The recurrent aphtous ulceration analyzed by cDNA microarray has a Th1 gene expression profile

Objetivo: Identificar o padrao de expressao genica da Ulceracao Aftosa Recorrente e classificar o tipo da resposta imune T-Helper a partir da comparacao da expressao genica da -doenca com controle normal, usando a tecnica de microarray de cDNA. Metodos: Vinte e nove pacientes portadores de ulceracao aftosa recorrente (Grupo Teste), com pelo menos uma crise a cada quinze dias, foram submetidos, na fase aguda da doenca, a biopsia da lesao ulcerada. Onze voluntarios saudaveis (Grupo Controle), livres da doenca, foram submetidos a biopsia de mucosa clinicamente normal. O RNA total das amostras foi isolado e dois pools de RNAs, representado os Grupos, foram formados, usando-se quantidades iguais do RNA de cada amostra. A sonda radioativa de cDNA foi preparada a partir de 17 gg do RNA total atraves da reacao de transcricao reversa. As sondas foram hibridadas com as membranas de microarray em duplicata. As imagens dos microarrays hibridados foram obtidas por varredura realizada em scaner a laser. Os alvos de cDNAs, marcados nas membranas, foram localizados e quantificados usando-se sofiware especifico. As normalizacoes dos pares de membranas foram executadas atraves do modelo linear de ajuste local, baseado no metodo de suavizacao conhecido como Lowess. Os genes foram considerados diferencialmente expressos quando o valor da media da diferenca de expressao das membranas, entre Teste e Controle, era maior do 2x e, o valor da media de expressao genita era maior do que o nivel do gene controle negativo da membrana. Para analisar o padrao da resposta imune, foram selecionados, no microarray, genes associados com atividades Th1 (n=36) ou Th2 (n=19) e, as medias da diferenca de expressao desses genes foram comparadas com a dos genes constitutivos (N=13). Resultados: Foram identificados vinte e um genes hiper-expressos e vinte Kipoexpressos no Grupo Teste. O nivel medio de expressao dos genes Th1 no Grupo Teste foi maior (p<0,05) do que no Grupo Controle. Por outro lado, a media dos genes Th2 foi a mesma nos dois Grupos. Conclusoes: O microarray de cDNA permitiu a identificacao dos padroes genitos da Ulceracao Aftosa Recorrente e da mucosa controle e possibilitou classificar a Ulceracao Aftosa Recorrente como doenca Th1BV UNIFESP: Teses e dissertaçõe

Proinflammatory and Anti-inflammatory Cytokines Present in the Acute Phase of Experimental Colitis Treated with Saccharomyces boulardii

To study the proinflammatory and anti-inflammatory cytokines present in the acute phase of trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis treated with Saccharomyces boulardii.Thirty male Wistar rats were divided into three groups: (1) treated group-received Saccharomyces boulardii for 14 days; (2) non-treated group-received sodium chloride solution for 14 days; (3) control group. Colitis was induced on the seventh day of the study in the treated and the non-treated groups using TNBS (10 mg) dissolved in 50% ethanol. Quantification of cytokines, including interleukin (IL)-1 beta (IL-1 beta), IL-6, transforming growth factor-beta (TGF-beta), IL-10 and tumor necrosis factor-alpha (TNF-alpha), in the serum and colonic tissue collected on day 14 were carried out using an enzyme-linked immunosorbent assay (ELISA).The mean concentrations of TGF-beta in both the serum and the colonic tissue of the treated group were statistically higher than that of the control group. the mean concentration of TGF-beta in the colonic tissue of the non-treated group was also statistically higher than the control group.The group treated with Saccharomyces boulardii showed increased amounts of TGF-beta, an anti-inflammatory cytokine, during the acute phase of colitis. There were no differences in the amount of TNF-alpha, IL-1 beta, IL-6, and IL-10 between the treated and the non-treated or the control groups during the acute phase of experimental colitis induced by TNBS.Sao Leopoldo Mand Dent Res Ctr, Campinas, SP, BrazilUniversidade Federal de São Paulo, Div Pediat Gastroenterol, São Paulo, BrazilUniversidade Federal de São Paulo, Div Pediat Gastroenterol, São Paulo, BrazilWeb of Scienc

cDNA microarray analysis of differentially expressed genes in penile tissue after treatment with tadalafil

