Eph/ephrins-mediated thymocyte–thymic epithelial cell interactions control numerous processes of thymus biology

Numerous studies emphasize the relevance of thymocyte–thymic epithelial cell (TECs) interactions for the functional maturation of intrathymic T lymphocytes. The tyrosine kinase receptors, Ephs (erythropoietin-producing hepatocyte kinases) and their ligands, ephrins (Eph receptor interaction proteins), are molecules known to be involved in the regulation of numerous biological systems in which cell-to-cell interactions are particularly relevant. In the last years, we and other authors have demonstrated the importance of these molecules in the thymic functions and the T-cell development. In the present report, we review data on the effects of Ephs and ephrins in the functional maturation of both thymic epithelial microenvironment and thymocyte maturation as well as on their role in the lymphoid progenitor recruitment into the thymus

Short communication: Effect of subclinical mastitis on reproductive performance of Holstein dairy cows in the Northwest of Spain

Aim of study: To investigate the effect of subclinical mastitis (SCM) before and after first artificial insemination (AI), characterized by a somatic cell count (SCC) higher than 200×103 cell/mL, on reproductive performance including first service conception rate (FSCR) and pregnancy loss (PL) in Holstein dairy cows. Area of study: The central area of Lugo, Galicia, Spain. Material and methods: This retrospective study was conducted on herd database of a population of 80 commercial Holstein dairy cow farms. A total number of 2053 lactations were included in this study. A binary logistic regression was carried out to analyse all data. Main results: The results of this study indicated that cows that registered a SCC lower than 200×103 cell/mL within 30 days after first AI were more likely to conceive pregnancy than cows with a higher SCC (31.2% and 25.1% FSCR, respectively; OR=1.285, 95% CI=1.000-1.653). Additionally, an increased SCC neither 30 days before nor 30 days after first AI had a negative effect on prevalence of PL in dairy cows. Research highlights: These findings revealed that SCM within 30 days after first AI negatively affected FSCR, whilst 30 days before first AI did not affect it. Therefore, it could be suggested that preventing subclinical mastitis after first AI, during a critical period of 30 days, is important to maximize the reproductive performance of dairy cows

Comparison between transrectal palpation, B-mode and Doppler ultrasonography to assess luteal function in Holstein cattle

IntroductionOver the years, the most common methods for monitoring reproductive health in cattle have varied from transrectal palpation to B-mode ultrasonography. Nowadays, some portable ultrasound equipment includes the Doppler mode. Therefore, the aim of this study was to compare the accuracy of the different methods to assess corpus luteum (CL) functionality.MethodsIn Experiment 1, 53 Holstein lactating cows undergoing a synchronization protocol were examined via transrectal palpation and B-mode scanning. Measurements for the largest diameter (LAD) and subjective size of CL (SCLS) were collected. Data were analyzed using correlation analysis and ROC Curves. In Experiment 2, 30 Holstein non-lactating cows with a CL were administered PGF2α and examined several times after injection, first in B-mode and then with Power Doppler. Measurements for LAD, CL area (CLA) and subjective and objective CL blood flow were collected. Blood samples were taken in both experiments to determine P4 concentration. Data were analyzed using correlation analysis and the GLM repeated measures test.ResultsResults for Experiment 1 showed that LAD was more accurate than SCLS. In Experiment 2, CLA was the best measurement to assess CL function, although both subjective and objective CL blood flow offer accurate information 24 h after PGF2α administration.DiscussionConsequently, ultrasonography provides more accurate information about CL function than transrectal palpation. Although CLA seems to be an earlier indicator of luteal function than blood flow, 24 h after the onset of luteolysis, both parameters are valid

Cyclin-dependent kinase 5 mediates pleiotrophin-induced endothelial cell migration

Pleiotrophin (PTN) stimulates endothelial cell migration through binding to receptor protein tyrosine phosphatase beta/zeta (RPTPβ/ζ) and ανβ3 integrin. Screening for proteins that interact with RPTPβ/ζ and potentially regulate PTN signaling, through mass spectrometry analysis, identified cyclin-dependent kinase 5 (CDK5) activator p35 among the proteins displaying high sequence coverage. Interaction of p35 with the serine/threonine kinase CDK5 leads to CDK5 activation, known to be implicated in cell migration. Protein immunoprecipitation and proximity ligation assays verified p35-RPTPβ/ζ interaction and revealed the molecular association of CDK5 and RPTPβ/ζ. In endothelial cells, PTN activates CDK5 in an RPTPβ/ζ- and phosphoinositide 3-kinase (PI3K)-dependent manner. On the other hand, c-Src, ανβ3 and ERK1/2 do not mediate the PTN-induced CDK5 activation. Pharmacological and genetic inhibition of CDK5 abolished PTN-induced endothelial cell migration, suggesting that CDK5 mediates PTN stimulatory effect. A new pyrrolo[2,3-α]carbazole derivative previously identified as a CDK1 inhibitor, was found to suppress CDK5 activity and eliminate PTN stimulatory effect on cell migration, warranting its further evaluation as a new CDK5 inhibitor. Collectively, our data reveal that CDK5 is activated by PTN, in an RPTPβ/ζ-dependent manner, regulates PTN-induced cell migration and is an attractive target for the inhibition of PTN pro-angiogenic properties

