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T helper lymphocytes become polarized upon antigen and cytokine stimuli received after their maturation in the thymus. Since the balance of Th1 and Th2 responses is critical in healthy and pathological immune responses, understanding the molecular base of T cell polarization still remained an important question. Using our Th0/Th1/Th2 hybridoma model system, we performed a comparative study on polarized Th1 and Th2 cells in terms of their membrane raft expression/composition, their TCR mediated activation signaling, and sensitivity to activation-induced cell death (AICD) using flow and image cytometric methods. We show here that the TCR stimulation induced more intense and sustained Ca2+-response in Th1 cells compared to Th2 ones correlates well with a shorter nuclear residence time of the Ca2+-dependent NFAT transcription factor in Th2 cells. In addition, NFAT translocation directly depended on lipid raft integrity/membrane cholesterol level. Expression pattern of raftophilic accessory proteins (CD4, CD59, and CD48) and lipids (GM1, cholesterol) were also different in the Th1 and Th2 hybridomas, similarly to differentiated spleen Th cells. The activation-induced, remarkably clustered and polarized membrane distribution of TCR/CD3 complex in Th1, but not in Th2 cells, together with an increased raft localization of Kv1.3 ion channels regulating the Ca2+-response, are consistent with the above properties of NFAT. Finally, the polarized Th cells, especially Th1, were more sensitive to AICD than their unpolarized Th0 precursor. These results suggest that the membrane microdomain organization—Ca2+-signaling—NFAT activation axis is an important determinant of polarized Th cell effector function and fate. © 2012 International Society for Advancement of Cytometry

Protein kinase C (PKC) isoforms have crucial roles in cutaneous signaling. Interestingly, we lack information about their involvement in human sebaceous gland biology. Therefore, in this current study, we investigated the functions of the PKC system in human immortalized SZ95 sebocytes. Using molecular biological approaches, imaging, and functional assays, we report that SZ95 sebocytes express the conventional cPKCα; the novel nPKCδ, ε, and η; and the atypical aPKCζ. Activation of the PKC system by phorbol 12-myristate 13-acetate (PMA) stimulated lipid synthesis (a hallmark of differentiation) and resulted in translocation and then downregulation of cPKCα and nPKCδ. In good accord with these findings, the effect of PMA was effectively abrogated by inhibitors and short interfering RNA-mediated “silencing” of cPKCα and nPKCδ. Of further importance, molecular or pharmacological inhibition of nPKCδ also prevented the lipogenic and apoptosis-promoting action of arachidonic acid. Finally, we also found that “knockdown” of the endogenous aPKCζ activity markedly increased basal lipid synthesis and apoptosis, suggesting its constitutive activity in suppressing these processes. Collectively, our findings strongly argue for the fact that certain PKCs have pivotal, isoform-specific, differential, and antagonistic roles in the regulation of human sebaceous gland–derived sebocyte biology.

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We report the synthesis, in vitro and in vivo investigation of folate-targeted, biocompatible, biodegradableself-assembled nanoparticles radiolabelled with 99mTc, as potential new SPECT or SPECT/CT imagingagent.Nanoparticles with hydrodynamic size in the range of 75?200 nm were prepared by self-assembly ofchitosan and folated poly

The transcriptional basis of sebocyte differentiation and lipid-producing mechanisms is mostly unclear. Peroxisome proliferator-activated receptor-gamma (PPAR?), a lipid activated transcription factor, has been implicated in lipid metabolism of various cell types. Here, we show that PPAR?? is differentially expressed in normal and pathological human sebocytes, and appears to have roles in their differentiation process. These were demonstrated in laser microdissected normal human sebaceous glands (SG) and SZ95 cells (immortalized sebocyte cell line) utilizing RT-qPCR, immunhistochemistry, for lipid analyses quantitative fluorimetry and mass spectrometry based analytical approaches. We have observed that PPAR? and its target genes, ADRP and PGAR, are expressed in and associated with the differentiation levels of sebocytes. Also, PPAR? is present in normal and hyperplastic SG, whereas its expression levels gradually decrease in cases obtained from SG adenoma and SG carcinoma cells, reflecting a maturation-linked expression pattern. Furthermore, in SZ95 sebocytes, natural lipids including arachidonic acid and arachidonic acid derivate keto-metabolites (e.g. 5-KETE, 12-KETE) appear to regulate PPAR? signaling pathways, which in turn modulate phospholipid biosynthesis and induce neutral lipid synthesis. Collectively, our findings highlight the importance of novel endogenous ligand-activated PPAR? signaling in human sebocyte biology and suggest that PPAR? might be a candidate target for the clinical management of SG disorders

