Human immune responses elicited by an intranasal inactivated H5 influenza vaccine

Intranasally administered influenza vaccines could be more effective than injected vaccines, because intranasal vaccination can induce virus-specific immunoglobulin A (IgA) antibodies in the upper respiratory tract, which is the initial site of infection. In this study, immune responses elicited by an intranasal inactivated vaccine of influenza A(H5N1) virus were evaluated in healthy individuals naive for influenza A(H5N1) virus. Three doses of intranasal inactivated whole-virion H5 influenza vaccine induced strong neutralizing nasal IgA and serum IgG antibodies. In addition, a mucoadhesive excipient, carboxy vinyl polymer, had a notable impact on the induction of nasal IgA antibody responses but not on serum IgG antibody responses. The nasal hemagglutinin (HA)-specific IgA antibody responses clearly correlated with mucosal neutralizing antibody responses, indicating that measurement of nasal HA-specific IgA titers could be used as a surrogate for the mucosal antibody response. Furthermore, increased numbers of plasma cells and vaccine antigen-specific Th cells in the peripheral blood were observed after vaccination, suggesting that peripheral blood biomarkers may also be used to evaluate the intranasal vaccine-induced immune response. However, peripheral blood immune cell responses correlated with neutralizing antibody titers in serum samples but not in nasal wash samples. Thus, analysis of the peripheral blood immune response could be a surrogate for the systemic immune response to intranasal vaccination but not for the mucosal immune response. The current study suggests the clinical potential of intranasal inactivated vaccines against influenza A(H5N1) viruses and highlights the need to develop novel means to evaluate intranasal vaccine-induced mucosal immune responses.     

Mono-(2-ethylhexyl) phthalate (MEHP) induces testicular alterations in male guinea pigs at prepubertal stage

We have recently shown that MEHP induces spermatogenic cell apoptosis in guinea pigs at prepubertal stage in vitro. To evaluate the effects of MEHP on the testicular tissues of guinea pigs in vivo, we conducted this research work. Five weeks old male guinea pigs were used in this experiment. They received a single oral dose of 2000 mg/ml of MEHP in corn oil by gavage at a volume equal to 4 ml/kg. Control group received a similar volume of corn oil vehicle. Vehicle- and MEHP-treated guinea pigs were sacrificed at the interval of 3, 6, and 9 h, and the testicular tissues were processed for histopathological studies. Distinct histopathological changes were recognized in testes. Detachment and displacement of spermatogenic cells, thin seminiferous epithelia, vacuolization of Sertoli cells were prominent at 6 h after MEHP treatment. The lumina of the efferent ductules were frequently occupied with sloughed seminiferous epithelia from 6 to 9 h after MEHP treatment. Apoptotic spermatogenic cells appeared at 3 h in the control group. The incidence of apoptotic spermatogenic cells significantly increased (*p<0.05) from 3 to 9 h, and the maximal increase of apoptotic spermatogenic cells were observed at 9 h after MEHP treatment. Time-dependent increases of apoptotic spermatogenic cells was recognized throughout the experimental period. It may be suggested here that MEHP also induces spermatogenic cell apoptosis in guinea pigs in vivo and guinea pigs may be considered as a useful animal model for sensitivity test of the reproductive toxicity to some phthalate esters at their earlier stage in vivo.     

Stem cells, vascular smooth muscle cells and atherosclerosis

Stem cells have the ability to differentiate into a variety of cells to replace dead cells or to repair tissue. Recently, accumulating evidence indicates that mechanical forces, cytokines and other factors can influence stem cell differentiation into vascular smooth muscle cells (SMCs). In developmental process, SMCs originate from several sources, which show a great heterogenicity in different vessel walls. In adult vessels, SMCs display a less proliferative nature, but are altered in response to risk factors for atherosclerosis. Traditional view on SMC origins in atherosclerotic lesions is challenged by the recent findings that stem cells and smooth muscle progenitors contribute to the development of atherosclerotic lesions. Vascular progenitor cells circulating in human blood and the presence of adventitia in animals are recent discoveries, but the source of these cells is still unknown. The present review gives an update on the progress of stem cell and SMC research in atherosclerosis, and discusses possible mechanisms of stem/progenitor cell differentiation that contribute to the disease process.     

Bioactive compounds and antioxidant activity exhibit high intraspecific variability in <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> mushrooms and correlate well with cultivation performance parameters

Experimental data related with oyster mushroom production and nutritional properties usually derive from the examination of only one strain, and hence their representativeness/usefulness is questionable. This work aims at assessing intraspecific variability in Pleurotus ostreatus by studying 16 strains, under the same conditions, in respect to essential cultivation and mushroom quality aspects, and by defining the impact of intrinsic/genetic factors on such parameters. Hence, mushroom yield, earliness, crop length, biological efficiency, productivity, and their content in selected macro and microconstituents (e.g. fatty acids, sterols, individual phenolic compounds, terpenic acids, glucans) as well as their antioxidant properties (i.e., antiradical activity, ferric reducing potential, inhibition of serum oxidation) were assayed. The effect of intrinsic/genetic factors was evident, especially as regards earliness, yield of each production flush and mushroom weight, whereas biological efficiency was not particularly influenced by the cultivated strain. Moreover, phenolics, ergosterol and antiradical activity demonstrated significant variability among strains in contrast to what was observed for fatty acids, β-glucans and ferric reducing potential. The observed heterogeneity reveals the limitations of using a low number of strains for evaluating mushroom production and/or their content in bioactive compounds, and as evidenced, it is valuable for breeding and commercial purposes.     

