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Vascular dysfunction has emerged as a significant risk factor for the development of cardio- and cerebrovascular diseases (CVDs), which are currently the leading cause of morbidity and mortality worldwide. T-lymphocytes (T-cells) have been shown to be important modulators of vascular function in primary aging and CVDs, likely by producing inflammatory cytokines and reactive oxygen species that influence vasoprotective molecules. This review summarizes the role of T-cells on vascular function in aging, hypertension, and atherosclerosis in animals and humans, and discusses potential T-cell targeted therapeutics to prevent, delay, or reverse vascular dysfunction.

Aging of the arteries is characterized by increased large artery stiffness and impaired endothelium-dependent dilation. We have previously shown that in old (22-24 month) mice T cells accumulate within aorta and mesentery. We have also shown that pharmacologic and genetic deletion of these T cells ameliorates age-related arterial dysfunction. These data indicate that T cells contribute to arterial aging; however, it is unknown if aged T cells alone can induce arterial dysfunction in otherwise young mice. To produce an aged-like T cell phenotype, mice were thymectomized at three-weeks of age or were left with their thymus intact. At 9 months of age, thymectomized mice exhibited greater proportions of both CD4 + and CD8 + memory T cells compared to controls in the blood. Similar changes were observed in the T cells accumulating in the aorta and mesentery. We also observed greater numbers of proinflammatory cytokine producing T cells in the aorta and mesentery. The phenotypic T cell changes in the blood, aorta and mesentery of thymectomized mice were similar to those observed when we compared young (4-6 month) to old thymus intact mice. Along with these alterations, compared to controls, thymectomized mice exhibited augmented large artery stiffness and greater aortic collagen deposition as well as impaired mesenteric artery endothelium dependent dilation due to blunted nitric oxide bioavailability. These results indicate that early life thymectomy results in arterial dysfunction and suggest that an aged-like T cell phenotype alone is sufficient to induce arterial dysfunction in otherwise young mice., (© 2023. The Author(s), under exclusive licence to American Aging Association.)

Background: Marked increases in serum muscle enzyme activity can occur in endurance horses but the diagnostic certainty in predicting cases of myopathy is unclear. Improved understanding of horse management effects on serum muscle enzyme activity as markers of muscle health would assist interpretation of serum muscle enzyme activity and guide management to reduce myopathy risk., Objectives: To investigate associations between serum muscle enzyme activity and management factors in endurance horses., Study Design: Cross-sectional study., Methods: One hundred endurance horses competing in four endurance events (offering distances of 20-120 km) in south-eastern Australia were observed. Data were collected from official horse logbooks, pre-and post-race serum samples, an owner questionnaire of pre-race and race management of horses and the Australian Endurance Riders Association results database. Multivariable linear regression modelling tested associations between management factors and serum muscle enzyme activity., Results: First leg speed, distance raced, number of rest days pre-race, and pre-race activity of aspartate aminotransferase (AST) and creatine kinase (CK) explained 47.3% of the variance in post-race CK. As first leg speed increased by 1 km/h, CK activity increased by 25.8% (95% CI 11%-35%). Race distances >80 km increased post-race CK activity by 124% (95% CI 116%-145%). Each additional pre-race rest day increased post-race CK activity by 30.5% (95% CI 11%-42%). Modelling a 10% increase in pre-race CK and pre-race AST activity was associated with post-race CK activity increasing by 7.3% (95% CI 3%-14.4%) and 8.5% (95% CI 0.3%-14.2%) respectively. Horses experiencing training distances >40 km and a greater number of rest days prior to race day developed increased pre-race AST and CK activity respectively., Main Limitations: Owner questionnaires may be subject to bias. Limited data were available to model ride terrain, horse fitness, ration detail and myopathy. Muscle biopsies were not used to confirm myopathy., Conclusions: Nearly half of the variation in post-race CK activity observed can be attributed to management factors unrelated to myopathy, suggesting increased CK activity may not be pathognomonic for myopathy. We advise caution in relying solely on serum muscle enzyme activity for diagnosis of myopathy until the strength of association between CK and myopathy is further ascertained in future studies., (© 2021 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Both glucose tolerance and adaptive immune function exhibit significant age-related alterations. The influence of the immune system on obesity-associated glucose intolerance is well characterized; however, whether the immune system contributes to age-related glucose intolerance is not as well understood. Here, we report that advancing age results in an increase in T cell infiltration in the epididymal white adipose tissue (eWAT), liver, and skeletal muscle. Subtype analyses show that both CD4+, CD8+ T cells are greater with advancing age in each of these tissues and that aging results in a blunted CD4 to CD8 ratio. Anti-CD3 F(ab')2 fragments depleted CD4+ and CD8+ cells in eWAT, CD4+ cells only in the liver, and did not deplete quadriceps T cells. In old mice, T cells producing both interferon-γ and tumor necrosis factor-α are accumulated in the eWAT and liver, and a greater proportion of skeletal muscle T cells produced interferon-γ. Aging resulted in increased proportion and numbers of T regulatory cells in eWAT, but not in the liver or muscle. Aging also resulted in greater numbers of eWAT and quadriceps CD206- macrophages and eWAT, liver and quadriceps B cells; neither cell type was altered by anti-CD3 treatment. Anti-CD3 treatment improved glucose tolerance in old mice and was accompanied by improved signaling related to liver and skeletal muscle insulin utilization and decreased gluconeogenesis-related gene expression in the liver. Our findings indicate a critical role of the adaptive immune system in the age-related metabolic dysfunction.

Graphite intercalation compounds (GICs) are often used to produce exfoliated or functionalised graphene related materials (GRMs) in a specific solvent. This study explores the formation of the Na-tetrahydrofuran (THF)-GIC and a new ternary system based on dimethylacetamide (DMAc). Detailed comparisons of in situ temperature dependent XRD with TGA-MS and Raman measurements reveal a series of dynamic transformations during heating. Surprisingly, the bulk of the intercalation compound is stable under ambient conditions, trapped between the graphene sheets. The heating process drives a reorganisation of the solvent and Na molecules, then an evaporation of the solvent; however, the solvent loss is arrested by restacking of the graphene layers, leading to trapped solvent bubbles. Eventually, the bubbles rupture, releasing the remaining solvent and creating expanded graphite. These trapped dopants may provide useful property enhancements, but also potentially confound measurements of grafting efficiency in liquid-phase covalent functionalization experiments on 2D materials., (© 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)

