RESUMO
INTRODUCTION: High-intensity visible light (HEV), also referred to as blue light, has a wavelength of 400-500 nm and accounts for approximately one-third of the visible light. Blue light is also emitted from electronic devices and artificial indoor lighting. Studies have shown that exposure of human skin cells to light emitted from electronic devices, even as short as 1 h, can cause an increase in reactive oxygen species (ROS), apoptosis and necrosis. Despite comprising a significant portion of the light spectrum, the effects of HEV light have not been studied as extensively. This is in part due to a lack of suitable in vitro testing methods. This work was conducted in order to develop a reproducible testing method for assessing the effects of blue light on the skin. METHODS: Testing was performed using a full thickness, 3D in vitro skin tissue model. Different exposure protocols were tested to (1) determine the biological effects of blue light on the skin and (2) to identify an appropriate exposure for routine testing of cosmetic materials that may protect the skin from blue light damage. Gene expression and protein biomarkers were measured using qPCR, ELISA and immunohistochemical (IHC) methods. RESULTS: Our work demonstrates that daily exposure to blue light produced dose-and-time-dependent changes in biomarkers associated with skin damage. Exposure to blue light for 6 h for 5 consecutive days (total intensity of 30 J/cm2 ) increased the expression of genes that regulate inflammation and oxidative stress pathways and decreased the expression of genes that maintain skin barrier and tissue integrity. Exposure to blue light significantly increased protein biomarkers associated with ageing, inflammation and tissue damage. IHC staining confirmed changes in collagen, filaggrin and NQO1 protein expression. Treatment with ascorbic acid inhibited the effects of blue light, demonstrating a role in protection from blue light. CONCLUSION: Our results showed that consistent blue light exposure produced skin damage via alterations in biological pathways that are associated with skin ageing. This work provides a new, reproducible in vitro testing method for assessing the effects of blue light on human skin using gene expression, protein ELISA and IHC staining.
INTRODUCTION: La lumière visible à haute énergie (VHE), également appelée lumière bleue, a une longueur d'onde de 400 à 500 nm et représente environ un tiers de la lumière visible. La lumière bleue est également émise par les appareils électroniques et l'éclairage intérieur artificiel. Des études ont montré que l'exposition des cellules cutanées humaines à la lumière émise par les appareils électroniques, même pour une période de seulement 1 h, peut entraîner une augmentation des dérivés réactifs de l'oxygène (DRO), de l'apoptose et de la nécrose. Bien qu'ils représentent une partie importante du spectre lumineux, les effets de la lumière VHE n'ont pas été étudiés aussi largement. Cela est en partie dû à un manque de méthodes de test in vitro appropriées. Ces travaux ont été réalisé afin de développer une méthode de test reproductible pour évaluer les effets de la lumière bleue sur la peau. MÉTHODES: Les tests ont été réalisés à l'aide d'un modèle de tissu cutané 3D in vitro de pleine épaisseur. Différents protocoles d'exposition ont été testés pour (1) déterminer les effets biologiques de la lumière bleue sur la peau et (2) identifier une exposition appropriée pour les tests de routine des produits cosmétiques susceptibles de protéger la peau des dommages causés par la lumière bleue. L'expression génique et les biomarqueurs protéiques ont été mesurés à l'aide des méthodes de PCR quantitative, de dosage par la méthode immuno-enzymatique ELISA et immunohistochimiques (IHC). RÉSULTATS: Nos travaux démontrent que l'exposition quotidienne à la lumière bleue a produit des modifications dépendantes de la dose et du temps dans les biomarqueurs associés aux lésions cutanées. L'exposition à la lumière bleue pendant 6 h au cours de 5 jours consécutifs (intensité totale de 30 J/cm2) a augmenté l'expression des gènes qui régulent l'inflammation et les voies du stress oxydatif, et a diminué l'expression des gènes qui maintiennent la barrière cutanée et l'intégrité tissulaire. L'exposition à la lumière bleue a significativement augmenté les biomarqueurs protéiques associés au vieillissement, à l'inflammation et aux lésions tissulaires. La coloration par IHC a confirmé les modifications de l'expression du collagène, de la filaggrine et de la protéine NQO1. Le traitement par acide ascorbique a inhibé les effets de la lumière bleue, démontrant un rôle dans la protection contre la lumière bleue. CONCLUSION: Nos résultats ont montré qu'une exposition continue à la lumière bleue produisait des lésions cutanées par le biais d'altérations des voies biologiques associées au vieillissement de la peau. Ces travaux fournissent une nouvelle méthode de test in vitro reproductible pour évaluer les effets de la lumière bleue sur la peau humaine à l'aide de l'expression des gènes, du test ELISA de détection de protéines et de la coloration IHC.
