RESUMO
High turnover and understaffing are significant issues plaguing the healthcare system. Some of the leading reasons of turnover include child-bearing and -rearing, stress related to working, and health concerns. With the onset of the coronavirus (COVID-19) pandemic, this problem of turnover worsened due to increased risk of infection and escalating workload. This study aimed to clarify and validate the effect of burnout on intention to leave among full-time nursing professionals and the structural relationship with sense of coherence (SOC) and striving for work-life balance (S-WLB). Secondary analyses of data obtained from a previous study was carried out; a hypothesized model was tested for goodness of fit and a final model was developed. Burnout directly affected intention to leave (P < .001). It also affected intention to leave through SOC and S-WLB (P < .01); SOC lessened the effect of burnout on S-WLB, therefore reducing its impact on intention to leave. Effective strategies need to be developed to improve the SOC and WLB of nurses to alleviate the effects of burnout and thus reduce the likelihood of turnover. Improving their ability to grasp and deal with emergencies and ambiguous situations, as well as providing emotional and tangible support can be other ways to retain nursing professionals.
Assuntos
Esgotamento Profissional , COVID-19 , Enfermeiras e Enfermeiros , Recursos Humanos de Enfermagem Hospitalar , Senso de Coerência , Humanos , Intenção , Equilíbrio Trabalho-Vida , Satisfação no Emprego , Recursos Humanos de Enfermagem Hospitalar/psicologia , Estudos Transversais , Esgotamento Profissional/etiologia , Esgotamento Profissional/psicologia , Inquéritos e QuestionáriosRESUMO
Lipid droplets are cytoplasmic organelles that store lipids for energy and membrane synthesis. The oleaginous yeast Lipomyces starkeyi is one of the most promising lipid producers and has attracted attention as a biofuel source. It is known that the expansion of lipid droplets is enhanced under nutrient-poor conditions. Therefore, we prepared a novel nitrogen-depleted medium (N medium) in which to culture L. starkeyi cells. Lipid accumulation was rapidly induced, and this was reversed by the addition of ammonium. In this condition, cell proliferation stopped, and cells with giant lipid droplets were arrested in G1 phase. We investigated whether cell cycle arrest at a specific phase is required for lipid accumulation. Lipid accumulation was repressed in hydroxyurea-synchronized S phase cells and was increased in nocodazole-arrested G2/M phase cells. Moreover, the enrichment of G1 phase cells seen upon rapamycin treatment induced massive lipid accumulation. From these results, we conclude that L. starkeyi cells store lipids from G2/M phase and then arrest cell proliferation in the subsequent G1 phase, where lipid accumulation is enhanced. Cell cycle control is an attractive approach for biofuel production.
Assuntos
Biocombustíveis , Lipomyces , Pontos de Checagem da Fase G1 do Ciclo Celular , Lipídeos , Lipomyces/metabolismo , LevedurasRESUMO
The oleaginous yeast Lipomyces starkeyi is a powerful lipid producer with great industrial potential. Recent studies have reported the isolation of mutant L. starkeyi cells with higher lipid producing capacity. Although genetic engineering strategies have been applied to L. starkeyi, classical genetic approaches are lacking. The development of tools that facilitate genetic crosses in L. starkeyi would not only make it possible to build improved lipid-producing strains but also facilitate molecular biological analysis of this species. In this study, I report a set of strains and approaches useful for performing genetic crosses with L. starkeyi. The homothallic L. starkeyi reportedly forms an ascus containing two to 20 spores. These spores were resistant to glusulase and could be dissected using a micromanipulator, suggesting that random spore and tetrad (spore dissection) analysis can be adapted for L. starkeyi. Additionally, to isolate a pair of heterothallic strains useful for genetic crosses, the homothallic strain was exposed to UV irradiation, and 10 self-sterile strains were crossed with one another. One of these combinations, Ls75 and Ls100, sporulated stably. Moreover, to detect genetic recombination, I introduced a different drug resistance marker into each strain and crossed them. The resulting progeny exhibited Mendelian segregation of the resistance markers. Altogether, the work reported here provides a powerful resource for genetic analysis in L. starkeyi.
