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1.
Nucleic Acids Res ; 50(20): 11470-11491, 2022 11 11.
Artigo em Inglês | MEDLINE | ID: mdl-36259644

RESUMO

Nonsense-mediated RNA decay (NMD) is a highly conserved and selective RNA turnover pathway that depends on the endonuclease SMG6. Here, we show that SMG6 is essential for male germ cell differentiation in mice. Germ-cell conditional knockout (cKO) of Smg6 induces extensive transcriptome misregulation, including a failure to eliminate meiotically expressed transcripts in early haploid cells, and accumulation of NMD target mRNAs with long 3' untranslated regions (UTRs). Loss of SMG6 in the male germline results in complete arrest of spermatogenesis at the early haploid cell stage. We find that SMG6 is strikingly enriched in the chromatoid body (CB), a specialized cytoplasmic granule in male germ cells also harboring PIWI-interacting RNAs (piRNAs) and the piRNA-binding protein PIWIL1. This raises the possibility that SMG6 and the piRNA pathway function together, which is supported by several findings, including that Piwil1-KO mice phenocopy Smg6-cKO mice and that SMG6 and PIWIL1 co-regulate many genes in round spermatids. Together, our results demonstrate that SMG6 is an essential regulator of the male germline transcriptome, and highlight the CB as a molecular platform coordinating RNA regulatory pathways to control sperm production and fertility.


Assuntos
Endorribonucleases , Grânulos de Ribonucleoproteínas de Células Germinativas , Espermatogênese , Transcriptoma , Animais , Masculino , Camundongos , Células Germinativas/metabolismo , RNA Interferente Pequeno/genética , Espermátides/metabolismo , Espermatogênese/genética , Endorribonucleases/metabolismo
2.
Nucleic Acids Res ; 46(D1): D246-D251, 2018 01 04.
Artigo em Inglês | MEDLINE | ID: mdl-29165655

RESUMO

Expression Atlas (http://www.ebi.ac.uk/gxa) is an added value database that provides information about gene and protein expression in different species and contexts, such as tissue, developmental stage, disease or cell type. The available public and controlled access data sets from different sources are curated and re-analysed using standardized, open source pipelines and made available for queries, download and visualization. As of August 2017, Expression Atlas holds data from 3,126 studies across 33 different species, including 731 from plants. Data from large-scale RNA sequencing studies including Blueprint, PCAWG, ENCODE, GTEx and HipSci can be visualized next to each other. In Expression Atlas, users can query genes or gene-sets of interest and explore their expression across or within species, tissues, developmental stages in a constitutive or differential context, representing the effects of diseases, conditions or experimental interventions. All processed data matrices are available for direct download in tab-delimited format or as R-data. In addition to the web interface, data sets can now be searched and downloaded through the Expression Atlas R package. Novel features and visualizations include the on-the-fly analysis of gene set overlaps and the option to view gene co-expression in experiments investigating constitutive gene expression across tissues or other conditions.


Assuntos
Bases de Dados Genéticas , Animais , Perfilação da Expressão Gênica , Humanos , Mamíferos/genética , Mamíferos/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Plantas/genética , Plantas/metabolismo , Proteômica , Análise de Sequência de RNA , Especificidade da Espécie , Interface Usuário-Computador
3.
Nucleic Acids Res ; 44(D1): D746-52, 2016 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-26481351

RESUMO

Expression Atlas (http://www.ebi.ac.uk/gxa) provides information about gene and protein expression in animal and plant samples of different cell types, organism parts, developmental stages, diseases and other conditions. It consists of selected microarray and RNA-sequencing studies from ArrayExpress, which have been manually curated, annotated with ontology terms, checked for high quality and processed using standardised analysis methods. Since the last update, Atlas has grown seven-fold (1572 studies as of August 2015), and incorporates baseline expression profiles of tissues from Human Protein Atlas, GTEx and FANTOM5, and of cancer cell lines from ENCODE, CCLE and Genentech projects. Plant studies constitute a quarter of Atlas data. For genes of interest, the user can view baseline expression in tissues, and differential expression for biologically meaningful pairwise comparisons-estimated using consistent methodology across all of Atlas. Our first proteomics study in human tissues is now displayed alongside transcriptomics data in the same tissues. Novel analyses and visualisations include: 'enrichment' in each differential comparison of GO terms, Reactome, Plant Reactome pathways and InterPro domains; hierarchical clustering (by baseline expression) of most variable genes and experimental conditions; and, for a given gene-condition, distribution of baseline expression across biological replicates.


