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1.
J Behav Med ; 46(4): 609-621, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36527573

RESUMO

The COVID-19 pandemic is associated with crucial changes in children's daily life including their physical activity (PA) and screen time (ST). Among preschool children, the family represents an important factor for sufficient PA levels by being the gatekeeper for PA. Thus, the aim of this study was to investigate the influence of the family environment, specifically SES, parental support, and having siblings on COVID-19-related changes of PA and ST behavior in 317 (170 boys, 147 girls) German preschool children using longitudinal data. Our results indicate a decline in total amount of sports-related PA, an increase in outdoor play, as well as an increase in leisure ST in preschool children. The changes in total amount of PA differed between children with different levels of parental support as well as in dependence on having siblings. Furthermore, levels of outdoor play and ST in preschool children were influenced by environmental factors like having access to their own garden. We conclude that the family environment (parental support as well as physical environment) is highly relevant for PA and ST levels in preschool children. To provide every child with PA opportunities during potential future lockdowns, restriction policies should be adapted and parents need sophisticated information about the importance of their support and thus the PA levels of their children.


Assuntos
COVID-19 , Pandemias , Masculino , Feminino , Humanos , Pré-Escolar , Comportamento Sedentário , Controle de Doenças Transmissíveis , Exercício Físico , Pais
2.
Biol Chem ; 403(5-6): 455-477, 2022 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-33759431

RESUMO

Antibody display technologies enable the successful isolation of antigen-specific antibodies with therapeutic potential. The key feature that facilitates the selection of an antibody with prescribed properties is the coupling of the protein variant to its genetic information and is referred to as genotype phenotype coupling. There are several different platform technologies based on prokaryotic organisms as well as strategies employing higher eukaryotes. Among those, phage display is the most established system with more than a dozen of therapeutic antibodies approved for therapy that have been discovered or engineered using this approach. In recent years several other technologies gained a certain level of maturity, most strikingly mammalian display. In this review, we delineate the most important selection systems with respect to antibody generation with an emphasis on recent developments.


Assuntos
Anticorpos , Biblioteca de Peptídeos , Animais , Anticorpos/genética , Anticorpos/uso terapêutico , Mamíferos/genética
3.
Am J Epidemiol ; 190(2): 220-229, 2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-33524119

RESUMO

Mental health (MH) and behavioral health are fundamental to a good quality of life. Only a few studies have investigated the association between behavioral health (e.g., physical activity (PA), screen time (ST)) and MH from childhood to adolescence. Therefore, we investigated the relationships of PA and ST with MH by sex in an 11-year longitudinal cohort study of German schoolchildren during 2003-2017. A subsample (n = 686; 55.2% female) of participants from the German Motorik-Modul (MoMo) Longitudinal Study who participated in all 3 measurement phases (mean ages: time 1 (baseline; 2003-2006), 5.57 (standard deviation (SD), 1.00) years; time 2 (wave 1; 2009-2012), 11.85 (SD, 1.03) years; time 3 (wave 2; 2014-2017), 16.86 (SD, 1.04) years) were analyzed with regard to PA, ST, and MH (emotional symptoms, conduct problems, hyperactivity/inattention, peer relationship problems, prosocial behavior, and overall strengths and difficulties). Path panel prediction models were fitted with time 1, time 2, and time 3 PA, ST, and MH indicators. PA predicted less television (TV)/video watching in females, and TV/video watching predicted personal computer (PC)/Internet use in both sexes. Behavior and MH results suggested that, for females, higher TV/video watching and PC/Internet use was related to higher MH challenges over the course of maturation. Some preadolescent males' MH challenges increased ST (TV/video watching and PC/Internet use) in adolescence. Researchers should explore innovative and effective methods for reducing childhood ST, especially among females with early signs of MH issues, and addressing preadolescent males' MH challenges.


Assuntos
Transtornos do Comportamento Infantil/epidemiologia , Exercício Físico , Saúde Mental/estatística & dados numéricos , Tempo de Tela , Adolescente , Criança , Pré-Escolar , Emoções , Feminino , Alemanha/epidemiologia , Humanos , Estudos Longitudinais , Masculino , Comportamento Sedentário , Fatores Sexuais , Fatores Socioeconômicos
4.
Mol Microbiol ; 113(3): 603-612, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31705780