Assuntos
Lipomyces , Cruzamentos Genéticos , Engenharia Genética , Lipomyces/genética , LevedurasRESUMO
The increase in the elderly population in need of healthcare services has led to a serious shortage in the nursing workforce. To retain a large nursing workforce, a strong work-life balance among nurses is needed along with a healthy work environment. This prospective study investigates the influence of work-life balance and sense of coherence on intention to leave among hospital nurses. A questionnaire survey was conducted with 2239 nurses as a baseline. The explanatory variables included striving for work-life balance behavior, a sense of coherence in terms of personal resources, and work-, organizational-, and individual-related factors. Using a cohort of 1368 valid responses, we measured intention to leave among 975 nurses with whom we were able to follow up 6 months after the baseline survey. We then performed multiple regression analysis. The behavior striving for work-life balance was shown to influence nurses' intention to leave. Nurses who exhibited less striving for work-life balance behavior showed higher intentions to leave. The sense of coherence was not identified as a factor affecting intention to leave. Securing a comfortable work-life balance would reduce the nurses' desire to quit the hospital. To reduce nurse turnover, nurse managers should develop support programs that can help nurses achieve a better work-life balance.
Assuntos
Enfermeiras e Enfermeiros , Recursos Humanos de Enfermagem Hospitalar , Senso de Coerência , Idoso , Estudos Transversais , Humanos , Intenção , Satisfação no Emprego , Estudos Prospectivos , Inquéritos e Questionários , Equilíbrio Trabalho-VidaRESUMO
AIM: This study aimed to develop and test a causal model focused on assertiveness, stress coping, and workplace environment as factors affecting burnout among novice nurses. DESIGN: Cross-sectional study was conducted with novice nurses of 17 hospitals. METHODS: The Novice Nurse Assertiveness Scale and the Japanese version of Maslach Burnout Inventory. RESULTS: Data from 645 female novice nurses were analysed. The mean age, Novice Nurse Assertiveness Scale and Maslach Burnout Inventory were 22.6 ± 3.0, 67.4 ± 10.3 and 13.7 ± 2.5. For the final model, the study adopted a model that includes indirect influences; inappropriate assertiveness and inappropriate coping affected the dissatisfaction with the job and then affected the burnout. The goodness of fit index was GFI = 0.94, AGFI = 0.91, RMSEA = 0.66, and R2 was .86. The findings validated this as a causal model of assertiveness, stress coping, and the work environment as factors affecting burnout for novice nurses.
Assuntos
Esgotamento Profissional , Enfermeiras e Enfermeiros , Adaptação Psicológica , Assertividade , Esgotamento Psicológico , Estudos Transversais , Feminino , Humanos , Satisfação no Emprego , Inquéritos e Questionários , Local de TrabalhoRESUMO
Blood arsenic has various toxicities including carcinogenicity, but urinary concentrations are often substituted to determine the exposure level. Since there is little information on the relation of urinary arsenic species to blood arsenic, the aim was to investigate relationships between blood total arsenic (T-As) and the urinary species adjusted by creatinine and specific gravity (SG). Blood and spot urine samples were collected from 109 Japanese subjects aged 18-66 years without occupational exposure. Positive correlations of blood T-As (median, 3.49 µg/L) with urinary creatinine-adjusted and SG-adjusted T-As and arsenobetaine were statistically significant and greater than those with the unadjusted ones. The magnitude of associations of blood T-As with creatinine-adjusted arsenic species was significantly larger than those with unadjusted or SG-adjusted ones. Most of the correlation coefficients among urinary arsenic species concentrations were significant in three adjustment methods, but there was not a significant correlation between monomethylarsonic acid and arsenobetaine after urinary creatinine and SG corrections. Given multiple regression analysis, plasma T-As concentrations showed significant relations to creatinine-adjusted T-As, dimethylarsinic acid, and arsenobetaine concentrations, but erythrocyte T-As did hardly reflect the variation of urinary arsenic species. In conclusion, creatinine-adjusted arsenic concentrations are suggested to be the most appropriate predictor of blood T-As; by contrast, use of the urinary unadjusted arsenic concentration may result in a misleading interpretation of inorganic arsenic toxicity because the associations between inorganic and organic arsenic species based on the unadjusted concentration were mutually close. Plasma T-As appeared to be the best indicator of low-level exposure in blood samples.
