Biomedical subjects

D J Buckley

Publications and source records attributed to D J Buckley.

At least 37 records · Page 2Linked to original sources

PubMed1998

Prolactin-regulated apoptosis of Nb2 lymphoma cells: pim-1, bcl-2, and bax expression.

Lactogen-dependent Nb2 lymphoma cells, widely employed for studying prolactin (PRL) mitogenic mechanisms, are also useful for investigations of apoptosis in T-lineage lymphocytes. Utilizing PRL-dependent Nb2-11 cultures, apoptosis-regulatory genes were evaluated for participation in dexamethasone- (DEX) provoked cell death or its inhibition by PRL. Treatment of lactogen-starved, G1-arrested Nb2-11 cells with DEX (100 nM) activated apoptosis within 12 h evaluated by flow cytometric analysis of fragmented DNA. This effect was not associated with altered expression of bcl-2, bax, or pim-1. PRL (10 ng/mL), coincubated with DEX-treated cells, completely blocked DEX-induced apoptosis. This inhibition was associated with increased expression of bcl-2 and pim-1 mRNAs, genes reported to suppress apoptosis, within 2-6 h after addition of the hormone. Moreover, the increased transcription of bcl-2 and pim-1 was coupled to increases in their protein levels. The results suggest that bcl-2, bax, and pim-1 do not play a critical role in DEX-induced apoptosis in Nb2 cells. However, expression of bcl-2, together with pim-1, may have a role in mediating the antiapoptotic actions of PRL.

Animals↗

PubMed1998

Prolactin receptor signaling: shared components with the T-cell antigen receptor in Nb2 lymphoma cells.

Previously, we reported that activation of the human prolactin receptor (PRLR) produced a protein phosphorylation pattern strikingly similar to that provoked by Concanavalin A (Con A), an activator of the T-cell antigen receptor (TCR). These results suggested that certain signaling components of the TCR may be shared by the activated PRLR. Additional studies here assessed the levels of TCR expression following PRLR stimulation and the effect of TCR activation on PRL-stimulated proliferation in lactogen-dependent pre-T Nb2-11 lymphoma cells. The results indicated that the TCR was expressed on the surface of approx 4% of exponentially proliferating and prolactin- (PRL) treated cells. In contrast, approx 45% of quiescent cells, cultured in the absence of PRL for 24 h, expressed the TCR at the cell surface, suggesting that lactogen withdrawal may up-regulate TCR cell-surface expression. Moreover, TCR activation with anti-CD3 antibodies attenuated PRL-stimulated Nb2-11 cell proliferation in a concentration-dependent manner. In other experiments, immunoprecipitation and immunoblotting of Nb2-11 lysates revealed that activation of the PRLR resulted in rapid tyrosyl phosphorylation of ZAP-70, a critical TCR-associated tyrosine kinase. In addition, ZAP-70 was found to associate transiently with the putative guanine nucleotide exchange factor and substrate, Vav, in PRL-treated cells. ZAP-70 was also found to associate constitutively with the PRLR; PRL stimulation provoked the transient recruitment of Vav to the complex. These observations suggest that PRL signaling reflects the transient formation of a PRLR-ZAP-70-Vav complex and its immunomodulatory actions involve diverse interactions that affect TCR expression and signaling mechanisms.

CD3 Complex↗

PubMed1997

Butyrate-induced reversal of dexamethasone resistance in autonomous rat Nb2 lymphoma cells.

The parental rat Nb2 lymphoma is a prolactin (PRL)-dependent T cell line. Exposure of a PRL-independent subline, Nb2-SFJCD1, to sodium butyrate (NaBT) causes transient reversal of their growth factor-independent proliferation in association with constitutive expression of protooncogenes pim-1 and c-myc. In the present study, we investigated the effect of NaBT treatment on the sensitivity of Nb2-SFJCD1 cells to dexamethasone (DEX)-induced apoptosis. Pretreatment with NaBT (2 mM, 72 h) partially reversed resistance to apoptosis in Nb2-SFJCD1 cells exposed to DEX (100 nM) for 12 h, assessed by flow cytometric analyses of DNA fragmentation. However, the cytolytic effect of DEX was abrogated by PRL in a time- and concentration-dependent manner. Evaluation of apoptosis-associated gene expression in NaBT-pre-treated cultures incubated with DEX or DEX+PRL indicated that the apoptosis resistance did not stem from altered bcl-2 or bax expression. However, there was a strong correlation between the resistance to DEX-activated apoptosis and their enhanced expression of pim-1 mRNA and protein. The results show that it is possible to reverse DEX-induced apoptosis of Nb2 pre-T cells and suggest the pim-1 gene product has an important role as a suppressor of this process, perhaps functioning as a mediator of PRL action.