Assuntos
Luz , Pele , Humanos , Espécies Reativas de Oxigênio/metabolismo , Apoptose , Biomarcadores/metabolismoRESUMO
Genomic reorganization, such as rearrangements and inversions, influences how genetic information is organized within the bacterial genomes. Inversions, in particular, facilitate genome evolution through gene gain and loss, and can alter gene expression. Previous studies have investigated the impact inversions have on gene expression induced inversions targeting specific genes or examine inversions between distantly related species. This fails to encompass a genome-wide perspective of naturally occurring inversions and their post-adaptation impact on gene expression. Here, we used bioinformatic techniques and multiple RNA-seq datasets to investigate the short- and long-range impact inversions have on genomic gene expression within Escherichia coli. We observed differences in gene expression between homologous inverted and non-inverted genes even after long-term exposure to adaptive selection. In 4% of inversions representing 33 genes, differential gene expression between inverted and non-inverted homologs was detected, with greater than two-thirds (71%) of differentially expressed inverted genes having 9.4-85.6-fold higher gene expression. The identified inversions had more overlap than expected with nucleoid-associated protein binding sites, which assist in the regulation of genomic gene expression. Some inversions can drastically impact gene expression, even between different strains of E. coli, and could provide a mechanism for the diversification of genetic content through controlled expression changes.
Assuntos
Inversão Cromossômica , Escherichia coli , Escherichia coli/genética , Expressão Gênica , Genoma Bacteriano , Genômica , Humanos , Ligação ProteicaRESUMO
Despite several studies on genetic markers and differentially expressed genes related to ribeye area (REA) and tenderness traits in beef cattle, there is divergence in the results regarding the genes associated with these traits. Thirteen genes associated with or exhibiting biological functions that might influence such phenotypes were included in this study. A total of five genes for REA (IGF-1, IGF-2, MSTN, NEDD4, and UBE4A) and eight genes for meat tenderness (CAPN1, CAPN2, CAST, HSPB1, DNAJA1, FABP4, SCD, and PRKAG3) were selected from previous studies on beef cattle. Genes and their respective proteins expression were validated in a commercial population of Nellore cattle using quantitative real-time PCR (RT-qPCR) and advanced mass spectrometry (LC/MS-MS) techniques, respectively. The MSTN gene was upregulated in animals with low REA. The CAPN1, CAPN2, CAST, HSPB1, and DNAJA1 genes were upregulated in animals with tough meat. The proteins translated by these genes were not differentially expressed. Our results confirm the potential of some of the studied genes as biomarkers for carcass and meat quality traits in Nellore cattle.
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Carne , Carne Vermelha , Animais , Bovinos/genética , Marcadores Genéticos , Carne/análise , Fenótipo , ProteômicaRESUMO
The effect of oxaprozin (OXP) on an experimental model of seizures in rats was investigated in this study. Seizures in Wistar rats (200-250 g) were induced by pentylenetetrazole (PTZ, 60 mg/kg). The anticonvulsant effect of OXP (100, 200, and 400 mg/kg, intraperitoneally) was evaluated in a seizure model. After behavioral tests, the animals underwent deep anesthesia and were put down painlessly. Animal serum was isolated for antioxidant assays (nitric oxide (NO) and glutathione (GSH)). The animals' brains were also isolated to gauge the relative expression of genes in the oxidative stress pathway (sirtuin 1 (Sirt1) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (Pgc1α)). Intraperitoneal injection of OXP increased the mean latency of myoclonic jerks and generalized tonic-clonic seizure (GTCS) and decreased the number of myoclonic jerks and GTCS duration compared with the PTZ group. Biochemical tests showed that pretreatment with OXP was able to restore GSH serum levels and reverse the augmented NO serum levels caused by PTZ induction to the normal level. The quantitative polymerase chain reaction results also revealed that OXP counteracts the negative effects of PTZ by affecting the expression of the Sirt1 and Pgc1α genes. Overall, this study suggests the potential neuroprotective effects of the nonsteroidal, anti-inflammatory OXP drug in a model of neural impairment caused by seizures via the mechanism of inhibition of the oxidative stress pathway.