RESUMO

Trans-acting small regulatory RNAs (sRNAs) are key players in the regulation of gene expression in bacteria. There are hundreds of different sRNAs in a typical bacterium, which in contrast to eukaryotic microRNAs are more heterogeneous in length, sequence composition, and secondary structure. The vast majority of sRNAs function post-transcriptionally by binding to other RNAs (mRNAs, sRNAs) through rather short regions of imperfect sequence complementarity. Besides, every single sRNA may interact with dozens of different target RNAs and impact gene expression either negatively or positively. These facts contributed to the view that the entirety of the regulatory targets of a given sRNA, its targetome, is challenging to identify. However, recent developments show that a more comprehensive sRNAs targetome can be achieved through the combination of experimental and computational approaches. Here, we give a short introduction into these methods followed by a description of two sRNAs, RyhB, and RsaA, to illustrate the particular strengths and weaknesses of these approaches in more details. RyhB is an sRNA involved in iron homeostasis in Enterobacteriaceae, while RsaA is a modulator of virulence in Staphylococcus aureus. Using such a combined strategy, a better appreciation of the sRNA-dependent regulatory networks is now attainable.


Assuntos
Biologia Computacional/métodos , Regulação Bacteriana da Expressão Gênica/genética , Pequeno RNA não Traduzido/genética , Bactérias/genética , Enterobacteriaceae/genética , Expressão Gênica/genética , Genes Bacterianos/genética , RNA Bacteriano/metabolismo , RNA Mensageiro/metabolismo , Pequeno RNA não Traduzido/metabolismo , Staphylococcus aureus/genética
5.
Oncologist ; 26(3): e492-e499, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33355968

RESUMO

BACKGROUND: Long-term colon cancer survivors present heterogeneous health-related quality of life (HRQOL) outcomes. We determined unobserved subgroups (classes) of survivors with similar HRQOL patterns and investigated their stability over time and the association of clinical covariates with these classes. MATERIALS AND METHODS: Data from the population-based PROFILES registry were used. Included were survivors with nonmetastatic (TNM stage I-III) colon cancer (n = 1,489). HRQOL was assessed with the Dutch translation of the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire C30 version 3.0. Based on survivors' HRQOL, latent class analysis (LCA) was used to identify unobserved classes of survivors. Moreover, latent transition analysis (LTA) was used to investigate changes in class membership over time. Furthermore, the effect of covariates on class membership was assessed using multinomial logistic regression. RESULTS: LCA identified five classes at baseline: class 1, excellent HRQOL (n = 555, 37.3%); class 2, good HRQOL with prevalence of insomnia (n = 464, 31.2%); class 3, moderate HRQOL with prevalence of fatigue (n = 213, 14.3%); class 4, good HRQOL with physical limitations (n = 134, 9.0%); and class 5, poor HRQOL (n = 123, 8.3%). All classes were stable with high self-transition probabilities. Longer time since the diagnosis, no comorbid conditions, and male sex were associated with class 1, whereas older age was associated with class 4. Clinical covariates were not associated with class membership. CONCLUSION: The identified classes are characterized by distinct patterns of HRQOL and can support patient-centered care. LCA and LTA are powerful tools for investigating HRQOL in cancer survivors. IMPLICATIONS FOR PRACTICE: Long-term colon cancer survivors show great heterogeneity in their health-related quality of life. This study identified five distinct clusters of survivors with similar patterns of health-related quality of life and showed that these clusters remain stable over time. It was also shown that these clusters do not significantly differ in tumor characteristics or received treatment. Cluster membership of long-term survivors can be identified by sociodemographic characteristics but is not predetermined by diagnosis and treatment.


Assuntos
Sobreviventes de Câncer , Neoplasias , Idoso , Colo , Humanos , Análise de Classes Latentes , Masculino , Qualidade de Vida , Sistema de Registros , Inquéritos e Questionários
6.
Bioinformatics ; 36(15): 4357-4359, 2020 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-32492127

RESUMO

MOTIVATION: The correct prediction of bacterial sRNA homologs is a prerequisite for many downstream analyses based on comparative genomics, but it is frequently challenging due to the short length and distinct heterogeneity of such homologs. GLobal Automatic Small RNA Search go (GLASSgo) is an efficient tool for the prediction of sRNA homologs from a single input query. To make the algorithm available to a broader community, we offer a Docker container along with a free-access web service. For non-computer scientists, the web service provides a user-friendly interface. However, capabilities were lacking so far for batch processing, version control and direct interaction with compatible software applications as a workflow management system can provide. RESULTS: Here, we present GLASSgo 1.5.2, an updated version that is fully incorporated into the workflow management system Galaxy. The improved version contains a new feature for extracting the upstream regions, allowing the search for conserved promoter elements. Additionally, it supports the use of accession numbers instead of the outdated GI numbers, which widens the applicability of the tool. AVAILABILITY AND IMPLEMENTATION: GLASSgo is available at https://github.com/lotts/GLASSgo/ under the MIT license and is accompanied by instruction and application data. Furthermore, it can be installed into any Galaxy instance using the Galaxy ToolShed.


