Regulon: An overview of plant abiotic stress transcriptional regulatory system and role in transgenic plants / Regulon: Uma visão geral do sistema regulador da transcrição do estresse abiótico em plantas e papel nas plantas transgênica

Population growth is increasing rapidly around the world, in these consequences we need to produce more foods to full fill the demand of increased population. The world is facing global warming due to urbanizations and industrialization and in this concerns plants exposed continuously to abiotic stresses which is a major cause of crop hammering every year. Abiotic stresses consist of Drought, Salt, Heat, Cold, Oxidative and Metal toxicity which damage the crop yield continuously. Drought and salinity stress severally affected in similar manner to plant and the leading cause of reduction in crop yield. Plants respond to various stimuli under abiotic or biotic stress condition and express certain genes either structural or regulatory genes which maintain the plant integrity. The regulatory genes primarily the transcription factors that exert their activity by binding to certain cis DNA elements and consequently either up regulated or down regulate to target expression. These transcription factors are known as masters regulators because its single transcript regulate more than one gene, in this context the regulon word is fascinating more in compass of transcription factors. Progress has been made to better understand about effect of regulons (AREB/ABF, DREB, MYB, and NAC) under abiotic stresses and a number of regulons reported for stress responsive and used as a better transgenic tool of Arabidopsis and Rice.

O crescimento populacional está aumentando rapidamente em todo o mundo, e para combater suas consequências precisamos produzir mais alimentos para suprir a demanda do aumento populacional. O mundo está enfrentando o aquecimento global devido à urbanização e industrialização e, nesse caso, plantas expostas continuamente a estresses abióticos, que é uma das principais causas do martelamento das safras todos os anos. Estresses abióticos consistem em seca, sal, calor, frio, oxidação e toxicidade de metais que prejudicam o rendimento da colheita continuamente. A seca e o estresse salino são afetados de maneira diversa pela planta e são a principal causa de redução da produtividade das culturas. As plantas respondem a vários estímulos sob condições de estresse abiótico ou biótico e expressam certos genes estruturais ou regulatórios que mantêm a integridade da planta. Os genes reguladores são principalmente os fatores de transcrição que exercem sua atividade ligando-se a certos elementos cis do DNA e, consequentemente, são regulados para cima ou para baixo para a expressão alvo. Esses fatores de transcrição são conhecidos como reguladores mestres porque sua única transcrição regula mais de um gene; nesse contexto, a palavra regulon é mais fascinante no âmbito dos fatores de transcrição. Progresso foi feito para entender melhor sobre o efeito dos regulons (AREB / ABF, DREB, MYB e NAC) sob estresses abióticos e uma série de regulons relatados como responsivos ao estresse e usados como uma melhor ferramenta transgênica de Arabidopsis e Rice.

Structure and character analysis of cotton response regulator genes family reveals that GhRR7 responses to draught stress

BACKGROUND: Cytokinin signal transduction is mediated by a two-component system (TCS). Two-component systems are utilized in plant responses to hormones as well as to biotic and abiotic environmental stimuli. In plants, response regulatory genes (RRs) are one of the main members of the two-component system (TCS). METHOD: From the aspects of gene structure, evolution mode, expression type, regulatory network and gene function, the evolution process and role of RR genes in the evolution of the cotton genome were analyzed. RESULT: A total of 284 RR genes in four cotton species were identified. Including 1049 orthologous/paralogous gene pairs were identified, most of which were whole genome duplication (WGD). The RR genes promoter elements contain phytohormone responses and abiotic or biotic stress-related cis-elements. Expression analysis showed that RR genes family may be negatively regulate and involved in salt stress and drought stress in plants. Protein regulatory network analysis showed that RR family proteins are involved in regulating the DNA-binding transcription factor activity (COG5641) pathway and HP kinase pathways. VIGS analysis showed that the GhRR7 gene may be in the same regulatory pathway as GhAHP5 and GhPHYB, ultimately negatively regulating cotton drought stress by regulating POD, SOD, CAT, H2O2 and other reactive oxygen removal systems. CONCLUSION: This study is the first to gain insight into RR gene members in cotton. Our research lays the foundation for discovering the genes related to drought and salt tolerance and creating new cotton germplasm materials for drought and salt tolerance.

