of life. Perhaps the most often studied stress-responsive variable has been blood corticosteroid concentrations. Factors such as age, gender, genetics, and degree of prior experience, can influence how an animal perceives and responds to a given stressor. Few studies have tried to control these variables, and accordingly, many conflicting results have been published regarding the impact of various stressors on cortisol response. We measured baseline plasma cortisol concentration over a 44-day study in Bos indicus and Bos taurus calves. Plasma cortisol values in Bos indicus calves were higher (32.60 +/- 0.66 ng/ml) than values in calves of Bos taurus (25.81 +/- 0.76) breeding. A precipitous decrease in cortisol concentration was observed 7 days after transport stress in all calves. Baseline cortisol concentration did not provide any indication of the intensity of the various stressors. However, significant differences were readily observed after ACTH administration. On the basis of cortisol secretion, stresses of transport and weaning were similar and were the most stressful to calves, regardless of genotype.

Adrenocorticotropic Hormone↗
PubMed1992

Lipopolysaccharide- and interferon-gamma-induced expression of hck and lyn tyrosine kinases in murine bone marrow-derived macrophages.

We have examined the role of tyrosine phosphorylation during the course of macrophage activation. Initial experiments indicated that vanadate, a known phosphotyrosine phosphatase inhibitor, enhanced the phorbol 12-myristate 13-acetate (PMA)-triggered respiratory burst and potentiated the priming effects of bacterial lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma), suggesting that tyrosine phosphorylation may be important in these end cell functions. As src-related kinases have been implicated in the activation of cells of other haemopoietic lineages, we examined the relationship between the activity of two such kinases, hck and lyn, and priming of the respiratory burst. We found that the level of hck and lyn is increased following exposure of bone marrow-derived macrophages (BMM) to LPS or IFN-gamma. The induction of both of these kinases follows similar kinetics with maximal activity occurring at 24-48 h. Interestingly, the kinetics of induction of hck and lyn kinase activity in BMM demonstrated a close temporal relationship with the priming effects of LPS and IFN-gamma on the macrophage respiratory burst. Collectively, these observations raise the possibility that modulation of expression of hck and lyn is involved in the regulation of the respiratory burst.

Animals↗
PubMed1991

Protein kinase C isotypes and signaling in neutrophils. Differential substrate specificities of a translocatable calcium- and phospholipid-dependent beta-protein kinase C and a phospholipid-dependent protein kinase which is inhibited by long chain fatty acyl coenzyme A.

Neutrophils possess a classical Ca2+, phosphatidyl serine (PS) and diglyceride (DG)-dependent protein kinase C (beta-PKC) which was translocatable from cytosol to membrane in response to elevated Ca2+ in the physiologic range or to pretreatment with phorbol myristate acetate (PMA). The translocatable beta-PKC was purified from neutrophil membranes prepared in the presence of Ca2+, eluted with EGTA and subjected to hydroxyapatite chromatography. An 80-kDa protein possessing Ca/DG/PS-dependent histone phosphorylating activity was recognized by a monoclonal antibody to beta-PKC but not to alpha-PKC or gamma-PKC. A cytosolic kinase activity remaining after Ca(2+)-induced translocation of beta-PKC was dependent on PS and DG but did not require Ca2+. This novel Ca(2+)-independent, PS/DG-dependent kinase, termed nPKC, eluted from hydroxyapatite between alpha-PKC and beta-PKC, ran as a 76-kDa band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and was reactive to a polyclonal consensus antibody but not to monoclonal antibodies to alpha-PKC, beta-PKC, or gamma-PKC. Long chain fatty acyl-CoA, but not the corresponding free fatty acids, inhibited nPKC in the 1-10 microM range. The chemotactic peptide fMet-Leu-Phe triggered prompt but transient increases in neutrophil long chain fatty acid acyl-CoA, suggesting that nPKC is regulated by fatty acyl-CoA as well as DG during neutrophil activation. Purified beta-PKC phosphorylated a number of cytosolic proteins in a Ca(2+)-dependent manner, including a major 47-kDa cytosolic protein, which may be implicated in superoxide anion generation. In contrast, nPKC did not phosphorylate the 47-kDa protein, but phosphorylated numerous cytosolic proteins in a Ca(2+)-independent manner, including a 66-kDa protein which was not phosphorylated by beta-PKC. Differences in location, substrate specificity, and cofactor dependence between nPKC and beta-PKC suggest these kinases may play selective roles in the activation sequence of the neutrophil.