Assuntos
Bases de Dados Genéticas , Perfilação da Expressão Gênica , Plantas/metabolismo , Proteínas/metabolismo , Proteômica , Animais , Linhagem Celular Tumoral , Humanos , Plantas/genética , Interface Usuário-Computador
4.
Mol Microbiol ; 99(1): 43-54, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26354085

RESUMO

In eubacteria, replacement of one σ factor in the RNA polymerase (RNAP) holoenzyme by another one changes the transcription pattern. Cyanobacteria are eubacteria characterized by oxygenic photosynthesis, and they typically encode numerous group 2 σ factors that closely resemble the essential primary σ factor. A mutant strain of the model cyanobacterium Synechocystis sp. PCC 6803 without functional group 2 σ factors (named as ΔsigBCDE) could not acclimate to heat, high salt or bright light stress, but in standard conditions ΔsigBCDE grew only 9% slower than the control strain. One-fifth of the genes in ΔsigBCDE was differently expressed compared with the control strain in standard growth conditions and several physiological changes in photosynthesis, and pigment and lipid compositions were detected. To directly analyze the σ factor content of RNAP holoenzyme in vivo, a His-tag was added to the γ subunit of RNAP in Synechocystis and RNAPs were collected. The results revealed that all group 2 σ factors were recruited by RNAP in standard conditions, but recruitment of SigB and SigC increased in heat stress, SigD in bright light, SigE in darkness and SigB, SigC and SigE in high salt, explaining the poor acclimation of ΔsigBCDE to these stress conditions.


Assuntos
Regulação Bacteriana da Expressão Gênica , Fator sigma/metabolismo , Synechocystis/genética , Synechocystis/fisiologia , RNA Polimerases Dirigidas por DNA/metabolismo , Perfilação da Expressão Gênica , Temperatura Alta , Luz , Metabolismo dos Lipídeos , Fotossíntese , Pigmentos Biológicos/metabolismo , Estresse Fisiológico , Synechocystis/crescimento & desenvolvimento , Synechocystis/efeitos da radiação
5.
Histochem Cell Biol ; 148(5): 545-555, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28712031

RESUMO

Early responses of stress-sensing proteins, muscle LIM protein (MLP), ankyrin repeat proteins (Ankrd1/CARP and Ankrd2/Arpp) and muscle-specific RING finger proteins (MuRF1 and MuRF2), along the titin molecule were investigated in the present experiment after submaximal exhaustive exercise. Ten healthy men performed continuous drop jumping unilaterally on a sledge apparatus with a submaximal height until complete exhaustion. Five stress-sensing proteins were analysed by mRNA measurements from biopsies obtained immediately and 3 h after the exercise from exercised vastus lateralis muscle while control biopsies were obtained from non-exercised legs before the exercise. Decreased maximal jump height and increased serum creatine kinase activities as indirect markers for muscle damage and HSP27 immunostainings on muscle biopsies as a direct marker for muscle damage indicated that the current exercised protocol caused muscle damage. mRNA levels for four (MLP, Ankrd1/CARP, MuRF1 and MuRF2) out of the five studied stress sensors significantly (p < 0.05) increased 3 h after fatiguing exercise. The magnitude of MLP and Ankrd2 responses was related to the proportion of type 1 myofibres. Our data showed that the submaximal exhaustive exercise with subject's own physical fitness level activates titin-based stretch-sensing proteins. These results suggest that both degenerative and regenerative pathways are activated in very early phase after the exercise or probably already during the exercise. Activation of these proteins represents an initial step forward adaptive remodelling of the exercised muscle and may also be involved in the initiation of myofibre repair.