Assuntos
Biologia Computacional , Software , Algoritmos , Genômica , Fluxo de Trabalho
7.
BMC Med Res Methodol ; 21(1): 269, 2021 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-34852769

RESUMO

BACKGROUND: Sleep apnea patients on CPAP therapy exhibit differences in how they adhere to the therapy. Previous studies have demonstrated the benefit of describing adherence in terms of discernible longitudinal patterns. However, these analyses have been done on a limited number of patients, and did not properly represent the temporal characteristics and heterogeneity of adherence. METHODS: We illustrate the potential of identifying patterns of adherence with a latent-class heteroskedastic hurdle trajectory approach using generalized additive modeling. The model represents the adherence trajectories on three aspects over time: the daily hurdle of using the therapy, the daily time spent on therapy, and the day-to-day variability. The combination of these three characteristics has not been studied before. RESULTS: Applying the proposed model to a dataset of 10,000 patients in their first three months of therapy resulted in nine adherence groups, among which 49% of patients exhibited a change in adherence over time. The identified group trajectories revealed a non-linear association between the change in the daily hurdle of using the therapy, and the average time on therapy. The largest difference between groups was observed in the patient motivation score. The adherence patterns were also associated with different levels of high residual AHI, and day-to-day variability in leakage. CONCLUSION: The inclusion of the hurdle model and the heteroskedastic model into the mixture model enabled the discovery of additional adherence patterns, and a more descriptive representation of patient behavior over time. Therapy adherence was mostly affected by a lack of attempts over time, suggesting that encouraging these patients to attempt therapy on a daily basis, irrespective of the number of hours used, could drive adherence. We believe the methodology is applicable to other domains of therapy or medication adherence.


Assuntos
Pressão Positiva Contínua nas Vias Aéreas , Apneia Obstrutiva do Sono , Humanos , Cooperação do Paciente , Apneia Obstrutiva do Sono/terapia
8.
BMC Nephrol ; 22(1): 337, 2021 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-34641801

RESUMO

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH) is a rare and life-threatening disease characterized by hyperactivation of the immune system that causes hypercytokinemia and potentially multi organ failure. HLH can occur in patients with underlying rheumatic or autoinflammatory disorders. Additionally, HLH can develop in patients during infections or malignancies without a known genetic predisposition. CASE PRESENTATION: We herein report a patient, who presented with fever, both acute kidney and liver injury, anemia, thrombocytopenia and HSV stomatitis. HLH was diagnosed based on clinical criteria and qPCR revealed an acute parvovirus B19 infection as potential underlying infectious trigger. Treatment was started with both IVIG and dexamethasone. Subsequently, kidney biopsy demonstrated TMA. CONCLUSIONS: In rare cases both HLH and aHUS can occur simultaneously in a patient as a consequence of viral infections. Insights from this unusual case might help physicians understand this complex symptom constellation.


Assuntos
Transplante de Rim , Linfo-Histiocitose Hemofagocítica/complicações , Infecções por Parvoviridae/complicações , Complicações Pós-Operatórias , Microangiopatias Trombóticas/complicações , Feminino , Humanos , Linfo-Histiocitose Hemofagocítica/diagnóstico , Pessoa de Meia-Idade , Infecções por Parvoviridae/diagnóstico , Complicações Pós-Operatórias/diagnóstico , Microangiopatias Trombóticas/diagnóstico
9.
BMC Public Health ; 21(1): 1716, 2021 09 21.
Artigo em Inglês | MEDLINE | ID: mdl-34548057