Journal Article↗

PubMed1997

Increased cystine uptake capability associated with malignant progression of Nb2 lymphoma cells.

Analysis of rat, pre-T cell 'Nb2 lymphoma' sublines, manifesting different degrees of malignant progression, can indicate phenotypic changes potentially useful as therapeutic targets. In this study, the prolactin (cytokine)-dependent Nb2-11 and autonomous Nb2-SFJCD1 sublines were compared for in vitro thiol growth requirements. Whereas Nb2-11 culture growth depended on 2-mercaptoethanol (2-ME; 33-100 microM), Nb2-SFJCD1 cells were 2-ME-independent. This difference stemmed from differential uptake of exogenous L-cystine, critically required for proliferation. Uptake of 35S-L-cystine (10 microCi/ml; 40 microM) showed Nb2-11 cells had low cystine uptake capability; 2-ME enhanced cystine uptake to growth-sustaining levels. Nb2-SFJCD1 cells did not require 2-ME due to intrinsic, 11-fold higher cystine uptake via the x(c)- cystine/glutamate transport system. In absence of 2-ME, monosodium glutamate abrogated Nb2-SFJCD1 proliferation by specifically inhibiting cystine uptake (85% at 10 mM). Elevated glutathione (GSH) levels were not essential for growth of either line as shown with L-buthionine-(S,R)-sulfoximine (0.1-4 mM) treatment. The cyst(e)ine requirement therefore did not primarily involve maintenance of normal GSH levels, reported critical for T lymphocyte replication. These and other results suggest increased cystine uptake capability constitutes another potential step in progression of T cell cancers which is not coupled to cytokine autonomy or metastatic ability development. The x(c)- transport system apparently provides a novel target for T cell cancer therapy. Its inhibition would suppress cystine uptake by certain progressed cells, and also interfere with cystine uptake, and subsequent cysteine release, by eg macrophages, thought to have a role in cysteine delivery to lymphoid cells.

Animals↗

PubMed1997

Determination of the influence of organic acids and nisin on shelf-life and microbiological safety aspects of fresh pork sausage.

The effect of replacing sulphur dioxide with organic acids and nisin to reduce the microbial counts in fresh pork sausage was examined. The potential of sodium citrate or sodium lactate, used singly or in combination with nisin, was also assessed in sausage inoculated with Staphylococcus aureus MMPR 3 and Salmonella kentucky AT 1. The results indicate that a combination of sodium lactate and nisin wa particularly effective in reducing total bacterial counts in this food product. It also appears that this combination provides an increased protection against common pathogenic contaminants of fresh pork sausage, i.e. Staph. aureus and Salmonella species.

Animals↗

PubMed1997

Tissue content of alpha-tocopherol and oxidative stability of broilers receiving dietary alpha-tocopheryl acetate supplement for various periods pre-slaughter.

1. The effects of dietary alpha-tocopheryl acetate on the alpha-tocopherol status of chicken plasma and tissues were investigated. The rate of iron-ascorbate-induced lipid peroxidation was also studied. 2. One hundred and forty four chicks were divided into 6 groups: one control group was fed a basal diet of 30 mg alpha-tocopheryl acetate/kg food for the duration of the trial. A supplemental diet of 200 mg alpha-tocopheryl acetate was fed to each of the other 5 groups for 1,2,3,4 or 5 weeks prior to slaughter. 3. Supplementation resulted in an increase in alpha-tocopherol in plasma and all tissues examined. Saturation levels of alpha-tocopherol were observed in plasma after 1 week of feeding and in tissues within 3 to 4 weeks of feeding. 4. Supplementation with alpha-tocopheryl acetate for up to 4 weeks pre-slaughter resulted in significant reductions in susceptibility to induced lipid peroxidation. 5. Overall, the results show that feeding 200 mg alpha-tocopheryl acetate/kg food to chicks for at least 4 weeks prior to slaughter is necessary to optimise muscle content and stability against lipid peroxidation.