Characteristics and Clinical Significance of Gene Mutation in Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

OBJECTIVE@#To investigate the characteristics of gene mutations in patients with myelodysplastic syndromes (MDS) and its prognostic significance.@*METHODS@#High-throughput sequencing was used to detect 34 blood tumor-related genes in 210 patients with MDS, and the relationship with the revised International Prognostic Scoring System (IPSS-R) and the impact on prognosis of the patients were analyzed.@*RESULTS@#Among the 210 MDS patients, 142 cases (67.6%) showed mutations, and the first six genes with the highest mutation detection rate were ASXL1(20.5%), TET2(17.1%), U2AF1(14.3%), DNMT3A (11.9%), TP53(10.5%) and RUNX1(10.0%). The gene mutation rate of the patients in IPSS-R relatively high-risk group was higher than those in relatively low-risk group (P=0.001). Both TP53 and BCOR genes showed higher mutation rates in the higher risk group than in the lower risk group (P<0.05). Survival time of the patients in TP53 mutant group was lower than those in non-mutant group (P<0.001), survival time of patients in SF3B1 mutant group was higher than those in non-mutant group (P=0.018). According to the number of gene mutations, the patients could be divided into groups with 0-1, 2 and ≥3 gene mutations, and the median OS of the three groups were not reached, 43 and 27 months, respectively (P=0.004). The Multivariate analysis showed that the increasing number of gene mutations and TP53 mutation was the independent risk factors affecting prognosis of the patients, while SF3B1 mutation was the independent protective factor for the prognosis of the patients.@*CONCLUSION@#The gene mutation rate was higher in MDS patients. And the increasing numbers of gene mutation, TP53 and SF3B1 were the influence factors of prognosis in the patients.

New Advane of Reaserch on Relationship between Long Noncoding RNA and Acute Myeloid Leukemia--Review / 中国实验血液学杂志

Acute myeloid leukemia（AML）is a myelopoietic stem/progenitor malignant disease. The exact etiology of this leukemia remains unclear, thus it is important to explore the pathogenesis of AML and to discover the new diagnostic markers and therapeutic targets. The long non coding RNA (lnc RNA) is a class of RNA molecules with transcripts over 200 nucleotides in eukaryotic cells which almost don't possess the ability to code proteins, but can regulate the expression of other genes at transcriptional and post-transcriptional levels, thereby participate in occurrence and development of varied tumors. Of late years, along with the deepening of study, the lncRNA roles played in the AML have been reported and confirmed. In this review, the relationships between the IncRNA (UCA1, ANRIL, H19, HOTAIR, CCAT1, ZFAS1, LINC00152, HOXA-A52, NEAT1, TUG1, IRAIN1, PANDAR, LINC00899, SNHG5, and KCNQ1OT1) and AML is summarized briefly, so as to provide the potential basis for the clinical diagnosis and therapy of AML.

Gene regulatory network inference and analysis of multidrug-resistant Pseudomonas aeruginosa

BACKGROUND Healthcare-associated infections caused by bacteria such as Pseudomonas aeruginosa are a major public health problem worldwide. Gene regulatory networks (GRN) computationally represent interactions among regulatory genes and their targets. They are an important approach to help understand bacterial behaviour and to provide novel ways of overcoming scientific challenges, including the identification of potential therapeutic targets and the development of new drugs. OBJECTIVES The goal of this study was to reconstruct the multidrug-resistant (MDR) P. aeruginosa GRN and to analyse its topological properties. METHODS The methodology used in this study was based on gene orthology inference using the reciprocal best hit method. We used the genome of P. aeruginosa CCBH4851 as the basis of the reconstruction process. This MDR strain is representative of the sequence type 277, which was involved in an endemic outbreak in Brazil. FINDINGS We obtained a network with a larger number of regulatory genes, target genes and interactions as compared to the previously reported network. Topological analysis results are in accordance with the complex network representation of biological processes. MAIN CONCLUSIONS The properties of the network were consistent with the biological features of P. aeruginosa. To the best of our knowledge, the P. aeruginosa GRN presented here is the most complete version available to date.

Expressão gênica e proteica de membros da família HOX no câncer de mama luminal / Gene and protein expression of HOX family members in luminal breast cancer