Acyl Coenzyme A↗
PubMed1991

Fusion of the upper cervical spine in children and adolescents. An analysis of 17 patients.

A retrospective review of 17 immature patients who underwent posterior spinal fusion of C1-C2 or C1-C3 was performed to establish the efficacy of the procedure, to determine the incidence of complications, and to identify those at high risk for complications. All had radiographic C1-C2 instability or dislocation. Etiologies included os odontoideum, fixed rotatory subluxation, atlantoaxial subluxation, type II dens fracture nonunion, and nonunion of a Hangman's fracture. Before surgery, 10 patients were neurologically intact, and seven had neurologic findings. Associated diagnosis included Down's syndrome in three and Ehler-Danhlos in one. Follow-up averaged 25 months. One neurologically intact patient had a C2 radiculopathy that resolved by 1 year. Three patients with Down's syndrome had complications: one with a preoperative Brown-Sequard syndrome had transient worsening in the immediate postoperative period, one with a preoperative myelopathy developed a late recurrence of a severe myelopathy that required odontectomy, and another sustained an intraoperative spinal cord contusion followed by postoperative quadriplegia and death due to respiratory failure. Of seven reporting neurologic symptoms before surgery, two had residual deficit in the late postoperative period. These two represented preventable technical errors. There were two nonunions, one of which required occiput to C2 fusion. In general, posterior spinal fusion of the upper cervical spine was found to be a reliable, safe, and predictable procedure, but extra caution should be employed when considering arthrodesis in patients with ongoing spinal cord compression, fixed dislocations, and inherited ligamentous laxity.

Adolescent↗
PubMed1991

Complications of fusion to the upper cervical spine.

Forty-seven operations for posterior fusion of the upper cervical spine were reviewed for complications. Alarmingly, only 11 patients had an entirely uncomplicated course. Most complications were minor, but there were four nonunions, one requiring reoperation. Although seven patients had increased neurologic deficits after surgery, only one was permanent. There was one death due to technical error. Patients with significant instability, myelopathy, prior failed fusions, or unreducible dislocations are at high risk for perioperative neurologic complications.

Adult↗
PubMed1991

Energy and protein supplementation of ammoniated wheat straw diets for growing steers.

Eighty-eight yearling beef steers (308 +/- 1.4 kg) were used in two separate trials to determine the protein-sparing value of the N added to wheat straw during the ammoniation process and to determine the effects of supplementing ammoniated straw diets with energy and ruminal escape protein. In Exp. 1, steers were fed untreated straw (US) with either 0, 150, or 500 g of soybean meal (SBM) for 88 d. The addition of SBM to US diets increased (P less than .01) straw intake and average daily gains (ADG), indicating that N was limiting. When ammoniated straw (AS) was substituted for US, the N in the AS was used as efficiently as 500 g of SBM for growth. In Exp. 2, steers had ad libitum access to AS with three levels of supplemental corn (0, 1.23, or 2.45 kg DM.animal-1.d-1) either with or without .41 kg DM of corn gluten meal (CGM) added. Straw intake decreased (P less than .01) as the amount of corn in the diet was increased, but ADG increased (P less than .01) with the addition of corn. Straw consumption was not altered by the addition of CGM, but ADG was increased (P less than .01) by an average .35 kg by CGM. Rumen and blood N components indicated that the N from AS was contributing to the ruminal N pool and that CGM was compensating for microbial protein deficiencies postruminally.

Ammonia↗