Introduction: The geographical vastness of Australia in rural and remote regions provides challenges for hospital nursing staff when a medical officer is not immediately available. Delaying treatment until a medical officer arrives can potentially impact on patient outcomes. Some Registered Nurses in rural and remote New South Wales have received advanced training and credentialing through the First Line Emergency Care Course (FLECC) enabling them to initiate treatment prior to the medical officer's arrival. Despite operating since 1990 the course and subsequent model of care has received limited evaluation., Purpose: The aim of this study was to evaluate whether undertaking the First Line Emergency Care Course (FLECC) improves confidence in rural and remote registered nurses' (RN) ability to provide and to initiate extended scope of practice for emergency care of critically ill adults in the absence of a medical officer., Procedure: A voluntary electronic cross sectional 19 question multi-centre survey was emailed to all nurses FLECC accredited since 2008 and employed in rural and remote Local Health District in New South Wales., Findings: A response rate of 60% was obtained (225/375). Since credentialing 93.3% nurses had commenced emergency treatments under the approved guidelines. The majority (97.4%) agreed that the course provided the required knowledge. The mean Likert nurse's self-assessed confidence score (1-5) for providing first line response rose significantly (Wilcoxon signed rank test V=375, p-value<0.0001) from 2.47 and up to 3.98 after the course. The majority (83.9%) now had "high" or "very high" confidence., Principle Conclusions: The FLECC provides registered nurses in rural and remote areas with skills, confidence, knowledge and ability to initiate potentially lifesaving treatment for critically ill or injured patients prior to the arrival of a medical officer. The FLECC model of care helps provide timely care delivery by nurses in rural and remote areas. Further research could examine the differences in outcome in patients treated at sites with FLECC nurses., (Copyright © 2019. Published by Elsevier Ltd.)

Grafting polymers onto single-walled carbon nanotubes (SWCNTs) usefully alters properties but does not typically yield stable, solvated species directly. Despite the expectation of steric stabilization, a damaging (re)dispersion step is usually necessary. Here, poly(vinyl acetate)s (PVAc's) of varying molecular weights are grafted to individualized, reduced SWCNTs at different concentrations to examine the extent of reaction and degree of solvation. The use of higher polymer concentrations leads to an increase in grafting ratio (weight fraction of grafted polymer relative to the SWCNT framework), approaching the limit of random sequentially adsorbed Flory "mushrooms" on the surface. However, at higher polymer concentrations, a larger percentage of SWCNTs precipitate during the reaction; an effect which is more significant for larger weight polymers. The precipitation is attributed to depletion interactions generated by ungrafted homopolymer overcoming Coulombic repulsion of adjacent like-charged SWCNTs; a simple model is proposed. Larger polymers and greater degrees of functionalization favor stable solvation, but larger and more concentrated homopolymers increase depletion aggregation. By using low concentrations (25 μM) of larger molecular weight PVAc (10 kDa), up to 65% of grafted SWCNTs were retained in solution (at 65 μg mL -1 ) directly after the reaction.

COX20/FAM36A encodes a mitochondrial complex IV assembly factor important for COX2 activation. Only one homozygous COX20 missense mutation has been previously described in two separate consanguineous families. We report four subjects with features that include childhood hypotonia, areflexia, ataxia, dysarthria, dystonia, and sensory neuropathy. Exome sequencing in all four subjects identified the same novel COX20 variants. One variant affected the splice donor site of intron-one (c.41A>G), while the other variant (c.157+3G>C) affected the splice donor site of intron-two. cDNA and protein analysis indicated that no full-length cDNA or protein was generated. These subjects expand the phenotype associated with COX20 deficiency.

We have discovered that wrapping single-walled carbon nanotubes (SWCNTs) with ionic surfactants induces a switch in the conductance-humidity behaviour of SWCNT networks. Residual cationic vs. anionic surfactant induces a respective increase or decrease in the measured conductance across the SWCNT networks when exposed to water vapour. The magnitude of this effect was found to be dependent on the thickness of the deposited SWCNT films. Previously, chemical sensors, field effect transistors (FETs) and transparent conductive films (TCFs) have been fabricated from aqueous dispersions of surfactant functionalised SWCNTs. The results reported here confirm that the electrical properties of such components, based on randomly orientated SWCNT networks, can be significantly altered by the presence of surfactant in the SWCNT layer. A mechanism for the observed behaviour is proposed based on electrical measurements, Raman and UV-Vis-NIR spectroscopy. Additionally, the potential for manipulating the sensitivity of the surfactant functionalised SWCNTs to water vapour for atmospheric humidity sensing was evaluated. The study also presents a simple method to establish the effectiveness of surfactant removal techniques, and highlights the importance of characterising the electrical properties of SWCNT-based devices in both dry and humid operating environments for practical applications.

Background: Pediatric arterial ischemic stroke remains incompletely understood. Population-based epidemiological data inform clinical trial design but are scant in this condition. We aimed to determine age-specific epidemiological characteristics of arterial ischemic stroke in neonates (birth to 28 days) and older children (29 days to 18 years)., Methods: We conducted a 16-year, prospective, national population-based study, the Canadian Pediatric Ischemic Stroke Registry, across all 16 Canadian acute care children's hospitals. We prospectively enrolled children with arterial ischemic stroke from January 1992 to December 2001 and documented disease incidence, presentations, risk factors, and treatments. Study outcomes were assessed throughout 2008, including abnormal clinical outcomes (stroke-related death or neurological deficit) and recurrent arterial ischemic stroke or transient ischemic attack., Results: Among 1129 children enrolled with arterial ischemic stroke, stroke incidence was 1.72/100,000/year, (neonates 10.2/100,000 live births). Detailed clinical and radiological information were available for 933 children (232 neonates and 701 older children, 55% male). The predominant clinical presentations were seizures in neonates (88%), focal deficits in older children (77%), and diffuse neurological signs (54%) in both. Among neonates, 44% had no discernible risk factors. In older children, arteriopathy (49% of patients with vascular imaging), cardiac disorders (28%), and prothrombotic disorders (35% of patients tested) predominated. Antithrombotic treatment increased during the study period (P < 0.001). Stroke-specific mortality was 5%. Outcomes included neurological deficits in 60% of neonates and 70% of older children. Among neonates, deficits emerged during follow-up in 39%. Overall, an initially decreased level of consciousness, a nonspecific systemic presentation, and the presence of stroke risk factors predicted abnormal outcomes. For neonates, predictors were decreased level of consciousness, nonspecific systemic presentation, and basal ganglia infarcts. For older children, predictors were initial seizures, nonspecific systemic presentation, risk factors, and lack of antithrombotic treatment. Recurrent arterial ischemic stroke or transient ischemic attack developed in 12% of older children and was predicted by arteriopathy, presentation without seizures, and lack of antithrombotic treatment. Emerging deficit was predicted by neonatal age at stroke and by cardiac disease., Conclusions: This national data set provides a population-based disease incidence rate and demonstrates the protective effect of antithrombotic treatment in older children, and frequent long-term emerging deficits in neonates and in children with cardiac disorders. Further clinical trials are required to develop effective age-appropriate treatments for children with acute arterial ischemic stroke., (Copyright © 2017. Published by Elsevier Inc.)

