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BACKGROUND: More than 50% of people who die by suicide have not been in contact with formal mental health services. The rate of people who fly 'under the radar' of mental health services is higher among men than women, indicating a need to improve engagement strategies targeted towards men who experience suicidal thoughts and/or behaviours. In Australia, a range of mental health support services exist, designed specifically for men, yet, a substantial proportion of men do not use these services. The aim of this study is to evaluate whether a brief online video-based messaging intervention is an effective approach for encouraging men with suicidal thoughts and/or behaviours to engage with existing support services. METHODS: Informed by a literature review, surveys, and consultation with men with a lived experience of suicidal thoughts and/or behaviours, we designed five video-based messages that will be used in this five-arm randomised controlled trial. A total of 380 (76 per arm) men aged 18 years or older with suicidal thoughts who are not currently accessing formal mental health services will be recruited online and randomly assigned to watch one of the five web-based video messages. After viewing the video, men will be presented with information about four existing Australian support services, along with links to these services. The primary outcome will be help-seeking, operationalised as a click on any one of the four support service links, immediately after viewing the video. Secondary outcomes include immediate self-reported help-seeking intentions in addition to self-reported use of the support services during a 1-week follow-up period. We will also use the Discrete Choice Experiment methodology to determine what aspects of support services (e.g. low cost, short appointment wait times) are most valued by this group of men. DISCUSSION: This study is the first to evaluate the effectiveness of a brief web-based video messaging intervention for promoting engagement with existing support services among men with suicidal thoughts who are not currently receiving formal help. If found to be effective, this would represent a scalable, cost-effective approach to promote help-seeking for this at-risk population. Limitations and strengths of this study design are discussed.
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Ideação Suicida , Prevenção do Suicídio , Humanos , Masculino , Intervenção Baseada em Internet , Gravação em Vídeo , Ensaios Clínicos Controlados Aleatórios como Assunto , Suicídio/psicologia , Internet , Resultado do Tratamento , Fatores de Tempo , Saúde Mental , Serviços de Saúde Mental , Aceitação pelo Paciente de Cuidados de Saúde , Fatores Sexuais , AustráliaRESUMO
BACKGROUND: The COVID-19 pandemic negatively impacted global mental health, with adolescents experiencing disproportionate effects. Limited research has explored the impact of different pandemic restrictions on adolescent mental health, and only a few studies have examined the longer-term impacts of the pandemic on adolescent mental health. These investigations are crucial for informing public health policies, particularly the integration of mental health care in future public health emergencies. METHODS: This study aimed to investigate the impact of lockdown duration and the impact of adolescents' subjective experiences of the pandemic on their wellbeing, internalising symptoms, and externalising symptoms. Australian adolescents (N = 1,001, mean age = 14.2 years) completed a baseline survey in 2021, shortly after pandemic lockdowns were lifted (Time 1), and a follow-up survey approximately 12 months later (Time 2). Predictors of interest were the total duration of COVID-19 lockdowns across 2020-2021, and adolescents' subjective experiences of the pandemic on their social connections, learning, technology use and family relationships. A range of covariates were included in analyses to examine subgroup differences. RESULTS: Linear mixed-effects models indicated that total duration of the lockdown was not associated with any of the outcomes at Time 1 or Time 2 (all ps > 0.017). Negative subjective experience of the pandemic on learning was associated with greater externalising symptoms at both Time 1 (t = 5.17, df = 980, p <.001) and Time 2 (t = 2.72, df = 708, p =.007). Negative subjective experience of the pandemic on social connection was associated with greater internalising symptoms at Time 2 only (t = 3.20, df = 709, p =.001). Negative subjective experience of the pandemic on family relationships or technology use was not associated with any of the outcomes at Time 1 or Time 2 (all ps > 0.017). CONCLUSIONS: Adolescents' negative subjective experience of the pandemic on learning and social connections was associated with greater internalising and externalising symptoms after the lockdown had been lifted. Duration of lockdowns was not associated with any of the primary outcomes. During future public health emergencies, mental health interventions should be tailored to assist adolescents to adapt to new learning environments, and to build and maintain social connections.