Tango de Slawomir Marozek

1 lám. (cartel) : col. ; 69 x 50 cm

Genetic inactivation of Pleiotrophin triggers amphetamine-induced cell loss in the substantia nigra and enhances amphetamine neurotoxicity in the striatum

Pleiotrophin (PTN) is a neurotrophic factor with important effects in survival and differentiation of dopaminergic neurons that has been suggested to play important roles in drug of abuse-induced neurotoxicity. To test this hypothesis, we have studied the effects of amphetamine (10 mg/kg, four times, every 2 h) on the nigrostriatal pathway of PTN genetically deficient (PTN-/-) mice. We found that amphetamine causes a significantly enhanced loss of dopaminergic terminals in the striatum of PTN-/- mice compared to wild type (WT+/+) mice. In addition, we found a significant decrease (~20%) of tyrosine hydroxylase (TH)-positive neurons only in the substantia nigra of amphetamine-treated PTN-/- mice, whereas this area of WT+/+ animals remained unaffected after amphetamine treatment. This effect was accompanied by enhanced amphetamine-induced astrocytosis in the substantia nigra of PTN-/- mice. Interestingly, we found a significant decrease in the phosphorylation levels of p42 extracellular-signal regulated kinase (ERK2) in both saline- and amphetamine-treated PTN-/- mice, whereas phosphorylation of p44 ERK (ERK1) was almost abolished in the striatum of PTN-/- mice compared to WT+/+ mice, suggesting that basal deficiencies in the phosphorylation levels of ERK1/2 could underlie the higher vulnerability of PTN-/- mice to amphetamine-induced neurotoxic effects. The data suggest an important role of PTN in the protection of nigrostriatal pathways against amphetamine insult. © 2010 IBRO.Peer Reviewe

Altered Maturation of Medullary TEC in EphB-Deficient Thymi Is Recovered by RANK Signaling Stimulation

In the present study, the relevance of EphB2 and EphB3 tyrosine kinase receptors for the maturation of medullary thymic epithelial cells (TECs) is analyzed. The absence of both molecules, but particularly that of EphB2, courses with altered maturation of medullary Cld3,4hiSSEA1+ epithelial progenitor cells, mature medulla epithelial cells, defined by the expression of specific cell markers, including UEA1, MHCII, CD40, CD80, and AIRE, and reduced expansion of medullary islets. In vivo assays demonstrate that these changes are a consequence of the absence of EphBs in both TECs and thymocytes. On the other hand, the changes, that remains in the adult thymus, correlated well with reduced proportions of E15.5 Vγ5+RANKL+ cells in EphB-deficient thymi that could result in decreased stimulation of RANK+ medullary TECs to mature, a fact that was confirmed by recovering of proportions of both CD40hiCD80+ and MHCIIhiUEA1+ mature medullary TECs of mutant E14.5 alymphoid thymic lobes by agonist anti-RANK antibody treatment. Accordingly, the effects of EphB deficiency on medullary TECs maturation are recovered by RANK stimulation

Induction of EROD activity by 1-phenylimidazole and beta-naphthoflavone in rainbow trout cultured hepatocytes: a comparative study

The classical pathway for induction of cytochrome P4501A (CYP1A) by xenobiotics is ligand binding to the aryl hydrocarbon receptor (AhR). However, several studies with mammalian cell systems point out a range of xenobiotics including imidazole derivatives, which are able to activate CYP1A through non-classical mechanisms. The objective of the present work is to compare induction of CYP1A (determined at the catalytic level as 7-ethoxyresorufin-O-deethylase, EROD) in rainbow trout (Oncorhynchus mykiss) hepatocytes by the prototypic AhR ligand, beta-naphthoflavone (betaNF), and by the imidazole derivative, 1-phenylimidazole (PIM). PIM was able to induce EROD activity although its potency was clearly lower than that of betaNF. In order to assess the relative importance of classical AhR ligand binding and alternative signaling pathways in CYP1A induction by PIM, co-exposure experiments with the partial AhR antagonist alpha-naphthoflavone (alphaNF) or with inhibitors of protein kinase C (staurosporine) and tyrosine kinases (genistein, herbimicine) were performed. alphaNF and herbimicin provoked a decrease of EROD induction both by betaNF and PIM, whereas staurosporine and genistein remained without effect. The overall similarities in the response of betaNF and PIM to the various inhibitors suggest that both compounds, in apparent contrast to the behaviour of some other imidazole derivatives, induce CYP1A following similar mechanisms