Chemical and electrochemical reduction methods allow the dispersion, processing, and/or functionalization of discrete sp 2 -hybridised nanocarbons, including fullerenes, nanotubes and graphenes. Electron transfer to the nanocarbon raises the Fermi energy, creating nanocarbon anions and thereby activating an array of possible covalent reactions. The Fermi level may then be partially or fully lowered by intended functionalization reactions, but in general, techniques are required to remove excess charge without inadvertent covalent reactions that potentially degrade the nanocarbon properties of interest. Here, simple and effective chemical discharging routes are demonstrated for graphenide polyelectrolytes and are expected to apply to other systems, particularly nanotubides. The discharging process is inherently linked to the reduction potentials of such chemical discharging agents and the unusual fundamental chemistry of charged nanocarbons.

Strategies for forming liquid dispersions of nanomaterials typically focus on retarding reaggregation, for example via surface modification, as opposed to promoting the thermodynamically driven dissolution common for molecule-sized species. Here we demonstrate the true dissolution of a wide range of important 2D nanomaterials by forming layered material salts that spontaneously dissolve in polar solvents yielding ionic solutions. The benign dissolution advantageously maintains the morphology of the starting material, is stable against reaggregation and can achieve solutions containing exclusively individualized monolayers. Importantly, the charge on the anionic nanosheet solutes is reversible, enables targeted deposition over large areas via electroplating and can initiate novel self-assembly upon drying. Our findings thus reveal a unique solution-like behaviour for 2D materials that enables their scalable production and controlled manipulation.

Carbon nanotube-based gas sensors can be used to detect harmful environmental pollutants such as NO 2 at room temperature. Although they show promise as low-powered, sensitive, and affordable monitoring devices, cross-sensitivity of functionalized carbon nanotubes to water vapor often obscures the detection of target molecules. This is a barrier to adoption for monitoring of airborne pollutants because of the varying humidity levels found in real world environments. Zeolites, also known as molecular sieves because of their selective adsorption properties, are used in this work to control the cross-sensitivity of single-walled carbon nanotube (SWCNT)-based sensors to water vapor. Zeolites incorporated into the sensing layer are found to reduce interference effects that would otherwise obscure the identification of NO 2 gas, permitting repeatable detection over a range of relative humidities. This significant improvement is found to depend on the arrangement of the SWCNT-zeolite layers in the sensing device, as well as the hydrophilicity of the chosen zeolite.

Objective: The Wagga Wagga Ambulatory Rehabilitation Service (WWARS) clinicians felt the Australian Modified Lawton's Instrumental Activities of Daily Living Scale (Lawton Scale) was unresponsive to changes detected in many patients following their individualised programs. This study examined the performance of the Goal Attainment Scaling (GAS) concomitantly with the Lawton Scale in ambulatory rehabilitation patients., Methods: Consecutive patients (n = 83) of WWARS were evaluated pre and post treatment with the Lawton Scale and GAS. The statistical properties, particularly responsiveness, of the scales were compared., Results: Statistically significant change (P < 0.001) was observed following treatment on both the Lawton Scale and GAS. Mean GAS scores changed by 38.7% compared with 8.3% for the Lawton Scale. GAS demonstrated a greater effect size (Cliff's δ) of 1.67 (95% confidence interval 1.51-1.91) than the Lawton Scale 0.83 (95% confidence interval 0.57-0.94). In 33.7% of patients, the Lawton Scale was invariant but improved with GAS., Conclusions: GAS was a more responsive measure than the Lawton Scale in rural ambulatory rehabilitation patients. Consequently, GAS is recommended as a performance outcome measure in the evaluation of ambulatory rehabilitation services to supplement standardised outcome measures such as the Lawton Scale.

Chemical charging of single-walled carbon nanotubes (SWCNTs) and graphenes to generate soluble salts shows great promise as a processing route for electronic applications, but raises fundamental questions. The reduction potentials of highly-charged nanocarbon polyelectrolyte ions were investigated by considering their chemical reactivity towards metal salts/complexes in forming metal nanoparticles. The redox activity, degree of functionalisation and charge utilisation were quantified via the relative metal nanoparticle content, established using thermogravimetric analysis (TGA), inductively coupled plasma atomic emission spectroscopy (ICP-AES) and X-ray photoelectron spectroscopy (XPS). The fundamental relationship between the intrinsic nanocarbon electronic density of states and Coulombic effects during charging is highlighted as an important area for future research.

Using the foraging movements of an insectivorous bat, Myotis mystacinus, we describe temporal switching of foraging behaviour in response to resource availability. These observations conform to predictions of optimized search under the Lévy flight paradigm. However, we suggest that this occurs as a result of a preference behaviour and knowledge of resource distribution. Preferential behaviour and knowledge of a familiar area generate distinct movement patterns as resource availability changes on short temporal scales. The behavioural response of predators to changes in prey fields can elicit different functional responses, which are considered to be central in the development of stable predator–prey communities. Recognizing how the foraging movements of an animal relate to environmental conditions also elucidates the evolution of optimized search and the prevalence of discrete strategies in natural systems. Applying techniques that use changes in the frequency distribution of movements facilitates exploration of the processes that underpin behavioural changes.

Reasons for Performing Study: Horse misbehaviour is an important cause of poor performance in Pony Club horses, is associated with horse-related rider injuries and has been implicated as a nonspecific presenting sign for musculoskeletal pain. Despite this, little is known about the incidence of and risk factors for misbehaviour in Pony Club horses., Objective: This study aimed to describe the incidence and types of misbehaviour in a cohort of Pony Club horses and to identify risk factors for misbehaviour during riding., Methods: A prospective longitudinal study was conducted with 84 Pony Club horses from 41 families belonging to 7 Pony Clubs in one inland region of Australia. Owners recorded misbehaviour events and kept daily records of horse housing, exercise, nutrition, healthcare and disease status. Horses were subjected to a monthly veterinary examination. Descriptive statistics were calculated to describe the incidence of misbehaviour, and multivariable logistic regression was used to assess putative risk factors., Results: Misbehaviour during riding occurred on 3% of days when horses were ridden. On 52% of days with misbehaviour, the misbehaviour was classified as dangerous. Risk of misbehaviour was independently increased on exercise days when the horse was competing, and in months when the horse was fat or obese, fed supplementary feed daily, grazed on paddocks with >50% of ground cover as green grass, exercised on 5 days per month or less, and ridden for a total of 12 h or more in the month. No significant relationship was detected between misbehaviour and back pain., Conclusions: In populations such as the study population the risk of misbehaviour is higher in fatter horses, in horses with access to pastures with greater green grass cover, in those fed daily supplements, in horses receiving exercise less frequently, and during competition., Potential Relevance: These results highlight the importance of considering horse body condition, nutrition and exercise in any investigation of horse misbehaviour. In addition, recommendations based on these results could be used by veterinarians assisting horse owners to prevent misbehaviour. From the perspective of recreational horse owners, behaviour is a key determinant of horse performance., (© 2012 EVJ Ltd.)

Solutions of negatively charged graphene (graphenide) platelets were produced by intercalation of nanographite with liquid potassium-ammonia followed by dissolution in tetrahydrofuran. The structure and morphology of these solutions were then investigated by small-angle neutron scattering. We found that >95 vol % of the solute is present as single-layer graphene sheets. These charged sheets are flat over a length scale of >150 Å in solution and are strongly solvated by a shell of solvent molecules. Atomic force microscopy on drop-coated thin films corroborated the presence of monolayer graphene sheets. Our dissolution method thus offers a significant increase in the monodispersity achievable in graphene solutions.

