Rosângela Vieira de Andrade

Extracellular Paracoccidioides brasiliensis phospholipase B involvement in alveolar macrophage interaction.

BackgroundPhospholipase B (PLB) has been reported to be one of the virulence factors for human pathogenic fungi and has also been described as necessary for the early events in infection. Based on these data, we investigated the role of PLB in virulence and modulation of the alveolar pulmonary immune response during infection using an in-vitro model of host-pathogen interaction, i.e. Paracoccidioides brasiliensis yeast cells infecting alveolar macrophage (MH-S) cells.ResultsThe effect of PLB was analyzed using the specific inhibitor alexidine dihydrochloride (0.25 μM), and pulmonary surfactant (100 μg mL-1), during 6 hours of co-cultivation of P. brasiliensis and MH-S cells. Alexidine dihydrochloride inhibited PLB activity by 66% and significantly decreased the adhesion and internalization of yeast cells by MH-S cells. Genes involved in phagocytosis (trl2, cd14) and the inflammatory response (nfkb, tnf-α, il-1β) were down-regulated in the presence of this PLB inhibitor. In contrast, PLB activity and internalization of yeast cells significantly increased in the presence of pulmonary surfactant; under this condition, genes such as clec 2 and the pro-inflammatory inhibitor (nkrf) were up-regulated. Also, the pulmonary surfactant did not alter cytokine production, while alexidine dihydrochloride decreased the levels of interleukin-10 (IL-10) and increased the levels of IL-12 and tumor necrosis factor-α (TNF-α). In addition, gene expression analysis of plb1, sod3 and icl1 suggests that P. brasiliensis gene re-programming is effective in facilitating adaptation to this inhospitable environment, which mimics the lung-environment interaction.ConclusionP. brasiliensis PLB activity is involved in the process of adhesion and internalization of yeast cells at the MH-S cell surface and may enhance virulence and subsequent down-regulation of macrophage activation.

Increased expression of genes after periodontal treatment with photodynamic therapy

BACKGROUND The current study was devised with the objective of using a split-mouth, controlled clinical trial to compare conventional mechanical debridement (scaling and root planing) treatment (T1) with conventional mechanical treatment followed by photodynamic therapy (PDT) (T2) in patients with severe periodontitis. METHODS Four PDT sessions were completed, and clinical parameters such as bleeding upon probing (BOP positive), plaque index (PI), probing pocket depth (PPD) and clinical attachment loss (CAL) were evaluated before and after the treatment series. In addition, gingival biopsies were collected at the start and finish of treatment, and were used for qPCR gene expression analysis of TNFA, IL1B, IL8, IL10, IL17, MMP13, FGF2, RANK, RANKL and OPG. RESULTS The clinical results showed a significant improvement in BOP with treatment T2 (p=0.03). The molecular data showed an up-regulation of FGF2, RANK and OPG gene expression after T2. The expression levels of the other genes were not significantly different between T1 and T2. PDT increased the expression of RANK and OPG, which could indicate a reduction in osteoclastogenesis. Furthermore, the use of PDT in conjunction with conventional treatment significantly increased the expression of FGF2, which has an important role in the periodontal repair process. CONCLUSIONS PDT technology could be a means to improve conventional periodontitis treatment. Our results suggest that PDT acts in part by controlling bone resorption and increasing the expression of genes important for tissue repair.

Analysis of genes that are differentially expressed during the Sclerotinia sclerotiorum-Phaseolus vulgaris interaction.

The fungus Sclerotinia sclerotiorum (Lib.) de Bary, one of the most important plant pathogens, causes white mold on a wide range of crops. Crop yield can be dramatically decreased due to this disease, depending on the plant cultivar and environmental conditions. In this study, a suppression subtractive hybridization cDNA library approach was used for the identification of pathogen and plant genes that were differentially expressed during infection of the susceptible cultivar BRS Pérola of Phaseolus vulgaris L. A total of 979 unigenes (430 contigs and 549 singletons) were obtained and classified according to their functional categories. The transcriptional profile of 11 fungal genes related to pathogenicity and virulence were evaluated by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Additionally, the temporal expression profile obtained by RT-qPCR was evaluated for the following categories of plant defense-related genes: pathogenesis-related genes (PvPR1, PvPR2, and PvPR3), phenylpropanoid pathway genes (PvIsof, PvFPS1, and 4CL), and genes involved in defense and stress-related categories (PvLox, PvHiprp, PvGST, PvPod, and PvDox). Data obtained in this study provide a starting point for achieving a better understanding of the pathosystem S. sclerotiorum–P. vulgaris.

Resistance training regulates gene expression of molecules associated with intramyocellular lipids, glucose signaling and fiber size in old rats

Sarcopenia is a complex multifactorial process, some of which involves fat infiltration. Intramyocellular lipid (IMCL) accumulation is postulated to play a role on sarcopenia during aging, which is believed to be due alterations in glucose homeostasis in the skeletal muscle. Sarcopenia, along with intramuscular lipids, is associated with physical inactivity. Resistance training (RT) has been indicated to minimize the age-induced muscle skeletal adaptations. Thus, we aimed to investigate the effects of RT on mRNA levels of regulatory components related to intramyocellular lipid, glucose metabolism and fiber size in soleus and gastrocnemius muscles of aged rats. Old male rats were submitted to RT (ladder climbing, progressive load, 3 times a week for 12 weeks). Age-induced accumulation of IMCL was attenuated by RT, which was linked to a PPARy-mediated mechanism, concomitant to enhanced regulatory components of glucose homeostasis (GLUT-4, G6PDH, Hk-2 and Gly-Syn-1). These responses were also linked to decreased catabolic (TNF-α, TWEAK/Fn14 axis; FOXO-1, Atrogin-1 and MuRF1; Myostatin) and increased anabolic intracellular pathways (IGF-1-mTOR-p70S6sk-1 axis; MyoD) in muscles of trained aged rats. Our results point out the importance of RT on modulation of gene expression of intracellular regulators related to age-induced morphological and metabolic adaptations in skeletal muscle.