Assuntos
Conectina/metabolismo , Exercício Físico , Fibras Musculares de Contração Lenta/metabolismo , Proteínas Musculares/metabolismo , Esforço Físico , Adolescente , Adulto , Humanos , Masculino , Adulto Jovem
6.
Histochem Cell Biol ; 139(5): 691-715, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23224298

RESUMO

The aim of the present study was to investigate leucocyte markers, CD11b, CD16, CD66b, CD68, myeloperoxidase and neutrophil elastase on skeletal muscle biopsies from biceps brachii after unaccustomed eccentric exercise followed by the second bout of exercise 3 weeks later. The subjects (10 subjects received COX-2 inhibitor (Celecoxib) and 13 subjects received placebo) were divided into three categories: mild, moderate and severe effect of eccentric exercise, according to the reduction and recovery of muscle force-generating capacity after performing 70 maximal eccentric actions with elbow flexors on an isokinetic dynamometer. The results showed that the CD66b antibody was applicable for localization of neutrophils in human skeletal muscle, whereas the other studied neutrophil markers recognized also other leucocytes than neutrophils. The number of CD66b positive cells in skeletal muscle was very low and was not affected by the exercise. The macrophage marker CD68 showed reactivity also against satellite cells and fibroblast-like cells in skeletal muscle and therefore cannot be applied as a quantitative value for inflammatory cells. Skeletal muscle fibre injury, shown as dystrophin negative fibres, was observed approximately in half of the biopsies at 4 and 7 days after the first exercise bout in the categories moderate and severe effect of eccentric exercise. These subjects represent the most prominent loss in muscle force-generating capacity both at the category and the individual levels. Furthermore, deformed skeletal muscle fibres were observed in five subjects in these categories after the second bout of exercise. The present results suggest that neutrophils are not involved in skeletal muscle fibre injury and the reduction in muscle force-generating capacity after a single bout of eccentric exercise is a good indirect indicator of muscle damage in humans. Furthermore, prolonged regeneration process could be one of the reasons for impaired peripheral muscle function after high-force eccentric exercise.


Assuntos
Exercício Físico , Inflamação/metabolismo , Músculo Esquelético/química , Músculo Esquelético/metabolismo , Adulto , Antígenos CD/análise , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/análise , Antígenos de Diferenciação Mielomonocítica/metabolismo , Biomarcadores/análise , Biomarcadores/metabolismo , Antígeno CD11b/análise , Antígeno CD11b/metabolismo , Celecoxib , Moléculas de Adesão Celular/análise , Moléculas de Adesão Celular/metabolismo , Inibidores de Ciclo-Oxigenase 2/administração & dosagem , Feminino , Proteínas Ligadas por GPI/análise , Proteínas Ligadas por GPI/metabolismo , Humanos , Elastase de Leucócito/análise , Elastase de Leucócito/metabolismo , Masculino , Contração Muscular , Peroxidase/análise , Peroxidase/metabolismo , Pirazóis/administração & dosagem , Receptores de IgG/análise , Receptores de IgG/metabolismo , Sulfonamidas/administração & dosagem
7.
FEBS Open Bio ; 13(4): 701-712, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36792971

RESUMO

Synechocystis sp. PCC 6803 is a model cyanobacterium, glucose-tolerant substrains of which are commonly used as laboratory strains. In recent years, it has become evident that 'wild-type' strains used in different laboratories show some differences in their phenotypes. We report here the chromosome sequence of our Synechocystis sp. PCC 6803 substrain, named substrain GT-T. The chromosome sequence of GT-T was compared to those of two other commonly used laboratory substrains, GT-S and PCC-M. We identified 11 specific mutations in the GT-T substrain, whose physiological consequences are discussed. We also provide an update on evolutionary relationships between different Synechocystis sp. PCC 6803 substrains.


Assuntos
Synechocystis , Synechocystis/genética , Mutação
8.
Heliyon ; 9(2): e13147, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36718152

RESUMO

Background: In coeliac disease (CoD), the role of B-cells has mainly been considered to be production of antibodies. The functional role of B-cells has not been analysed extensively in CoD. Methods: We conducted a study to characterize gene expression in B-cells from children developing CoD early in life using samples collected before and at the diagnosis of the disease. Blood samples were collected from children at risk at 12, 18, 24 and 36 months of age. RNA from peripheral blood CD19+ cells was sequenced and differential gene expression was analysed using R package Limma. Findings: Overall, we found one gene, HNRNPL, modestly downregulated in all patients (logFC -0·7; q = 0·09), and several others downregulated in those diagnosed with CoD already by the age of 2 years. Interpretation: The data highlight the role of B-cells in CoD development. The role of HNRPL in suppressing enteroviral replication suggests that the predisposing factor for both CoD and enteroviral infections is the low level of HNRNPL expression. Funding: EU FP7 grant no. 202063, EU Regional Developmental Fund and research grant PRG712, The Academy of Finland Centre of Excellence in Molecular Systems Immunology and Physiology Research (SyMMyS) 2012-2017, grant no. 250114) and, AoF Personalized Medicine Program (grant no. 292482), AoF grants 292335, 294337, 319280, 31444, 319280, 329277, 331790) and grants from the Sigrid Jusélius Foundation (SJF).