Objective: To assess the impact of the opening of an after-hours general practice clinic on the number of daily low-urgency presentations to the nearby emergency department., Design, Participants and Setting: Retrospective time series analysis of emergency presentation data, from the New South Wales Health Emergency Department Information System, for all patients presenting to the emergency department of Wagga Wagga Base Hospital between January 1998 and October 2008., Main Outcome Measures: Daily emergency department presentations, before and after the March 2003 opening of the after-hours clinic, of patients triaged as Australasian Triage Scale (ATS) category 4 or 5 (at any time of day, and during the hours of operation of the clinic), and of patients triaged as ATS category 1, 2 or 3 (at any time of day)., Results: After adjusting for long-term trends and weekly and annual cycles, the opening of the after-hours clinic was associated with a daily reduction of 7.04 patients (95% CI, 5.39-8.70) in emergency department presentations with an ATS category of 4 or 5. This represented an 8.2% reduction in total presentations (95% CI, 6.2%-10.2%). Presentations of ATS category 1, 2 or 3 patients rose by 1.36 patients a day (95% CI, 0.36-2.35), representing 1.6% of total presentations (95% CI, 0.4%-2.7%). The impact of the after-hours clinic was best modelled by a gradual permanent change., Conclusion: An after-hours general practice clinic was associated with a reduction in low-urgency presentations to the emergency department in Wagga Wagga.

In adults newly diagnosed with epilepsy, treatment with the first prescribed antiepileptic drug fails for approximately one half. In two studies that addressed this question in children, the failure rates were 20% and 40%. The present study used a detailed chart review of children newly diagnosed with epilepsy over a 4-year span in a major childhood epilepsy referral clinic to assess (1) the percentage of children for whom first-line antiepileptic drug treatment failed and (2) the reasons for the treatment failure. Charts were reviewed for 95 children who were diagnosed with epilepsy, started on their first antiepileptic drug, and then monitored for approximately 5 years. Of these 95 children, 48 were classified as having idiopathic epilepsy (50.5 %), 30 as having cryptogenic epilepsy (31.6%), and 17 as having symptomatic epilepsy (17.9%). The two main antiepileptic drugs used were valproic acid (43.2% of patients) and carbamazepine (38.9% of patients). Treatment with the first antiepileptic drug failed in 30/95 children (31.6%). Treatment failure was due to adverse effects in 12/30 children (40.0%), due to lack of efficacy in 11/30 (37.9%), and due to both adverse effects and lack of efficacy in 7/30 (24.1%). Also examined was the effect on treatment failure of patient age at diagnosis, antiepileptic drug choice, maximum drug dose, etiology of epilepsy, and particular epilepsy syndromes on treatment failures; there was no statistically significant effect of any of these variables on first-line treatment outcome. In this population, approximately one third of children newly diagnosed with epilepsy experienced treatment failure with the first antiepileptic drug used. Lack of efficacy and unacceptable adverse effects contributed equally to these treatment failures.

Previously we observed that the antiestrogens tamoxifen and 4-hydroxytamoxifen (4OHT) induce CYP3A4 in primary human hepatocytes and activate human pregnane X receptor (PXR) in cell-based reporter assays. Given the complex cross-talk between nuclear receptors, tissue-specific expression of CYP3A4, and the potential for tamoxifen and 4OHT to interact with a myriad of receptors, this study was undertaken to gain mechanistic insights into the inductive effects of tamoxifen and 4OHT. First, we observed that transfection of the primary cultures of human hepatocytes with PXR-specific small interfering RNA reduced the PXR mRNA expression and the extent of CYP3A4 induction by tamoxifen and 4OHT by 50%. Second, in LS174T colon carcinoma cells, which were observed to have significantly lower PXR expression relative to human hepatocytes, neither tamoxifen nor 4OHT induced CYP3A4. Third, N-desmethyltamoxifen, which did not induce CYP3A4 in human hepatocytes, also did not activate PXR in LS174T cells. We then used cell-based reporter assay to evaluate the effects of other receptors such as glucocorticoid receptor GR alpha and estrogen receptor ER alpha on the transcriptional activation of PXR. The cotransfection of GR alpha in LS174T cells augmented PXR activation by tamoxifen and 4OHT. On the other hand, the presence of ER alpha inhibited PXR-mediated basal activation of CYP3A4 promoter, possibly via competing for common cofactors such as steroid receptor coactivator 1 and glucocorticoid receptor interacting protein 1. Collectively, our findings suggest that the CYP3A4 induction by tamoxifen and 4OHT is primarily mediated by PXR but the overall stoichiometry of other nuclear receptors and transcription cofactors also contributes to the extent of the inductive effect.

We previously reported the identification of HRPAP20 (hormone-regulated proliferation-associated protein 20), a novel hormone-regulated, proliferation-associated protein. In tumor cell lines, constitutive HRPAP20 expression enhanced proliferation and suppressed apoptosis, characteristics frequently associated with malignant progression. Here, we report that highly invasive breast cancer cell lines and human breast tumor specimens express elevated HRPAP20, which in transfection experiments in MCF-7 and MDA-MB-231 cells, increased invasion. Results from mechanistic studies revealed that HRPAP20 bound to calmodulin (CaM) via a conserved CaM-binding motif. Transfection of MCF-7 breast cancer cells with HRPAP20 harboring a mutated CaM-binding motif (HRPAP20K73A) inhibited its interaction with CaM and failed to increase invasion. Other experiments revealed that transfection with HRPAP20, but not HRPAP20K73A, increased secretion of matrix metalloproteinase-9 (MMP-9). Moreover, knockdown of HRPAP20 with small interfering RNA in MCF-7/HRPAP20 transfectants and wild-type MDA-MB-231 cells reduced invasion and inhibited secretion of MMP-9. Together these observations suggest that HRPAP20 may be an important regulator of breast tumor cell invasion by a CaM-mediated mechanism that leads to increased MMP-9 secretion. We conclude that dysregulation of HRPAP20 expression in tumor cells may contribute to the observed phenotypic changes associated with breast cancer progression.

Background: We previously showed that the anti-inflammatory drug, sulfasalazine (salicylazosulfapyridine, SASP), can arrest proliferation of MCF-7 and MDA-MB-231 mammary cancer cells by inhibiting uptake of cystine via the x(c-) cystine/glutamate antiporter. Here we examined SASP with regard to reduction of cellular glutathione (GSH) levels and drug efficacy-enhancing ability., Methods: GSH levels were measured spectrophotometrically. Cellular drug retention was determined with 3H-labeled methotrexate, and drug efficacy with a colony formation assay., Results: Incubation of the mammary cancer cells with SASP (0.3-0.5 mM) led to reduction of their GSH content in a time- and concentration-dependent manner. Similar to MK-571, a multidrug resistance-associated protein inhibitor, SASP increased intracellular accumulation of methotrexate. Preincubation of cells with SASP (0.3 mM) significantly enhanced the potency of the anticancer agent doxorubicin (2.5 nM)., Conclusions: SASP-induced reduction of cellular GSH levels can lead to growth arrest of mammary cancer cells and enhancement of anticancer drug efficacy., (Copyright 2007 S. Karger AG, Basel.)

