RESUMO
BACKGROUND: Migrant mothers in high-income countries often encounter more complications during pregnancy, delivery, and the postpartum period. To enlighten health care providers concerning potential barriers, the objective of this study was to explore positive and negative experiences with maternal health services in the University Hospitals of Geneva and Zurich and to describe barriers to maternity services from a qualitative perspective. METHODS: In this qualitative study, six focus groups (FGs) were conducted involving 33 women aged 21 to 40 years. All FG discussions were audio-recorded and later transcribed. Data were analysed using a thematic analysis approach assisted by the Atlas.ti qualitative data management software. RESULTS: Positive experiences included not only the availability of maternity services, especially during emergency situations and the postpartum period, but also the availability of specific maternity services for undocumented migrants in Geneva. Negative experiences were classified into either personal or structural barriers. On the personal level, the main barriers were a lack of social support and a lack of health literacy, whereas the main themes on the structural level were language barriers and a lack of information. CONCLUSION: Structural adaptation is necessary to meet the needs of the extremely diverse population. The needs include (1) the provision of specific information for migrant women in multiple languages, (2) the availability of trained interpreters who are easily accessible to health care providers, (3) specifically trained nurses or social assistance providers to guide migrants through the health system, and (4) a cultural competence-training programme for health care providers.
Assuntos
Barreiras de Comunicação , Conhecimentos, Atitudes e Prática em Saúde , Pessoal de Saúde/normas , Serviços de Saúde Materna/normas , Mães/psicologia , Parto/psicologia , Migrantes/psicologia , Adulto , Competência Cultural , Feminino , Grupos Focais , Acessibilidade aos Serviços de Saúde , Humanos , Serviços de Saúde Materna/organização & administração , Período Pós-Parto , Gravidez , Pesquisa Qualitativa , Suíça , Adulto JovemRESUMO
Genital prolapse is frequent and can be found in about 50% of parous women. Its etiology is complex and multifactorial. Predisposing factors include: genetics (connective tissue disorders, family history); general state (age, parity, weight, smoking, obstructive pulmonary disease); trauma (carrying heavy loads, intense physical exercise); or iatrogenic (post hysterectomy). Treatment can be conservative or surgical and depends mainly on the severity of symptoms. Developments in surgical techniques and synthetic material in the last 20 years enabled us to use minimally invasive procedures with improved post operative course and decreased recurrence rates.
Assuntos
Laparoscopia , Telas Cirúrgicas , Prolapso Uterino/cirurgia , Feminino , Seguimentos , Humanos , Pessoa de Meia-Idade , Complicações Pós-OperatóriasRESUMO
RNA-binding proteins are essential for spermatogenesis: they are required in the nucleus of germ cells, for the production of specific mRNA isoforms, and in the cytoplasm - where proteins required for chromatin condensation and for changes in cell morphology are translated long after transcription ceases. Some of the RNA targets and the RNA-binding proteins themselves have been identified recently. Both nuclear and cytoplasmic proteins are affected in examples of azoospermia in men.
Assuntos
Infertilidade Masculina/fisiopatologia , Proteínas de Ligação a RNA/fisiologia , Espermatogênese/fisiologia , Animais , Citoplasma/metabolismo , Humanos , Masculino , Proteínas Nucleares/fisiologiaAssuntos
Abscesso Abdominal/microbiologia , Doenças dos Anexos/microbiologia , Bacteriemia/microbiologia , Doenças do Prematuro/microbiologia , Infecções Pneumocócicas/microbiologia , Infecção Puerperal/microbiologia , Streptococcus pneumoniae/isolamento & purificação , Abscesso Abdominal/diagnóstico , Abscesso Abdominal/terapia , Doenças dos Anexos/diagnóstico , Doenças dos Anexos/terapia , Adulto , Feminino , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Masculino , Infecções Pneumocócicas/congênito , Período Pós-Parto , Gravidez , Infecção Puerperal/diagnóstico , Infecção Puerperal/terapiaRESUMO
Efficient splicing of the 5'-most intron of pre-mRNA requires a 5' m7G(5')ppp(5')N cap, which has been implicated in U1 snRNP binding to 5' splice sites. We demonstrate that the cap alters the kinetic profile of U1 snRNP binding, but its major effect is on U6 snRNA binding. With two alternative wild-type splice sites in an adenovirus pre-mRNA, the cap selectively alters U1 snRNA binding at the site to which cap-independent U1 snRNP binding is stronger and that is used predominantly in splicing; with two consensus sites, the cap acts on both, even though one is substantially preferred for splicing. However, the most striking quantitative effect of the 5' cap is neither on U1 snRNP binding nor on the assembly of large complexes but on the replacement of U1 snRNP by U6 snRNA at the 5' splice site. Inhibition of splicing by a cap analogue is correlated with the loss of U6 interactions at the 5' splice site and not with any loss of U1 snRNP binding.
