RESUMO
This study examined the effects of constant current electrical stimulation (CCES) compared to constant voltage electrical stimulation (CVES), when applied within the same beef carcass (n = 79), on longissimus thoracis et lumborum (LTL) quality and palatability. There was a stimulation method × time interaction for pH, with CCES reducing the 3 h post-mortem pH, but increasing the 72 h post-mortem pH compared to CVES (P < 0.001). The CCES decreased the meat subjective Japanese Meat Grading Agency (JMGA) colour scores (P < 0.05) and increased the objective Lâ (P < 0.01), aâ (P < 0.05) and bâ (P < 0.05) colour values at 3 d post-mortem and Lâ and bâ values (P < 0.05) during retail display compared to CVES, although the objective values from both stimulation methods were above established consumer acceptability thresholds. Additionally, CCES reduced the purge (P < 0.05) and drip (P < 0.01) losses, and tended to reduce shear force values (P = 0.089) compared to CVES, although these did not translate into differences in juiciness or tenderness evaluated by trained panelists (P > 0.1). Regarding flavour, the CCES meat had greater bloody/serumy flavour (P < 0.05) and corn aroma (P < 0.05), less unidentified aroma (P < 0.05), and tended to have greater corn flavour (P = 0.077) and less barnyard aroma (P = 0.079) than CVES meat. There were also increased concentrations of flavour-related volatile compounds including 2-methyl-butanal, 3-methyl-butanal and 2-5-dimethyl pyrazine levels (P < 0.05) with CCES. Overall, the CCES system slightly improved meat quality and flavour compared to CVES when applied to the same beef carcasses. Further consumer studies would be warranted to determine whether these differences translate into more acceptable meat.
Assuntos
Cor , Comportamento do Consumidor , Estimulação Elétrica , Carne Vermelha , Paladar , Animais , Bovinos , Carne Vermelha/análise , Estimulação Elétrica/métodos , Humanos , Concentração de Íons de Hidrogênio , Músculo Esquelético/química , Masculino , Feminino , Adulto , Manipulação de Alimentos/métodosRESUMO
This study evaluated the effects of a constant current electrical stimulation (CCES) system and hormonal growth-promoting (HGP) implants on the quality and palatability of the longissimus thoracis et lumborum (LTL) from yearling-finished steers. The experiment used a total of 46 Angus cross steers, which were either non-implanted (n = 20) or implanted with trenbolone acetate and estradiol benzoate (n = 26). The CCES was applied to one side of each carcass during the slaughter process, whereas the other side remained unstimulated. Regardless of the application of HGP implants, the CCES reduced pH at 3 and 72 h post-mortem and shear force at all ageing times (P < 0.05), improved colour at 72 h post-mortem and during the retail display (P < 0.05), increased initial and overall tenderness (P < 0.01), and decreased the amount of perceived connective tissue and the proportion of trained panelists detecting spongy texture (P < 0.05) compared to meat from unstimulated carcass sides. Although CCES increased meat purge losses and reduced moisture content (P < 0.05), this did not affect meat juiciness (P > 0.10). CCES interacted with HGP to prevent increase in drip loss (P > 0.10), increase frequency of panelists detecting bloody/serumy flavour and typical texture, and reduce the proportion of panelists detecting rubbery texture in meat (P < 0.05). Regardless of stimulation treatment, meat from implanted animals had a more pronounced pH decline at 72 h post-mortem (P < 0.05) and a higher proportion of panelists finding no off-flavours (P < 0.05) or bloody/serumy flavour (P < 0.01) than non-implanted cattle. The CCES system tested in this study improved LTL quality and palatability of heavier beef carcasses.