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Anticonvulsivantes , Mioclonia , Oxaprozina , Convulsões , Animais , Anticonvulsivantes/uso terapêutico , Modelos Animais de Doenças , Glutationa/metabolismo , Mioclonia/tratamento farmacológico , Oxaprozina/uso terapêutico , Estresse Oxidativo , Pentilenotetrazol/efeitos adversos , Coativador 1-alfa do Receptor gama Ativado por Proliferador de Peroxissomo/metabolismo , Ratos , Ratos Wistar , Convulsões/induzido quimicamente , Convulsões/tratamento farmacológico , Convulsões/metabolismo , Sirtuína 1/metabolismoRESUMO
Topical steroids (TS) have been widely prescribed since the 1950s. This study investigated for the first time the transgenerational effects of TS on the antioxidant mechanism of the hypothalamus-pituitary-adrenal (HPA) axis, both in prenatal and infancy. Three generations (F1, F2, and F3) and prenatal group (P) were investigated in both sexes with two different time points; P45th and P75th day were accepted as puberty and early adulthood, respectively. Clobetasol propionate 0.05% was used as TS. Quantitative real-time PCR was performed to expressional analyses of Sod1, Sod2, and Sod3 genes in the HPA tissues. The Sod mRNA expression of the HPA belonging to P and F1 groups revealed similar results in both genders. The downregulation in the adrenal Sod level was determined in P and F1, F2, and F3 generations in both genders, especially in females (p < 0.05). The Sod activities in the pituitary of all groups were downregulated in female rats (p < 0.05). Interestingly, in male rats, Sod2 and Sod3 were not expressed in the pituitary compared with the control on the day P45, while Sod2 and Sod3 expressions were determined in all the groups on day P75. Sod1 overexpression was found in pituitary and hypothalamus of males in the F3 generation. This study showed that TS applied in infancy had a transgenerational adverse effect on antioxidant defense mechanisms, especially in the adrenal gland.
Assuntos
Antioxidantes , Maturidade Sexual , Animais , Antioxidantes/metabolismo , Feminino , Sistema Hipotálamo-Hipofisário/metabolismo , Hipotálamo , Masculino , Sistema Hipófise-Suprarrenal/metabolismo , Gravidez , Ratos , Esteroides/metabolismo , Esteroides/farmacologia , Superóxido Dismutase-1/metabolismo , Superóxido Dismutase-1/farmacologiaRESUMO
It has long been acknowledged that changes in the regulation of gene expression may account for major organismal differences. However, we still do not fully understand how changes in gene expression evolve and how do such changes influence organisms' differences. We are even less aware of the impact such changes might have in restricting gene flow between species. Here, we focus on studies of gene expression and speciation in the Drosophila model. We review studies that have identified gene interactions in post-mating reproductive isolation and speciation, particularly those that modulate male gene expression. We also address studies that have experimentally manipulated changes in gene expression to test their effect in post-mating reproductive isolation. We highlight the need for a more in-depth analysis of the role of selection causing disrupted gene expression of such candidate genes in sterile/inviable hybrids. Moreover, we discuss the relevance to incorporate more routinely assays that simultaneously evaluate the potential effects of environmental factors and genetic background in modulating plastic responses in male genes and their potential role in speciation.
Assuntos
Drosophila , Expressão Gênica , Especiação Genética , Animais , Drosophila/genética , Fluxo Gênico , Masculino , Isolamento ReprodutivoRESUMO
Chloroplast ribonucleoproteins (cpRNPs) are implicated in splicing, editing, and stability control of chloroplast RNAs as well as in regulating development and stress tolerance. To facilitate a comprehensive understanding of their functions, we carried out a genome-wide identification, curation, and phylogenetic analysis of cpRNP genes in Oryza sativa (rice) and Arabidopsis thaliana (Arabidopsis). Ten cpRNP genes were identified in each of Arabidopsis and rice genomes based on the presence of two RRM (RNA-recognition motif) domains and an N-terminal chloroplast targeting signal peptide in the predicted proteins. These proteins are localized to chloroplasts. Gene expression analysis revealed that cpRNP genes have differential tissue expression patterns and some cpRNP genes are induced by abiotic stresses such as cold, heat, and drought. Taken together, our study provides a comprehensive annotation of the cpRNP gene family and their expression patterns in Arabidopsis and rice which will facilitate further studies on their roles in plant growth and stress responses.