RESUMO

BACKGROUND: Outdoor play, sedentary behavior (SB), and moderate-to-vigorous physical activity (MVPA) are related to youth's health, however, there are research gaps regarding 1) associations between outdoor play, SB, and MVPA across a broad pediatric age range (6-17 years), and 2) longitudinal associations between outdoor play, SB, and MVPA across childhood and adolescence. Two studies were conducted to address those research gaps: Study 1 aimed to investigate relationships between outdoor play and accelerometer-assessed SB and MVPA in a cross-sectional nationwide sample of children and adolescents in Germany. Study 2 aimed to investigate prospective associations between outdoor play and self-reported screen-time SB and MVPA and in a sample of children with three measurement timepoints across 11 years. METHODS: Data were obtained of the German national representative Motorik-Modul (MoMo) Study and the German Health Interview and Examination Survey for Children and Adolescents (KiGGS). In Study 1, N = 2278 participants (6-17 years) were included with self-reported outdoor play and accelerometer-assessed SB and MVPA. Associations were examined via multiple linear regressions. In Study 2, N = 570 participants (baseline: 4-7 years) were included in the longitudinal analysis with follow-ups six and 11 years later. Screen-time SB (TV watching and PC/Gaming), MVPA, and outdoor play were self-reported. Associations were investigated through a path prediction model. RESULTS: Study 1 showed that compared to <1 h outdoor play, higher engagement in daily outdoor play was related to lower SB (1-2 h: - 9.75 min/day, P = 0.017; ≥2 h: - 17.78 min/day, P < 0.001) and higher MVPA (≥2 h: + 3.87 min/day, P = 0.001). The cross-sectional relationship between MVPA and outdoor play was moderated by sex (in favor of males) and age (in favor of younger children). Study 2 showed that outdoor play in early childhood negatively predicted PC use/Gaming in later childhood, but was unrelated to MVPA. CONCLUSION: In Study 1, outdoor play was negatively related to SB cross-sectionally. In Study 2, outdoor play in early childhood was negatively related to PC and Gaming time in later childhood. Thus, providing outdoor play opportunities, especially during early childhood, has potential to prevent SB. Future research should investigate longitudinal relationships using device-based assessments for SB and MVPA.


Assuntos
Exercício Físico , Comportamento Sedentário , Acelerometria , Adolescente , Criança , Pré-Escolar , Estudos de Coortes , Humanos , Masculino , Tempo de Tela , Autorrelato
10.
BMC Public Health ; 21(1): 1081, 2021 06 05.
Artigo em Inglês | MEDLINE | ID: mdl-34090411

RESUMO

BACKGROUND: As children show a more complex but less structured movement behavior than adults, assessment of their many spontaneous and impulsive movements is a challenge for physical activity (PA) assessment. Since neither questionnaires nor accelerometers enable optimal detection of all facets of PA, a multimodal, combined approach of self-reported and device-based methods is recommended. Based on the number of days on which the participants reached the physical activity (PA) values given in the WHO guideline, this study examines 1) the difference between self-reported and device-based, measured PA and 2) whether PA differences between age and gender groups obtained by two methods are comparable. METHODS: Participants aged 6-17 years were randomly chosen and data were collected representatively at 167 sample points throughout Germany within the Motorik-Modul Study. PA of n = 2694 participants (52.3% female) was measured using the ActiGraph accelerometer (ACC) and a physical activity questionnaire (PAQ). The sample was divided into three age groups (6-10 yrs. n = 788, 11-13 yrs. n = 823, 14-17 yrs. n = 1083). Numbers of days per week with at least 60 min moderate to vigorous PA (MVPA) were analyzed for both methods. RESULTS: Only every 25th respondent (4%) reaches the WHO standard of 60 min MVPA every day if measured with ACC. Self-reported PA was slightly higher (9%) (meanPAQ = 3.82 days; meanACC = 2.34 days; Fmethod = 915.85; p = <.001; fCohen = .64). The differences between the methods are significantly smaller in younger children than in the older age groups (Fage = 264.2, p < .001; fCohen = .48). The older the subjects are, the lower is the proportion of those who meet the WHO guideline on each day, with girls meeting the guideline less frequently than boys in all age groups. CONCLUSION: Children and adolescents living in Germany show a very low adherence to the WHO guideline on PA. While younger children are much more active with their free play, especially children over 10 years of age and especially girls should be the target of programs to increase PA.


Assuntos
Etnicidade , Exercício Físico , Acelerometria , Adolescente , Adulto , Idoso , Criança , Feminino , Alemanha , Humanos , Masculino , Autorrelato , Inquéritos e Questionários
11.
Eur J Public Health ; 31(2): 373-378, 2021 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-33011779