Assuntos
Anabolizantes , Músculo Esquelético , Bovinos , Animais , Músculo Esquelético/fisiologia , Carne , Acetato de Trembolona/farmacologia , Anabolizantes/farmacologia , Estimulação ElétricaRESUMO
An automated method of estimating the spatial distribution of piglets within a pen was used to assess huddling behaviour under normal conditions and during a febrile response to vaccination. The automated method was compared with a manual assessment of clustering activity. Huddling behaviour was partly related to environmental conditions and clock time such that more huddling occurred during the night and at lower ambient air temperatures. There were no positive relationships between maximum pig temperatures and environmental conditions, suggesting that the narrow range of air temperatures in this study was not a significant factor for pig temperature. Spatial distribution affected radiated pig temperature measurements by IR thermography. Higher temperatures were recorded in groups of animals displaying huddling behaviour. Huddling behaviour was affected by febrile responses to vaccination with increased huddling occurring 3 to 8 h post-vaccination. The automated method of assessing spatial distribution from an IR image successfully identified periods of huddling associated with a febrile response, and to changing environmental temperatures. Infrared imaging could be used to quantify temperature and behaviour from the same images.
Assuntos
Comportamento Animal , Febre/veterinária , Comportamento Social , Suínos/fisiologia , Termografia/veterinária , Vacinação/veterinária , Animais , Temperatura Corporal , Meio Ambiente , Suínos/imunologia , Termografia/métodosRESUMO
An automated, non-invasive system for monitoring of thermoregulation has the potential to mitigate swine diseases through earlier detection. Measurement of radiated temperature of groups of animals by infrared thermography (IRT) is an essential component of such a system. This study reports on the feasibility of monitoring the radiated temperature of groups of animals as a biomarker of immune response using vaccination as a model for febrile disease. In Study A, weaned pigs were either treated with an intramuscular vaccine (FarrowSure Gold), a sham injection of 0.9% saline or left as untreated controls. An infrared thermal camera (FLIR A320) was fixed to the ceiling directly above the pen of animals, and recorded infrared images of the treatment groups at 5 min intervals. The effect on temperature of the spatial distribution of pigs within the pen was significant, with higher temperatures recorded when pigs were grouped together into a single cluster. A higher frequency of clustering behaviour was observed in vaccinated animals compared with controls during a period of the afternoon ~4 to 7 h post-vaccination. The daily mean of the maximum image temperature was significantly higher in vaccinated animals compared with control and sham-treated animals. In the vaccination treated group, the 24 h mean of the maximum temperature was significantly higher during the post-vaccination period compared with the 24 h period before vaccination. Increased temperature in the vaccinated animals occurred from ~3 h, peaked at ~10 h, and remained elevated for up to 20 h post-vaccination. In Study B, the effect of prevalence was tested in terms of the difference in maximum temperature between control and vaccination days. A thermal response to vaccination was detected in a pen of 24 to 26 animals when <10% of the animals were vaccinated. The results support the concept of radiated temperature measurements of groups of animals by IRT as a screening tool for febrile diseases in pig barns.
Assuntos
Febre/veterinária , Doenças dos Suínos/diagnóstico , Suínos/imunologia , Termografia/veterinária , Vacinação/veterinária , Animais , Comportamento Animal , Temperatura Corporal , Febre/diagnóstico , Raios Infravermelhos , Injeções Intramusculares/veterinária , Doenças dos Suínos/prevenção & controle , Termografia/métodosRESUMO
We describe the synthesis of cellulose fibers densely grafted with the cationic polymer poly[2-(methacryloyloxy)ethyl]-trimethylammoniumchloride (PMeDMA) through aqueous ATRP. The hydroxyl groups present on the cellulose surface were exploited to initiate the ATRP polymerization of MeDMA. We first grafted a bromide initiator, known to be an efficient initiator for ATRP, on the cellulose surface from which the polymer was then directly grown. The resulting fibers/PMeDMA complex was analyzed with infra-red, XPS and SEM techniques and present clear evidences that the polymer is present on the cellulose surface. In order to better characterize the polymer, sacrificial initiators were also added in the mixture and subsequently recovered for analysis. Size exclusion chromatography shows that the polymerization in this heterogeneous medium was controlled. Finally, we show that the mechanical properties of test hand sheets made from modified pulp are markedly improved by the grafting of the cationic PMeDMA.