Objective: Current evidence supports the conclusion that prolactin (PRL) is not an obligate immunoregulatory hormone and influences the immune system predominantly during stress conditions. In this study, we examined the impact of PRL on the psychogenic stress-induced responses of myeloid cells., Methods: Seven-week-old PRL+/- (normal) and PRL-/- (deficient) mice were exposed to a predator for 1 h/day on 3 consecutive days. Another group of PRL-deficient mice received either 1 pituitary graft (hyperprolactinemic) or sham surgery at 5 weeks of age, while PRL-normal mice only received sham surgery. Two weeks later, these mice were also subjected to predator exposure. One day after the last predator exposure session, all mice were killed and the bone marrow and blood harvested., Results: Significant differences in the myeloid cells between PRL-normal and PRL-deficient mice only occurred in stressed conditions. The median serum corticosterone levels were consistently higher in PRL-deficient mice. The implantation of a pituitary graft lowered the corticosterone levels to those observed in PRL-normal mice. The absolute number of immature neutrophils as well as the numbers of granulocyte macrophage, monocyte/macrophage and granulocyte colonies were significantly higher in the stressed PRL-deficient mice; however, only the increased number of immature neutrophils was reversed by pituitary grafting., Conclusions: Our findings support previous observations that PRL influences myeloid cells of the bone marrow most profoundly in stressed conditions. However, the mechanism by which PRL influences bone marrow myeloid cells during stress cannot be explained solely by its effect on serum corticosterone., ((c) 2008 S. Karger AG, Basel.)

Pork loins were pumped to 110% of original weight with solutions containing 5.5% salt and 3.3% sodium tripolyphosphate, 5.5% salt and 3.3% β-lactoglobulin (β-lg) enriched fraction or 5.5% salt and 3.3% whey protein concentrate 80% (WPC80) for comparison with non-enhanced control loins. The enhancement of pork loins significantly increased (p<0.001) the tenderness and juiciness. Warner-Bratzler shear force values were lower (p<0.001) in enhanced then in non-enhanced control chops. The enhanced pork chops had a slightly higher overall flavour and overall acceptability to the control pork chops. Sensory analysis showed that β-lg enriched fraction and WPC80 can be used as replacement ingredients to reduce the amount of phosphate used in enhancement solutions, as they were comparable to salt/sodium tripolyphosphate (salt/STPP) enhancement solution. Consumers rated the tenderness, juiciness and taste of the enhanced chops significantly (p<0.001) higher then the control chops.

Infantile spasms associated with brain tumors have been reported. A focal cortical lesion can induce infantile spasms by triggering the brainstem and basal ganglia in this vulnerable age group. We report the case of a female infant with a low-grade glioma in the right basal ganglia, spreading to the cortical area. She presented at the age of five months with left hemiparesis and partial seizures. She developed infantile spasms at the age of 12 months. This is the first video clip report of partial seizures triggering symmetrical spasms in series, secondary to a basal ganglia glioma extending to the cortex. [Published with video sequences].

Four Beta-lactoglobulin (β-lg) enriched fractions containing different mineral contents were prepared and evaluated in frankfurters. Frankfurters were assessed for cook loss, water holding capacity (WHC), textural and sensory characteristics. The addition of the β-lg fractions reduced the cook loss (p<0.001) in comparison to the control (6.63% vs 3.98%). The fractions (β-lg 1 and 2) with the lowest calcium level significantly reduced WHC (p<0.01). The β-lg fractions had no detrimental effect on the sensory characteristics (p>0.05). All of the fractions increased the TPA value of hardness in comparison to the control (p<0.001) while the springiness decreased in the fractions (p<0.001) with the lowest mineral level. This study showed that the mineral composition of the β-lactoglobulin fractions affected cook loss, tenderness and hardness (TPA) of the frankfurters and the addition of the β-lactoglobulin enriched fraction did not affect the organoleptic quality of frankfurters in comparison to the control. This study shows the potential for next generation whey protein fractions and their application in meat products.

Patients with refractory inflammatory bowel disease (IBD) exhibit increased expression of intestinal P-glycoprotein (P-gp) as well as elevated luminal IFN-gamma and nitric oxide (NO) levels. Using the in vitro Caco-2 cell culture model, we investigated whether these pathological mediators associated with the etiology of IBD affect functional activity of intestinal efflux systems. IFN-gamma reduced cellular uptake of cyclosporin A (CysA) but not methotrexate (MTX) in a time- and concentration-dependent manner. Simultaneously, P-gp expression increased by approximately twofold. Coincubation with the inducible NO synthase inhibitor l-N(6)-(1-iminoethyl)lysine (l-NIL) dramatically reduced production of intracellular NO in response to IFN-gamma stimulus. The presence of l-NIL also abrogated the cytokine-mediated increase in P-gp expression and function suggesting that NO is required for IFN-gamma-mediated activation of this efflux system. Exposure of Caco-2 cells to the chemical NO donor S-nitroso-N-acetylpenicillamine (SNAP) produced a concentration-dependent decrease in intracellular CysA accumulation that was paralleled by an increase in P-gp expression. Both IFN-gamma and SNAP enhanced DNA binding of NF-kappaB, whereas inclusion of l-NIL dramatically decreased this cytokine-induced effect on NF-kappaB binding. These results suggest that NO mediates IFN-gamma-induced increase in expression and function of intestinal P-gp in the human Caco-2 cell culture model by altering DNA binding of NF-kappaB, which may enhance transcription of the ABCB1 gene encoding for this efflux system.

The objective of this experiment was to determine the contribution of some biochemical processes of postmortem muscle to the variation in tenderness of beef from Belgian Blue bull cross Holstein-Friesian steers (n=10). These animals were managed optimally from conception to consumption with the aim of reducing tenderness variation. The M. longissimus dorsi (LD) from the left hand side (LHS) and the right hand side (RHS) were analysed for variation in tenderness using Bartletts test. The quality measurements included pH, temperature, Warner Bratzler shear force, sensory tenderness, chemical composition and sarcomere length. Biochemical measurements included myofibrilar proteolysis, glycolytic potential, adenine/inosine ratio and collagen content. No difference for variances or means were observed between LHS and RHS for chemical, quality or biochemical attributes. Biochemical variation was greater than the variation observed in most of the quality attributes measured. Proteolysis was the main biochemical contributor to the variation in shear force tenderness after 2 and 7 as postmortem, but not sensory tenderness. Glycolysis levels and adenine/inosine ratio explained much of the variation in sensory tenderness, but not WBSF. Collagen content in the LD muscles did not explain variation in shear force or sensory tenderness. This would suggest biochemical variation is one of the main contributors to variation in tenderness of beef managed optimally pre- and post-slaughter.