open access - http://www.biomedcentral.com/1471-2415/13/40

Objective: to explore the relationship among health-related quality of life (HRQoL), clinical variables, anthropometric measures, physical activity and cardiorespiratory fitness in children and adolescents with type 1 diabetes. Furthermore, we aimed to find predictors of HRQoL and metabolic control. Methods: A total of 106 patients (sex ratio: 1:1) with mean HbA1c of 8.55 (? 1.44) % and diabetes duration of 5.15 (? 3.24) years were assessed. The average age was 13.22 (? 3.08) years. Results: We observed statistically significant negative medium correlation between HbA1c and VO2max (r = ?0.343; p = 0,000). There was statistically significant small positive correlation between the HRQoL and the maximal oxygen consumption (r = 0.208; p = 0.032). We found no significant correlation between the HbA1c and the patients' HRQoL. In the multiple linear regression analysis both the better metabolic control and the HRQoL was predicted by the VO2max, other variables had no effect. Physical activity level did not explain the HRQoL. Boys had significantly better HRQoL and less skinfold thickness than girls. Conclusion: Better cardiorespiratory fitness associated with both favourable metabolic control and better HRQoL of diabetic youths. Regular aerobic exercise improves the young patients' physical fitness and overall health status, and perception of health-related quality of life, respectively

Rheological blood parameters of neonates are different form those of adults. Many authors have studied changes in blood rheology in neonates in different clinical disorders. To-date, no one set the normal values for blood rheological parameters in healthy neonates. The aim of this study is to set the norm for rheological blood parameters in healthy newborns and to describe the changes in those parameters in common clinical disorders that affect the newborns. We recruited all the neonates born to mothers experiencing un eventful pregnancies, blood was taken from the umbilical cord right after the delivery. In this time period we recruited 4985 neonate. From this huge database we were able to set the standards for blood rheology in neonates, namely plasma viscosity of 1.06 ? 0.072 mPa, erythrocyte aggregation at stasis of 2.41 ? 2.74 s-1 and erythrocyte aggregation under low shear forces of 8.51 ? 6.38 s-1. These values changed significantly in some diseased neonates. This is the largest study investigating normal rheological parameters and deviations from the norm in common clinical disorders occurring in this early stage of life

In this study the emergence of rotavirus A genotype G12 in children

The removal of ferric ions from aqueous solutions by a nanoparticle-enhanced ultrafiltration techniquewas investigated. Biodegradable poly-gamma-glutamic acid (c-PGA), a linear biopolymer, and itscross-linked nanoparticles were used to complex the metal ions by forming nanosized spherical particleswith more or less deformability. These polymer?metal ion particles were then removed by membraneseparation. Two ultrafiltration techniques were studied with the aim of developing a nanoparticleenhancedseparation process for the efficient removal of ferric ions from model solutions. The influenceof parameters such as the feed c-PGA and Fe3+ concentrations, their proportions and the extent of crosslinkingof c-PGA were studied. It was observed that c-PGA could bind and remove ferric ions to producewater with low Fe3+ concentration. These studies show that optimal nano-membrane separation technologycan be performed, depending on the environment and quality of treatable water by the combinationof the filtration parameters and type of membrane techniques shown here

1. The irritant fungal terpenoid isovelleral caused protective eye-wiping movements in the rat upon intraocular instillation and showed cross-tachyphylaxis with capsaicin, the pungent principle in hot pepper. 2. Isovelleral induced a dose-dependent calcium uptake by rat dorsal root ganglion neurones cultured in vitro with an EC50 of 95 nM, which was fully inhibited by the competitive vanilloid receptor antagonist capsazepine. 3. Isovelleral inhibited specific binding of [3H]-resiniferatoxin (RTX), an ultrapotent capsaicin analogue, to rat trigeminal ganglion or spinal cord preparations with an IC50 of 5.2 microM; in experiments in which the concentration of [3H]-RTX was varied, isovelleral changed both the apparent affinity (from 16 pM to 37 pM) and the co-operativity index (from 2.1 to 1.5), but not the Bmax. 4. The affinity of isovelleral for inducing calcium uptake or inhibiting RTX binding was in very good agreement with the threshold dose (2.2. nmol) at which it provoked pungency on the human tongue. 5. For a series of 14 terpenoids with an unsaturated 1,4-dialdehyde, a good correlation was found between pungency on the human tongue and affinity for vanilloid receptors on the rat spinal cord. 6. The results suggest that isovelleral-like compounds produce their irritant effect by interacting with vanilloid receptors on capsaicin-sensitive sensory neurones. Since these pungent diterpenes are structurally distinct from the known classes of vanilloids, these data provide new insights into structure-activity relations and may afford new opportunities for the development of drugs targeting capsaicin-sensitive pathways.