Assuntos
Celulose/síntese química , Metacrilatos/química , Cátions/química , Celulose/química , Tamanho da Partícula , Propriedades de Superfície , Água/químicaRESUMO
UNLABELLED: Fifty to eighty-five percent of schizophrenic patients are impaired on ocular pursuit paradigms. However, results regarding the relatives are more discordant. The aim of this study was to investigate whether eye movement disorders could be a vulnerability marker of schizophrenia. METHOD: Twenty-one schizophrenic patients (DSM-IV), 31 first-degree relatives of those patients without schizophrenic spectrum disorders, and two groups of healthy controls matched by age and sex were included. Three oculomotor tasks (smooth pursuit, reflexive saccades and antisaccades) were used. RESULTS: Patients had a lower averaged gain (P= 0.035) during smooth pursuit than controls, made less correct visually guided saccades (P< 0.001) and more antisaccades errors (P= 0.002) than controls. In contrast, none of the comparison between the relatives and their controls was significant. CONCLUSION: Schizophrenic patients were impaired on smooth pursuit and antisaccade paradigms. None of these impairments was, however, observed in their first-degree relatives. Our results suggest that the eye movement parameters tested could not be considered as vulnerability markers for schizophrenia.
Assuntos
Transtornos da Motilidade Ocular/genética , Esquizofrenia/genética , Adulto , Suscetibilidade a Doenças , Feminino , Marcadores Genéticos/genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Transtornos da Motilidade Ocular/diagnóstico , Acompanhamento Ocular Uniforme/genética , Reflexo/genética , Movimentos Sacádicos/genética , Esquizofrenia/diagnósticoRESUMO
Familial idiopathic basal ganglia calcification (FIBGC) is a rare condition and its pathophysiology has not so far been elucidated. We report the results of a clinical study in two patients of a family affected with FIBGC. Brain imaging with 18-FDG-PET was performed in one. Psychiatric and cognitive troubles were the main clinical symptoms. Basal ganglia calcifications were associated with white matter lesions. The PET study performed in one patient revealed a striatal and a posterior cingulate hypometabolism. Posterior cingulate gyrus is involved in episodic memory processing, and could be involved in episodic memory deficit observed in this patient. These results suggest that a cortical dysfunction could be associated to the disease. The underlying mechanism, that could be a neuronal loss, a cortical deafferentation or an alteration of synaptic transmission, remains to be elucidated.
Assuntos
Encefalopatias/genética , Calcinose/genética , Giro Denteado/patologia , Globo Pálido/patologia , Neostriado/patologia , Adulto , Idoso , Química Encefálica , Encefalopatias/diagnóstico por imagem , Encefalopatias/metabolismo , Calcinose/diagnóstico por imagem , Calcinose/metabolismo , Giro Denteado/diagnóstico por imagem , Família , Feminino , Fluordesoxiglucose F18 , Globo Pálido/diagnóstico por imagem , Humanos , Masculino , Pessoa de Meia-Idade , Neostriado/diagnóstico por imagem , Linhagem , Tomografia Computadorizada de Emissão , Tomografia Computadorizada de Emissão de Fóton Único , Tomografia Computadorizada por Raios XAssuntos
Processamento Alternativo , Núcleo Celular/metabolismo , Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B/química , Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B/metabolismo , Precursores de RNA/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Éxons , Ribonucleoproteína Nuclear Heterogênea A1 , Humanos , Íntrons , Dados de Sequência MolecularRESUMO
BACKGROUND: Familial idiopathic basal ganglia calcification (FIBGC) is generally associated with neurological and psychiatric symptoms. An association between FIBGC and schizophrenia has been described but it remains uncertain. We studied the relationship between the presence and extent of basal ganglia calcification and schizophrenia in a multiply affected family. METHOD: Symmetrical basal ganglia calcifications (BGC) were detected on computerized tomography (CT) in a schizophrenic proband and led us to carry out CTs and standardized psychiatric evaluations (SADS--Endicott & Spitzer, 1978) in all available first-degree relatives (mother and six siblings). RESULTS: Five subjects had BGC, including three subjects diagnosed as schizophrenic. Three subjects had no BGC and none of them was diagnosed as schizophrenic. We subdivided the BGC into three groups: massive (pallidum, striatum and dentate nuclei affected); medium (pallidum and striatum); and mild (pallidum only). The two subjects with massive BGC and one of the two with medium BGC had schizophrenia. The subject with mild BGC had no psychotic symptoms. CONCLUSION: Our results are consistent with the hypothesis that BGC favours the occurrence of a schizophrenia-like syndrome and that the risk of occurrence of this syndrome is proportional to the extent of calcification. These findings support the hypothesis that schizophrenia is determined by a disruption of thalamo-cortico-striatal circuits.