Assuntos
Arsênio , Exposição Ocupacional , Adolescente , Adulto , Idoso , Ácido Cacodílico , Exposição Ambiental , Humanos , Pessoa de Meia-Idade , Urinálise , Adulto JovemRESUMO
Centromeres function as a platform for the assembly of multiple kinetochore proteins and are essential for chromosome segregation. An active centromere is characterized by the presence of a centromere-specific histone H3 variant, CENP-A. Faithful centromeric localization of CENP-A is supported by heterochromatin in almost all eukaryotes; however, heterochromatin proteins have been lost in most Saccharomycotina. Here, identification of CENP-A (CENP-AL.s.) and heterochromatin protein 1 (Lsw1) in a Saccharomycotina species, the oleaginous yeast Lipomyces starkeyi, is reported. To determine if these proteins are functional, the proteins in S. pombe, a species widely used to study centromeres, were ectopically expressed. CENP-AL.s. localizes to centromeres and can be replaced with S. pombe CENP-A, indicating that CENP-AL.s. is a functional centromere-specific protein. Lsw1 binds at heterochromatin regions, and chromatin binding is dependent on methylation of histone H3 at lysine 9. In other species, self-interaction of heterochromatin protein 1 is thought to cause folding of chromatin, triggering transcription repression and heterochromatin formation. Consistent with this, it was found that Lsw1 can self-interact. L. starkeyi chromatin contains the methylation of histone H3 at lysine 9. These results indicated that L. starkeyi has a primitive heterochromatin structure and is an attractive model for analysis of centromere heterochromatin evolution.
Assuntos
Proteína Centromérica A/genética , Proteínas Cromossômicas não Histona/genética , Proteínas Fúngicas/genética , Lipomyces/genética , Proteína Centromérica A/metabolismo , Homólogo 5 da Proteína Cromobox , Proteínas Cromossômicas não Histona/metabolismo , Clonagem Molecular , Proteínas Fúngicas/metabolismo , Ligação Proteica , Domínios Proteicos , Schizosaccharomyces , Homologia de SequênciaRESUMO
AIM: Burnout is a common feature among healthcare professionals; however, little systematic research exists on burnout among nurses who are raising children. The burnout-related factors among female nurses with children under the age of 3 years were identified in order to ascertain potential burnout prevention methods. METHODS: In total, 1681 nurses with children who worked at nine city hospitals in Kanagawa Prefecture, Japan, were sent the Maslach Burnout Inventory-Human Services Survey; 1173 nurses responded in June 2014. They were divided according to their sex and children's ages. A data analysis was undertaken for those female nurses with children who were aged under 3 years who provided valid responses (n = 158). RESULTS: A number of factors related to burnout in female nurses with children aged under 3 years was found via a multiple regression analysis: irritation at being unable to attend to their own affairs, over 4-6 h of overtime work per week, having a child aged under 3 years as the first or second child, little sense of work fulfillment, using a childcare facility outside the workplace, dissatisfaction with their salary, feeling ill-qualified as a parent, and a sense of inadequate support. CONCLUSIONS: Child care occurs during a limited period and appropriate support is needed. A workplace environment with no overtime work, a childcare facility in the workplace, and mental health support to reduce "feelings of irritation" and "feeling ill-qualified as a parent" could help to prevent burnout in female nurses with toddlers and infants.
Assuntos
Esgotamento Profissional , Mães/psicologia , Recursos Humanos de Enfermagem Hospitalar/psicologia , Adulto , Esgotamento Profissional/prevenção & controle , Pré-Escolar , Emoções , Feminino , Humanos , Japão , Inquéritos e Questionários , Local de TrabalhoRESUMO
Histone gene expression is regulated in a cell cycle-dependent manner, with a peak at S phase, which is crucial for cell division and genome integrity. However, the detailed mechanisms by which expression of histone genes are tightly regulated remain largely unknown. Fission yeast Ams2, a GATA-type zinc finger motif-containing factor, is required for activation of S phase-specific core histone gene transcription. Here we report the molecular characterisation of Ams2. We show that the zinc finger motif in Ams2 is necessary to bind the histone gene promoter region and to activate histone gene transcription. An N-terminal region of Ams2 acts as a self-interaction domain. Intriguingly, N-terminally truncated Ams2 binds to the histone gene promoters, but does not fully activate histone gene transcription. These observations imply that Ams2 self-interactions are required for efficient core histone gene transcription. Moreover, we show that Ams2 interacts with Teb1, which itself binds to the core histone gene promoters. We discuss the relationships between Ams2 domains and efficient transcription of the core histone genes in fission yeast.