9.
Life (Basel) ; 12(2)2022 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-35207450

RESUMO

Acclimation of cyanobacterium Synechocystis sp. PCC6803 to suboptimal conditions is largely dependent on adjustments of gene expression, which is highly controlled by the σ factor subunits of RNA polymerase (RNAP). The SigB and SigD σ factors are close homologues. Here we show that the sigB and sigD genes are both induced in high light and heat stresses. Comparison of transcriptomes of the control strain (CS), ΔsigB, ΔsigD, ΔsigBCE (containing SigD as the only functional group 2 σ factor), and ΔsigCDE (SigB as the only functional group 2 σ factor) strains in standard, high light, and high temperature conditions revealed that the SigB and SigD factors regulate different sets of genes and SigB and SigD regulons are highly dependent on stress conditions. The SigB regulon is bigger than the SigD regulon at high temperature, whereas, in high light, the SigD regulon is bigger than the SigB regulon. Furthermore, our results show that favoring the SigB or SigD factor by deleting other group 2 σ factors does not lead to superior acclimation to high light or high temperature, indicating that all group 2 σ factors play roles in the acclimation processes.

10.
Dev Cell ; 57(20): 2350-2364.e7, 2022 10 24.
Artigo em Inglês | MEDLINE | ID: mdl-36283390

RESUMO

Ductal carcinoma in situ (DCIS) is a pre-invasive stage of breast cancer. During invasion, the encapsulating DCIS basement membrane (BM) is compromised, and tumor cells invade the surrounding stroma. The mechanisms that regulate functional epithelial BMs in vivo are poorly understood. Myosin-X (MYO10) is a filopodia-inducing protein associated with metastasis and poor clinical outcome in invasive breast cancer (IBC). We identify elevated MYO10 expression in human DCIS and IBC, and this suggests links with disease progression. MYO10 promotes filopodia formation and cell invasion in vitro and cancer-cell dissemination from progressively invasive human DCIS xenografts. However, MYO10-depleted xenografts are more invasive. These lesions exhibit compromised BMs, poorly defined borders, and increased cancer-cell dispersal and EMT-marker-positive cells. In addition, cancer spheroids are dependent on MYO10-filopodia to generate a near-continuous extracellular matrix boundary. Thus, MYO10 is protective in early-stage breast cancer, correlating with tumor-limiting BMs, and pro-invasive at later stages, facilitating cancer-cell dissemination.


Assuntos
Neoplasias da Mama , Carcinoma Ductal de Mama , Carcinoma Intraductal não Infiltrante , Humanos , Feminino , Carcinoma Intraductal não Infiltrante/metabolismo , Carcinoma Intraductal não Infiltrante/patologia , Pseudópodes/metabolismo , Neoplasias da Mama/patologia , Miosinas/metabolismo , Membrana Basal/metabolismo , Carcinoma Ductal de Mama/metabolismo
11.
J Physiol ; 586(12): 3005-16, 2008 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-18420709