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Arabidopsis/genética , Cloroplastos/genética , Cloroplastos/metabolismo , Regulação da Expressão Gênica de Plantas , Oryza/genética , Ribonucleoproteínas/genética , Ribonucleoproteínas/metabolismo , Proteínas de Cloroplastos/genética , Proteínas de Cloroplastos/metabolismo , Secas , Genes de Plantas/genética , Genoma de Planta , Filogenia , Proteínas de Plantas/genética , Splicing de RNA , Ribonucleoproteínas/classificação , Estresse Fisiológico/genéticaRESUMO
WUSCHEL-related homeobox (WOX) proteins are plant-specific transcription factors that are profoundly involved in regulation of plant development and stress responses. In this study, we totally identified 11 WOX transcription factor family members in cucumber (Cucumis sativus, CsWOX) genome and classified them into three clades with nine subclades based on phylogenetic analysis results. Alignment of amino acid sequences revealed that all WOX members in cucumber contained the typical homeodomain, which consists of 60-66 amino acids and is folded into a helix-turn-helix structure. Gene duplication event analysis indicated that CsWOX1a and CsWOX1b were a segment duplication pair, which might affect the number of WOX members in cucumber genome. The expression profiles of CsWOX genes in different tissues demonstrated that the members sorted into the ancient clade (CsWOX13a and CsWOX13b) were constitutively expressed at higher levels in comparison to the others. Cis-element analysis in promoter regions suggested that the expression of CsWOX genes was associated with phytohormone pathways and stress responses, which was further supported by RNA-seq data. Taken together, our results provide new insights into the evolution of cucumber WOX genes and improve our understanding about the biological functions of the CsWOX gene family.
Assuntos
Cucumis sativus , Genes de Plantas , Família Multigênica , Fatores de Transcrição , Cucumis sativus/genética , Regulação da Expressão Gênica de Plantas , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
In the absence of a vaccine, the treatment of SARS-CoV2 has focused on eliminating the virus with antivirals or mitigating the cytokine storm syndrome (CSS) that leads to the most common cause of death: respiratory failure. Herein we discuss the mechanisms of antiviral treatments for SARS-CoV2 and treatment strategies for the CSS. Antivirals that have shown in vitro activity against SARS-CoV2, or the closely related SARS-CoV1 and MERS-CoV, are compared on the enzymatic level and by potency in cells. For treatment of the CSS, we discuss medications that reduce the effects or expression of cytokines involved in the CSS with an emphasis on those that reduce IL-6 because of its central role in the development of the CSS. We show that some of the medications covered influence the activity or expression of enzymes involved in epigenetic processes and specifically those that add or remove modifications to histones or DNA. Where available, the latest clinical data showing the efficacy of the medications is presented. With respect to their mechanisms, we explain why some medications are successful, why others have failed, and why some untested medications may yet prove useful.
Assuntos
Antivirais/uso terapêutico , Tratamento Farmacológico da COVID-19 , Síndrome da Liberação de Citocina/tratamento farmacológico , Síndrome da Liberação de Citocina/virologia , Citocinas , Epigênese Genética , Expressão Gênica , Humanos , Interleucina-6 , SARS-CoV-2/efeitos dos fármacosRESUMO
The tall wheatgrass species Thinopyrum elongatum carries on the long arm of chromosome 7E, a locus that contributes strongly to resistance to fusarium head blight (FHB), a devastating fungal disease affecting wheat crops in all temperate areas of the world. Introgression of Th. elongatum 7E chromatin into chromosome 7D of wheat was induced by the ph1b mutant of CS. Recombinants between chromosome 7E and wheat chromosome 7D, induced by the ph1b mutation, were monitored by a combination of molecular markers and phenotyping for FHB resistance. Progeny of up to five subsequent generations derived from two lineages, 64-8 and 32-5, were phenotyped for FHB symptoms and genotyped using published and novel 7D- and 7E-specific markers. Fragments from the distal end of 7EL, still carrying FHB resistance and estimated to be less than 114 and 66 Mbp, were identified as introgressed into wheat chromosome arm 7DL of progeny derived from 64-8 and 32-5, respectively. Gene expression analysis revealed variation in the expression levels of genes from the distal ends of 7EL and 7DL in the introgressed progeny. The 7EL introgressed material will facilitate the use of the 7EL FHB resistance locus in wheat breeding programs.