Os genes homeobox são fatores de transcrição que regulam a expressão de múltiplos genes que influenciam o crescimento celular e fazem a mediação entre o epitélio e o estroma, regulando a diferenciação específica de cada tecido. Sua expressão é comumente desregulada em tumores, e estudos indicam que estes genes atuam como oncogenes, promovendo crescimento celular e invasão, ou como supressores tumorais, devido à sua atuação nos processos de morfogênese. No caso do câncer de mama, alguns genes homeobox tem expressão aumentada e outros, reduzida, e em geral não há ocorrência de mutações. Objetivos: Este trabalho procurou estudar a expressão de membros da família HOX em carcinomas luminais da mama, e correlacionar essa expressão com características clínico-patológicas, sobrevida global e livre de doença; avaliar a correlação entre a expressão de HOXA1 e Receptor de Progesterona; avaliar a correlação da expressão de HOXB7 e de MYC, bem como comparar os resultados da expressão gênica por imunohistoquímica e RT-qPCR. Métodos: Foram selecionados 260 pacientes com Carcinoma Mamário Luminal (CML) diagnosticados entre 2007 e 2010, 29 pacientes com amostras de CML congelados no Biobanco do AC Camargo Cancer Center, todos pareados com as respectivas amostras para imunohistoquímica, e 4 casos de tecido mamário normal congelado oriundos das pacientes doadoras de amostras de tumor congeladas. A expressão gênica foi pesquisada através dos métodos de imunohistoquímica e reação em cadeia da polimerase da transcrição reversa em tempo real. A técnica imunohistoquímica e os ensaios de expressão gênica foram realizados para os genes HOXA1, HOXA5, HOXA9, HOXB7, HOXB9, HOXB13, HOXC13 e HOXD3. Para a técnica de imunohistoquímica, o número de células positivas foi quantificado para cada marcador através do sistema de morfometria e escaneamento de lâminas Aperio ScanScope XT. A quantificação relativa de expressão gênica, na técnica de RT-qPCR foi realizada utilizando o software Sequence Detection System (Applied Biosystems), e calculada pelo modelo matemático descrito por Pfaffl. Todos os testes estatísticos foram realizados utilizando os softwares Excel (Microsoft 2011) e IBM SPSS, versão 24. Resultados: Houve associação com entre a expressão de HOXB7 e estadiamento patológico T ao diagnóstico (p=0,015) e entre a expressão de HOXC13 e estadiamento N ao diagnóstico (p=0,002; OR=2,61). Aumento da expressão de HOXA9 foi associado com redução da sobrevida global (p=0,031; RR: 2,331, IC95% [1,054-5,157], P=0,037). Não houve associação entre a expressão de HOXA1 e Receptor de Progesterona e/ou entre a expressão de HOXB7 e MYC. A correlação entre a expressão gênica por imunohistoquímica e RT-qPCR foi inexistente, negativa fraca ou positiva muito fraca. Conclusões: Aumento da expressão de HOXB7 é associado com pior estadiamento patológico T ao diagnóstico. Aumento da expressão de HOXC13 é associado com maior ocorrência de metástases linfonodais. Aumento da expressão de HOXA9 reduz a sobrevida global

Homeobox genes are transcription factors that regulate the expression of multiple genes which affect cell growth and make the mediation between the epithelium and the stroma, so regulating the differentiation of each specific tissue. Its expression is commonly deregulated in tumors, and studies indicate that these genes act as oncogenes, promoting cell growth and invasion, or act as tumor suppressors genes, due to its role in morphogenesis processes. In breast cancer, homeobox genes can have their increased or decreased expression, and generally there are no ocurrence of mutations. Objectives: This work aimed to study the expression of HOX family members in luminal carcinomas of the breast and to correlate this expression with clinical-pathological characteristics, global and disease-free survival; to evaluate the correlation between the expression of HOXA1 and Progesterone Receptor; to evaluate the correlation of HOXB7 and MYC expression, as well as to compare the results of the gene expression by immunohistochemistry and RT-qPCR. Methods: We selected 260 patients with Luminal Mammary Carcinoma (CML) diagnosed between 2007 and 2010, 29 patients with frozen CML samples in the AC Camargo Cancer Center Biobank, all paired with the respective samples for immunohistochemistry, and 4 cases of normal breast tissue frozen from donor patients with frozen tumor samples. Gene expression was investigated through the immunohistochemistry and reverse transcription polymerase chain reaction in real time. Immunohistochemical technique and gene expression assays were performed for the genes HOXA1, HOXA5, HOXA9, HOXB7, HOXB9, HOXB13, HOXC13 and HOXD3. For the immunohistochemical technique, the number of positive cells was quantified for each marker through the morphometry and scanning system of Aperio ScanScope XT slides. The relative quantification of gene expression in the RT-qPCR technique was performed using the Sequence Detection System (Applied Biosystems) software, and calculated by the mathematical model described by Pfaffl. Results: There was an association between the expression of HOXB7 and pathological staging T at the diagnosis (p = 0.015) and between the expression of HOXC13 and staging N at diagnosis (p = 0.002, OR = 2.61). Increased expression of HOXA9 was associated with a reduction in overall survival (p = 0.031, RR = 2.331, 95% CI [1.054-5.157], P = 0.037). There was no association between the expression of HOXA1 and Progesterone Receptor and / or between the expression of HOXB7 and MYC. The correlation between the gene expression by immunohistochemistry and RT-qPCR was non-existent, negative negative or very weak positive. Conclusions: Increased expression of HOXB7 is associated with poorer T staging at diagnosis. Increased expression of HOXB13 is associated with increased occurrence of lymph node metastases. Increased expression of HOXA9 reduces overall survival

Síndrome de Chediak-Higashi em fase acelerada: um relato de caso / Accelerated Chediak-Higashi syndrome: case report

A síndrome de Chediak-Higashi (CHS) é um distúrbio genético autossômico recessivo decorrente de uma mutação no gene regulador do transporte lisossomal (LYST ou CHS1). Os sintomas da síndrome são resultado de alterações funcionais de melanócitos, plaquetas, neutrófilos e células natural killer, e incluem albinismo parcial, fotossensibilidade, infecções recorrentes, principalmente bacterianas, linfocitose hemofagocítica, sangramentos e manifestações neurológicas, como neuropatia central e periférica, perda de sensibilidade, fraqueza muscular, ataxia cerebelar e déficit cognitivo. Aproximadamente 85% dos casos se apresentam como a forma avançada, caracterizada por pancitopenia, hemofagocitose e infiltrado linfocítico em todos os órgãos, determinando falência múltipla dos órgãos. Nesse estudo é relatado o caso de uma paciente diagnosticada com a síndrome aos 8 anos de idade, apresentando a doença já em fase avançada, além de uma rápida revisão bibliográfica sobre a doença em questão.