Assuntos
Capuzes de RNA/genética , Precursores de RNA/genética , Splicing de RNA/genética , RNA Nuclear Pequeno/genética , Ribonucleoproteína Nuclear Pequena U1/genética , Processamento Alternativo/genética , Núcleo Celular/genética , Reagentes de Ligações Cruzadas/metabolismo , Fosfatos de Dinucleosídeos/genética , Ficusina/metabolismo , Células HeLa , Humanos , Cinética , Ribonuclease H/metabolismo , Transcrição Gênica/genéticaRESUMO
The human alpha-tropomyosin gene hTMnm has two mutually exclusive versions of exon 5 (NM and SK), one of which is expressed specifically in skeletal muscle (exon SK). A minigene construct expresses only the nonmuscle (NM) isoform when transfected into COS-1 cells and both forms when transfected into myoblasts. Twenty-four mutants were produced to determine why the SK exon is not expressed in COS cells. The results showed that exons NM and SK are not in competition for splicing to the flanking exons and that there is no intrinsic barrier to splicing between the exons. Instead, exon SK is skipped whenever there are flanking introns. Splicing of exon SK was induced when the branch site sequence 70 nucleotides upstream of the exon was mutated to resemble the consensus and when the extremities of the exon itself were changed to the corresponding NM sequence. Precise swaps of the NM and SK exon sequences showed that the exon sequence effect was dominant to that of intron sequences. The mechanism of regulation appears to be unlike that of other tropomyosin genes. We propose that exclusion of exon SK arises because its 3' splicing signals are weak and are prevented by an exon-specific repressor from competing for splice site recognition.
Assuntos
Éxons , Músculos/metabolismo , Splicing de RNA , Tropomiosina/genética , Sequência de Bases , Linhagem Celular , DNA , Regulação da Expressão Gênica , Humanos , Dados de Sequência Molecular , Mutação , Transcrição GênicaRESUMO
The first component known to recognize and discriminate among potential 5' splice sites (5'SSs) in pre-mRNA is the U1 snRNP. However, the relative levels of U1 snRNP binding to alternative 5'SSs do not necessarily determine the splicing outcome. Strikingly, SF2/ASF, one of the essential SR protein-splicing factors, causes a dose-dependent shift in splicing to a downstream (intron-proximal) site, and yet it increases U1 snRNP binding at upstream and downstream sites simultaneously. We show here that hnRNP A1, which shifts splicing towards an upstream 5'SS, causes reduced U1 snRNP binding at both sites. Nonetheless, the importance of U1 snRNP binding is shown by proportionality between the level of U1 snRNP binding to the downstream site and its use in splicing. With purified components, hnRNP A1 reduces U1 snRNP binding to 5'SSs by binding cooperatively and indiscriminately to the pre-mRNA. Mutations in hnRNP A1 and SF2/ASF show that the opposite effects of the proteins on 5'SS choice are correlated with their effects on U1 snRNP binding. Cross-linking experiments show that SF2/ASF and hnRNP A1 compete to bind pre-mRNA, and we conclude that this competition is the basis of their functional antagonism; SF2/ASF enhances U1 snRNP binding at all 5'SSs, the rise in simultaneous occupancy causing a shift in splicing towards the downstream site, whereas hnRNP A1 interferes with U1 snRNP binding such that 5'SS occupancy is lower and the affinities of U1 snRNP for the individual sites determine the site of splicing.