Abattoirs↗

PubMed1997

Influence of dietary vitamin E and oxidised sunflower oil on the storage stability of cooked chicken muscle.

1. The effects of oxidised dietary sunflower oil and dietary alpha-tocopheryl acetate supplementation on alpha-tocopherol concentrations in broiler muscle and on the storage stability of refrigerated, cooked, minced muscle were determined. Broiler chicks were fed on diets containing fresh sunflower oil and 30 (FS30) or 200 (FS200) mg alpha-tocopheryl acetate/kg, or oxidised sunflower oil and 0 (OS0), 30 (OS30) or 200 (OS200) mg alpha-tocopheryl acetate/kg. 2. Inclusion of oxidised sunflower oil significantly reduced dietary and hence, muscle alpha-tocopherol concentrations. 3. Oxidised oil increased oxidation in raw and cooked muscle, and reduced the oxidative stability of muscle during refrigerated and frozen storage. 4. Supplementation with alpha-tocopheryl acetate improved the stability of muscle, with stability increasing as muscle alpha-tocopherol concentrations increased, when fresh or oxidised oil was fed. Supplementation with 200 mg alpha-tocopheryl acetate/kg offset the effects of oxidised oil in breast, but not in thigh. 5. The results show that the prooxidising effects of oxidised oils in muscle foods can be overcome, but alpha-tocopherol content needs to be adequately adjusted to compensate for increased oxidative stress. Supplementation with 200 to 400 mg alpha-tocopheryl acetate/kg may be necessary to achieve an optimum muscle alpha-tocopherol concentration.

Animals↗

PubMed1997

Low doses of 105AD7 cancer vaccine preferentially stimulate anti-tumor T-cell immunity.

Clinical studies with the human anti-idiotypic antibody 105AD7 have clearly shown that 791Tgp72 is a good target antigen for cell-mediated immunity. No antibody-related toxicity was observed in any of the 135 patients entered into phase I/II clinical trials of 105AD7, whereas both helper and cytotoxic T-cell responses were induced. The helper responses were exemplified by induction of interleukin-2 (IL-2), antigen-specific blastogenesis, and enhanced natural killer (NK) activity. Anti-tumor cytotoxicity was measured directly and was supported by activation of circulating CD8 cells. In this study, it is shown that a 100-microgram injection of 105AD7 was more effective than the 200-microgram dose. Enhanced IL-2 production was observed following 15/19 injections of 100 micrograms of 105AD7 whereas only 4/11 injections of 200 micrograms of 105AD7 induced responses (p < 0.02). Similarly, time to progression was significantly (p < 0.05) slower (median 6 m) in patients injected with 100 micrograms than patients receiving the higher dose, suggesting that 100 micrograms or lower may be the optimal dose. The standard dose for hepatitis vaccination is 10 micrograms. In vitro blastogenesis assays on naive donors have shown that a dose of 105AD7, which is either too high or too low, fails to activate T cells. The optimal dose in vitro is 10 ng.

Adult↗

PubMed1997

Dietary vegetable oils and alpha-tocopherol reduce lipid oxidation in rabbit muscle.