RESUMO

Women are at greater risk than men of sustaining certain kinds of injury and diseases of collagen-rich tissues. To determine whether a high level of oestradiol has an acute influence on collagen synthesis in tendons at rest and in response to exercise, one-legged kicking exercise was performed for 60 min at 67% of maximum power by healthy, young oral contraceptive (OC) users when circulating synthetic (ethinyl) oestradiol was high (n = 11, HE-OC) and compared to similar women who had never used OCs when circulating endogenous oestrogen was low (n = 12, LE-NOC). Interstitial fluid was collected 24 h post-exercise through microdialysis catheters placed anterior to the patellar tendon in both legs and subsequently analysed for the amino-terminal propeptide of type I collagen (PINP), a marker of tendon collagen synthesis. To determine the long-term effect of OC usage, patellar tendon cross-sectional area (CSA) was measured by magnetic resonance imaging (MRI). A lower exercise-induced increase in tendon collagen synthesis was observed in HE-OC than in LE-NOC (DeltaPINP (mean +/- s.e.m.) 1.5 +/- 5.3 versus 24.2 +/- 9.4 ng ml(-1), P < 0.05). Furthermore, serum and the interstitial peritendinous tissue concentrations of insulin-like growth factor I (IGF-I) and IGF-binding proteins showed a reduced bioavailability in HE-OC compared with results in LE-NOC. No difference in patellar tendon CSA was observed between groups. In conclusion, the selective increase in tendon collagen synthesis in LE-NOC but not HE-OC 24 h post-exercise is consistent with the hypothesis that oestradiol inhibits exercise-induced collagen synthesis in human tendon. The mechanism behind this is either a direct effect of oestradiol, or an indirect effect via a reduction in levels of free IGF-I. However, the data did not indicate any long-term effect on tendon size associated with chronic OC use.


Assuntos
Colágeno/biossíntese , Etinilestradiol/farmacologia , Exercício Físico/fisiologia , Esforço Físico/fisiologia , Tendões/fisiologia , Adulto , Regulação para Baixo , Feminino , Humanos , Esforço Físico/efeitos dos fármacos , Tendões/efeitos dos fármacos
12.
Matrix Biol ; 27(2): 86-95, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17931846

RESUMO

Whether adjacent collagen fascicles transmit force in parallel is unknown. The purpose of the present study was to examine the magnitude of lateral force transmission between adjacent collagen fascicles from the human patellar and Achilles tendon. From each sample two adjacent strands of fascicles (phi 300-530 mum) enclosed in a fascicular membrane were dissected. The specimen was deformed to approximately 3% strain in three independent load-displacement cycles in a small-scale tensile testing device. Cycle 1: the fascicles and the fascicular membrane were intact. Cycle 2: one fascicle was transversally cut while the other fascicle and the fascicular membrane were kept intact. Cycle 3: both fascicles were cut in opposite ends while the fascicular membrane was left intact. A decline in peak force of 45% and 55% from cycle 1 to cycle 2, and 93% and 92% from cycle 2 to cycle 3 was observed in the patellar and Achilles tendon fascicles, respectively. A decline in stiffness of 39% and 60% from cycle 1 to cycle 2, and of 93% and 100% from cycle 2 to cycle 3 was observed in the patellar and Achilles tendon fascicles, respectively. The present data demonstrate that lateral force transmission between adjacent collagen fascicles in human tendons is small or negligible, suggesting that tendon fascicles largely act as independent structures and that force transmission principally takes place within the individual fascicles.


Assuntos
Colágenos Fibrilares/fisiologia , Ligamento Patelar/fisiologia , Tendões/fisiologia , Tendão do Calcâneo/fisiologia , Adulto , Fenômenos Biomecânicos , Tecido Conjuntivo/anatomia & histologia , Elasticidade , Colágenos Fibrilares/ultraestrutura , Humanos , Masculino , Microscopia Eletrônica de Transmissão , Microscopia de Interferência , Ligamento Patelar/anatomia & histologia , Ligamento Patelar/ultraestrutura , Estresse Mecânico , Tendões/anatomia & histologia , Tendões/ultraestrutura , Resistência à Tração/fisiologia
13.
J Appl Physiol (1985) ; 105(1): 180-6, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18420714