Assuntos
Doenças dos Gânglios da Base/genética , Doenças dos Gânglios da Base/psicologia , Calcinose/genética , Esquizofrenia/genética , Adulto , Doenças dos Gânglios da Base/complicações , Calcinose/complicações , Feminino , Humanos , Masculino , Linhagem , Esquizofrenia/etiologia , Tomografia Computadorizada por Raios XRESUMO
The hnRNP A1 protein and a shortened derivative (UP1) promote telomere elongation in mammalian cells. In support of a direct role for A1 in telomere biogenesis, we have shown that the recombinant UP1 protein binds to telomeric DNA sequences in vitro, and pulls down telomerase activity from a cell extract. Here we show that A1/UP1 can interact directly with the RNA component of human telomerase (hTR). A portion of A1/UP1 that contains RNA recognition motif 2 (RRM2) is sufficient for an interaction with the first 208 nt of hTR. Given that the portion of A1/UP1 that contains RRM1 is sufficient for binding to a telomeric DNA oligonucleotide, we have tested whether A1/UP1 can interact simultaneously with both nucleic acids. Using a chromatography assay, we find that A1/UP1 bound to hTR can interact with telomeric DNA. Notably, these interactions are sufficiently robust to withstand incubation in a cell extract. Our results suggest that hnRNP A1 may help recruit telomerase to the ends of chromosomes.
Assuntos
DNA/metabolismo , Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B , RNA/metabolismo , Ribonucleoproteínas/metabolismo , Telomerase/genética , Telômero/genética , Sítios de Ligação , DNA/genética , Ribonucleoproteína Nuclear Heterogênea A1 , Ribonucleoproteínas Nucleares Heterogêneas , Humanos , Conformação de Ácido Nucleico , Ligação Proteica , Estrutura Terciária de Proteína , RNA/química , Ribonucleoproteínas/químicaRESUMO
Using a protein truncation assay, we have identified a new mutation in the neurofibromatosis type 1 (NF1) gene that causes a severe defect in NF1 pre-mRNA splicing. The mutation, which consists of a G to A transition at position +1 of the 5' splice site of exon 12a, is associated with the loss of both exons 11 and 12a in the NF1 mRNA. Through the use of in vivo and in vitro splicing assays, we show that the mutation inactivates the 5' splice site of exon 12a, and prevents the definition of exon 12a, a process that is normally required to stimulate the weak 3' splice site of exon 12a. Because the 5' splice site mutation weakens the interaction of splicing factors with the 3' splice site of exon 12a, we propose that exon 11/exon 12a splicing is also compromised, leading to the exclusion of both exons 11 and 12a. Our results provide in vivo support for the importance of the exon definition model during NF1 splicing, and suggest that the NF1 region containing exons 11 and 12a plays an important role in the activity of neurofibromin.