Assuntos
Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B , Proteínas Nucleares/metabolismo , Sítios de Splice de RNA , Ribonucleoproteína Nuclear Pequena U1/metabolismo , Ribonucleoproteínas/metabolismo , Processamento Alternativo , Sítios de Ligação , Ligação Competitiva , Ribonucleoproteína Nuclear Heterogênea A1 , Ribonucleoproteínas Nucleares Heterogêneas , Modelos Biológicos , Proteínas Nucleares/genética , Precursores de RNA/genética , Precursores de RNA/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Ribonucleoproteína Nuclear Pequena U1/genética , Ribonucleoproteínas/genética , Fatores de Processamento de Serina-ArgininaRESUMO
Alternative pre-mRNA splicing of two terminal exons (alpha and beta) regulates the expression of the human DNA ligase III gene. In most tissues, the alpha exon is expressed. In testes and during spermatogenesis, the beta exon is used instead. The alpha exon encodes the interaction domain with a scaffold DNA repair protein, XRCC1, while the beta exon-encoded C-terminal does not. Sequence elements regulating the alternative splicing pattern were mapped by in vitro splicing assays in HeLa nuclear extracts. Deletion of a region beginning in the beta exon and extending into the downstream intron derepressed splicing to the beta exon. Two silencing elements were found within this 101 nt region: a 16 nt exonic splicing silencer immediately upstream of the beta exon polyadenylation signal and a 45 nt intronic splicing silencer. The exonic splicing silencer inhibited splicing, even when the poly-adenylation signal was deleted or replaced by a 5' splice site. This element also enhanced polyadenylation under conditions unfavourable to splicing. The splicing silencer partially inhibited assembly of spliceo-somal complexes and functioned in an adenoviral pre-mRNA context. Silencing of splicing by the element was associated with cross-linking of a 37 kDa protein to the RNA substrate. The element exerts opposite functions in splicing and polyadenylation.
Assuntos
DNA Ligases/genética , Éxons , Inativação Gênica , Splicing de RNA , Testículo/enzimologia , Sequência de Bases , Núcleo Celular/enzimologia , DNA Ligase Dependente de ATP , Primers do DNA , Células HeLa , Humanos , Masculino , Mutagênese , Proteínas de Ligação a Poli-ADP-Ribose , Proteínas de XenopusRESUMO
Current methods for measuring the efficiency of splicing in mammalian cells rely on either direct analysis of the RNA, which does not lend itself to rapid assays, or on single reporter functions that are subject to numerous intrinsic variables. If two protein activities are encoded within a single reading frame but on separate exons, with an intervening sequence containing termination codons, then the expression of the second activity is dependent on removal of the intervening sequence by pre-mRNA splicing. Thus, the ratio of the activities encoded by exon 2 to exon 1 reflects the ratio of expression from spliced mRNA to the total expression of spliced and unspliced RNA. This provides a rapid and convenient assay for the effects on splicing efficiency of trans-acting factors or of alterations in the sequences of the intron and surrounding exon sequences.
Assuntos
Genes Reporter , Técnicas Genéticas , Splicing de RNA , RNA Mensageiro/análise , Animais , Linhagem Celular , Humanos , Luciferases/análise , Luciferases/genética , Mamíferos , RNA Mensageiro/metabolismo , Proteínas Recombinantes de Fusão/análise , beta-Galactosidase/análise , beta-Galactosidase/genéticaRESUMO
The effects of DNA gyrase and quinolone drugs on in vitro transcription of a template containing a preferred gyrase cleavage site have been investigated. We have found that gyrase-quinolone complexes with DNA lead to blocking of transcription by Escherichia coli and bacteriophage T7 RNA polymerases. Either gyrase or quinolone alone has no effect on transcription. With DNA gyrase containing a point mutation in the gyrase A protein, known to confer quinolone resistance, blocking was found to occur only at much higher concentrations of the drug. Other agents that inhibit gyrase-catalysed supercoiling (novobiocin and 5'-adenylyl-beta,gamma-imidodiphosphate) do not arrest transcription in the presence of gyrase. Mapping of the transcription termination sites in the presence of gyrase and quinolones shows that blocking occurs about 10 to 20 base-pairs upstream of the gyrase cleavage site. Analysis of transcription in the absence of drug suggests that RNA polymerase does not displace gyrase from the template. These results are discussed in the light of models for the bactericidal effects of quinolone drugs.