Assuntos
Resistência à Doença/genética , Fusarium , Doenças das Plantas/genética , Poaceae , Triticum , Mapeamento Cromossômico , Cromossomos de Plantas/genética , DNA de Plantas , Fusarium/patogenicidade , Expressão Gênica , Marcadores Genéticos , Melhoramento Vegetal , Doenças das Plantas/microbiologia , Poaceae/genética , Triticum/genéticaRESUMO
We previously demonstrated that genome reorganization, through chromosome territory repositioning, occurs concurrently with significant changes in gene expression in normal primary human fibroblasts treated with the drug rapamycin, or stimulated into quiescence. Although these events occurred concomitantly, it is unclear how specific changes in gene expression relate to reorganization of the genome at higher resolution. We used computational analyses, genome organization assays, and microscopy, to investigate the relationship between chromosome territory positioning and gene expression. We determined that despite relocation of chromosome territories, there was no substantial bias in the proportion of genes changing expression on any one chromosome, including chromosomes 10 and 18. Computational analyses identified that clusters of serum deprivation and rapamycin-responsive genes along the linear extent of chromosomes. Chromosome conformation capture (3C) analysis demonstrated the strengthening or loss of specific long-range chromatin interactions in response to rapamycin and quiescence induction, including a cluster of genes containing Interleukin-8 and several chemokine genes on chromosome 4. We further observed that the LIF gene, which is highly induced upon rapamycin treatment, strengthened interactions with up- and down-stream intergenic regions. Our findings indicate that the repositioning of chromosome territories in response to cell stimuli, this does not reflect gene expression changes occurring within physically clustered groups of genes.
Assuntos
Cromatina/química , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Soro/metabolismo , Sirolimo/farmacologia , Núcleo Celular/genética , Proliferação de Células , Coloração Cromossômica , Cromossomos Artificiais Bacterianos , Cromossomos Humanos Par 10 , Cromossomos Humanos Par 18 , Análise por Conglomerados , Biologia Computacional , Perfilação da Expressão Gênica , Biblioteca Gênica , Genoma Humano , Humanos , Hibridização in Situ Fluorescente , Interleucina-8/metabolismo , Família MultigênicaRESUMO
Lysophosphatidic acid (LPA) is an important lipid molecule for signal transduction in cell proliferation. Although the effects of LPA on vascular smooth muscle (VSM) cell growth have been reported previously, the underlying mechanisms of its action are not fully understood. The present study was undertaken to investigate the effects of some inhibitors of different protein kinases and other molecular targets on LPA-induced DNA synthesis as well as gene expression in the aortic VSM cells. The DNA synthesis was studied by the [3H]thymidine incorporation method and the gene expression was investigated by the real-time PCR technique. It was observed that the LPA-induced DNA synthesis was attenuated by inhibitors of protein kinase C (PKC) (staurosporine, calphostin C, and bisindolylmaleimide), phosphoinositide 3-kinase (PI3K) (wortmannin and LY294002), and ribosomal p70S6 kinase (p70S6K) (rapamycin). The inhibitors of guanine protein coupled receptors (GPCR) (pertussis toxin), phospholipase C (PLC) (U73122 and D609), and sodium-hydrogen exchanger (NHE) (amiloride and dimethyl amiloride) were also shown to depress the LPA-induced DNA synthesis. Furthermore, gene expressions for PLC ß1 isoform, PKC δ and ε isoforms, casein kinase II ß isoform, and endothelin-1A receptors were elevated by LPA. These results suggest that the LPA-induced proliferation of VSM cells is mediated through the activation of GPCR and multiple protein kinases as well as gene expressions of some of their specific isoforms.