Assuntos
Processamento Alternativo/genética , Éxons/genética , Mutação/genética , Proteínas do Tecido Nervoso/genética , Neurofibromatose 1/genética , Sequência de Bases , Southern Blotting , Linhagem Celular Transformada , Análise Mutacional de DNA , Feminino , Humanos , Modelos Genéticos , Proteínas do Tecido Nervoso/química , Neurofibromina 1 , Sítios de Splice de RNA/genética , Deleção de Sequência , Células Tumorais CultivadasRESUMO
Depending on the cell lines and cell types, dimethyl sulfoxide (Me2SO) can induce or block cell differentiation and apoptosis. Although Me2SO treatment alters many levels of gene expression, the molecular processes that are directly affected by Me2SO have not been clearly identified. Here, we report that Me2SO affects splice site selection on model pre-mRNAs incubated in a nuclear extract prepared from HeLa cells. A shift toward the proximal pair of splice sites was observed on pre-mRNAs carrying competing 5'-splice sites or competing 3'-splice sites. Because the activity of recombinant hnRNP A1 protein was similar when added to extracts containing or lacking Me2SO, the activity of endogenous A1 proteins is probably not affected by Me2SO. Notably, in a manner reminiscent of SR proteins, Me2SO activated splicing in a HeLa S100 extract. Moreover, the activity of recombinant SR proteins in splice site selection in vitro was improved by Me2SO. Polar solvents like DMF and formamide similarly modulated splice site selection in vitro but formamide did not activate a HeLa S100 extract. We propose that Me2SO improves ionic interactions between splicing factors that contain RS-domains. The direct impact of Me2SO on alternative splicing may explain, at least in part, the different and sometimes opposite effects of Me2SO on cell differentiation and apoptosis.
Assuntos
Processamento Alternativo/efeitos dos fármacos , Dimetil Sulfóxido/farmacologia , Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B , Moléculas de Adesão de Célula Nervosa/genética , Ribonucleoproteínas/metabolismo , Regiões 3' não Traduzidas/efeitos dos fármacos , Regiões 3' não Traduzidas/genética , Animais , Linhagem Celular , Éxons , Formamidas/farmacologia , Genes Reporter , Glutationa Transferase/genética , Células HeLa , Ribonucleoproteína Nuclear Heterogênea A1 , Ribonucleoproteínas Nucleares Heterogêneas , Humanos , Cinética , Camundongos , Precursores de RNA/genética , Precursores de RNA/metabolismo , Proteínas Recombinantes/metabolismoRESUMO
UNLABELLED: Several studies have confirmed the existence of genetic factors in schizophrenia. However, the genotype predisposing for the disease is not known yet. Nevertheless, those genetic factors in the families of schizophrenic patients urge us to search for genetic vulnerability markers of schizophrenia. Ocular pursuit disorders, in particular, could be one of those vulnerability markers. Eye movements have been often tested in schizophrenia. Most of the schizophrenic patients have eye-tracking disorders and their biological relatives demonstrate an increased prevalence of eye-tracking impairments. The aim of the study was to research if smooth pursuit eye movements could be a vulnerability marker of schizophrenia. In order to have an indication about this hypothesis, impairments of smooth pursuit eye movements were researched in both schizophrenics and their parents. METHODS: Fifteen DSM IV schizophrenic patients stabilized at the time of the inclusion and not treated with lithium, benzodiazepines, barbiturates, or chloral hydrate; 19 parents without history of schizophrenic spectrum disorders (SADSLA and IPDE), and 2 groups of healthy subjects matched in age and sex with probands and with the parents, were included in the study. Parents only were included (fathers or mothers) in order to have an homogeneous population for the genetic risk and age. The eye-tracking paradigm used was a smooth pursuit task. The stimulus was a sinusoidal wave form moving on a horizontal line, with a frequency of 0.4 Hz and an amplitude of 30 degrees. Different parameters were measured: gain (ratio between the eye velocity and the target velocity) and saccades frequencies (catch-up saccades, back-up saccades, anticipatory saccades and square-wave-jerks). For each parameter, analysis of covariance (ANCOVA) with age as covariable was carried out. For the results reaching the significance of 0.05, the Bonferroni correction was applied (level of significance 0.016). The effect size of the parameter was calculated ((the mean of the subjects minus the mean of the matched controls) divided by standard deviation of the two groups). According to Cohen, 0.20 indicates a small effect size, 0.50 indicates a medium effect size and 0.80 indicates a large effect size. RESULTS: Comparison between patients and matched controls: the means of global gain, of gain for the movements to the left and of gain for the movements to the right did not differ significantly between patients and their matched controls. The size effects are 0.31 for the global gain, 0.20 for the movements to the left and 0.41 for the movements to the right. The frequencies of total saccades, catch-up saccades, back-up saccades, anticipatory saccades and square-wave-jerks did not differ significantly