Chediak-Higashi syndrome (CHS) is an autosomal recessive genetic disorder caused by a mutation in the lysosomal trafficking regulator gene (LYST or CHS1). Symptoms of the syndrome result from functional abnormalities in melanocytes, platelets, neutrophils and natural killer cells and include partial albinism, photosensitivity, recurrent infections (mainly bacterial), hemophagocytic lymphohistiocytosis, bleeding and neurological manifestations such as central and peripheral neuropathy, loss of sensitivity, muscle weakness, cerebellar ataxia and cognitive deficit. Approximately 85% of the cases present the advanced form, characterized by pancytopenia, hemophagocytosis and lymphocyte infiltration of all organs, determining multiple organ failure. This study reports the case of a patient diagnosed with the syndrome at 8 years of age, already at an advanced stage. A brief review of the literature available on the condition is presented.

Diallyl Trisulfide Suppresses the Production of Lipopolysaccharide-induced Inflammatory Mediators in BV2 Microglia by Decreasing the NF-κB Pathway Activity Associated With Toll-like Receptor 4 and CXCL12/CXCR4 Pathway Blockade

BACKGROUND: Diallyl trisulfide (DATS), a garlic-derived organosulfuric compound, has been documented for potential anti-inflammatory effects. However, the mechanism in microglia remains unknown. In this study, we investigated the anti-inflammatory effects of DATS in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. METHODS: The effects of DATS on LPS-induced pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2) were assessed under conditions not in the cytotoxicity of DATS. The protein expression of inflammation regulatory genes was measured by Western blot analysis. RESULTS: DATS significantly inhibited the LPS-induced secretion of NO and PGE2, which was associated with the suppression of their regulatory genes, inducible NO synthase and COX-2. DATS had been shown to inhibit nuclear translocation of NF-κB by destroying the degradation and phosphorylation of IκB-α inhibitors in the cytoplasm. In addition, DATS effectively inhibited the expression of LPS-induced toll-like receptor 4 (TLR4) and myeloid differentiation factor 88. Furthermore, DATS markedly reduced the LPS-induced expression of chemokine (CXC motif) ligand (CXCL) 12 and CXC receptor (CXCR) 4, demonstrating its capacity to block chemo-attractive activity. CONCLUSIONS: These results indicate that DATS inhibits the activation of the CXCL12/CXCR4 axis associated with antagonizing effect on TLR4 and blocks NF-κB signaling, thus demonstrating anti-inflammatory effects against LPS stimulation.

Research on Ralation of Long Non-coding RNA with Diffuse Large B-cell Lymphomas -Review / 中国实验血液学杂志

Diffuse large B-cell lymphoma (DLBCL) accounts for approximately 30% of the non-Hodgkin's lymphoma patients. The underlying molecular mechanism of its pathogenesis is not well defined and the survival rate of DLBCL patients is very low. Moreover, the annual incidence and mortality of DLBCL is still rising. Accordingly, identification and characterization of new molecular pathways of DLBCL will lead to the development of novel diagnostic markers and molecular therapeutic targets. Long non-coding RNAs (LncRNA) are non-coding RNAs with a length greater than 200 bp in eukaryotic cells, which can regulate the expression of their target genes at the transcriptional and post transcriptional levels. The function of LncRNAs is involved in the initiation, progression, invasion and metastasis of many cancers. Recently, the role of LncRNAs in DLBCL has been identified and intensely studied. This review summarizes the recent discoveries in the expression and function of LncRNAs including HULC,PEG10,LincRNA-p21,HOTAIR,LUNAR1,MALAT1 and SubSigLnc-17 in DLBCL, so as to find potential diagnostic biomarkers and therapeutic targets for DLBCL.