This experiment was conducted to study the effect of dietary vegetable oil on lipid oxidation in rabbit muscle. A control diet with no added fat and two diets with olive or sunflower oil (30 g/kg) were used. Within each treatment, one group was fed a low level of alpha-tocopheryl acetate (10 mg/kg diet), and the other a supplemental level (200 mg/kg). Rabbits were fed experimental diets from weaning (20 d) to slaughter (69 d). The supplemental level of dietary alpha-tocopheryl acetate produced higher alpha-tocopherol concentration in muscle (P < 0.006) and lower lipid oxidation (P < 0.004). Rabbits that received sunflower oil had higher concentrations of thiobarbituric acid reactive substances than rabbits that consumed olive oil (P < 0.05). Moreover, a significant effect due to fat inclusion in the diet was found. Muscles from rabbits fed diets not enriched with fat had higher susceptibility to lipid oxidation (P < 0.005) and higher concentration of (n-3) fatty acids in polar lipids (P < 0.04) than those from rabbits fed fat-enriched diets. A second experiment was conducted and confirmed the higher lipid oxidation in the muscle of rabbits fed diets not enriched with fat than in that of rabbits fed diets containing sunflower oil (28 g/kg) (P < 0.003) as well as in diets with identical digestible energy. In this experiment, alpha-tocopheryl acetate was at the lower level (10 mg/kg feed). Inclusion of oils rich in oleic (olive oil) or linoleic acid (sunflower oil) in rabbit diets reduces lipid oxidation in muscles.

Animals↗

PubMed1996

Oxidative stability and alpha-tocopherol retention in turkey burgers during refrigerated and frozen storage as influenced by dietary alpha-tocopheryl acetate.

1. The effect of vitamin E (alpha-tocopheryl acetate) in turkey diets on the oxidative stability of raw and cooked turkey burgers and on the retention of alpha-tocopherol during refrigerated (4 degrees C) or frozen (-20 degrees C) storage was investigated. One hundred and two, one-day-old T-8s turkey poults were divided at random into 3 groups of 34 animals each and fed on either a basal diet (normal commercial turkey diet) supplemented with 20 mg alpha-tocopheryl acetate/kg (control) or fed an alpha-tocopherol supplemented diet containing 300 (E300) or 600 (E600) mg alpha-tocopheryl acetate/kg for 21 weeks. 2. Dietary supplementation with alpha-tocopheryl acetate significantly reduced TBARS numbers in both raw and cooked burgers during refrigerated and frozen storage. 3. The mean values of alpha-tocopherol in raw and cooked burgers stored at 4 degrees C did not change during storage. 4. In the case of both raw and cooked samples stored at -20 degrees C, the alpha-tocopherol values decreased from 5.67 to 3.54 and from 3.56 to 2.30 micrograms/g in the raw burgers from turkeys from the E600 and E300 treatments, respectively, after 4 months storage. The values decreased from 5.60 to 2.88 and from 3.29 to 1.85 micrograms/g in cooked burgers from turkeys from the E600 and E300 treatments, respectively, after 5 months storage.

Animals↗

PubMed1996

Midazolam in the treatment of refractory neonatal seizures.

Midazolam, a potent short-acting benzodiazepine, is a safe and highly effective agent for the control of status epilepticus. Its efficacy in the control of neonatal seizures, however, has not been determined. Six neonates (aged 1-9 days; gestation, 30-41 weeks) developed seizures from a variety of causes. In each case, seizures persisted for > 12 h despite high-dose phenobarbital therapy with or without the addition of phenytoin. Midazolam was then administered by continuous intravenous infusion (0.1-0.4 mg/kg/h) for 1 to 3 days. Within 1 h of initiation of midazolam, seizures were controlled in all six neonates. Electroencephalographic seizures were abolished in four of six neonates; however, two neonates continued to have electrographic seizures (without clinical accompaniment) for a further 12 h. Blood pressure and pulse rate were not changed after the initiation of midazolam, and adverse reactions were not observed in any neonate. Because more than one third of all neonatal seizures are refractory to high-dose phenobarbital and phenytoin, midazolam administered by continuous intravenous infusion may be a valuable adjunctive therapy.

Anticonvulsants↗

PubMed1996

Rapid modulation of the apoptosis regulatory genes, bcl-2 and bax by prolactin in rat Nb2 lymphoma cells.