Assuntos
DNA Topoisomerases Tipo II/metabolismo , RNA Polimerases Dirigidas por DNA/antagonistas & inibidores , DNA/metabolismo , Quinolonas/metabolismo , Transcrição Gênica , Adenilil Imidodifosfato/farmacologia , Bacteriófago T7/enzimologia , Sequência de Bases , RNA Polimerases Dirigidas por DNA/metabolismo , Escherichia coli/enzimologia , Dados de Sequência Molecular , Novobiocina/farmacologia , Plasmídeos , Regiões Terminadoras Genéticas , Transcrição Gênica/efeitos dos fármacos , Proteínas ViraisRESUMO
The relationship between preferences among alternative 5' splice sites and their sequences has been investigated for 37 sequences by assessing their use in splicing relative to the 5' splice site of IVS-2 of rabbit beta-globin. There are strong correlations between the intrinsic strength of a 5' splice site and both the extent to which it resembles the consensus sequence and the calculated stability of its interactions with U1 small nuclear RNA. However, present methods of calculating either of the latter values do not allow predictions to be made of the relative preferences among a small number of sequences.
Assuntos
Globinas/genética , Splicing de RNA , RNA Mensageiro/genética , RNA Nuclear Pequeno/genética , Animais , Sequência de Bases , Clonagem Molecular , Genes , Células HeLa/metabolismo , Humanos , Dados de Sequência Molecular , Sondas de Oligonucleotídeos , Precursores de RNA/genética , Coelhos , Ratos , Endonucleases Específicas para DNA e RNA de Cadeia SimplesRESUMO
Single nucleotide changes to the sequence between two alternative 5' splice sites, separated by 25 nucleotides in a beta-globin gene derivative, caused substantial shifts in pre-mRNA splicing preferences, both in vivo and in vitro. An activating sequence for splicing was located. Models for the recognition by U1 small nuclear ribonucleoproteins (snRNPs) of competing 5' splice sites were tested by altering the distance separating the two sites. Use of the upstream splice site declined sharply when it was separated from the downstream (natural) site by distances of 40 nucleotides or more. This effect was reversed in vivo, but not in vitro, by altering the upstream sequence to that of a consensus 5' splice site sequence. Dilution of an extract used for splicing in vitro shifted preferences when the sites were close towards the downstream site. We conclude that the mechanism of selection depends on the distance apart of the potential splice sites and that with close sites steric interference between factors bound to both sites may impede splicing and affect splicing preferences.
Assuntos
Splicing de RNA , RNA Mensageiro/genética , Ribonucleoproteínas/fisiologia , Animais , Sequência de Bases , Clonagem Molecular , Análise Mutacional de DNA , Globinas/genética , Técnicas In Vitro , Dados de Sequência Molecular , Coelhos , Ribonucleoproteínas Nucleares Pequenas , Relação Estrutura-AtividadeRESUMO
Several groups have investigated the role of T cells in the pathogenesis of psoriasis by determination of T-cell receptor (TCR) B-chain variable (V) region usage, both in chronic plaque (psoriasis vulgaris) and guttate forms, with various results. Because there are no data on TCR expression in early psoriasis vulgaris, when specific cellular immune events may be expected to be most pronounced, we have analyzed early lesions (less than 3 wk old) of ten patients, with highly reproducible results. We have developed a highly controlled anchored polymerase chain reaction (PCR) method in which TCR beta chain species are all amplified with the same primer pair and products are quantified by dot blot hybridization with BV family-specific oligonucleotide probes. Overexpression of certain TCR BV genes was observed in the majority of lesional biopsies, but in samples in which the expanded BV family formed more than 10% of total lesional BV (half of the samples analyzed), BV2 and BV6 predominated. The consistency of overexpression of these BV species between patients was much less than in previous studies of TCRBV usage in established chronic plaque psoriasis lesions. Complementarity-determining region 3 (CDR3) size spectratyping demonstrated evidence for selective clonal T cell accumulation in less than half of the lesional samples showing BV expansion. These results indicate that selective amplification of TCRBV species occurs in early psoriasis vulgaris but is not essential to the pathogenic process and may be more important in the maintenance or expansion of chronic lesions.