MicroRNA expression profiles in human CD3 + T cells following stimulation with anti-human CD3 antibodies

BackgroundAnti-CD3 therapy can induce immunosuppression by several non mutually exclusive mechanisms that have been proposed to explain the therapeutic effect the administration anti-CD3 mAb, but its immunoregulatory mechanism is still not completely clear. In T cells, microRNAs (miRNAs) regulate several pathways, including those associated with immune tolerance. Here, we report changes in miRNA expression in T cells following treatment with anti-human CD3 antibodies. Peripheral blood mononuclear cells were cultured in the presence of the monoclonal antibody OKT3 or a recombinant fragment of humanized anti-CD3. Following these treatments, the expression profiles of 31 miRNA species were assessed in T cells using TaqMan arrays.ResultsEight of the tested miRNAs (miR-155, miR-21, miR-146a, miR-210, miR-17, miR-590-5p, miR-106b and miR-301a) were statistically significantly up- or down-regulated relative to untreated cells.ConclusionsStimulation of T cells with anti-human CD3 antibodies alters miRNA expression patterns, including of miRNA species associated with immune regulatory pathways.

Use of microRNAs in directing therapy and evaluating treatment response in colorectal cancer

ABSTRACT Colorectal cancer is the third most common cancer worldwide. Survival and prognosis depend on tumor stage upon diagnosis, and in more than 50% of cases, the tumor has already invaded adjacent tissues or metastasis has occurred. Aiming to improve diagnosis, clinical prognosis and treatment of patients with colorectal cancer, several studies have investigated microRNAs as molecular markers of the disease due to their potential regulatory functions on tumor suppressor genes and oncogenes. This review aimed to summarize the main topics related to the use of microRNAs in diagnosis, clinical prognosis and evaluating treatment response in colorectal cancer.

Cryptosporidium genotyping and land use mapping for hazard identification and source tracking in a small mixed rural–urban watershed in Southeastern Brazil

Cryptosporidium, faecal indicator organisms and physical and chemical water quality variables were monitored in a small mixed rural–urban watershed in southeastern Brazil. Cryptosporidium was present in 43% of 117 water samples analysed by microscopy. Concentrations varied from nondetects to 14 oocysts L . All samples were further analysed by nested-PCR, and Cryptosporidium spp. were detected in 24% (28) of them. Sequencing at the 18S rRNA locus gave high quality sequences in eight samples, revealing the presence of Cryptosporidium parvum. Cryptosporidium was not correlated with faecal indicator organisms (total coliforms, Escherichia coli, Enterococcus and coliphages), nor with physical and chemical water quality variables (e.g. turbidity, electrical conductivity and chemical oxygen demand), but it was with farm animal density (number of animals per ha). Land use mapping reinforced the suggestions from Cryptosporidium genotyping that both animals (livestock) and humans are potential sources to environmental contamination with oocysts within the watershed. doi: 10.2166/wh.2018.143 s://iwaponline.com/jwh/article-pdf/17/1/149/564835/jwh0170149.pdf Rosane C. Andrade Ministério da Saúde. Departamento de Saúde Ambiental e Saúde do Trabalhador, Coordenação Geral de Vigilância em Saúde Ambiental, Brasília-DF, 70723-040, Brazil Rafael K. X. Bastos (corresponding author) Departamento de Engenharia Civil, Universidade Federal de Viçosa, Viçosa-MG, 36570-000, Brazil E-mail: rkxb@ufv.br Paula D. Bevilacqua Departamento de Veterinária, Universidade Federal de Viçosa, Viçosa-MG, 36570-000, Brazil Rosângela V. Andrade Programa de Pós-graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, Brasília-DF, 70790-160, Brazil

Effect of Resistance Training on Extracellular Matrix Adaptations in Skeletal Muscle of Older Rats

Accumulation of connective tissue, particularly extracellular matrix (ECM) proteins, has been observed in skeletal muscles with advancing age. Resistance training (RT) has been widely recommended to attenuate age-induced sarcopenia, even though its effects on the components that control ECM turnover in skeletal muscles remain to be elucidated. Thus, the aim of this study was to determine the effects of RT on connective tissue content and gene expression of key components of ECM in the skeletal muscles of aged rats. Young (3 mo.) and older (21 mo.) adult male Wistar rats were submitted to a RT protocol (ladder climbing with 65, 85, 95, and 100% load), 3 times a week for 12 weeks. Forty-eight hours post-training, the soleus (SOL) and gastrocnemius (GAS) muscles were dissected for histological and mRNA analysis. RT mitigated the age-associated increase of connective tissue content in both muscles, even though mRNA levels of COL-1 and−3 were elevated in older trained rats. Overall, RT significantly elevated the gene expression of key components of connective tissue deposition (TGFβ and CTGF; MMP-2 and-9; TIMP-1 and−2) in the GAS and SOL muscles of older rats. In conclusion, RT blunted the age-induced accumulation of connective tissue concomitant to the upregulation of genes related to synthesis and degradation of the ECM network in the SOL and GAS muscles of older rats. Although our findings indicate that RT plays a crucial role reducing connective tissue accumulation in aged hindlimb muscles, key components of ECM turnover were paradoxically elevated. The phenotypic responses induced by RT were not accompanied by the gene expression of those components related to ECM turnover.