To evaluate differential gene expression in penile tissue after treatment with the phosphodiesterase 5 (PDE5) inhibitor tadalafil, as of the three clinically available PDE5 inhibitors (sildenafil, tadalafil, and vardenafil) used for the treatment of erectile dysfunction (ED), tadalafil has a long half-life and low incidence of side-effects. In all, 32 adult rats were divided into two groups. The control group received 0.5 mL of drinking water alone, while the tadalafil group was treated with tadalafil at a dose of 0.27 mg/kg. At 4 h after treatment with water or tadalafil the rats were killed and the penile tissue was removed. The total RNA was isolated from the penile tissue from both groups and differentially expressed genes were identified by cDNA microarray analysis. To validate the expression data from the microarray analysis, quantitative real-time polymerase chain reaction (PCR) and immunohistochemistry were used. In all, 153 genes were differentially expressed between the control group and the tadalafil group. We validated the microarray results by quantitative PCR for the insulin-like growth factor binding protein 6 (IGFBP-6) gene and the neuronal calcium sensor 1 (NCS-1) gene, both of which were up-regulated in the tadalafil group, and for the natriuretic peptide receptor 1 (NPR-1) gene that was down-regulated in this group. Immunohistochemistry showed localization of the NCS-1 protein in sinusoid trabeculae of the corpus cavernosum in control and tadalafil-treated rats. There was differential expression in 153 genes after tadalafil treatment. Some of these genes such as IGFBP-6, NPR-1 and NCS-1, might result in new targets in the treatment of ED

Response of peripheral blood mononuclear cells to conditioned medium from cultured oral squamous cell carcinomas

The current study investigated the capacity for tumor factors secreted by head and neck squamous cell carcinoma (HNSCC) cell lines, KB, KB16, and HEP, to induce the secretion of various cytokines from peripheral blood mononuclear cells (PBMCs). PBMCs were isolated from blood samples collected from six healthy volunteers and these cells were incubated for 6, 24, 48, or 72 hours in the presence of 50% conditioned medium collected from cultured cell lines pretreated with, or without, stimulants such as phytohemagglutinin (PHA) or lipopolysaccharides (LPS). Aliquots of each supernatant were then assayed for levels of IFN-&#915;, vascular endothelial growth factor (VEGF), TNF-&#945;, and IL-4 using enzyme linked immunosorbent assays (ELISAs). Data collected were analyzed using Student's t-test, an ANOVA test followed by Tukey's test, and tests of Pearson's Correlation. PBMCs cultured with KB16-conditioned medium produced the highest levels of IFN-&#915;. VEGF was also detected in conditioned media collected from all of the squamous cell carcinoma (SCC) cell lines used, and a significant difference in VEGF levels between control and KB- or KB16-conditioned media was observed. TNF-&#945; was secreted by all PBMC groups within 6 hours of receiving conditioned media, and these levels increased up to the 24 hour timepoint, after which levels of TNF-&#945; stabilized. In contrast, none of the supernatant samples contained detectable levels of IL-4. In combination, these data suggest that direct contact between fresh human PBMCs and conditioned media from tumor cells induces the secretion of TNF-&#945; and VEGF by PBMCs, and this represents an initial angiogenic response

Toll-like receptor activity in Recurrent Aphthous Ulceration

Toll-like receptors (TLR) are membrane proteins that recognize conserved molecules derived from bacterial, virus, fungal or host tissues. Activation of TLRs causes the production of cytokines that mediate inflammatory responses and drive T helper (Th) 1 and 2 cell development. As an exaggerated Th1 immune response is supposed to be involved in pathogenesis of Recurrent Aphthous Ulceration (RAU), we suggest that RAU patients may have an imbalance in TLR pathways. To study the function of TLR activation ex vivo, peripheral blood mononuclear cells (PBMCs) from RAU patients (n = 17) and controls (n = 17) were exposed to TLR2 [lipoteichoic acid (LTA), heat-killed Listeria monocytogenes (HKLM) and PamC3CSK4], TLR3 [Poly(I:C)], TLR4 [lipopolysaccharide (LPS)], TLR5 (flagellin) and TLR7 (imiquimod) ligands, and the time course of supernatant tumor necrosis factor-alpha (TNF-alpha) levels was quantified by enzyme-linked immunosorbent assay. In addition, serological and salivary TNF-alpha and soluble CD14 levels were quantified. The TNF-alpha produced by PBMCs in contact with each TLR ligand and autologous serum or saliva at the same time was also investigated. The data were analyzed by statistical multivariate tests. The control group had a higher response to LTA, whereas RAU had a higher response to HKLM. LTA and LPS interfered with the salivary stimulation of the RAU PBMC and HKLM with the stimulation of the control. Autologous serum was capable of inhibiting TLR2 responsiveness to LTA and enhancing LPS stimulation. Salivary and serological levels of sCD14 and TNF-alpha were not significantly different. Recurrent Aphthous Ulceration patients have an anomalous activity of the TLR2 pathway that probably influences the stimulation of an abnormal Th1 immune response.Fundo de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[05/59738-2

Involvement of GDF-9, leptin, and IGF1 receptors associated with adipose tissue transplantation on fertility restoration in obese anovulatory mice