The Nb2 rat lymphoma represents a useful model for investigation of molecular events coupled to PRL-induced proliferation. Moreover, recent evidence has also demonstrated the utility of this system to study mechanisms linked to programmed cell death (apoptosis). Thus, glucocorticosteroids activate apoptosis in lactogen-dependent Nb2 cells, whereas the addition of PRL abrogates this effect. The present study was conducted to determine whether PRL stimulation in lactogen-dependent Nb2-11 or autonomous Nb2-SFJCD1 cultures alters expression of bcl-2 or bax, genes that suppress or facilitate apoptosis, respectively. We demonstrate that PRL stimulation in stationary Nb2-11 cultures significantly increased the level of bcl-2 messenger RNA (mRNA) within 3 h (15-fold) and its protein product by 6 h, time points previously shown to correspond with G1 cell cycle progression. In Nb2-SFJCD1 cells, bcl-2 mRNA was found to be constitutively present. Addition of PRL to these lactogen-independent cultures further enhanced its expression at the mRNA and protein levels with a kinetic pattern similar to that observed in the PRL-dependent line. Results from stability studies indicated that increased bcl-2 mRNA evoked by PRL in Nb2 cell lines was most likely not attributable to increased stability of its transcripts. Furthermore, the rapid increase in its expression in PRL-stimulated Nb2-11 cells was not altered by inhibition of protein synthesis suggesting a direct action of the hormone. PRL also increased bax mRNA by 8 h in Nb2-11 cultures. However, hormone stimulation markedly attenuated the level of the Bax protein from 2-6 h. In contrast, bax expression in Nb2-SFJCD1 cultures remained unaltered by the addition of the hormone. These results demonstrate that altered expression of bcl-2 and bax are each associated with PRL-stimulated cell cycle progression in Nb2 cells. Moreover, bcl-2 appears to be an immediate-early gene induced by PRL in the hormone-dependent line and may represent an important regulator of early G1 cell cycle progression most likely by suppressing cell death mechanisms.

Animals↗

PubMed1996

Alterations in pim-1 and c-myc expression associated with sodium butyrate-induced growth factor dependency in autonomous rat Nb2 lymphoma cells.

Whereas Nb2-11 lymphoma cells critically require prolactin (PRL) for growth, Nb2-SFJCD1 subline cells are growth factor independent. Treatment with the differentiating agent, sodium butyrate (NaBT), has been demonstrated previously to lead to growth arrest of Nb2-SFJCD1 cells and a transient reversion to PRL growth requirement following removal of NaBT. In the present study, the relation of NaBT-induced growth arrest to the cell cycle was examined using flow cytometry, and the effect of PRL on expression of the immediate-early proto-oncogenes, pim-1 and c-myc, in NaBT-pretreated cultures was evaluated. Treatment of Nb2-SFJCD1 cells with 2 mM NaBT for 72 h caused growth arrest in the majority of the cells in the G1 phase of the cell cycle, an effect similar to that produced by lactogen deprivation in PRL-dependent Nb2 cultures. The addition of PRL stimulated a concentration-dependent re-entry into the cell cycle. In other experiments, NaBT treatment significantly reduced the steady-state levels of pim-1 and c-myc mRNA. Stimulation with PRL induced a rapid and concentration-dependent biphasic accumulation of each mRNA with similar kinetics. Maximal expression of both proto-oncogenes occurred within 2-4 h and after 12 h. Results from mRNA stability studies suggest that the observed increases in expression of pim-1 and c-myc most likely do not reflect increased stability of the transcripts. The results indicate that NaBT-induced differentiation in autonomous Nb2-SFJCD1 causes growth arrest of the cells in the G1 phase of the cell cycle and reduces the basal levels of pim-1 and c-myc mRNAs. Mitogenic stimulation with PRL reinitiates cell cycle progression characterized by biphasic expression of each proto-oncogene. It is suggested that NaBT is a useful tool for investigation of the malignant progression from growth factor dependency to autonomy in the Nb2 lymphoma paradigm.

Animals↗

PubMed1995

A novel heat shock response in prolactin-dependent Nb2 node lymphoma cells.