Assuntos
Fatores de Transcrição GATA/genética , Histonas/genética , Proteínas de Schizosaccharomyces pombe/genética , Schizosaccharomyces/genética , Transcrição Gênica/genética , Regiões Promotoras Genéticas/genética , Fase S/genética , Dedos de Zinco/genéticaRESUMO
CONTEXT: Plant-based diets (PBDs) are a healthy alternative to westernized diets. A semivegetarian diet, a PBD, has been shown to prevent a relapse in Crohn disease. However, there is no way to measure adherence to PBDs. OBJECTIVE: To develop a simple way of evaluating adherence to a PBD for Japanese patients with inflammatory bowel disease (IBD). DESIGN: PBD scores were assigned according to the frequency of consumption provided on a food-frequency questionnaire, obtained on hospitalization for 159 patients with ulcerative colitis and 70 patients with Crohn disease. Eight items considered to be preventive factors for IBD were scored positively, and 8 items considered to be IBD risk factors were scored negatively. The PBD score was calculated from the sum of plus and minus scores. Higher PBD scores indicated greater adherence to a PBD. The PBD scores were evaluated on hospitalization and 2 years after discharge for 22 patients with Crohn disease whose dietary pattern and prognosis were established. MAIN OUTCOME MEASURE: Plant-Based Diet score. RESULTS: The PBD scores differed significantly, in descending order, by dietary type: pro-Japanese diet, mixed type, and pro-westernized diet (Wilcoxon/Kruskal-Wallis test). The PBD scores in the ulcerative colitis and Crohn disease groups were 10.9 ± 9.5 and 8.2 ± 8.2, respectively. For patients with Crohn disease, those with long-term remission and normal C-reactive protein concentration were significantly more likely to have PBD scores of 25 or greater than below 25 (χ2). CONCLUSION: The PBD score is a valid assessment of PBD dietary adherence.
Assuntos
Doença de Crohn/dietoterapia , Dieta/normas , Comportamento Alimentar , Cooperação do Paciente , Plantas , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteína C-Reativa/metabolismo , Criança , Doença de Crohn/sangue , Dieta Vegetariana , Fabaceae , Feminino , Frutas , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Indução de Remissão , Inquéritos e Questionários , Verduras , Adulto JovemRESUMO
Overweight and obesity are global health concerns. Various effective weight-loss diets have been developed, including the Atkins diet. The Atkins diet is known as an extreme low-carbohydrate diet. This diet reduces body weight and has gained widespread popularity. However, the metabolite profiles of such a diet have been shown to be detrimental to colonic health. Therefore, a concern for the long-term health effects of this diet exists. We encountered a case in which ulcerative colitis developed while the patient was following the Atkins diet.A man, 172 cm in height and weighing 72 kg, at age 36 years followed a low-carbohydrate weight-loss diet. His weight decreased to 66 kg as desired. Thereafter he noticed bloody stool. Colonoscopy revealed diffuse inflammation limited to the rectum, and he was diagnosed with ulcerative colitis. He underwent an educational hospitalization for ulcerative colitis. A plant-based/semivegetarian diet was provided during hospitalization. Bloody stool disappeared during hospitalization and he achieved remission without medication for inflammatory bowel disease.This case indicates that an onset of ulcerative colitis can be an adverse event to a low-carbohydrate weight-loss diet.
Assuntos
Colite Ulcerativa/dietoterapia , Dieta com Restrição de Carboidratos/efeitos adversos , Dieta Redutora , Dieta Vegetariana , Adulto , Colite Ulcerativa/diagnóstico , Colite Ulcerativa/etiologia , Humanos , MasculinoRESUMO
AIM: This prospective cohort study aims to clarify the factors affecting burnout in female nurses who have preschool-age children. METHODS: The subjects were 2151 female nurses who have preschool-age children and work at 70 city hospitals across Japan. The questionnaires were completed by 1644 female nurses with preschool-age children in October 2010, and they were divided into a cohort to observe the incidence of burnout, which was investigated in October 2011. RESULTS: At the baseline, the authors obtained responses from 1802 subjects (83.8%) who had agreed to join the study. The subjects the authors were able to investigate totaled 523, of whom 117 (22.4%) had experienced burnout. Multiple logistic regression analysis showed that influencing or predictive factors in burnout were years at the present workplace, will to continue work, assertiveness, overtime work, and spanking children. CONCLUSION: Female nurses who have preschool-age children tend to burnout easily if they have been at their present workplace for less than 3 years, wish to quit working, respond to a child's misbehavior with spanking, are "very low" in assertiveness, or work 4-6 h of overtime per week.