between patients and their controls. The size effects for those parameters were 0.09, 0.03, 0.00, 0.39 and 0.63 respectively. Comparison between parents and matched controls: the means of global gain, of gain for the movements to the left and of gain for the movements to the right did not differ significantly between the two groups. The size effects for those parameters were 0.00, 0.05 and 0.17 respectively. The frequency of total saccades did not differ significantly between the groups whereas the size effect was 0.63. The frequency of catch-up saccades was significantly more important in parents than in controls (p = 0.006) and the size effect was 0.80. The other saccadic parameters did not differ significantly between groups, their size effects were 0.24 for the back-up saccades, 0.21 for the anticipatory saccades and 0.00 for the square-wave-jerks. Whereas the gain of the patients had a tendency to be lower than the gain of their controls, no significant difference was observed between patients and their controls. Only a size effect of 0.63 for the frequency of square-wave-jerks was obtained. This large effect size suggests that the difference between patients and controls might be significant in a larger sample. The catch-up saccades frequency between parents and controls was significant. The differences between our study and the previous studies could be due to several factors. The paradigms used were different between the studies and our sample was small (only 15 patients and 19 relatives). Moreover, some patients in the previous studies were treated by lithium, drug well known to modify ocular pursuit and, finally the relatives in the other studies were 10 years older than ours and age is known to alter ocular pursuit. Since an impairment of the smooth pursuit was observed in the relatives of schizophrenic patients but not in the probands, this study does not support the hypothesis that eye-tracking disorders could be considered as a marker of vulnerability of schizophrenia.
Assuntos
Pais , Movimentos Sacádicos/fisiologia , Esquizofrenia/genética , Esquizofrenia/fisiopatologia , Adulto , Feminino , Humanos , Masculino , Prevalência , Esquizofrenia/epidemiologiaRESUMO
Alternative splicing of exon 7B in the hnRNP A1 pre-mRNA produces mRNAs encoding two proteins: hnRNP A1 and the less abundant A1B. We have reported the identification of several intron elements that contribute to exon 7B skipping. In this study, we report the activity of a novel element, conserved element 9 (CE9), located in the intron downstream of exon 7B. We show that multiple copies of CE9 inhibit exon 7B-exon 8 splicing in vitro. When CE9 is inserted between two competing 3' splice sites, a single copy of CE9 decreases splicing to the distal 3' splice site. Our in vivo results also support the conclusion that CE9 is a splicing modulator. First, inserting multiple copies of CE9 into an A1 minigene compromises the production of fully spliced products. Second, one copy of CE9 stimulates the inclusion of a short internal exon in a derivative of the human beta-globin gene. In this case, in vitro splicing assays suggest that CE9 decreases splicing of intron 1, an event that improves splicing of intron 2 and decreases skipping of the short internal exon. The ability of CE9 to act on heterologous substrates, combined with the results of a competition assay, suggest that the activity of CE9 is mediated by a trans-acting factor. Our results indicate that CE9 represses the use of the common 3' splice site in the hnRNP A1 alternative splicing unit.
Assuntos
Processamento Alternativo/fisiologia , Éxons/genética , Proteínas de Ligação ao GTP/química , Inativação Gênica , Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B , Íntrons/genética , Sequências Reguladoras de Ácido Nucleico , Ribonucleoproteínas/genética , Sequência de Bases , Proteínas de Ligação ao GTP/genética , Proteínas de Ligação ao GTP/metabolismo , Regulação da Expressão Gênica , Células HeLa , Ribonucleoproteína Nuclear Heterogênea A1 , Ribonucleoproteínas Nucleares Heterogêneas , Humanos , Dados de Sequência Molecular , Precursores de RNA/genética , Precursores de RNA/metabolismo , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Transativadores/fisiologiaRESUMO
The heterogeneous nuclear ribonucleoprotein A1 protein and a shortened derivative (UP1) promote telomere elongation in mammalian cells. To gain insights into the function of A1/UP1 in telomere biogenesis, we have investigated the binding properties of recombinant A1/UP1 and derivatives to single-stranded DNA oligonucleotides. Our results indicate that UP1 prefers to bind to DNA carrying single-stranded telomeric extensions at the 3' terminus. The RNA recognition motif 1 is sufficient for strong and specific binding to oligomers carrying vertebrate telomeric repeats. We find that the binding of A1/UP1 protects telomeric sequences against degradation by endo- and exonucleases. Moreover, A1/UP1 binding prevents extension by telomerase and terminal deoxynucleotidyltransferase and inhibits rNTP-dependent DNA synthesis in vitro. These observations are consistent with the hypothesis that A1/UP1 is a telomere end-binding protein that plays a role in the maintenance of long 3' overhangs.