Virtually all cells respond to heat stress by increased expression or induction of one or more of the highly conserved cellular stress response proteins, heat shock proteins (HSPs). Here, we report the unusual property of rat Nb2-11 cells, a prolactin-dependent pre-T-cell line, to display reduced HSP expression following exposure to elevated temperature. After heat stress (41 degrees C, 1 h), there was no evidence of inducible members of the 70 kDa HSP family, a response common to other cell culture and tissue systems. Moreover, expression of constitutive members of the HSP70 and HSP90 families decreased during the heat stress, apparently reflecting a decrease in mRNA stability. Gel shift assays revealed that heat shock factor (HSF) was activated in spite of the lack of expression of inducible HSP70 transcripts, although its DNA binding rapidly deteriorated. Immunoblotting, using an antibody specific to HSF1, indicated that proteolysis of HSF1 may be responsible for this rapid termination of heat shock element binding. CCAAT binding, a component of constitutive HSP70 expression, was also reduced by heat stress in Nb2-11 cells and may account for the decline in constitutive HSP70 expression. Prolactin pretreatment prevented the fragmentation of HSF1, protected heat shock element and CCAAT binding, prevented the decline in constitutive HSP70 and HSP90 expression, and restored a modest expression of inducible HSP70 following heat treatment. Results of this study describe a unique regulatory defect in HSP expression in Nb2-11 cells, possibly a common characteristic of other hormone-dependent tumors.

Animals↗

PubMed1995

The nuclear prolactin receptor: a 62-kDa chromatin-associated protein in rat Nb2 lymphoma cells.

Previously we demonstrated that lactogen-dependent Nb2 cells express a nuclear prolactin (PRL) receptor. Thus, the nuclear receptor expressed in PRL-dependent Nb2-11 and -independent Nb2-SFJCD1 cells was characterized. Initially, the potential proteolytic processing of internalized 125I-rPRL was investigated. Radiolabeled hormone eluted from a Sephadex G-100 column with a retention time identical to that found for stock hormone, indicating that nuclear PRL was intact. Experiments to investigate the nuclear distribution of the hormone demonstrated that 90% of 125I-rPRL bound to chromatin; the remaining was distributed between "sap-protein" and nucleoplasmic fractions. Chromatin-bound PRL was resistant to high salt and detergent extraction indicating a tight association. Immunoprecipitation and immunoblot analysis revealed the PRL receptor to be 62 kDa in each cell line. Affinity crosslinking experiments and immunoprecipitation demonstrated that 125I-rPRL complexed with a protein(s) of similar M(r) in intact cells. 125I-rPRL binding was saturable and of high affinity (Kds of 180 and 170 pM, for Nb2-11 and Nb2-SFJCD1 lines, respectively). PRL binding was competitively inhibited by ovine and bovine PRLs and hGH, but not by rat GH, and by monoclonal antibodies (McAbs) which recognize the lactogen binding site. These results demonstrate that: (1) 125I-rPRL translocates intact to the Nb2 cell nucleus and tightly associates with chromatin; (2) the chromatin receptor specifically binds 125I-rPRL with high affinity; and (3) the chromatin receptor is essentially identical to its membrane counterpart with respect to mass, binding characteristics, and McAb recognition.

Animals↗

PubMed1995

Nuclear translocation of prolactin: collaboration of tyrosine kinase and protein kinase C activation in rat Nb2 node lymphoma cells.