RESUMO

Although the negative effects of bed rest on muscle strength and muscle mass are well established, it still remains a challenge to identify effective methods to restore physical capacity of elderly patients recovering from hospitalization. The present study compared different training regimes with respect to muscle strength, muscle fiber size, muscle architecture, and stair walking power in elderly postoperative patients. Thirty-six patients (60-86 yr) scheduled for unilateral hip replacement surgery due to hip osteoarthritis were randomized to either 1) resistance training (RT: 3/wk x 12 wk), 2) electrical stimulation (ES: 1 h/day x 12 wk), or 3) standard rehabilitation (SR: 1 h/day x 12 wk). All measurements were performed at baseline, at 5 wk and 12 wk postsurgery. After 12 wk of resistance training, maximal dynamic muscle strength increased by 30% at 60 degrees /s (P < 0.05) and by 29% at 180 degrees /s (P < 0.05); muscle fiber area increased for type I (+17%, P < 0.05), type IIa (+37%, P < 0.05), and type IIx muscle fibers (+51%, P < 0.05); and muscle fiber pennation angle increased by 22% and muscle thickness increased by 15% (P < 0.05). Furthermore, stair walking power increased by 35% (P < 0.05) and was related to the increase in type II fiber area (r = 0.729, P < 0.05). In contrast, there was no increase in any measurement outcomes with electrical stimulation and standard rehabilitation. The present study is the first to demonstrate the effectiveness of resistance training to induce beneficial qualitative changes in muscle fiber morphology and muscle architecture in elderly postoperative patients. In contrast, rehabilitation regimes based on functional exercises and neuromuscular electrical stimulation had no effect. The present data emphasize the importance of resistance training in future rehabilitation programs for elderly individuals.


Assuntos
Idoso/fisiologia , Músculo Esquelético/anatomia & histologia , Músculo Esquelético/fisiologia , Aptidão Física/fisiologia , Levantamento de Peso/fisiologia , Idoso de 80 Anos ou mais , Artroplastia de Quadril , Índice de Massa Corporal , Peso Corporal/fisiologia , Estimulação Elétrica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fibras Musculares Esqueléticas/fisiologia , Fibras Musculares Esqueléticas/ultraestrutura , Força Muscular/fisiologia , Osteoartrite/reabilitação , Osteoartrite/cirurgia , Caminhada/fisiologia
14.
Connect Tissue Res ; 49(3): 165-8, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18661335

RESUMO

The connective tissue of tendon and skeletal muscle is a crucial structure for force transmission. A dynamic adaptive capacity of these tissues in healthy individuals is evident from reports of altered gene expression and protein levels of the fibrillar and network-forming collagens, when subjected to mechanical loading. While it appears that the fibroblast is a key player in sensing and responding to loading, the issue of how these signals are converted into changed gene expression is not fully understood. It is clear, however, that the loading-induced response involves a variety of growth factors, in particular TGF-beta-1, and matrix remodelling enzymes such as MMP-2. Furthermore, it is under hormonal influence. In skeletal muscle, the extracellular matrix demonstrates its potential for cross-talk by regulating the activity of cells with which it is in contact. Taken together, the studies highlighted in this article provide strong evidence for the highly adaptable nature of connective tissue in muscle and tendon.


Assuntos
Adaptação Fisiológica , Colágeno/biossíntese , Matriz Extracelular/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Músculo Esquelético/fisiologia , Tendões/fisiologia , Animais , Tecido Conjuntivo/metabolismo , Feminino , Fibroblastos/metabolismo , Humanos , Masculino , Metaloproteinase 14 da Matriz/metabolismo , Caracteres Sexuais , Estresse Mecânico
15.
Sci Rep ; 8(1): 10305, 2018 07 09.
Artigo em Inglês | MEDLINE | ID: mdl-29985458

RESUMO

We show that the formation of the RNAP holoenzyme with the primary σ factor SigA increases in the ΔsigBCDE strain of the cyanobacterium Synechocystis sp. PCC 6803 lacking all group 2 σ factors. The high RNAP-SigA holoenzyme content directly induces transcription of a particular set of housekeeping genes, including ones encoding transcription and translation machineries. In accordance with upregulated transcripts, ΔsigBCDE contain more RNAPs and ribosomal subunits than the control strain. Extra RNAPs are fully active, and the RNA content of ΔsigBCDE cells is almost tripled compared to that in the control strain. Although ΔsigBCDE cells produce extra rRNAs and ribosomal proteins, functional extra ribosomes are not formed, and translation activity and protein content remained similar in ΔsigBCDE as in the control strain. The arrangement of the RNA polymerase core genes together with the ribosomal protein genes might play a role in the co-regulation of transcription and translation machineries. Sequence logos were constructed to compare promoters of those housekeeping genes that directly react to the RNAP-SigA holoenzyme content and those ones that do not. Cyanobacterial strains with engineered transcription and translation machineries might provide solutions for construction of highly efficient production platforms for biotechnical applications in the future.