Assuntos
DNA Helicases/fisiologia , Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B , Ribonucleoproteínas/fisiologia , Telômero/fisiologia , Hormônios do Timo/fisiologia , Sequência de Bases , Células HeLa , Ribonucleoproteína Nuclear Heterogênea A1 , Ribonucleoproteínas Nucleares Heterogêneas , Humanos , Oligodesoxirribonucleotídeos , Proteínas Recombinantes/metabolismo , Telomerase/antagonistas & inibidores , Telomerase/metabolismoRESUMO
In order to test the hypothesis that an excess of summer births is a risk factor for deficit syndrome, the month of birth was studied in 53 deficit schizophrenic patients compared to 158 non-deficit patients. No significant difference in terms of month of birth or season of birth was observed between deficit and non-deficit patients, suggesting that summer births might not be a risk factor for deficit schizophrenia.
Assuntos
Trabalho de Parto , Esquizofrenia/diagnóstico , Adulto , Feminino , Humanos , Masculino , Gravidez , Fatores de Risco , Psicologia do Esquizofrênico , Estações do AnoRESUMO
The neural cell adhesion molecule (NCAM) gene contains an 801 nt exon that is included preferentially in neuronal cells. We have set up an in vitro splicing system that mimics the neuro-specific alternative splicing profile of NCAM exon 18. Splicing regulation is observed using model pre-mRNAs that contain competing 5' or 3' splice sites, suggesting that distinct pathways regulate NCAM 5' and 3' splice site selection. While inclusion of exon 18 is the predom-inant choice in neuronal cells, an element in the 5' common exon 17 improves exon 17/exon 19 splicing in a neuronal cell line. A similar behavior is observed in vitro as the element can stimulate the 5' splice site of exon 17 or a heterologous 5' splice site. The minimal 32 nt sequence of the exon 17 enhancer consists of purine stretches and A/C motifs. Mutations in the purine stretches compromise the binding of SR proteins and decreases splicing stimulation in vitro. Mutations in the A/C motifs do not affect SR protein binding but reduce enhancing activity. Our results suggest that the assembly of an enhancer complex containing SR proteins in a 5' common exon ensures that NCAM mRNAs lacking exon 18 are made in neuronal cells.
Assuntos
Processamento Alternativo , Moléculas de Adesão de Célula Nervosa/genética , Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , Animais , Sequência de Bases , Éxons/genética , Células HeLa , Humanos , Camundongos , Dados de Sequência Molecular , Mutação , Precursores de RNA/metabolismo , Splicing de RNA , Proteínas de Ligação a RNA , Homologia de Sequência do Ácido Nucleico , Fatores de Processamento de Serina-Arginina , Células Tumorais CultivadasRESUMO
The RNA-binding protein hnRNP A1 is a splicing regulator produced by exclusion of alternative exon 7B from the A1 pre-mRNA. Each intron flanking exon 7B contains a high-affinity A1-binding site. The A1-binding elements promote exon skipping in vivo, activate distal 5' splice site selection in vitro and improve the responsiveness of pre-mRNAs to increases in the concentration of A1. Whereas the glycine-rich C-terminal domain of A1 is not required for binding, it is essential to activate the distal 5' splice site. Because A1 complexes can interact simultaneously with two A1-binding sites, we propose that an interaction between bound A1 proteins facilitates the pairing of distant splice sites. Based on the distribution of putative A1-binding sites in various pre-mRNAs, an A1-mediated change in pre-mRNA conformation may help define the borders of mammalian introns. We also identify an intron element which represses the 3' splice site of exon 7B. The activity of this element is mediated by a factor distinct from A1. Our results suggest that exon 7B skipping results from the concerted action of several intron elements that modulate splice site recognition and pairing.