Longissmus dorsi muscles were removed from Suffolk cross-breed lambs (aged 4-9 months) and cut into steaks. Lamb steaks were over-wrapped on trays and placed in vacuum pack bags. Bags were divided into 3 groups and flushed with gas mixtures containing 100:0, 90:10 or 80:20/CO(2):N(2). Mother packed lamb bags were stored for 4 days (T2) and 7 days (T3), respectively, in darkness at 4 °C, prior to retail display. The effect of aerobic packaging alone on lamb meat quality was used as the control (T1). Under retail display, all over-wrapped trays were held under refrigerated conditions (4 °C, 616 lx) for up to 8 days. Steaks were assessed for microbial growth, oxidative and colour stability as well as pH every 2 days. Mother-packing in 100:0/CO(2):N(2) was the most effective way of extending the storage life of retail ready lamb prior to display, particularly over longer storage periods. TVCs for T3 lamb meat using all gas compositions remained below 2.0×10(6) CFUs/g meat up until day 6 compared to day 4 in both T1 and T2 lamb. Lipid oxidation in lamb mother-packed for 7 days occurred at a faster comparative rate than discolouration and microbial growth and was the major determinant of shelf-life. However, under simulated retail display in aerobic packages, TBARS values did not increase significantly. There was no significant difference between Hunter 'a' values for T3 lamb meat and the control, but T3 meat mother-packed in 100:0/CO(2):N(2) had higher 'a' values than those of the control and T3 meat packed in other gas compositions. Lamb steaks in T3 previously mother-packed in 100:0/CO(2):N(2) were also significantly (p<0.05) higher than those of T2 on day 0. T3 meat also maintained initial colour values over those of the control.

Longissmus dorsi loins were removed from Suffolk cross-breed lambs (4-9 months) and hoggets (15-20 months). The effect of package gas composition was investigated by packaging loins with gas mixtures containing 80:20:0, 60:20:20 and 60:40:0/O(2):CO(2):N(2) with a 2:1 headspace to meat volume ratio. The most effective gas mixture for prolonging shelf-life was used to study the effect of different headspace to meat volume ratios. Loins were packaged with a headspace to meat volume ratio of 2:1, 1.5:1 or 1:1. All modified atmosphere (MA) packs were held under refrigerated display conditions (4 °C, 616 lx) for 12 days. Loins were assessed for microbial, oxidative and colour stability and headspace composition every 3 days. The 80:20:0/O(2):CO(2):N(2) gas composition and the 2:1 headspace to meat volume ratio was the most effective packaging combination at maintaining and prolonging the attractive red colour of MA packaged lamb and hogget meat. 80:20:0/O(2):CO(2):N(2) resulted in significantly (p<0.01) higher Hunter a values in lamb. The 2:1 ratio gave higher visual assessment values in lamb and higher Hunter `a' values for hogget meat throughout the trial. The 2:1 ratio was the most effective at decreasing Pseudomonas and increasing the numbers of lactic acid bacteria in the total microbial load in both lamb and hogget meat. Lipid oxidation in lamb and hogget meat occurred at a slower comparative rate than discolouration or microbial growth and was not the major determinant of shelf-life. The 2:1 headspace to meat volume ratio was most effective at maintaining the initial gas mix in both lamb and hogget MA packs.

The antiretroviral agent efavirenz enhances the systemic clearance of coadministered drugs that are cytochrome P450 (CYP) 3A4 substrates. The mechanism of the apparent increase in CYP3A4 activity by efavirenz and the magnitude of change relative to other known inducers are not known. The authors tested the hypothesis that increased enzymatic activity by efavirenz entails CYP3A4 induction and activation of the human pregnane X receptor (hPXR), a key transcriptional regulator of CYP3A4. Employing primary cultures of human hepatocytes, they compared the CYP3A4 inductive effects of efavirenz (1-10 microM) to rifampin (10 microM) and phenobarbital (2 mM). A cell-based reporter assay was employed to assess hPXR activation. The authors observed that efavirenz caused a concentration-dependent CYP3A4 induction and hPXR activation. Based on the CYP3A4 activity assay, the average magnitude of induction by efavirenz (5-10 microM) was approximately 3- to 4-fold. In comparison, phenobarbital (2 mM) and rifampin (10 microM) caused a 5- and 6-fold induction, respectively.

Purpose: The induction of cytochrome P450 (CYP) 3A4 by drugs and other xenobiotics is a common cause of serious drug interactions. The aim of this study was to comparatively examine the effects of paclitaxel and docetaxel, two structurally related taxane anticancer agents, on the activity and expression of hepatic CYP3A4., Methods: Employing primary cultures of human hepatocytes from multiple donors, we investigated the differences in the magnitude of CYP3A4 induction and relative accumulation of paclitaxel and docetaxel. The CYP3A4 activity of intact hepatocytes was measured as the rate of testosterone 6beta-hydroxylation. The CYP3A4-specific immunoreactive protein and mRNA levels were measured employing Western blot and Northern blot analysis, respectively. Furthermore, employing cell-based reporter gene assay in CV-1 cells, we evaluated the capacity of paclitaxel and docetaxel to activate human pregnane X receptor (hPXR), an orphan nuclear receptor that plays a key role in the transcriptional regulation of CYP3A4., Results: In concurrence with previous reports, we observed that paclitaxel potently induced CYP3A4 activity and expression in hepatocytes treated for 48-96 h. However, docetaxel did not increase the activity or the CYP3A4 immunoreactive protein levels for treatment periods up to 96 h. A marginal increase in the CYP3A4 mRNA levels was observed in cells treated with higher levels (5 and 10 microM) of docetaxel. Furthermore, while paclitaxel effectively activated hPXR (the half-maximal effective concentration, EC50, being about 5.2 microM), docetaxel weakly activated hPXR, and moreover the activation occurred only at high concentrations relative to paclitaxel. A comparison of the cellular concentrations of paclitaxel and docetaxel, in the cell culture models employed for evaluating CYP3A4 induction and hPXR activation, revealed that the intracellular paclitaxel levels were three-fold higher than that of docetaxel. Thus, it appears that both pharmacokinetic (drug concentration) and pharmacodynamic differences (hPXR activation) may account for the observed differences in CYP3A induction by paclitaxel and docetaxel., Conclusion: Our studies suggest that docetaxel has markedly reduced propensity to cause drug interactions that may entail hepatic CYP3A4 induction.

Eating quality and variation within eating quality attributes of beef from young bull progeny of a Charolais sire of average conformation heritability (CF44) (n=14) and young bull progeny of a Charolais sire of good conformation heritability (IC27) (n=16) were examined. The M. longissimus dorsi (up to 12th and/or 13th ribs) was excised 24 h post-slaughter and eating quality attributes analysed at 2, 7 and 14 days postmortem. While progeny muscularity and carcass weight reflected that of each sire, in general no variation was observed in the quality attributes. In addition no significant difference in mean values was evident between sire progenies for carcass and meat quality attributes examined. Significant variation was observed in colour after 2 days ageing, but this was not evident after 7 or 14 days ageing. Average sarcomere length did differ significantly (p<0.05) between progeny of both sire types (CF44=1.87 μm and IC27=1.77 μm), but did not appear to impact on tenderness. The similarity between the progeny of the average or good conformation sires examined in this experiment suggests such sires have no effect on the eating quality of their young bull beef progeny.