Background and methods: The association between nutritional status, antioxidant human paraoxonase-1 (PON1) activity and low grade inflammation in hemodialized (HD) patients with chronic kidney disease (CKD) is unclear. The aim of this study was to determine PON1 paraoxonase and lactonase activities, ADMA, adiponectin and leptin concentrations, and to clarify the relationship between paraoxonase activity and a set of cardiovascular risk factors in malnourished, normal weight and obese HD patients; 114 HD patients with end-stage renal failure were enrolled. Results: Leptin levels were significantly higher and PON1 paraoxonase activities were significantly lower in obese patients compared to the other groups. Plasma adiponectin concentration was significantly lower in obese subjects compared to malnourished patients. Paraoxonase activity was negatively correlated with CRP level in HD and malnourished patients. Furthermore, we found significant inverse correlation between paraoxonase activity and BMI in the whole patient group. In multiple regression analysis, PON1 lactonase activity, CRP level and leptin concentration proved to be independent predictors of paraoxonase activity.Conclusion: Despite the previous findings of reverse epidemiology for the mortality rate of HD patients, further studies are needed to clarify the effects of nutritional state on atherosclerosis in obese and malnourished patients with end-stage renal failure

Pályázati azonosító:- OTKA K101591- Baross Gábor projekt #EA_SPIN_06-DIABDIAG- Research Fund Management and Research Exploitation KMA 0149/3.0 projekt- Bioincubator House projekt

Growing number of clinical and experimental data reflect to the gender differences of hemorheological parameters.However, little is known about the potential hemorheological effect of gonadectomy and consequent changes in sex hormoneconcentration. Adult, same-aged male and female rats were involved in the study. In control male and female group no surgicalintervention was performed. In gonadectomized (GoE) male and female groups bilateral orchidectomy or ovariectomy werecompleted. Body weight measurement and blood sampling were carried out in the 1st, 2nd and 3rd postoperative months.The GoE females had significant bodyweight augmentation and their plasma estrogen concentration decreased by 40?45%by the 1st postoperative month, while in males the testosterone level was not detectable after gonadectomy. Leukocyte andplatelet counts moderately increased in GoE males. Elongation index values of erythrocytes slightly decreased in both gendersafter gonadectomy, showing converging values. Erythrocyte aggregation index values of GoE females significantly raised bythe 2nd month. It can be concluded that gonadectomy in rats resulted in alteration (dominantly impairment) of blood microrheologicalparameters, by different manner in males and females. Supposedly decrease in estrogen can cause more expressedhemorheological changes than the cessation of testosterone

Introduction We aimed to determine the neutralization of Neisseria meningitidis outer membrane vesicles (blebs) by humoral and cellular elements of whole blood. Methods The interaction of FITC-labeled blebs with monocytes was studied by spectrofluorometry. Blebs are able to induce an oxidative burst in neutrophils, and we evaluated the inhibitory effect of plasma on this process. Results Human plasma reduced the priming activity of blebs containing 1?3 ng/ml lipopolysaccharide (LPS) by 50?60% and bactericidal permeability increasing protein(BPI) reduced priming to background levels. A complete neutralization of LPS and blebs by plasma and BPI was measured using the limulus amebocyte lysate (LAL) assay. Furthermore, only 3% of blebs were cell-associated, while the remainder were in the supernatant. Conclusions Plasma and BPI are able to neutralize blebs, with phagocytosis playing only a minor role. As such, we conclude that blebs do not behave like particles but more like free LPS

Mesenchymal stem cell like (MSCl) cells were generated from human embryonic stem cells (hESC) through embryoid body formation, and isolated by adherence to plastic surface. MSCl cell lines could be propagated without changes in morphological or functional characteristics for more than 15 passages. These cells, as well as their fluorescent protein expressing stable derivatives, efficiently supported the growth of undifferentiated human embryonic stem cells as feeder cells. The MSCl cells did not express the embryonic (Oct4, Nanog, ABCG2, PODXL, or SSEA4), or hematopoietic (CD34, CD45, CD14, CD133, HLA-DR) stem cell markers, while were positive for the characteristic cell surface markers of MSCs (CD44, CD73, CD90, CD105). MSCl cells could be differentiated toward osteogenic, chondrogenic or adipogenic directions and exhibited significant inhibition of mitogen-activated lymphocyte proliferation, and thus presented immunosuppressive features. We suggest that cultured MSCl cells can properly model human MSCs and be applied as efficient feeders in hESC cultures