RESUMO

BACKGROUND: Inactivity in children and adolescents is a global issue requiring interventions that target different domains of physical activity, such as active transport. The aim of this study was to assess the prevalence, historical trends and socio-demographic correlates of active commuting to school (ACS) in a nationwide sample of girls and boys from Germany. METHODS: Data of commuting behaviour and socio-demographic factors were collected, covering three measurement points from 2003 to 2017. The MoMo Study derived its data from a representative sample of children and adolescents aged 4-17 years who answered a questionnaire (N = 11 387). Statistically significant differences between Baseline, Wave 1 and Wave 2 were determined via 95% confidence intervals (CIs) for complex samples. RESULTS: Overall, ACS decreased from 84.4% at the first measurement point to 78.3% in the third measurement point. The proportion of cases in which children opted for passive modes of commuting increased predominantly in those aged 4-5 years, in children with a low-to-medium socio-economic status, and in children residing in small- or medium-sized towns. No gender differences were found in active commuting. The results of multinomial logistic regression identified age, migration background and residential area as correlates of walking for boys. For girls, the likelihood of walking, cycling and taking public transport instead of opting for motorized transport increased with age. CONCLUSIONS: Intervention programmes to increase active transport in children and adolescents should target different age groups and also consider parental influence in determining the child's choice of transport mode.


Assuntos
Instituições Acadêmicas , Meios de Transporte , Adolescente , Ciclismo , Criança , Estudos Transversais , Feminino , Alemanha , Humanos , Masculino , Caminhada
12.
Nucleic Acids Res ; 46(W1): W25-W29, 2018 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-29788132

RESUMO

The Freiburg RNA tools webserver is a well established online resource for RNA-focused research. It provides a unified user interface and comprehensive result visualization for efficient command line tools. The webserver includes RNA-RNA interaction prediction (IntaRNA, CopraRNA, metaMIR), sRNA homology search (GLASSgo), sequence-structure alignments (LocARNA, MARNA, CARNA, ExpaRNA), CRISPR repeat classification (CRISPRmap), sequence design (antaRNA, INFO-RNA, SECISDesign), structure aberration evaluation of point mutations (RaSE), and RNA/protein-family models visualization (CMV), and other methods. Open education resources offer interactive visualizations of RNA structure and RNA-RNA interaction prediction as well as basic and advanced sequence alignment algorithms. The services are freely available at http://rna.informatik.uni-freiburg.de.


Assuntos
Sequência de Bases/genética , Internet , RNA/genética , Software , Algoritmos , Conformação de Ácido Nucleico , RNA/química , Alinhamento de Sequência/instrumentação , Análise de Sequência de RNA/instrumentação , Relação Estrutura-Atividade
13.
BMC Med Inform Decis Mak ; 20(1): 277, 2020 10 27.
Artigo em Inglês | MEDLINE | ID: mdl-33109175

RESUMO

BACKGROUND: The increased availability of patient reported outcome data makes it feasible to provide patients tailored risk information of cancer treatment side effects. However, it is unclear how such information influences patients' risk interpretations compared to generic population-based risks, and which message format should be used to communicate such individualized statistics. METHODS: A web-based experiment was conducted in which participants (n = 141) read a hypothetical treatment decision-making scenario about four side effect risks of adjuvant chemotherapy for advanced colon cancer. Participants were cancer patients or survivors who were recruited from an online Dutch cancer patient panel. All participants received two tailored risks (of which the reference class was based on their age, gender and tumor stage) and two generic risks conveying the likelihood of experiencing the side effects. The risks were presented either in words-only ('common' and 'very common'), or in a combination of words and corresponding numerical estimates ('common, 10 out of 100' and 'very common, 40 out of 100'). Participants' estimation of the probability, accuracy of their estimation, and perceived likelihood of occurrence were primary outcomes. Perceived personal relevance and perceived uncertainty were secondary outcomes. RESULTS: Tailored risks were estimated as higher and less accurate than generic risks, but only when they were presented in words; Such differences were not found in the verbal and numerical combined condition. Although tailoring risks did not impact participants' perceived likelihood of occurrence, tailored risks were perceived as more personally relevant than generic risks in both message formats. Finally, tailored risks were perceived as less uncertain than generic risks, but only in the verbal-only condition. CONCLUSIONS: Considering current interest in the use of personalized decision aids for improving shared decision-making in oncology, it is important that clinicians consider how tailored risks of treatment side effects should be communicated to patients. We recommend both clinicians who communicate probability information during consultations, and decision aid developers, that verbal descriptors of tailored risks should be supported by numerical estimates of risks levels, to avoid overestimation of risks.