Assuntos
Ribonucleoproteínas Nucleares Heterogêneas Grupo A-B , Ribonucleoproteínas/genética , Ribonucleoproteínas/metabolismo , Processamento Alternativo , Sequência de Bases , Sítios de Ligação/genética , Primers do DNA/genética , Éxons , Glicina/química , Células HeLa , Ribonucleoproteína Nuclear Heterogênea A1 , Ribonucleoproteínas Nucleares Heterogêneas , Humanos , Íntrons , Modelos Biológicos , Dados de Sequência Molecular , Precursores de RNA/genética , Precursores de RNA/metabolismo , Ribonucleoproteínas/química , TransfecçãoRESUMO
According to the neurodevelopmental hypothesis, antenatal aggressions (hypoxia and seasonal viral infections) could increase the risk of schizophrenia in adulthood as shown by an excess of obstetric complications and births in winter--spring in schizophrenic patients. As schizoid and schizotypal personality disorders are genetically linked to schizophrenia, we wanted to verify whether such disorders in the premorbid period in schizophrenic patients could be markers of a more genetic and less environmental sub-type of schizophrenia. Therefore, the aim of this study was to assess schizoid and schizotypal premorbid personality disorders (PPD) in 60 schizophrenic patients, and to assess the weight of familial and environmental factors according to the diagnosis of PPD. 41.7% of patients (25/60) had a schizoid or schizotypal PPD. Compared with patients without PPD, patients with PPD had more often schizophrenia spectrum disorders in first degree relatives (33.3% vs 14.7%, NS), less often obstetric complications (20.8% vs 50.0%, p < 0.05) and were less often born in the first half-year (44.0% vs 68.6%, p = 0.05). So, we showed a non significant positive association between schizoid--schizotypal PPD and family history of schizophrenia spectrum disorders, and a significant negative association between PPD and environmental factors: obstetric complications (OC) and birth in winter-spring. So, the absence of PPD could enable us to identify a sub-group of patients in whom environmental factors play a major role. Moreover, the relations between genetic factors and PPD seem to be complicated. Nevertheless, the notion of PPD could give information about the kind of genetic factors implicated in schizophrenia.
Assuntos
Desenvolvimento da Personalidade , Efeitos Tardios da Exposição Pré-Natal , Esquizofrenia/genética , Transtorno da Personalidade Esquizotípica/genética , Meio Social , Adolescente , Adulto , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Escalas de Graduação Psiquiátrica , Fatores de Risco , Esquizofrenia/diagnóstico , Transtorno da Personalidade Esquizotípica/diagnóstico , Estações do AnoRESUMO
Telomeric DNA of mammalian chromosomes consists of several kilobase-pairs of tandemly repeated sequences with a terminal 3' overhang in single-stranded form. Maintaining the integrity of these repeats is essential for cell survival; telomere attrition is associated with chromosome instability and cell senescence, whereas stabilization of telomere length correlates with the immortalization of somatic cells. Telomere elongation is carried out by telomerase, an RNA-dependent DNA polymerase which adds single-stranded TAGGGT repeats to the 3' ends of chromosomes. While proteins that associate with single-stranded telomeric repeats can influence tract lengths in yeast, equivalent factors have not yet been identified in vertebrates. Here, it is shown that the heterogeneous nuclear ribonucleoprotein A1 participates in telomere biogenesis. A mouse cell line deficient in A1 expression harbours telomeres that are shorter than those of a related cell line expressing normal levels of A1. Restoring A1 expression in A1-deficient cells increases telomere length. Telomere elongation is also observed upon introduction of exogenous UP1, the amino-terminal fragment of A1. While both A1 and UP1 bind to vertebrate single-stranded telomeric repeats directly and with specificity in vitro, only UP1 can recover telomerase activity from a cell lysate. These findings establish A1/UP1 as the first single-stranded DNA binding protein involved in mammalian telomere biogenesis and suggest possible mechanisms by which UP1 may modulate telomere length.