Assuntos
Psoríase/genética , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Adulto , Biópsia , Células Clonais , Feminino , Humanos , Região Variável de Imunoglobulina/biossíntese , Região Variável de Imunoglobulina/sangue , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , Masculino , Pessoa de Meia-Idade , Psoríase/sangue , Receptores de Antígenos de Linfócitos T alfa-beta/biossíntese , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Reprodutibilidade dos Testes , Pele/patologia , Linfócitos T/citologiaRESUMO
Previous studies have come to different conclusions about the presence of RNA in particles known variously as prosomes, proteasomes or multicatalytic proteinase (MCP). To determine the reason for this, MCP was isolated from rat liver by 4 different purification protocols. One major band of RNA, about 80 nucleotides in length, co-purified in all preparations. The amount of RNA detected was less than one molecule per MCP particle suggesting that there may be more than one population of MCP in rat liver cells.
Assuntos
Cisteína Endopeptidases/química , Fígado/enzimologia , Complexos Multienzimáticos/química , RNA/análise , Animais , Peso Molecular , Complexo de Endopeptidases do Proteassoma , Ratos , Ribonucleoproteínas/químicaRESUMO
OBJECTIVES: To evaluate the technique of laparoscopic lateral colpo-uterine suspension using a mesh to treat pelvic organ prolapse, with a sufficient follow-up. PATIENTS AND METHODS: The technique consists of two steps. First, the lateral suspension of the vaginal vault and of the uterus is performed using a polypropylene mesh placed in the vesicovaginal septum as a transversal hammock. The second step is the application of a polypropylene patch to the posterior surface of the vagina and the rectovaginal septum. The transversal hammock is placed laterally by the tension-free fixation of the mesh to the lateral abdominal wall above the iliac crests. Between January 2004 and December 2007, 218 patients were treated. It is a continuous series including all the patients needing a surgical procedure to treat a genital prolapse. We excluded, from the study, the patients with a severe cardiorespiratory disease and the cases of isolated rectocele. RESULTS: We observed a recurrence of the prolapse in thirty patients (13.76%). A reoperation was performed in 10 patients (4.6%). One complication was related to the technique of lateral suspension (bladder injury immediately sutured 0.46%). A mesh erosion was noted in 13 cases (5.96%), nine were treated by vaginal excision of the mesh (4.12%). CONCLUSIONS: The laparoscopic lateral colpo-uterine suspension using a mesh corrects the pelvic organ prolapse with a predominant cystocele or rectocele. It is an interesting alternative to the other procedures because of the low risk of complications and the satisfactory results.
Assuntos
Laparoscopia/métodos , Prolapso de Órgão Pélvico/cirurgia , Telas Cirúrgicas , Idoso , Feminino , Humanos , Complicações Intraoperatórias/epidemiologia , Laparoscopia/efeitos adversos , Laparoscopia/instrumentação , Pessoa de Meia-Idade , Complicações Pós-Operatórias/epidemiologia , Recidiva , Resultado do Tratamento , Incontinência Urinária/cirurgia , Útero , VaginaAssuntos
DNA Mitocondrial , Aminoácidos/análise , Animais , Sequência de Bases , Bovinos , Códon , Replicação do DNA , Genes , Humanos , RNA Ribossômico , RNA de TransferênciaRESUMO
A method is described for the preparation of single-stranded DNA from clones in bacteriophage M13 vectors. This procedure allows multiples of 96 clones to be processed at once, utilizing filtration to remove host cells and simplifying the treatment of bacteriophage pellets. The DNA produced can be used for sequencing of mutagenesis.