The objective of this experiment was to determine the effectiveness of current "best-practise" management of steers pre- and post-slaughter in reducing variation in the eating quality of beef. Steers sired by one Belgian Blue bull from Holstein-Friesian cows were managed optimally from birth to slaughter. Animals were slaughtered at target body weights and subcutaneous fat scores of 620 kg and 4L (LH) (n=23) or 720 kg and 4H (HH) (n=24). On each slaughter occasion, commercial steers with similar carcass weights and classification scores to the homogenous steers were selected from the factory lairage; n=19 for light commercial steers (LC) and n=20 for heavy commercial steers (HC). Carcasses were hung by the pubic bone at 10 °C for 10 h and 2 °C until 24 h postmortem, when M. longissimus dorsi and M. semimembranosus muscles were excised. Following ageing for 2, 7 and/or 14 days postmortem, eating quality was assessed. Muscle from HH steers was more variable in terms of tenderness, protein, moisture and water-holding capacity compared to muscle from LH steers within LD muscle. Muscle from HC steers was more variable in terms of tenderness, redness colour, protein and intramuscular fat compared to muscle from LC steers within LD muscle. Applying best practice management to the homogenous and commercial steers in the present experiment reduced variances in Warner Bratzler shear force (25.69 and 23.9, respectively) compared to variance (154.9) of previous research carried out by the present authors.

Response surface methodology (RSM) was employed for simultaneous analysis of the effects of added surimi (0-40%), fat (5-30%) and water (10-35%), on the physical, textural and sensory characteristics of fresh breakfast pork sausages. Experimental design allowed for evaluation of potential interactive effects between these ingredients. Sausages were evaluated for texture, colour, water holding capacity (WHC) and sensory attributes. Three optimum recipes, R1 (25.3% surimi, 22.2% fat, 12.7% water, 25.3% pork), R2 (12.2% surimi, 5.5% fat, 38.7% water, 33.2% pork) and R3 (25.3% surimi, 6.3% fat, 28.5% water, 25.3% pork), were determined and these were evaluated against a full-fat commercial control (R4). Force values of R1 were not significantly different to R4, however, force values for R2 and R3 were lower (P<0.001). No significant differences were observed between R1, R3 and R4 for visual colour or sensory acceptability scores throughout the study, whereas scores for R2 were lower. Sensory analysis indicated that R2 had lower scores for texture (P<0.01), chewiness (P<0.01), acceptability (P<0.01), flavour (P<0.05) and preference (P<0.01). Results from this study suggest that it is possible to successfully replace pork meat with functional fish proteins in the manufacture of sausage type products.

The aim of this investigation was to quantify the scale of variation in the eating quality of two commercial beef muscles, M. longissimus dorsi (LD) and M. semimembranosus (Sm). Both the LD and Sm were excised from steers (n=81) and heifers (n=81) (classification grade O4H, O4L, R4H, R4L) within 48 h postmortem, vacuum packaged and stored at 4 °C until tested for eating quality at 14 days postmortem. Quality measurements analysed were: pH, Warner Bratzler shear force, sensory attributes, sarcomere length, Hunter L a b muscle and subcutaneous fat colour and chemical composition. Extent of variation in many eating quality measurements, with the exception of most sensory attributes and subcutaneous fat colour, depended on gender, classification grade or a combination of both. The LD was more variable than the Sm for most quality attributes and heifers were more variable than steers. No one carcass grade was more variable over all attributes analysed; with different grades causing higher or lower variances within certain attributes. Knowledge of the current scale of variation in the eating quality of beef is required by the meat industry, and is one that requires further research.

The prolactin (PRL)-dependent rat Nb2 T lymphoma is a valuable model for investigation of molecular mechanisms that underlie tumor progression in hormone-dependent cancers. mRNA differential display was used to screen for novel gene products expressed in hormone-stimulated or differentiating agent-treated Nb2 sublines. From numerous transcripts identified, DNA sequencing and GenBank analysis revealed a novel 289-bp fragment. Using 5'-rapid amplification of complementary ends-PCR, this fragment was used to clone a unique 2117-bp cDNA, designated HRPAP20 (hormone-regulated proliferation-associated protein), in rat lymphoma cells. Computer-assisted sequence analysis revealed a single open reading frame that encoded a putative 20.2-kDa protein. The effect of hormone stimulation to alter expression of HRPAP20 was evaluated by Northern blot analysis of total RNA obtained from PRL-stimulated, lactogen-dependent Nb2-11 cells. Quiescent cells, synchronized in the G(0)-G(1) phase of cell cycle, exhibited reduced HRPAP20 expression compared with exponentially proliferating cultures. The addition of mitogenic concentrations of PRL to stationary cells increased HRPAP20 mRNA accumulation within 4-6 h, corresponding to G(1) cell cycle progression. Immunoblot analysis showed that PRL also increased HRPAP20 protein levels within 4 h. In addition, PRL stimulated serine phosphorylation of the HRPAP20 protein with a similar kinetic pattern. Stable transfection of the HRPAP20 cDNA into Nb2-11 cells significantly (P < 0.01) increased proliferation in the absence of hormonal stimulation and inhibited apoptosis induced by lactogen deprivation (P < 0.001). In the hormone-independent and highly malignant Nb2-SFJCD1 subline, the constitutive expression of HRPAP20 was markedly reduced by exposure of the cells to dietary differentiating agents (butyrate, retinoic acid, and vitamin D(3)). After removal of these substances, PRL stimulated its expression in a manner similar to that observed in PRL-dependent Nb2-11 cells. HRPAP20 expression was also evaluated in MCF-7 cells. Its expression was detectable in quiescent cultures; addition of PRL significantly (P < 0.05) increased HRPAP20 during G(1) cell cycle progression. Exposure of the cells to butyrate or retinoic acid reduced HRPAP20 expression, similar to the effects of these substances in the malignant rat lymphoma. Stable transfection of HRPAP20 into MCF-7 cells significantly (P < 0.006) increased proliferation in the absence of hormone stimulation and augmented survival in the absence of serum (P < 0.05). We conclude that HRPAP20 is a phosphoprotein that is required for proliferation and survival of hormone-dependent tumor cells.