Assuntos
Proteínas de Bactérias/metabolismo , Fator sigma/metabolismo , Synechocystis/metabolismo , Proteínas de Bactérias/genética , RNA Polimerases Dirigidas por DNA/metabolismo , Regiões Promotoras Genéticas , Biossíntese de Proteínas/genética , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Ribossomos/metabolismo , Fator sigma/genética , Synechocystis/genética , Ativação Transcricional , Regulação para Cima
16.
Genome Announc ; 6(17)2018 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-29700151

RESUMO

We report here the nearly complete Illumina-sequenced consensus genome sequences of six isolates of echovirus 7 (E7), including oncolytic virotherapy virus RIGVIR and the Wallace prototype. Amino acid identities within the coding region were highly conserved across all isolates, ranging from 95.31% to 99.73%.

17.
Rejuvenation Res ; 10(4): 485-500, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17985945

RESUMO

The loss of muscle mass and strength with aging is well characterized, but our knowledge of the molecular mechanisms underlying the development of sarcopenia remains incomplete. Although menopause is often accompanied with first signs of age-associated changes in muscle structure and function, the effects of hormone replacement therapy (HRT) or menopause-related decline in estrogen production in the muscles of postmenopausal women is not well understood. Furthermore the knowledge of the global transcriptional changes that take place in skeletal muscle in relation to estrogen status has thus far been completely lacking. We used a randomized double-blinded study design together with an explorative microarray experiment to characterize possible effects of continuous, combined HRT and estrogen deprivation on the skeletal muscle of fifteen women. Here, we report the differential response of both Gene Ontology-annotated biological processes and some individual genes responding differentially to the use or non-use of HRT. Our results revealed transcription level changes in, for example, muscle protein and energy metabolism. In particular, the ubiquitine-proteosome system was found to be effected at several levels. HRT seemed to partially counteract the postmenopause-related transcriptional changes. Our results suggest that during the early postmenopausal years, when there is no counteracting medication available, muscle transcriptome changes notably, whereas HRT appears to slow down this phenomenon and could therefore aid in maintaining proper muscle mass and function after menopause.


Assuntos
Terapia de Reposição de Estrogênios , Perfilação da Expressão Gênica , Músculo Esquelético/metabolismo , Pós-Menopausa/metabolismo , Feminino , Regulação da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/análise , Receptores de Estrogênio/genética
18.
J Appl Physiol (1985) ; 103(2): 425-31, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17463304

RESUMO

The consumption of nonsteroidal anti-inflammatory drugs (NSAIDs) is widespread among athletes when faced with muscle soreness or injury, but the effects of NSAIDs on satellite cell activity in humans are unknown. To investigate this, 14 healthy male endurance athletes (mean peak oxygen consumption 62 ml x kg(-1) x min(-1)) volunteered for the study, which involved running 36 km. They were divided into two groups and received either 100 mg indomethacin per day or placebo. Muscle biopsies collected before the run and on days 1, 3, and 8 afterward were analyzed for satellite cells by immunohistochemistry with the aid of neural cell adhesion molecule (NCAM) and fetal antigen-1 (FA1) antibodies. Muscle biopsies were also collected from untrained individuals for comparison. Compared with preexercise levels, a 27% increase in the number of NCAM+ cells was observed on day 8 postexercise in the placebo group (P < 0.05), while levels remained similar at all time points in the NSAID group. No change was seen in the proportion of FA1+ cells, although lower levels were found in the muscle of endurance-trained athletes compared with untrained individuals (P < 0.05). These results suggest that ingestion of anti-inflammatory drugs attenuates the exercise-induced increase in satellite cell number, supporting the role of the cyclooxygenase pathway in satellite cell activity.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Inibidores de Ciclo-Oxigenase/farmacologia , Exercício Físico/fisiologia , Células Satélites de Músculo Esquelético/efeitos dos fármacos , Células Satélites de Músculo Esquelético/fisiologia , Adulto , Anti-Inflamatórios não Esteroides/uso terapêutico , Biomarcadores/metabolismo , Biópsia , Proteínas de Ligação ao Cálcio , Proliferação de Células/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase/uso terapêutico , Feminino , Humanos , Indometacina/farmacologia , Indometacina/uso terapêutico , Peptídeos e Proteínas de Sinalização Intercelular/fisiologia , Masculino , Proteínas de Membrana/fisiologia , Músculo Esquelético/citologia , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/fisiologia , Moléculas de Adesão de Célula Nervosa/fisiologia , Prostaglandina-Endoperóxido Sintases/fisiologia , Regeneração/fisiologia , Corrida , Células Satélites de Músculo Esquelético/citologia
19.
Matrix Biol ; 23(4): 259-64, 2004 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-15296940