Recent evidence has suggested that prolactin (PRL), internalized by lactogen-dependent Nb2 lymphoma cells, is actively translocated to the nucleus where it binds to PRL receptors. Moreover, the mitogenic action of PRL in these cells has been separately linked to protein tyrosyl phosphorylation and activation of protein kinase C (PKC). Therefore, the coupling of PRL internalization and nuclear translocation to the activation of these signal transduction pathways was investigated. Results from control experiments indicated that 30% of internalized and 5% total cell-associated 125I-rat PRL could be recovered within nuclei obtained from Nb2 cells previously incubated with the radiolabel for 3 h at 37 degrees C. Furthermore, internalized PRL was found to be intact and not associated with any carrier proteins. Addition of tyrosine kinase (TK) antagonists, genistein or tyrphostin, significantly reduced cell surface binding, internalization, and nuclear translocation of 125I-rat PRL. In contrast, neither the level of cell-associated nor internalized hormone differed between cells treated with the PKC antagonists, staurosporine or calphostin C, and control cultures. Instead, PKC inhibition significantly reduced nuclear PRL translocation. The inhibitory effects of the TK and PKC antagonists on PRL internalization and nuclear translocation in intact Nb2 cells were verified by immunofluorescence microscopy in parallel experiments. In other experiments, each of the kinase inhibitors blocked PRL-stimulated Nb2 cell proliferation in a concentration-dependent manner. It is concluded that activated TK and PKC collaborate in the process of PRL internalization and translocation to the nucleus. TK activation may participate in PRL receptor binding or hormone internalization while activation of PKC appears to be required for its nuclear targeting. Since TK and PKC activation are required for lactogen-stimulated Nb2 cell proliferation, we suggest that a component of the mitogenic pathway in these cells is a direct nuclear interaction of PRL.

Animals↗

PubMed1995

Rapid induction of pim-1 expression by prolactin and interleukin-2 in rat Nb2 lymphoma cells.

The lactogen-dependent Nb2 lymphoma line (Nb2-11) represents a useful pre-T cell model for investigation of early molecular events coupled to PRL-stimulated cell cycle progression. Expression of pim-1, a protooncogene that encodes a conserved cytosolic serine/threonine protein kinase, is rapidly induced in hematopoietic cells upon mitogen stimulation and is thought to be important for lymphocyte activation. The present study was conducted to determine whether mitogen stimulation in Nb2-11 or lactogen-independent Nb2-SFJCD1 cells provokes pim-1 gene expression. The pim-1 transcript was undetectable in control growth-arrested Nb2-11 cultures; however, PRL rapidly stimulated its expression in a biphasic manner. Peak expression occurred within 2-4 h (> 40-fold) and was followed by a second elevation at 12 h. The effect of PRL and IL-2 to induce pim-1 at 2 h was concentration dependent and not inhibited by cycloheximide. In Nb2-SFJCD1 cells, pim-1 messenger RNA was expressed in control cultures and augmented by PRL stimulation. Results from stability studies indicated that the t1/2 values for the pim-1 transcript were 79 and 81 min in PRL-stimulated Nb2-11 cells at 2 and 12 h. However, in the lactogen-treated Nb2-SFJCD1 line, it was nearly 3-fold more stable (219 min) at 2 h compared to that determined at either 12 h or in unstimulated cultures. In other experiments, PRL-stimulated expression of the pim-1 protein was evaluated in [35S]methionine-labeled cells by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In Nb2-11 cells, enhanced [35S]pim-1 expression paralleled its messenger RNA transcription through 8 h. Elevated [35S]pim-1 was detected within 1 h and peaked by 2-4 h. Therefore, pim-1 represents an immediate early gene induced by PRL stimulation in Nb2-11 cells. Its initial peak of transcription occurs early during G1 cell cycle progression, whereas a second elevation is coincident with the G1/S transition. These results demonstrate that mitogen-induced expression of pim-1 is a rapid event in Nb2 lymphoma cells and suggest that it may be associated with cell cycle progression.

Animals↗

PubMed1995

Influence of dietary vitamin E on the oxidative stability and quality of pig meat.

Oxidation of lipids is a major cause of deterioration in the quality of muscle foods and can directly affect many quality characteristics such as flavor, color, texture, nutritive value, and safety of the food. Lipid oxidation in muscle systems is initiated at the membrane level in the intracellular phospholipid fractions. In the processing of muscle foods, one of the most important questions concerns the methods used to delay the initiation of oxidation and loss of quality. Vitamin E is a major lipid-soluble antioxidant, and one of its primary functions is to maintain and protect biological membranes against lipid peroxidation. Dietary vitamin E supplementation above requirement levels is effective in reducing lipid oxidation. This review focuses on the antioxidant function of vitamin E and how supplementation of the diet of pigs with vitamin E influences the rate of lipid peroxidation, color, water-holding capacity, and cholesterol oxidation in pig meat.

Animals↗