Assuntos
Antineoplásicos/efeitos adversos , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Comunicação em Saúde/normas , Neoplasias/tratamento farmacológico , Educação de Pacientes como Assunto/métodos , Antineoplásicos/uso terapêutico , Tomada de Decisão Compartilhada , Feminino , Humanos , Masculino , Medição de Risco , Fatores de Risco
14.
Biol Chem ; 400(3): 383-393, 2019 02 25.
Artigo em Inglês | MEDLINE | ID: mdl-30465712

RESUMO

Antibodies can be successfully engineered and isolated by yeast or phage display of combinatorial libraries. Still, generation of libraries comprising heavy chain as well as light chain diversities is a cumbersome process involving multiple steps. Within this study, we set out to compare the output of yeast display screening of antibody Fab libraries from immunized rodents that were generated by Golden Gate Cloning (GGC) with the conventional three-step method of individual heavy- and light-chain sub-library construction followed by chain combination via yeast mating (YM). We demonstrate that the GGC-based one-step process delivers libraries and antibodies from heavy- and light-chain diversities with similar quality to the traditional method while being significantly less complex and faster. Additionally, we show that this method can also be used to successfully screen and isolate chimeric chicken/human antibodies following avian immunization.


Assuntos
Cadeias Pesadas de Imunoglobulinas/análise , Cadeias Leves de Imunoglobulina/análise , Saccharomyces cerevisiae/química , Animais , Galinhas , Clonagem Molecular , Humanos , Cadeias Pesadas de Imunoglobulinas/imunologia , Cadeias Leves de Imunoglobulina/imunologia , Biblioteca de Peptídeos , Engenharia de Proteínas , Saccharomyces cerevisiae/imunologia , Propriedades de Superfície
15.
BMC Microbiol ; 19(1): 147, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31262257

RESUMO

BACKGROUND: The presence and activity of CRISPR-Cas defense systems is a hallmark of many prokaryotic microorganisms. Here, the distribution of sequences related to the highly iterated palindrome 1 (HIP1) element and the DNA methylation of CGATCG motifs embedded within HIP1 as a vital part of the CRISPR1 repeat sequence was analyzed in the cyanobacterium Synechocystis sp. PCC 6803. Previously suggested functions of HIP1 include organization of chromosomal structure, DNA recombination or gene regulation, all of which could be relevant in CRISPR-Cas functionality. RESULTS: The CRISPR1 repeat-spacer array contains more than 50 CGATCG elements that are double-methylated (5mCG6mATCG) by the enzymes M.Ssp6803I and M.Ssp6803III. Hence, more than 200 possible methylation events cluster over a stretch of 3600 bp of double-stranded DNA. Bisulfite sequencing showed that these motifs were highly methylated at the m5CGATCG positions whereas specific motifs within the CRISPR1 cas genes were hypomethylated suggesting a lowered accessibility for the DNA methylase to these regions. Assays for conjugation and CRISPR1-mediated DNA interference revealed a 50% drop in conjugation efficiency in the mutant lacking the 5mC methylation of CGATCG motifs, while the highly efficient DNA interference activity was not affected by the lack of m5CGATCG DNA-methylation, nor was the capability to differentiate between self and non-self targets based on the protospacer adjacent motifs (PAMs) GTA and GTC versus the non-PAM AGC. A third DNA methylation mediated by M.Ssp6803II modifies the first cytosine in the motif GGCC yielding GGm4CC. We found a remarkable absence of GGCC motifs and hence the corresponding methylation over an 11 kb stretch encompassing all the cas genes involved in interference and crRNA maturation but not adaptation of the CRISPR1 system. CONCLUSIONS: The lack of GGCC tetranucleotides along the CRISPR1 interference and maturation genes supports the reported hybrid character of subtype I-D CRISPR-Cas systems. We report tight and very high 5mC methylation of the CRISPR1 repeat sequences. Nevertheless, cells lacking the 5mC methylation activity were unaffected in their CRISPR1-mediated interference response but the efficiency of conjugation was reduced by 50%. These results point to an unknown role of m5CGATCG DNA-methylation marks in conjugation and DNA transformation.


Assuntos
Proteínas Associadas a CRISPR/genética , Sistemas CRISPR-Cas/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Metilação de DNA , Synechocystis/genética , DNA , DNA (Citosina-5-)-Metiltransferases , DNA Bacteriano/genética , Motivos de Nucleotídeos , Recombinação Genética , Análise de Sequência de DNA
16.
Nucleic Acids Res ; 45(W1): W560-W566, 2017 07 03.
Artigo em Inglês | MEDLINE | ID: mdl-28582575