Spermidine Protects against Oxidative Stress in Inflammation Models Using Macrophages and Zebrafish

Spermidine is a naturally occurring polyamine compound that has recently emerged with anti-aging properties and suppresses inflammation and oxidation. However, its mechanisms of action on anti-inflammatory and antioxidant effects have not been fully elucidated. In this study, the potential of spermidine for reducing pro-inflammatory and oxidative effects in lipopolysaccharide (LPS)-stimulated macrophages and zebrafish was explored. Our data indicate that spermidine significantly inhibited the production of pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2), and cytokines including tumor necrosis factor-α and interleukin-1β in RAW 264.7 macrophages without any significant cytotoxicity. The protective effects of spermidine accompanied by a marked suppression in their regulatory gene expression at the transcription levels. Spermidine also attenuated the nuclear translocation of NF-κB p65 subunit and reduced LPS-induced intracellular accumulation of reactive oxygen species (ROS) in RAW 264.7 macrophages. Moreover, spermidine prevented the LPS-induced NO production and ROS accumulation in zebrafish larvae and was found to be associated with a diminished recruitment of neutrophils and macrophages. Although more work is needed to fully understand the critical role of spermidine on the inhibition of inflammation-associated migration of immune cells, our findings clearly demonstrate that spermidine may be a potential therapeutic intervention for the treatment of inflammatory and oxidative disorders.

Estimation of the Genetic Substitution Rate of Hanwoo and Holstein Cattle Using Whole Genome Sequencing Data

Despite the importance of mutation rate, some difficulties exist in estimating it. Next-generation sequencing (NGS) data yields large numbers of single-nucleotide polymorphisms, which can make it feasible to estimate substitution rates. The genetic substitution rates of Hanwoo and Holstein cattle were estimated using NGS data. Our main findings was to calculate the gene's substitution rates. Through estimation of genetic substitution rates, we found: diving region of altered substitution density exists. This region may indicate a boundary between protected and unprotected genes. The protected region is mainly associated with the gene ontology terms of regulatory genes. The genes that distinguish Hanwoo from Holstein in terms of substitution rate predominantly have gene ontology terms related to blood and circulatory system. This might imply that Hanwoo and Holstein evolved with dissimilar mutation rates and processes after domestication. The difference in meat quality between Hanwoo and Holstein could originate from differential evolution of the genes related to these blood and circulatory system ontology terms.

Fluvastatin inhibits advanced glycation end products-induced proliferation, migration, and extracellular matrix accumulation in vascular smooth muscle cells by targeting connective tissue growth factor

Connective tissue growth factor (CTGF) is a novel fibrotic mediator, which is considered to mediate fibrosis through extracellular matrix (ECM) synthesis in diabetic cardiovascular complications. Statins have significant immunomodulatory effects and reduce vascular injury. We therefore examined whether fluvastatin has anti-fibrotic effects in vascular smooth muscle cells (VSMCs) and elucidated its putative transduction signals. We show that advanced glycation end products (AGEs) stimulated CTGF mRNA and protein expression in a time-dependent manner. AGE-induced CTGF expression was mediated via ERK1/2, JNK, and Egr-1 pathways, but not p38; consequently, cell proliferation and migration and ECM accumulation were regulated by CTGF signaling pathway. AGE-stimulated VSMC proliferation, migration, and ECM accumulation were blocked by fluvastatin. However, the inhibitory effect of fluvastatin was restored by administration of CTGF recombinant protein. AGE-induced VSMC proliferation was dependent on cell cycle arrest, thereby increasing G1/G0 phase. Fluvastatin repressed cell cycle regulatory genes cyclin D1 and Cdk4 and augmented cyclin-dependent kinase inhibitors p27 and p21 in AGE-induced VSMCs. Taken together, fluvastatin suppressed AGE-induced VSMC proliferation, migration, and ECM accumulation by targeting CTGF signaling mechanism. These findings might be evidence for CTGF as a potential therapeutic target in diabetic vasculature complication.

Inhibitory effects on the production of inflammatory mediators and reactive oxygen species by Mori folium in lipopolysaccharide-stimulated macrophages and zebrafish

ABSTRACT Mori folium, the leaf of Morus alba L. (Moraceae), has been traditionally used for various medicinal purposes from ancient times to the present. In this study, we examined the effects of water extract of Mori folium (WEMF) on the production of inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), and reactive oxygen species (ROS) in lipopolysaccharide (LPS)-stimulated murine RAW 264.7 macrophages. Our data indicated that WEMF significantly suppressed the secretion of NO and PGE2 in RAW 264.7 macrophages without any significant cytotoxicity. The protective effects were accompanied by a marked reduction in their regulatory gene expression at the transcription level. WEMF attenuated LPS-induced intracellular ROS production in RAW 264.7 macrophages. It inhibited the nuclear translocation of the nuclear factor-kappa B p65 subunit and the activation of mitogen-activated protein kinases in LPS-treated RAW 264.7 macrophages. Furthermore, WEMF reduced LPS-induced NO production and ROS accumulation in zebrafish. Although more efforts are needed to fully understand the critical role of WEMF in the inhibition of inflammation, the findings of the present study may provide insights into the approaches for Mori folium as a potential therapeutic agent for inflammatory and antioxidant disorders.