Assuntos
Lisofosfolipídeos/farmacologia , Músculo Liso Vascular/citologia , Animais , Caseína Quinase II/genética , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Endotelina-1/genética , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Proteína Quinase C/genética , RNA Mensageiro/genética , Ratos , Receptores de Ácidos Lisofosfatídicos/metabolismo , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Fosfolipases Tipo C/genéticaRESUMO
Candidiasis caused by multidrug-resistant Candida species continues to be difficult to eradicate. The use of live probiotic bacteria has gained a lot of interest in the treatment of candidiasis; however, whole-cell probiotic use can often be associated with a high risk of sepsis. Strategies manipulating cell-free methods using probiotic strains could lead to the development of novel antifungal solutions. Therefore, we evaluated the effect of three probiotic cell-free extracts (CFEs) on the growth, virulence traits, and drug efflux pumps in C. albicans. On the basis of its minimum inhibitory concentration, Lactobacillus rhamnosus was selected and assessed against various virulence traits and drug resistance mechanisms. The results showed that L. rhamnosus CFE significantly inhibited hyphae formation and reduced secretion of proteinases and phospholipases. Moreover, L. rhamnosus inhibited the drug efflux proteins in resistant C. albicans strains thus reversing drug resistance. Gene expression data confirmed downregulation of genes associated with microbial virulence and drug resistance following treatment of C. albicans with L. rhamnosus CFE. Through gas chromatography - mass spectrometry chemical characterization, high contents of oleic acid (24.82%) and myristic acid (13.11%) were observed in this CFE. Collectively, our findings indicate that L. rhamnosus may potentially be used for therapeutic purposes to inhibit C. albicans infections.
Assuntos
Antibiose/fisiologia , Candida albicans/efeitos dos fármacos , Candidíase/microbiologia , Farmacorresistência Fúngica/fisiologia , Lacticaseibacillus rhamnosus/fisiologia , Probióticos , Antifúngicos/farmacologia , Biofilmes , Candida albicans/patogenicidade , Humanos , Lacticaseibacillus rhamnosus/química , Virulência/efeitos dos fármacosRESUMO
The hormone auxin is involved in many biological processes throughout a plant's lifecycle. However, genes in the GH3 (Gretchen Hagen3) family, one of the three major auxin-responsive gene families, have not yet been identified in oilseed rape (Brassica napus). In this study, we identified 63 BnaGH3 genes in oilseed rape using homology searches. We analyzed the chromosome locations, gene structures, and phylogenetic relationships of the BnaGH3 genes, as well as the cis-elements in their promoters. Most BnaGH3 genes are located on chromosomes A03, A09, C02, C03, and C09, each with 4-7 members. In addition, we analyzed the expression patterns of BnaGH3 genes in seven tissues by transcriptome sequencing and quantitative RT-PCR analysis of plants under exogenous IAA treatment. The BnaGH3 genes showed different expression patterns in various tissues. BnaA.GH3.2-1 and BnaC.GH3.2-1 were expressed in the seed and seed coat during development and in response to IAA treatment. These results shed light on the possible roles of the GH3 gene family in oilseed rape.
Assuntos
Brassica napus/genética , Genoma de Planta , Mapeamento Cromossômico , Cromossomos de Plantas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ácidos Indolacéticos/farmacologia , Família Multigênica , Sequências Reguladoras de Ácido NucleicoRESUMO
Heat shock transcription factors (Hsfs) are important regulators of biotic and abiotic stress responses in plants. Currently, the Hsf gene family is not well understood in cassava, an important tropical crop. In the present study, 32 MeHsf genes were identified from the cassava genome database, which were divided into three types based on functional domain and motif distribution analyses. Analysis of the differential expression of the genes belonging to the Hsf family in cassava was carried out based on published cassava transcriptome data from tissues/organs (leaf blade, leaf midvein, lateral buds, organized embryogenic structures, friable embryogenic callus, fibrous roots, storage roots, stem, petiole, shoot apical meristem, and root apical meristem) under abiotic stress (cold, drought) or biotic stress (mealybugs. cassava brown streak disease, cassava bacterial blight). The results show the expression diversity of cassava Hsfs genes in various tissues/organs. The transcript levels of MeHsfB3a, MeHsfA6a, MeHsfA2a, and MeHsfA9b were upregulated by abiotic and biotic stresses, such as cold, drought, cassava bacterial blight, cassava brown streak disease, and mealybugs, indicating their potential roles in mediating the response of cassava plants to environment stresses. Further interaction network and co-expression analyses suggests that Hsf genes may interact with Hsp70 family members to resist environmental stresses in cassava. These results provide valuable information for future studies of the functional characterization of the MeHsf gene family.