RESUMO

RNA-based regulation has become a major research topic in molecular biology. The analysis of epigenetic and expression data is therefore incomplete if RNA-based regulation is not taken into account. Thus, it is increasingly important but not yet standard to combine RNA-centric data and analysis tools with other types of experimental data such as RNA-seq or ChIP-seq. Here, we present the RNA workbench, a comprehensive set of analysis tools and consolidated workflows that enable the researcher to combine these two worlds. Based on the Galaxy framework the workbench guarantees simple access, easy extension, flexible adaption to personal and security needs, and sophisticated analyses that are independent of command-line knowledge. Currently, it includes more than 50 bioinformatics tools that are dedicated to different research areas of RNA biology including RNA structure analysis, RNA alignment, RNA annotation, RNA-protein interaction, ribosome profiling, RNA-seq analysis and RNA target prediction. The workbench is developed and maintained by experts in RNA bioinformatics and the Galaxy framework. Together with the growing community evolving around this workbench, we are committed to keep the workbench up-to-date for future standards and needs, providing researchers with a reliable and robust framework for RNA data analysis. AVAILABILITY: The RNA workbench is available at https://github.com/bgruening/galaxy-rna-workbench.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , RNA/química , Análise de Sequência de RNA/métodos , Software , Biologia Computacional , Internet , Conformação de Ácido Nucleico , RNA/metabolismo , RNA não Traduzido/química , Fluxo de Trabalho
17.
BMC Med Res Methodol ; 17(1): 164, 2017 Dec 08.
Artigo em Inglês | MEDLINE | ID: mdl-29221434

RESUMO

BACKGROUND: The bias caused by drop-out is an important factor in large population-based epidemiological studies. Many studies account for it by weighting their longitudinal data, but to date there is no detailed final approach for how to conduct these weights. METHODS: In this study we describe the observed longitudinal bias and a three-step longitudinal weighting approach used for the longitudinal data in the MoMo baseline (N = 4528, 4-17 years) and wave 1 study with 2807 (62%) participants between 2003 and 2012. RESULTS: The most meaningful drop-out predictors were socioeconomic status of the household, socioeconomic characteristics of the mother and daily TV usage. Weighting reduced the bias between the longitudinal participants and the baseline sample, and also increased variance by 5% to 35% with a final weighting efficiency of 41.67%. CONCLUSIONS: We conclude that a weighting procedure is important to reduce longitudinal bias in health-oriented epidemiological studies and suggest identifying the most influencing variables in the first step, then use logistic regression modeling to calculate the inverse of the probability of participation in the second step, and finally trim and standardize the weights in the third step.


Assuntos
Pacientes Desistentes do Tratamento/estatística & dados numéricos , Adolescente , Viés , Criança , Feminino , Humanos , Modelos Logísticos , Estudos Longitudinais , Masculino
18.
Eur J Pediatr ; 176(1): 99-107, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27888413

RESUMO

To study oxygen saturation (SpO2) targeting before and after training and guideline implementation of manual oxygen titration, two cohorts of preterm infants <30 weeks of gestation needing respiratory support and oxygen therapy were compared. The percentage of the time spent with SpO2 within the target range (85-95%) was calculated (%SpO2-wtr). SpO2 was collected every minute when oxygen is >21%. ABCs where oxygen therapy was given were identified and analyzed. After training and guideline implementation the %SpO2-wtr increased (median interquartile range (IQR)) 48.0 (19.6-63.9) % vs 61.9 (48.5-72.3) %; p < 0.005, with a decrease in the %SpO2 > 95% (44.0 (27.8-66.2) % vs 30.8 (22.6-44.5) %; p < 0.05). There was no effect on the %SpO2 < 85% (5.9 (2.8-7.9) % vs 6.2 (2.5-8) %; ns) and %SpO2 < 80% (1.9 (1.0-3.0) % vs 1.7 (0.8-2.6) %; ns). In total, 186 ABCs with oxygen therapy before and 168 ABCs after training and guideline implementation occurred. The duration of SpO2 < 80% reduced (2 (1-2) vs 1 (1-2) minutes; p < 0.05), the occurrence of SpO2 > 95% did not decrease (73% vs 64%; ns) but lasted shorter (2 (0-7) vs 1 (1-3) minute; p < 0.004). CONCLUSION: Training and guideline implementation in manual oxygen titration improved SpO2 targeting in preterm infants with more time spent within the target range and less frequent hyperoxaemia. The durations of hypoxaemia and hyperoxaemia during ABCs were shorter. What is Known: • Oxygen saturation targeting in preterm infants can be challenging and the compliance is low when oxygen is titrated manually. • Hyperoxaemia often occurs after oxygen therapy for oxygen desaturation during apnoeas. What is New: • Training and implementing guidelines improved oxygen saturation targeting and reduced hyperoxaemia. • Training and implementing guidelines improved manual oxygen titration during ABC.