Gene regulation network based analysis associated with TGF-beta stimulation in lung adenocarcinoma cells

Transforming growth factor [TGF]-beta is over-expressed in a wide variety of cancers such as lung adenocarcinoma. TGF-beta plays a major role in cancer progression through regulating cancer cell proliferation and remodeling of the tumor micro-environment. However, it is still a great challenge to explain the phenotypic effects caused by TGF-beta stimulation and the effect of TGF-beta stimulation on tumor micro-environment

Objectives: To address this issue, in the present study we used two time-course microarray data in human lung adenocarcinoma cells and applied bioinformatics methods to explore the gene regulation network responding to TGF-beta stimulation in lung adenocarcinoma cells

Materials and Methods: The time-dependent reverse-engineering method, protein-protein interaction network analyses, and calculation of the similarity measures between the links were used to construct gene regulatory network and to extract gene clusters

Results: Utilizing the constructed gene regulation network, we predicted NEFL and LUC7A show the opposite and the same change with C21orf90 if HAND2 is knocked-out after treatment with TGF-beta[1] for 4 hours and for 12 hours respectively. FGG and HSPC009 are predicted to display the opposite change with NEFL if CSMD1 is knocked out after treatment with TGF-beta[1] for 12 hours. Additionally, by integrating two datasets, we specially identified several nested clusters which included those genes regulated by TGF-beta stimulation in lung adenocarcinoma cells

Conclusions: Our analysis can help a better understanding regarding how TGF-beta stimulation causes the expression change of a number of the genes and provide a novel insight into TGF-beta stimulation effect on lung adenocarcinoma cells

The effect of methyl jasmonate on Zea mays tassel development / O efeito de methyl jasmonato sobre o devolvimento do pendão de Zea mays

Methyl jasmonate (MeJA) is a lipid-derived plant hormone that mediates diverse biological phenomena. Application of MeJA onto rice spikelet could exhibit abnormal floral organ development. Although jasmonic acid (JA) has been proved to be involved in maize tassel sex determination process, the roles of JA and its precursor MeJA in maize tassel development still remain obscure. In this study, we found that tassel development was decelerated by application of 2 mM MeJA. Exogenous MeJA also influenced the number of palea and stamens of tassel spikelets. Exogenous MeJA increased the expression level of some key regulator genes, which may responsible for the phenotypic change in MeJA-treated tassel, and may mediate the crosstalk between MeJA and other hormones.

jasmonate (MeJA) é um derivado lipídico vegetal hormônio que medeia Diversos fenómenos biológicos.Aplicação de MeJA para arroz spikelet poderá apresentar Desenvolvimento anormal DOS órgãos florais.Apesar de Ácido jasmónico (Ja), precursor de MeJA, mostrou ser envolvido no processo de determinação do sexo de milho tassel, OS papéis DOS dois compostos de milho tassel Desenvolvimento ainda permanecem obscuros.No presente estudo, descobrimos que o Desenvolvimento FOI desacelerada pelo pedido do tassel de 2 mm MeJA.MeJA exógeno também influenciou o número de palea e estames de tassel spikelets.MeJA aumentou o nível de expressão exógena, um regulador chave genes, o que Pode o responsável PELA alteração fenotípica EM MeJA tratados tassel, e podem mediar a interferência entre MeJA e outras hormonas.

Expressão gênica de metaloproteinases e de seus reguladores em neoplasias mieloproliferativas: associação com biomarcadores de angiogênese e status mutacional / Gene expression of metalloproteinases and theirs regulators myeloproliferative neoplasms: association with angiogenesis markers and mutational status