Assuntos
Resposta ao Choque Frio , Proteínas de Choque Térmico/genética , Manihot/genética , Proteínas de Plantas/genética , Transcriptoma , Secas , Regulação da Expressão Gênica de Plantas , Proteínas de Choque Térmico/metabolismo , Manihot/microbiologia , Manihot/parasitologia , Proteínas de Plantas/metabolismoRESUMO
Idiopathic pulmonary arterial hypertension (IPAH) is a rare and devastating condition. There is no known cure for IPAH, and current treatment options are not always effective. Autologous myeloid angiogenic cells (MACs) have been explored as a novel therapy for IPAH, but preliminary data from clinical trials show limited beneficial effects. A complete understanding of IPAH MAC function remains elusive. This study was designed to comprehensively compare cell function between IPAH MACs and healthy control MACs. MACs were procured through the culture of peripheral blood mononuclear cells in endothelial selective medium for 7 days. Compared with healthy MACs, IPAH MACs exhibited (1) significantly lower levels of endothelial markers as shown by fluorescence microscopy; (2) a markedly higher rate of apoptosis under both normal culture condition and serum starvation as shown by the TUNEL assay; (3) significantly decreased migration towards vascular endothelial growth factor as shown by a modified Boyden chamber migration assay; and (4) similar vascular endothelial growth factor and endothelial nitric oxide synthase mRNA levels as shown by reverse transcription quantitative PCR. In conclusion, various aspects of IPAH MAC function are impaired. To achieve greater therapeutic benefits, pharmacologic and (or) genetic manipulations to improve IPAH MAC function, particularly to promote cell survival and migration, are warranted.
Assuntos
Apoptose , Movimento Celular , Células Endoteliais/metabolismo , Hipertensão Pulmonar Primária Familiar/patologia , Regulação da Expressão Gênica , Células Mieloides/patologia , Neovascularização Fisiológica , Adulto , Biomarcadores/metabolismo , Estudos de Casos e Controles , Proliferação de Células , Hipertensão Pulmonar Primária Familiar/metabolismo , Hipertensão Pulmonar Primária Familiar/fisiopatologia , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The aims of this study were to evaluate the presence of genes associated with adhesion (cadF), invasion (ciaB), and cytotoxin production (cdtA, cdtB, and cdtC) among Campylobacter jejuni isolates from a poultry slaughterhouse and to investigate the effect of different temperatures on the expression of these virulence-associated genes. A total of 88 C. jejuni isolates from cecum, liver, chicken carcasses, chilled water, and scalding water were submitted to PCR assay for detection of virulence genes. Representative isolates were selected for gene expression evaluation at 37 and 42 °C, according to their virulence gene profile and genotypic typing. All C. jejuni isolates carried the five virulence-associated genes, which play an important role in the infectious process. Differential gene expression by RT-qPCR was observed among C. jejuni isolates at 37 and 42 °C. The expression levels at 37 °C showed upregulation of the ciaB, cdtA, cdtB, and cdtC genes in five isolates, with the exception of ciaB for isolate 4. At 42 °C, upregulation was observed for ciaB and cdtC, cdtA and cdtB, and cadF in four, three, and two isolates, respectively. The C. jejuni isolates expressed the virulence genes evaluated, and the expression is gene- and isolate-dependent and varied according the temperature.