Purpose: In clinical studies, topiramate (TPM) was shown to cause a dose-dependent increase in the clearance of ethinyl estradiol. We hypothesized that this interaction results from induction of hepatic cytochrome P450 (CYP) 3A4 by TPM. Accordingly, we investigated whether TPM induces CYP3A4 in primary human hepatocytes and activates the human pregnane X receptor (hPXR), a nuclear receptor that serves as a regulator of CYP3A4 transcription., Methods: Human hepatocytes were treated for 72 h with TPM (10, 25, 50, 100, 250, and 500 microM) and known inducers, phenobarbital (PB; 2 mM), and rifampicin (10 microM). The rate of testosterone 6beta-hydroxylation by hepatocytes served as a marker for CYP3A4 activity. The CYP3A4-specific protein and mRNA levels were determined by using Western and Northern blot analyses, respectively. The hPXR activation was assessed with cell-based reporter gene assay., Results: Compared with controls, TPM (50-500 microM)-treated hepatocytes exhibited a considerable increase in the CYP3A4 activity (1. 6- to 8.2-fold), protein levels (4.6- to 17.3-fold), and mRNA levels (1.9- to 13.3-fold). Comparatively, rifampicin (10 microM) effected 14.5-, 25.3-, and a 20.3-fold increase in CYP3A4 activity, immunoreactive protein levels, and mRNA levels, respectively. TPM (50-500 microM) caused 1.3- to 3-fold activation of the hPXR, whereas rifampicin (10 microM) caused a 6-fold activation., Conclusions: The observed induction of CYP3A4 by TPM, especially at the higher concentrations, provides a potential mechanistic explanation of the reported increase in the ethinyl estradiol clearance by TPM. It also is suggestive of other potential interactions when high-dose TPM therapy is used.

Background: Malignant progression of lymphoma cells is associated with acquisition of the cystine/glutamate antiporter, xc-, enhancing cystine uptake. Recently, we showed that sulfasalazine (SASP) is a specific xc- inhibitor. Here, we investigated xc- in mammary cancer cell lines., Materials and Methods: Expression and function of xc- were evaluated by RT-PCR and 35S-cystine uptake analysis., Results: Xc- expression was elevated 4-fold (p < 0.001) in cells of the most malignant line, MDA-MB-231, associated with increased 35S-cystine uptake (p < 0.001). Proliferation was inhibited by 0.2-0.5 mM SASP. 2-Mercaptoethanol (60 microM), a cystine uptake enhancer, completely prevented SASP-mediated growth inhibition in MDA-MB-231 cultures, but only partially in 184A1 and MCF-7 cultures. SASP-induced growth arrest was reversible and not cell cycle-specific., Conclusion: The results suggest: (i) malignant progression of human mammary cancer may be associated with acquisition of xc- expression potentially leading to increased growth autonomy and drug resistance, (ii) xc- may act as a therapeutic target.

This study prospectively correlated the level of expression of CD55 on tumours with 7-year survival in 136 colorectal cancer patients. Patients with tumours expressing high levels of CD55 had a significantly worse survival (24%) than patients with low CD55 levels (50%, p<0.02). A similar difference was seen for patients (Duke's B or C) with a high risk of recurrence (29% vs 58%, p<0.05). Furthermore, there was a progressive deterioration in prognosis with increasing antigen expression ( p=0.01). It remains unclear if CD55 is overexpressed by tumours to protect them from complement or if it is related to the recent observation that CD55 is a ligand for the T-cell activation antigen CD97. However, it is a marker of aggression, as colorectal cancer patients whose tumours overexpress CD55 have a significantly reduced 7-year survival.

The conditions of PSE (pale, soft, exudative) and DFD (dark, firm, and dry) are significant causes for downgrading of pigmeat resulting in financial loss to the slaughter plant. The objective of this study was to document the variation in quality of pigmeat from one slaughter plant over a 15-month period, based on colour, appearance and pH. During 4 days of most weeks, 30 pigs were randomly selected from the slaughter line, resulting in a total number of 4560 pigs. Muscle pH between the third and fourth rib was measured 45 min, 90 min and 24 h after slaughter. Using a Minolta colorimeter (Hunter L, a, b scale), colour of the longissimus dorsi muscle was measured 24 h after slaughter. Muscle colour was also visually scored on a scale of 1-6. Carcass pH at 24 h post-mortem varied significantly with month of slaughter (P<0.04) as did colour (Hunter L, P<0.001; Hunter a, P<0.001; Hunter b, P<0.001). It was concluded that meat quality as measured by 24 h pH and colour was poorest during the months of November and December. While this may be at least partly related to weather, it is probable that the increased slaughtering rates at this time and variable resting period before slaughter had the biggest impact.

Meat quality comparisons were carried out on hams produced from pork legs showing evidence of PSE and normal pork legs. Quality measurements included drip loss, sliceability, cooking loss, TBARS analysis and colour evaluation (Hunter L, a, b). A total of 40 pork legs were selected from a commercial slaughtering plant, 20 from carcasses showing PSE and 20 from normal carcasses based on L, a, b, and pH values. Carcass pH taken between the third/fourth rib was measured 45 min, 90 min, and 24 h after slaughter. Using a Minolta colorimeter (Hunter L, a, b,), colour of the Longissimus dorsi muscle was measured 24 h after slaughter. PSE had an adverse effect on cooked hams in comparison to normal cooked hams with respect to driploss (P<0.01), cooking loss (P<0.01), sliceability (P<0.01), colour (P<0.05) and lipid oxidation (P<0.05). Cooked hams manufactured from severe PSE pork in this study had an estimated financial loss of 50% (IRL£4.90) in comparison to those manufactured from normal pork (IRL£9.80).

The hypothesis that prolactin (PRL) functions as an immunomodulator was based on studies showing lymphocyte PRL receptors, and its effects on growth, differentiation, and apoptosis in lymphoid cells. However, studies of PRL (PRL-/-) and PRL receptor knockout mice indicated that PRL was not required for immune system development or function under basal conditions. Because PRL maintains survival in glucocorticoid (GC)-treated Nb2-T lymphocytes in vitro, and PRL and GCs are elevated during stress, we investigated whether PRL protected T cells in vivo from GC-induced apoptosis. Adrenalectomized mice [PRL -/-, undetectable PRL; pituitary grafted PRL-/- (PRL-/-Graft), elevated PRL; and PRL+/-, normal PRL] were treated with dexamethasone (DEX) or PBS. Thymocytes and splenocytes were isolated and annexin V labeling of phosphatidylserine, DNA fragmentation, and caspase-3 activation were assessed as indices of apoptosis. Total thymocytes and CD4+ and CD8+ T cells obtained from DEX-treated PRL-/- mice exhibited significantly increased annexin V binding. In contrast, binding was not altered by DEX in PRL-/-Graft thymocytes. In addition, DEX induced classic DNA fragmentation in PRL-/- thymocytes. Elevated serum PRL reduced this effect. Thymocytes from DEX-treated PRL-/- mice exhibited increased caspase-3 activation, which was inhibited in cells from PRL-/-Graft mice. Finally, elevated expression of X-linked inhibitor of apoptosis, XIAP, was observed in thymi from DEX-treated PRL -/-Graft mice. This is the first demonstration that elevated PRL antagonizes apoptosis in thymocytes exposed to GCs in vivo. These observations suggest that, under conditions of increased GCs, such as during stress, elevated PRL functions physiologically to maintain survival and function of T-lymphocytes.