The aim was to analyze the effect of adipose tissue transplantation on growth differentiation factor-9 (GDF-9), insulin growth factor 1 receptor (IGF1R), and leptin receptor (LEPR) protein expression in ovaries of obese anovulatory mice. Leptin-deficient female (ob/ob) and wild-type mice were divided into untreated ob/ob mice and gonadal white adipose tissue transplanted ob/ob mice, with evaluation after 7, 15, and 45 days and compared to control wild-type mice. the corporal weight and glycemia levels increased in the obese group concomitant with polymicrocyst formation and abundant estrone, mimicking anovulatory disease. in the treated group after 45 days, glycemia, weight, ovarian size, and number of follicles were decreased and corpora lutea were decreased. the analysis of GDF-9 revealed that, whereas control ovaries presented follicular localization, the obese ovary lacked this protein. On the other hand, obese ovaries showed elevated expression of IGF1R that was normalized after the transplantation. Finally, LEPR was reduced in obese ovaries, and adipose tissue transplantation was efficient in returning it to normal levels. in conclusion, the adipose tissue transplantation, especially after 45 days, seems to stimulate ovulation, supported by the fact that several proteins involved in ovulation returned to basal levels.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Gynecol, BR-04039032 São Paulo, BrazilSao Leopoldo Mand, Postgrad Program, Campinas, SP, BrazilUniversidade Federal de São Paulo, Dept Morphol & Genet, BR-04039032 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Gynecol, BR-04039032 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Morphol & Genet, BR-04039032 São Paulo, BrazilFAPESP: 07/55629-0Web of Scienc

Differential gene expression assessed by cDNA microarray analysis in breast cancer tissue under tamoxifen treatment

Our purpose was to identify tamoxifen (TAM) responsive genes after 30 days of TAM treatment in tumor tissues obtained from women with breast cancer using microarray expression analysis. In our study, we identified 12 candidates to be considered as tamoxifen-modulated genes. Among them, we selected two candidates the TEGT BI-1 (testis enhanced gene transcript Bax Inhibitor-1) and the CD63 gene in order to further confirm their differential expression under tamoxifen effects. We observed that both were down-regulated in tumor tissues of patients during TAM treatment. TEGT is able to inhibit the expression of Bax, which is known to promote apoptosis. On the other hand, CD63 encodes a cell membrane protein and it seems to be involved in mechanisms of platelet activation. cell adhesion and cell motility. We therefore hypothesize that TAM would be able to modulate tumor growth by down-regulating genes involved in mechanisms such as cell cycle control, tumor invasion and metastasis.Univ Fed Sao Paulo, Dept Gynecol, Mol Gynecol Lab, Paulista Sch Med, Sao Paulo, SP, BrazilLudwig Inst Canc Res, Sao Paulo Branch, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Gynecol, Mol Gynecol Lab, Paulista Sch Med, Sao Paulo, SP, BrazilWeb of Scienc

Ag Nanoparticles/AgX (X=Cl, Br and I) Composites with Enhanced Photocatalytic Activity and Low Toxicological Effects

This is the pre-peer reviewed version of the following article: M. Assis, F. C. Groppo Filho, D. S. Pimentel, T. Robeldo, A. F. Gouveia, T. F. D. Castro, H. C. S. Fukushima, C. C. de Foggi, J. P. C. da Costa, R. C. Borra, J. Andrés, E. Longo. Ag Nanoparticles/AgX (X=Cl, Br and I) Composites with Enhanced Photocatalytic Activity and Low Toxicological Effects, which has been published in final form at https://doi.org/10.1002/slct.202000502. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.Periodic structures induced by electron irradiation are a unique phenomenon when electron beams irradiate on the surface of some materials. These periodic structures have potential for technological applications. However, the fuzzy nature of the electron‐induced structuring hinders its further exploration in such applications. In this paper, novel Ag nanoparticle/AgX (X=Cl, Br and I) composites, with enhanced photocatalytic activity and low toxicological effects, were prepared, for the first time, using electron beam irradiation. The remarkable advantage of this approach is that the Ag nanoparticles/AgX composites can be easily prepared in one‐step without the need for high‐pressure conditions, surfactants, ionic liquids, or reducing agents. Furthermore, our method does not involve any toxic substances, which makes the as‐synthesized samples highly applicable for technological applications. The structure, morphology and physicochemical properties of the Ag nanoparticles/AgX composites were studied using various characterization techniques. Using first‐principles calculations based on density functional theory and the quantum theory of atoms in molecules, we reveal how the concentration of excess electrons in the AgX materials induces the formation of the Ag nanoparticles under electron beam irradiation. These results extend the fundamental understanding of the atomic process underlying the mechanism of Ag−X bond rupture observed during the transformation induced via electron irradiation of the AgX crystals by increasing the total number of electrons in the bulk structure. Thus, our findings provide viable guidance for the realization of new materials for the degradation of contaminated wastewater with low toxicity