RESUMO

Increases in procollagen processing within skeletal muscle have previously been reported in small animal models only. While indirect measurements in humans have suggested an increase procollagen processing, no intra-skeletal muscle measurements have confirmed these findings. In this study, eight young healthy male subjects performed a single bout of unaccustomed high intensity eccentric exercise on one leg, with the contralateral leg being the control. A significant increase in the muscle interstitial concentration of the N-terminal propeptide of procollagen type I (PINP) was observed (day 0: 1.96 +/- 0.44 ng ml(-1), day 2: 1.94 +/- 0.32 ng ml(-1), day 4: 3.90 +/- 1.03 ng ml(-1), day 8: 7.23 +/- 2.34 ng ml(-1)*, *P < 0.05 vs. basal and control) with no change being noted in the control leg. By day 2 post-exercise, an increase in the histological immunoreactivity of PINP and the N-terminal propeptide of procollagen type III (PIIINP) was also shown in the exercising leg only. Further, from day 2 post-exercise, immunoreactivity for tenascin C and reactive macrophages (CD68+ cells) was seen within the perimysial and endomysial connective tissue of the exercising leg only, indicating a high mechanical load and inflammation. This study shows that following a single bout of high intensity eccentric exercise there is an increase in procollagen processing within skeletal muscle in humans.


Assuntos
Exercício Físico/fisiologia , Músculo Esquelético/metabolismo , Pró-Colágeno/metabolismo , Adulto , Proteínas da Matriz Extracelular/metabolismo , Humanos , Perna (Membro) , Macrófagos/citologia , Masculino , Monócitos/citologia , Músculo Esquelético/citologia , Fragmentos de Peptídeos/metabolismo , Tenascina/metabolismo
20.
J Am Geriatr Soc ; 52(6): 939-44, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15161458

RESUMO

OBJECTIVES: To investigate the association between muscle size, density, and fiber composition; body composition; maximal isometric knee extension strength (KES); and lower-body muscle power in healthy postmenopausal women. DESIGN: Cross-sectional analysis of baseline data from a 1-year randomized controlled experiment. SETTING: University-based research laboratory. PARTICIPANTS: Seventy-eight healthy postmenopausal women aged 50 to 57. MEASUREMENTS: Maximal lower-body muscle power was assessed using vertical jump height (VJH). Maximal isometric KES was measured on a dynamometer chair. Computed tomography scans were used to determine lean-tissue cross-sectional area and density of the thigh and lower leg muscles. Relative area occupied by type I, IIa, IIax, and IIx muscle fibers was assessed from the vastus lateralis muscle. lean body mass and total body fat mass were assessed using bioelectrical impedance. RESULTS: High VJH was associated with low body fat mass, high KES, and high density of thigh and lower leg muscles. Multivariate linear regression modeling revealed that high thigh muscle density (beta=0.242; P=.019), relative area occupied by the fastest muscle fiber types (IIax+IIx; beta=0.246; P=.007), KES (beta=0.247; P=.007), and low body fat mass (beta=-0.455; P<.001) were independently associated with high VJH, accounting for 45% of the variability in VJH. CONCLUSION: This study showed that thigh muscle composition, muscle strength, and body fat mass are important determinants of lower-body muscle power production during weight-bearing activity in healthy postmenopausal women.


Assuntos
Perna (Membro)/fisiologia , Músculo Esquelético/fisiologia , Pós-Menopausa/fisiologia , Idoso , Biópsia por Agulha , Composição Corporal , Estatura , Índice de Massa Corporal , Estudos Transversais , Distrofina/análise , Impedância Elétrica , Feminino , Humanos , Imuno-Histoquímica , Contração Isométrica , Articulação do Joelho/fisiologia , Pessoa de Meia-Idade , Análise Multivariada , Fibras Musculares Esqueléticas/química , Fibras Musculares Esqueléticas/citologia , Músculo Esquelético/diagnóstico por imagem , Tomografia Computadorizada por Raios X
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