Assuntos
Esgotamento Profissional , Recursos Humanos de Enfermagem Hospitalar/psicologia , Adaptação Psicológica , Adulto , Pré-Escolar , Feminino , Humanos , JapãoRESUMO
Cryopreservation of mature oocytes and embryos has provided numerous benefits in reproductive medicine. Although successful cryopreservation of germinal-vesicle stage (GV) oocytes holds promise for further advances in reproductive biology and clinical embryology fields, reports regarding cryopreservation of immature oocytes are limited. Oocyte survival and maturation rates have improved since vitrification is being performed at the GV stage, but the subsequent developmental competence of GV oocytes is still low. The purpose of this study was to evaluate the effects of supplementation of the maturation medium with cyclic adenosine monophosphate (cAMP) modulators on the developmental competence of vitrified-warmed GV bovine oocytes. GV oocytes were vitrified-warmed and cultured to allow for oocyte maturation, and then parthenogenetically activated or fertilized in vitro. Our results indicate that addition of a cAMP modulator forskolin (FSK) or 3-isobutyl-1-methylxanthine (IBMX) to the maturation medium significantly improved the developmental competence of vitrified-warmed GV oocytes. We also demonstrated that vitrification of GV oocytes led to a decline in cAMP levels and maturation-promoting factor (MPF) activity in the oocytes during the initial and final phases of maturation, respectively. Nevertheless, the addition of FSK or IBMX to the maturation medium significantly elevated cAMP levels and MPF activity during IVM. Taken together, our results suggest that the cryopreservation-associated meiotic and developmental abnormalities observed in GV oocytes may be ameliorated by an artificial increase in cAMP levels during maturation culture after warming.
Assuntos
AMP Cíclico/metabolismo , Fertilização in vitro , Técnicas de Maturação in Vitro de Oócitos , Oócitos/crescimento & desenvolvimento , 1-Metil-3-Isobutilxantina/administração & dosagem , Animais , Bovinos , Colforsina/administração & dosagem , Criopreservação , Feminino , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Vitrificação/efeitos dos fármacosRESUMO
Hydroxypropyl cellulose (HPC) was investigated as a replacement for serum substitute supplement (SSS) for use in cryoprotectant solutions for embryo vitrification. Mouse blastocysts from inbred (n = 1056), hybrid (n = 128) strains, and 121 vitrified blastocysts donated by infertile patients (n = 102) were used. Mouse and human blastocysts, with or without zona pellucida, were vitrified and warmed in either 1% or 5% HPC or in 5% or 20% SSS-supplemented media using the Cryotop (Kitazato BioPharma Co. Ltd, Fuji, Japan) method, and the survival and oxygen consumption rates were assessed. Viscosity of each vitrification solution was compared. Survival rates of mouse hybrid blastocysts and human zona pellucida-intact blastocysts were comparable among the groups. Mouse and human zona pellucida-free blastocysts, which normally exhibit poor cryoresistance, showed significantly higher survival rates in 5% HPC than 5% SSS (P < 0.05). The 5% HPC-supplemented vitrification solution showed a significantly higher viscosity (P < 0.05). The blastocysts were easily detached from the Cryotop strip during warming when HPC-supplemented vitrification solution was used. The oxygen consumption rates were similar between non-vitrified and 5% HPC groups. The results suggest possible use of HPC for supplementation of cryoprotectant solutions and provide useful information to improve vitrification protocols.