As neoplasias mieloproliferativas (NMPs) BCR-ABL1 negativas compreendem a mielofibrose primária (PMF), trombocitemia essencial (TE) e a policitemia vera (PV). A patogênese e progressão dessas NMPs não estão completamente elucidadas. As metaloproteinases de matriz (MMPs) degradam a matriz extracelular, ativando citocinas e fatores de crescimento que, por sua vez, participam da tumorigênese e angiogênese. O objetivo deste estudo foi avaliar a relação da expressão gênica das MMPs, TIMPs, HIF1-α e SPARC com os marcadores angiogênicos bFGF e VEGFA em pacientes com MF e TE, considerando o status mutacional; bem como avaliar a regulação desses genes em camundongos submetidos à hipóxia, e em modelos HIF1-α(-/-) e VHL(-/-). Foram incluídos 21 pacientes com MF, 21 com MF pós-TE, 6 com MF pós-PV, 23 com TE e 78 indivíduos controle. As análises realizadas foram: dosagem sérica e expressão de RNAm de MMP2, MMP9, TIMP1, TIMP2 e SPARC, hemograma, determinação da proteína C reativa ultrassensível, determinação das concentrações de VEGFA e bFGF e avaliação das mutações nos genes JAK2, cMPL e CALR. A avaliação da densidade microvascular da medula óssea foi feita em 30 dos pacientes incluídos. Os pacientes com MFP, MFPTE e TE apresentaram maior expressão de MMP2, SPARC, TIMP1, TIMP2 e bFGF quando comparados aos seus controles (P<0,05), enquanto MMP9 foi mais expressa nos pacientes com MFPTE e TE (P= 0,011 e P=0,047, respectivamente). Os pacientes com TE apresentaram maior expressão de HIF1-α e VEGFA em relação ao grupo controle (P<0,05). Pacientes com MF JAK2V617F positivos apresentaram maiores concentrações de MMP9, TIMP2, bFGF e VEGFA quando comparados aos pacientes portadores de mutações na CALR (P<0,05). Os pacientes com TE JAK2V617F positivos apresentaram maiores concentrações de MMP2 e TIMP2 (P=0,049 e P=0,020, respectivamente). As concentrações das proteínas estudadas não apresentaram correlação com a carga alélica de JAK2V617F e nem com a densidade microvascular da medula óssea. Células de medula óssea de camundongos submetidos à hipóxia apresentaram maior expressão de MMP2 e TIMP1 comparados aos camundongos em normóxia. Camundongos VHL(-/-) apresentaram aumento na expressão dos genes MMP2, MMP9, TIMP1, TIMP2 e VEGFA. Diferentemente, embriões HIF1-α(-/-) não foram considerados um bom modelo para este estudo devido ao envolvimento das MMPs na embriogênese/organogênese. Frente aos resultados encontrados, pode-se sugerir que a maior expressão de MMP2, SPARC e de bFGF estão associadas às NMPs. A mutação JAK2V617F foi associada a maiores concentrações de MMPs, TIMP2 VEGFA e bFGF. HIF1-α foi mais expresso na PV e na TE, sugerindo uma possível regulação da expressão das MMPs e TIMPs nessas doenças

Myeloproliferative neoplasms (MPNs) BCR-ABL1-negative include primary myelofibrosis (PMF), essential thrombocythemia (ET) and polycythemia vera (PV). The mechanisms underlying the pathology and disease progression in MPN are not completely elucidated. The matrix metalloproteinases (MMPs) cleave extracellular matrix, activating cytokines and growth factors that, in turn, regulate tumorigenesis and angiogenesis. The aim of this study was to evaluate the relationship of MMPs, TIMPs, HIF1-α and SPARC gene expression with angiogenic markers bFGF and VEGFA in patients with MPN considering their mutational status; as well as to assess the regulation of these genes in animal models HIF1-α and VHL knockouts. Twenty-one MF, 21 MF post-ET, 6 MF post-PV, 23 ET patients and 78 controls were enrolled. The analysis performed in peripheral blood were: serum and mRNA expression of MMP2, MMP9, TIMP1, TIMP2 and SPARC, blood count, high-sensitivity C-reactive protein determination and VEGFA and bFGF measurements in plasma. We also evaluate mutations in JAK2, MPL and CALR. The assessment of microvascular density (MVD) in bone marrow was performed in 30 patients. Patients with MFP, MFPET and ET presented higher expression of MMP2, SPARC, TIMP1, TIMP2 and bFGF compared to their controls (P <0.05), while MMP9 expression was higher in patients with MFPET and ET (P=0.011 and P=0.047, respectively). Higher expression of HIF1-α and VEGFA was found in ET patients compared to the controls (P <0.05). PMF JAK2V617F patients had higher concentrations of MMP9, TIMP2, bFGF and VEGFA compared to CALR mutated ones (P <0.05). ET patients JAK2V617F positive had higher levels of MMP2 and TIMP2 (P=0.049 and P=0.020, respectively). The JAK2V617F allele burden was not associated with MVD in the bone marrow. Bone marrow cells from mice in hypoxia condition showed higher MMP2 and TIMP1 expression compared to the control. VHL(-/-) mice exhibited increased expression of MMP2, MMP9, TIMP1, TIMP2 and VEGFA. In contrast, the HIF1-α(-/-) embryos were not considered an applicable model for this study due to MMPs role in embryogenesis/organogenesis. In view of these findings, we can conclude that increased expression of MMP2, SPARC and bFGF are associated with MPN. The JAK2V617F mutation was associated with higher concentrations of MMPs, TIMP2 VEGFA and bFGF. HIF1-α is upregulated in PV and ET and perhaps regulate the MMPs and TIMPs expression in these diseases

Aberrant Promoter Methylation at CpG Cytosines Induce the Upregulation of the E2F5 Gene in Breast Cancer / 한국유방암학회지