Assuntos
Doenças do Prematuro/enfermagem , Terapia Intensiva Neonatal/métodos , Enfermagem Neonatal/educação , Oxigenoterapia/enfermagem , Oxigênio/sangue , Guias de Prática Clínica como Assunto , Feminino , Idade Gestacional , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Masculino , Oximetria , Oxigênio/efeitos adversos , Estudos Prospectivos , Estudos Retrospectivos
19.
Br J Anaesth ; 111(2): 178-84, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23485882

RESUMO

BACKGROUND: In this study, we tested the reliability of a non-invasive finger-cuff-based continuous arterial blood pressure monitoring device (Nexfin, BMEYE, Amsterdam, NL) in critically ill surgical patients. METHODS: Invasive intra-arterial and non-invasive arterial pressure measurements from 25 patients during a 4-h period were compared at five time points. Correlation and linear regression analysis were used and mean bias, precision [sd of bias] and limits of agreement (LOA) [bias (2.0 sd)] were calculated using the Bland-Altman method. RESULTS: Eight data pairs were excluded because of error message from the non-invasive technique, and thus a total of 117 data pairs were analysed. Overall, correlation between mean arterial pressure (MAP) was r(2)=0.50. Bias, precision, and LOA between invasive and non-invasive MAP were 6 (12) and -18 to +30 mm Hg. In patients requiring norepinephrine (83 data pairs), correlation was r(2)=0.28 and bias, precision, and LOA were 6 (13) and -20 to +32 mm Hg, whereas in patients not receiving norepinephrine (34 data pairs) r(2) was 0.80 and mean bias, precision, and LOA were 6 (11) and -16 to +28 mm Hg. In patients with peripheral oedema (49 data pairs), r(2) was 0.40 and mean bias, precision and LOA were 7 (15) and -23 to +37 mm Hg. In patients without oedema (64 data pairs), r(2) was 0.66 and mean bias, precision, and LOA were 5 (9) and -13 to +23 mm Hg. CONCLUSIONS: Non-invasive blood pressure monitoring with Nexfin does not seem to be sufficiently accurate to replace intra-arterial invasive blood pressure measurements in critically ill patients.


Assuntos
Pressão Arterial/fisiologia , Monitores de Pressão Arterial , Cateterismo Periférico/estatística & dados numéricos , Estado Terminal , Monitorização Fisiológica/instrumentação , Monitorização Fisiológica/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Determinação da Pressão Arterial/instrumentação , Determinação da Pressão Arterial/métodos , Cateterismo Periférico/métodos , Feminino , Dedos/irrigação sanguínea , Humanos , Masculino , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Adulto Jovem
20.
PLoS One ; 18(10): e0293555, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37883524

RESUMO

BACKGROUND & AIMS: Body composition (BC) changes with age and is associated with morbidity and mortality. A physically active lifestyle influences BC and represents an important predictor of successful aging. To emphasize this, the World Health Organization established activity recommendations for all age groups. We describe BC during adulthood using a cross-sectional sample from a German community and investigate the associations between physical activity (PA), physical fitness (PF), and BC. METHODS: Data from 329 men and women aged 35 to 86 years were analyzed. PA was measured by questionnaire and classified into sport activity and habitual activity. PF was measured through physical performance tests and BC by bioelectrical impedance analysis. Fat mass index (FMI) and fat-free mass index (FFMI) were calculated to represent height-adjusted BC. Associations between PA, PF, and BC were analyzed using linear regression models. RESULTS: For both sexes, strength was positively associated with FFMI (♂: ß = 0.313; ♀: ß = 0.213) and phase angle (♂: ß = 0.357; ♀: ß = 0.409). For FMI, a significant negative association with strength was found only in women (ß = -0.189). Cardiorespiratory fitness showed a negative association with FMI (ß = -0.312) and FFMI (ß = -0.201) for men, while in women a positive association was found for FFMI (ß = 0.186). For coordination, a significant association with FMI was observed only in women (ß = -0.190). Regarding PA only one significant relationship between sport activity and FMI among women (ß = -0.170) was found. CONCLUSIONS: In our sample, PF was closer related to BC than PA. Strength and cardiorespiratory fitness were the strongest predictors for BC. This supports the World Health Organization's activity recommendations to include both resistance and endurance training in the weekly sports program to maintain a healthy BC.


Assuntos
Composição Corporal , Aptidão Física , Masculino , Humanos , Adulto , Feminino , Estudos Transversais , Exercício Físico , Alemanha , Índice de Massa Corporal
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