Assuntos
Celulose/análogos & derivados , Crioprotetores , Embrião de Mamíferos , Técnicas de Reprodução Assistida , Animais , Blastocisto/citologia , Adesão Celular , Sobrevivência Celular , Celulose/administração & dosagem , Meios de Cultura , Transferência Embrionária , Feminino , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Consumo de Oxigênio , SoluçõesRESUMO
AIM: This study aimed to clarify conditions of women experiencing post-partum anemia and identify factors that work against the alleviation of anemia. METHODS: This was a retrospective longitudinal study, involving 246 women giving birth at five participating institutions, diagnosed with anemia on day 3 post-partum, and given a blood sample at the medical examination 1 month after the birth. With answers about alleviation of anemia during the 1 month post-partum period as an objective variable, and explanatory variables, multiple logistic regression analysis was performed. The explanatory variables included demographic data of the participants, information about anemia, following the nutrition instruction advice for anemia alleviation, family structure and others assisting the participants, and self-management skills (measured by the Self-Management Skill [SMS] scale). RESULTS: The present authors collected 211 valid responses; the average age was 32.6 years. The mean hemoglobin values on the 3rd and 30th days post-partum were 9.6 and 12.0 g/dL, respectively. One fifth of the participants (21.3%) showed no anemia alleviation. The mean value on the SMS scale was 28.35, and the anemia alleviated group (30.78) was significantly higher than the non-alleviated group (19.38). Results of the multiple logistic regression analysis showed a strong relationship between anemia risk and self-management skills. The absence of anemia alleviation increased 2.51 times as the total score of the SMS decreased 1 point. CONCLUSION: There is an urgent need to develop an intervention program to alleviate post-partum anemia focusing on the low score items because self-management skills strongly affect alleviation improvement.
Assuntos
Anemia/etiologia , Período Pós-Parto , Complicações Hematológicas na Gravidez/etiologia , Adulto , Feminino , Humanos , Estudos Longitudinais , Pessoa de Meia-Idade , Gravidez , Estudos RetrospectivosRESUMO
BACKGROUND: Many individuals with autism spectrum disorders (ASD) have difficulty with verbal communication, which might be due to a lack of spontaneous orientation toward social auditory stimuli. Previous studies have shown that a single dose of oxytocin improves speech comprehension in autism. The primary aim of this study was to investigate whether the orientation behaviors toward human sounds are different for neurotypical (NT) adults and adults with ASD and whether oxytocin has an effect on their orientation behaviors toward human sounds. METHODS: This was a randomized, placebo-controlled, within-subject, crossover design study of intranasal oxytocin versus placebo in 13 NT adults and 16 adults with ASD. Subjects were randomized to 24 IU intranasal oxytocin or placebo on different days, and they were blind to the treatment. The participants then listened passively to human and non-human affective sounds while their skin conductance responses (SCRs) and the changes in peripheral blood vessel constriction were monitored as an indicator of spontaneous orientation. The monitored data were analyzed by a mixed-design ANOVA. RESULTS: Oxytocin enhanced the difference between the SCRs to human and non-human sounds in both the NT and ASD groups (F(1,56) = 6.046, p = 0.017). Further correlation coefficient analysis showed significant correlations between this SCR difference and the scores in the autism spectrum quotient 'attention to detail' and 'social skill' subscales and interpersonal reactivity index and social functioning scale in the ASD group. Oxytocin was well tolerated, and no serious adverse effects were reported. CONCLUSIONS: The difference in SCRs implies that oxytocin nasal spray may enhance orientation behaviors toward human sounds in the presence of other environmental sounds in both ASD and NT adults. TRIAL REGISTRATION: UMIN-CTR Clinical Trial, Unique trial number: UMIN000005809.
RESUMO
Various plants possess non-photosynthetic, hydrophilic chlorophyll (Chl) proteins called water-soluble Chl-binding proteins (WSCPs). WSCPs are categorized into two classes; Class I (photoconvertible type) and Class II (non-photoconvertible type). Among Class II WSCPs, only Lepidium virginicum WSCP (LvWSCP) exhibits a low Chl a/b ratio compared with that found in the leaf. Although the physicochemical properties of LvWSCP have been characterized, its molecular properties have not yet been documented. Here, we report the characteristics of the LvWSCP gene, the biochemical properties of a recombinant LvWSCP, and the intracellular localization of LvWSCP. The cloned LvWSCP gene possesses a 669-bp open reading frame. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis revealed that the precursor of LvWSCP contains both N- and C-terminal extension peptides. RT-PCR analysis revealed that LvWSCP was transcribed in various tissues, with the levels being higher in developing tissues. A recombinant LvWSCP and hexa-histidine fusion protein (LvWSCP-His) could remove Chls from the thylakoid in aqueous solution and showed an absorption spectrum identical to that of native LvWSCP. Although LvWSCP-His could bind both Chl a and Chl b, it bound almost exclusively to Chl b when reconstituted in 40 % methanol. To clarify the intracellular targeting functions of the N- and C-terminal extension peptides, we constructed transgenic Arabidopsis thaliana lines expressing the Venus protein fused with the LvWSCP N- and/or C-terminal peptides, as well as Venus fused at the C-terminus of LvWSCP. The results showed that the N-terminal peptide functioned in ER body targeting, while the C-terminal sequence did not act as a trailer peptide.