Assuntos
Bacteriófagos/análise , DNA Viral/isolamento & purificação , Sequência de Bases , DNA Viral/análise , Eletroforese em Gel de Poliacrilamida , Filtração , MutaçãoRESUMO
The neural cell adhesion molecule (N-CAM) is an important mediator of calcium independent cell-cell interactions. Variations in the primary structure of the protein are due to alternative splicing of pre-mRNA in the region encoding the extracellular, trans-membrane and cytoplasmic domains. In order to identify the patterns of exon usage during development of skeletal muscle and brain of the mouse, a coupled reverse-transcriptase/polymerase chain reaction was used to identify the murine homologues of the muscle-specific domain (MSD), located between exons 12 and 13 in human N-CAM mRNA. The cDNAs produced have been cloned and sequenced, or analysed directly. The amplification reactions were shown to maintain the concentration ratios of the initial cDNAs. The results indicate that the mouse homologue to exon MSD1a is under tissue and developmental regulation that is independent of exons MSD1b and MSD1c. The inclusion of the triplet exon AAG is also regulated in a cell- and stage-specific manner, which is independent of the other alternatively spliced exons of this domain.
Assuntos
Moléculas de Adesão Celular Neuronais/genética , Éxons/genética , Desenvolvimento Muscular , Splicing de RNA/fisiologia , Animais , Sequência de Bases , Encéfalo/crescimento & desenvolvimento , Galinhas , Clonagem Molecular , Regulação da Expressão Gênica/fisiologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Músculos/química , Reação em Cadeia da Polimerase , DNA Polimerase Dirigida por RNA/metabolismo , Homologia de Sequência do Ácido NucleicoRESUMO
We have carried out a systematic investigation of the efficiency of misincorporation by Avian Myeloblastosis Virus reverse transcriptase with all possible combinations of dNTP substrate, template nucleotide, and the nucleotide at the 3' terminus of the primer. A series of synthetic oligonucleotide primers were annealed to single stranded M13 DNA templates, and a single dNTP was misincorporated at the primer 3' end using AMV reverse transcriptase. The proportion and pattern of misincorporation and incorporation in all 64 situations was assayed using [5'-32p] labelled primers, and the products were separated on denaturing polyacrylamide gels. Correct incorporations occurred more readily than misincorporations. The efficiency of misincorporation depended on the individual primer, but, comparing primers, a clear dependence on the template nucleotide was observed for the preferential misincorporation of different dNTPs. The exact combination of template and dNTP was important; although purine:pyrimidine (dNTP substrate:template nucleotide) and pyrimidine:purine misincorporations occurred comparatively readily, some pyrimidine:pyrimidine and purine:purine reactions were equally efficient and yet others were never seen to occur. Some misincorporations were facilitated by subsequent correct incorporations, but despite this our results suggest that the level of misincorporation is limited by the rate of reaction and enzyme inactivation rather than by exonuclease activity.
Assuntos
DNA Polimerase Dirigida por RNA/metabolismo , Vírus da Mieloblastose Aviária/enzimologia , Sequência de Bases , Desoxirribonucleotídeos/metabolismo , Cinética , Moldes GenéticosRESUMO
The nucleotide sequence spanning the ribosomal RNA (rRNA) genes of cloned human mitochondrial DNA reveals an extremely compact genome organization wherein the putative tRNA genes are probably 'butt-jointed' around the two rRNA genes. The sequences of the rRNA genes are significantly homologous in some regions to eukaryotic and prokaryotic sequences, but distinctive; the tRNA genes also have unusual nucleotide sequences. It seems that human mitochondria did not originate from recognizable relatives of present day organisms.