Endothelial Lineage Differentiation from Induced Pluripotent Stem Cells Is Regulated by MicroRNA-21 and Transforming Growth Factor β2 (TGF-β2) Pathways

Finding a suitable cell source for endothelial cells (ECs) for cardiovascular regeneration is a challenging issue for regenerative medicine. In this paper, we describe a novel mechanism regulating induced pluripotent stem cells (iPSC) differentiation into ECs, with a particular focus on miRNAs and their targets. We first established a protocol using collagen IV and VEGF to drive the functional differentiation of iPSCs into ECs and compared the miRNA signature of differentiated and undifferentiated cells. Among the miRNAs overrepresented in differentiated cells, we focused on microRNA-21 (miR-21) and studied its role in iPSC differentiation. Overexpression of miR-21 in predifferentiated iPSCs induced EC marker up-regulation and in vitro and in vivo capillary formation; accordingly, inhibition of miR-21 produced the opposite effects. Importantly, miR-21 overexpression increased TGF-β2 mRNA and secreted protein level, consistent with the strong up-regulation of TGF-β2 during iPSC differentiation. Indeed, treatment of iPSCs with TGFβ-2 induced EC marker expression and in vitro tube formation. Inhibition of SMAD3, a downstream effector of TGFβ-2, strongly decreased VE-cadherin expression. Furthermore, TGFβ-2 neutralization and knockdown inhibited miR-21-induced EC marker expression. Finally, we confirmed the PTEN/Akt pathway as a direct target of miR-21, and we showed that PTEN knockdown is required for miR-21-mediated endothelial differentiation. In conclusion, we elucidated a novel signaling pathway that promotes the differentiation of iPSC into functional ECs suitable for regenerative medicine applications.     

Bacteroides fragilis Toxin Coordinates a Pro-carcinogenic Inflammatory Cascade via Targeting of Colonic Epithelial Cells

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Effects of 17 alpha-methyltestosterone on uterine morphology and heat shock protein expression are mediated through estrogen and androgen receptors

Testosterone and the synthetic androgen, 17 alpha-methyltestosterone (MT), have been shown to increase uterine weights and alter uterine morphology. However, whereas the mechanism of action of testosterone in the uterus has been studied, it is not known if the actions of MT are mediated through androgen (AR) or estrogen (ER) receptors. In the present study, we have shown that MT, at 0.5 or 10 mg/kg per day, increases uterine weight and alters uterine morphology in a dose-dependent manner. Co-administration of the anti-androgen, flutamide, or the anti-estrogen, ICI 182,780, with MT revealed that the effects of the low dose of MT are mediated through the ER, whereas those of the high dose are mediated through both the ER and AR. In addition, we have studied the effects of MT on uterine heat shock proteins (hsps), a group of estrogen-regulated proteins whose levels increase in response to growth signals and protein damage. MT increased levels of hsp90 alpha, hsp72, and grp94. All effects on uterine hsp levels were antagonized by the anti-estrogen and not the anti-androgen. Collectively, the results of the present study indicate that the effects of MT in the uterus are mediated through the AR and ER.     

Analysis of dialkyl phosphate metabolites in hair using gas chromatography-mass spectrometry: A biomarker of chronic exposure to organophosphate pesticides

The aim of our study was to develop and validate an analytical approach for the quantitative determination of three dialkyl phosphate (DAP) metabolites, dimethyl phosphate (DMP), dimethyl thiophosphate (DMTP) and diethyl phosphate (DEP), of organophosphate pesticides (OPs) in hair samples. The proposed methodology comprises a decontamination step, solid–liquid extraction, followed by liquid–liquid extraction, pentafluorobenzyl bromide derivatization, clean-up on Florisil/PSA column and analysis by gas chromatography–mass spectrometry (GC-MS). Extraction recovery, obtained from 50?mg hair samples spiked at two concentration levels, ranged from 56.1 to 107.9% and the within-day precision ranged from 13.5 to 17.5%. Limits of detection (LODs) ranged from 0.02 to 0.10?ng mg^?1. The results obtained from the analysis of hair samples of 30 agricultural workers show the suitability of the proposed method for monitoring people occupationally exposed to OPs. The most frequently detected compound was DEP followed by DMP. This is the first report on the detection of dialkyl phosphates in human hair which reflects the ability of hair testing to assess chronic exposure to OPs.