Assuntos
Proteínas de Ligação à Clorofila/genética , Proteínas de Ligação à Clorofila/metabolismo , Lepidium/genética , Lepidium/metabolismo , Sequência de Aminoácidos , Arabidopsis/genética , Arabidopsis/metabolismo , Sequência de Bases , Clorofila/metabolismo , Clorofila A , Proteínas de Ligação à Clorofila/química , Clonagem Molecular , DNA de Plantas/genética , Retículo Endoplasmático/metabolismo , Genes de Plantas , Filogenia , Plantas Geneticamente Modificadas , Ligação Proteica , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Homologia de Sequência de Aminoácidos , SolubilidadeRESUMO
Spontaneous DNA damage may occur nonrandomly in the genome, especially when genome maintenance mechanisms are undermined. We developed single-strand DNA (ssDNA)-associated protein immunoprecipitation followed by sequencing (SPI-seq) to map genomic hotspots of DNA damage. We demonstrated this method with Rad52, a homologous recombination repair protein, which binds to ssDNA formed at DNA lesions. SPI-seq faithfully detected, in fission yeast, Rad52 enrichment at artificially induced double-strand breaks (DSBs) as well as endogenously programmed DSBs for mating-type switching. Applying Rad52 SPI-seq to fission yeast mutants defective in DNA helicase Pfh1 or histone H3K56 deacetylase Hst4, led to global views of DNA lesion hotspots emerging in these mutants. We also found serendipitously that histone dosage aberration can activate retrotransposon Tf2 and cause the accumulation of a Tf2 cDNA species bound by Rad52. SPI-seq should be widely applicable for mapping sites of DNA damage and uncovering the causes of genome instability.
Assuntos
Imunoprecipitação da Cromatina , Mapeamento Cromossômico , Dano ao DNA , DNA de Cadeia Simples , Sequenciamento de Nucleotídeos em Larga Escala , Imunoprecipitação da Cromatina/métodos , Quebras de DNA de Cadeia Dupla , DNA Helicases/genética , Replicação do DNA , Genômica , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Histona Desacetilases/genética , Histonas/genética , Histonas/metabolismo , Ligação Proteica , Proteína Rad52 de Recombinação e Reparo de DNA/metabolismo , Schizosaccharomyces/genética , Schizosaccharomyces/metabolismo , Proteínas de Schizosaccharomyces pombe/genética , Proteínas de Schizosaccharomyces pombe/metabolismoRESUMO
AIM: The aim of this study was to examine the relationship between dual-task performance and neurocognitive measures in community-dwelling older people with mild cognitive impairment (MCI). METHODS: A total of 98 subjects (mean age 74.8 years, 52.0% female) participated in the study. We compared 36 participants with amnestic MCI (aMCI) with 62 participants with non-amnestic MCI (non-aMCI) on dual-task performance as measured by reaction time responses. The relationships between dual-task performance and multiple domains of neurocognitive functions, including general cognitive function, visual memory, working memory, executive function and processing speed, were examined. RESULTS: Although there were no statistically significant group differences in simple reaction times (P = 0.734), the aMCI group showed significantly slower dual-task reaction times than the non-aMCI group (P = 0.012). Using multiple regression analysis, we found that there was a significant relationship between executive function and dual-task reaction times (ß = 0.298, P = 0.006). CONCLUSION: These results showed that aMCI subjects showed a specific deficit in dual-task performance compared with non-aMCI subjects, and poor dual-task performance was associated with declines in executive function in older people with MCI. Future longitudinal and interventional studies should investigate the use of dual-task testing with varying levels of cognitive demand in older adults at risk of dementia.