Assuntos
Antígenos de Bactérias/genética , Aderência Bacteriana/genética , Proteínas da Membrana Bacteriana Externa/genética , Toxinas Bacterianas/genética , Campylobacter jejuni/genética , Proteínas de Transporte/genética , Regulação Bacteriana da Expressão Gênica/fisiologia , Virulência/genética , Animais , Infecções por Campylobacter/microbiologia , Infecções por Campylobacter/veterinária , Campylobacter jejuni/isolamento & purificação , Campylobacter jejuni/patogenicidade , Galinhas , DNA Bacteriano/genética , Genes Bacterianos , Reação em Cadeia da Polimerase , Doenças das Aves Domésticas/microbiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Temperatura , Fatores de VirulênciaRESUMO
MicroRNAs (miRNAs) are small non-coding RNAs that play important roles in a variety of biological processes. Dysregulation of miRNAs is tightly associated with the malignancy of cancers. Aberrant expression of miR-378 has been observed in human cancers; however, the function of miR-378 in osteosarcoma (OS) remains largely unknown. Here, we showed that miR-378 was highly expressed in human OS tissues and cell lines. Overexpression of miR-378 significantly promoted the cell proliferation of OS cells. Molecular studies identified Kruppel-like factor-9 (KLF9) as a functional downstream target of miR-378. MiR-378 directly bound to the mRNA 3'-UTR region of KLF9 and suppressed the expression of KLF9. Highly expressed KLF9 reversed the promoting effect of miR-378 on the proliferation of OS cells. The expression level of miR-378 was negatively correlated with that of KLF9 in OS tissues. Collectively, our results demonstrated the molecular interaction between miR-378 and KLF9, indicating the therapeutic potential of miR-378 for OS.
Assuntos
Neoplasias Ósseas/metabolismo , Proliferação de Células , Regulação para Baixo , Regulação Neoplásica da Expressão Gênica , Fatores de Transcrição Kruppel-Like/biossíntese , MicroRNAs/metabolismo , Proteínas de Neoplasias/biossíntese , Osteossarcoma/metabolismo , RNA Neoplásico/metabolismo , Neoplasias Ósseas/genética , Neoplasias Ósseas/patologia , Linhagem Celular Tumoral , Humanos , Fatores de Transcrição Kruppel-Like/genética , MicroRNAs/genética , Proteínas de Neoplasias/genética , Osteossarcoma/genética , Osteossarcoma/patologia , RNA Neoplásico/genéticaRESUMO
The diversity of mating systems among animals is astounding. Importantly, similar mating systems have evolved even across distantly related taxa. However, our understanding of the mechanisms underlying these convergently evolved phenotypes is limited. Here, we examine on a genomic scale the neuromolecular basis of social organization in cichlids of the tribe Ectodini from Lake Tanganyika. Using field-collected males and females of four closely related species representing two independent evolutionary transitions from polygyny to monogamy, we take a comparative transcriptomic approach to test the hypothesis that these independent transitions have recruited similar gene sets. Our results demonstrate that while lineage and species exert a strong influence on neural gene expression profiles, social phenotype can also drive gene expression evolution. Specifically, 331 genes (â¼6% of those assayed) were associated with monogamous mating systems independent of species or sex. Among these genes, we find a strong bias (4:1 ratio) toward genes with increased expression in monogamous individuals. A highly conserved nonapeptide system known to be involved in the regulation of social behavior across animals was not associated with mating system in our analysis. Overall, our findings suggest deep molecular homologies underlying the convergent or parallel evolution of monogamy in different cichlid lineages of Ectodini.
Assuntos
Ciclídeos/genética , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Reprodução/genética , Transcriptoma , Animais , Ciclídeos/classificação , Feminino , Genômica/métodos , Lagos , Masculino , Filogenia , Especificidade da Espécie , TanzâniaRESUMO
SQUAMOSA promoter-binding protein-like (SPL) is a class of plant-specific transcription factors that play critical roles in regulating plant growth and development. However, little systematic research on SPL genes has been conducted in strawberry. In this study, 14 SPL genes were identified in the genome of woodland strawberry (Fragaria vesca), one of the model plants of the family Rosaceae. Chromosome localization analysis indicated that the 14 FvSPL genes were unevenly distributed on six chromosomes. Phylogenetic analysis indicated that the FvSPL proteins could be clustered into six groups (G1 to G6). Genes with similar structure were classified into the same group, implying their functional redundancy. In addition, nine out of the 14 FvSPL genes, belonging to G1, G2, and G5, were found to be the putative targets of FvmiR156 genes. Expression analysis indicated FvSPL genes exhibited highly diverse expression patterns in the tissues and organs examined. The transcript levels of most FvmiR156-targeted FvSPL genes in fruit were lower than those non-miR156-targeted genes. In addition, the expression of the FvmiR156-targeted FvSPL genes decreased during fruit ripening, whereas the expression of FvmiR156 genes increased in fruit during this process. The results provide a foundation for future functional analysis of FvSPL genes in strawberry growth and development.