PURPOSE: The promoter methylation status of cell cycle regulatory genes plays a crucial role in the regulation of the eukaryotic cell cycle. CpG cytosines are actively subjected to methylation during tumorigenesis, resulting in gain/loss of function. E2F5 gene has growth repressive activities; various studies suggest its involvement in tumorigenesis. This study aims to investigate the epigenetic regulation of E2F5 in breast cancer to better understand tumor biology. METHODS: The promoter methylation status of 50 breast tumor tissues and adjacent normal control tissues was analyzed. mRNA expression was determined using SYBR® green quantitative polymerase chain reaction (PCR), and methylation-specific PCR was performed for bisulfite-modified genomic DNA using E2F5-specific primers to assess promoter methylation. Data was statistically analyzed. RESULTS: Significant (p<0.001) upregulation was observed in E2F5 expression among tumor tissues, relative to the control group. These samples were hypo-methylated at the E2F5 promoter region in the tumor tissues, compared to the control. Change in the methylation status (Δmeth) was significantly lower (p=0.022) in the tumor samples, indicating possible involvement in tumorigenesis. Patients at the postmenopausal stage showed higher methylation (75%) than those at the premenopausal stage (23.1%). Interestingly, methylation levels gradually increased from the early to the advanced stages of the disease (p<0.001), which suggests a putative role of E2F5 methylation in disease progression that can significantly modulate tumor biology at more advanced stage and at postmenopausal age (Pearson's r=0.99 and 0.86, respectively). Among tissues with different histological status, methylation frequency was higher in invasive lobular carcinoma (80.0%), followed by invasive ductal carcinoma (46.7%) and ductal carcinoma in situ (20.0%). CONCLUSION: Methylation is an important epigenetic factor that might be involved in the upregulation of E2F5 gene in tumor tissues, which can be used as a prognostic marker for breast cancer.

Shikonin Isolated from Lithospermum erythrorhizon Downregulates Proinflammatory Mediators in Lipopolysaccharide-Stimulated BV2 Microglial Cells by Suppressing Crosstalk between Reactive Oxygen Species and NF-kappaB

According to the expansion of lifespan, neuronal disorder based on inflammation has been social problem. Therefore, we isolated shikonin from Lithospermum erythrorhizon and evaluated anti-inflammatory effects of shikonin in lipopolysaccharide (LSP)-stimulated BV2 microglial cells. Shikonin dose-dependently inhibits the expression of the proinflammatory mediators, nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor-alpha (TNF-alpha) as well as their main regulatory genes and products such as inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-alpha in LPS-stimulated BV2 microglial cells. Additionally, shikonin suppressed the LPS-induced DNA-binding activity of nuclear factor-kappaB (NF-kappaB) to regulate the key regulatory genes of the proinflammatory mediators, such as iNOS, COX-2, and TNF-alpha, accompanied with downregulation of reactive oxygen species (ROS) generation. The results indicate that shikonin may downregulate the expression of proinflammatory genes involved in the synthesis of NO, PGE2, and TNF-alpha in LPS-treated BV2 microglial cells by suppressing ROS and NF-kappaB. Taken together, our results revealed that shikonin exerts downregulation of proinflammatory mediators by interference the ROS and NF-kappaB signaling pathway.

Improvement of natamycin production in an industrial strain by heterologous expression of the afsRS(cla) global regulatory genes / 生物工程学报

The afsRS(cla) global regulatory genes from Streptomyces clavuligerus activate the production of two antibiotics in Streptomyces lividans. In this study, we gained an increase of 38% in the production of natamycin (3.56 g/L) in an industrial strain Streptomyces gilvosporeus TZ1401 through the integration of pHL851 that bears the afsRS(cla) global regulatory genes into its genome. We discovered by quantitive real-time reverse transcription PCR (qRT-PCR) that the expression of 6 genes of the natamycin biosynthetic gene cluster were improved from 1.9 to 2.7 times. This suggests that afsRS(cla) improve the production of natamycin through increased transcription. This study provides a good example for applying afsRS(cla) in high yield breeding of industrial antibiotic producers.

The accessory gene regulator (agr) controls Staphylococcus aureus virulence in a murine intracranial abscesses model

Background: Intracranial abscesses are associated with high mortality. Staphylococcus aureus is one of the main pathogens that cause intracranial infection. Until now, there is no report to identify the key effectors of S. aureus during the intracranial infection. Methods: The murine intracranial abscesses model induced by S. aureus was constructed. The vital sign and survival rate of mice were observed to evaluate the infection. Histological examination was used to diagnose the pathological alterations of mouse tissues. The sensitivity of S. aureus to whole blood was evaluated by whole-blood killing assay. Results: In murine intracranial abscesses model, it was shown that the mortality caused by the accessory gene regulator (agr) locus deficient strain was significant decreased compared with its parent strain. Moreover, we found that RNAIII, the effector of agr system, was essential for the intracranial infection caused by S. aureus. In the further investigation, it was shown that restoration the expression of α-toxin in agr deficient strain could partially recover the mortality in the murine intracranial abscesses model. Conclusion: Our data suggested that the agr system of S. aureus is an important virulence determinant in the induction and mortality of intracranial abscesses in mice. .