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PURPOSE: Postoperative shoulder pain is a common problem after laparoscopic surgery. This study aimed to investigate the association between operative side and postoperative shoulder pain following urologic laparoscopic surgery performed in the lateral recumbent position. METHODS: This was a retrospective cohort study conducted at a single tertiary care center. A total of 506 patients who underwent urologic laparoscopic surgery (including adrenalectomy, radical nephrectomy, partial nephrectomy, and pyeloplasty) between January 2010 and December 2019 were included. Patients who underwent total nephroureterectomy or resection of other organs were excluded. The primary outcome was the incidence of postoperative shoulder pain. A multivariable logistic regression analysis investigated the association between the operative side and postoperative shoulder pain. RESULTS: Among the 506 included patients, there were an equal number of surgeries on the left and right sides. Eighty-eight patients had postoperative shoulder pain. The incidence of postoperative shoulder pain in the left-side group was significantly higher than that in the right-side group (21.3% [54/253] versus 13.4% [34/253], crude odds ratio = 1.75, 95% confidence interval [CI] 1.07-2.89). After adjustment for potential confounders (age, sex, body mass index, operation duration, operative technique, epidural block, peripheral nerve block, American Society of Anesthesiologists physical status classification, and intraoperative rocuronium dose), the left operative side was found to be associated with postoperative shoulder pain (adjusted odds ratio = 1.89, 95% CI 1.15-3.09). CONCLUSION: The left operative side is associated with an increased incidence of postoperative shoulder pain after urologic laparoscopic surgery performed in the lateral recumbent position.
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Laparoscopia , Dor Pós-Operatória , Dor de Ombro , Procedimentos Cirúrgicos Urológicos , Humanos , Masculino , Feminino , Laparoscopia/métodos , Laparoscopia/efeitos adversos , Estudos Retrospectivos , Dor de Ombro/epidemiologia , Dor de Ombro/etiologia , Pessoa de Meia-Idade , Dor Pós-Operatória/epidemiologia , Dor Pós-Operatória/etiologia , Procedimentos Cirúrgicos Urológicos/métodos , Procedimentos Cirúrgicos Urológicos/efeitos adversos , Idoso , Estudos de Coortes , Incidência , Adulto , Posicionamento do Paciente/métodos , Posicionamento do Paciente/efeitos adversosRESUMO
BACKGROUND: Depression is common during adolescence and is associated with adverse educational, employment, and health outcomes in later life. Digital programs are increasingly being implemented in schools to improve and protect adolescent mental health. Although digital depression prevention programs can be effective, there is limited knowledge about how contextual factors influence real-world delivery at scale in schools. OBJECTIVE: The purpose of this study was to examine the contextual factors that influence the implementation of the Future Proofing Program (FPP) from the perspectives of school staff. The FPP is a 2-arm hybrid type 1 effectiveness-implementation trial evaluating whether depression can be prevented at scale in schools, using an evidence-based smartphone app delivered universally to year 8 students (13-14 years of age). METHODS: Qualitative interviews were conducted with 23 staff from 20 schools in New South Wales, Australia, who assisted with the implementation of the FPP. The interviews were guided by our theory-driven logic model. Reflexive thematic analysis, using both deductive and inductive coding, was used to analyze responses. RESULTS: Staff perceived the FPP as a novel ("innovative approach") and appropriate way to address an unmet need within schools ("right place at the right time"). Active leadership and counselor involvement were critical for planning and engaging; teamwork, communication, and staff capacity were critical for execution ("ways of working within schools"). Low student engagement and staffing availability were identified as barriers for future adoption and implementation by schools ("reflecting on past experiences"). CONCLUSIONS: Four superordinate themes pertaining to the program, implementation processes, and implementation barriers were identified from qualitative responses by school staff. On the basis of our findings, we proposed a select set of recommendations for future implementation of digital prevention programs delivered at scale in schools. These recommendations were designed to facilitate an organizational change and help staff to implement digital mental health programs within their schools. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): RR2-10.1136/bmjopen-2020-042133.
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BACKGROUND: When COVID-19 spread to Australia in January 2020, many communities were already in a state of emergency from the Black Summer bushfires. Studies of adolescent mental health have typically focused on the effects of COVID-19 in isolation. Few studies have examined the impact of COVID-19 and other co-occurring disasters, such as the Black Summer bushfires in Australia, on adolescent mental health. METHODS: We conducted a cross-sectional survey to examine the impact of COVID-19 and the Black Summer bushfires on the mental health of Australian adolescents. Participants (N = 5866; mean age 13.61 years) answered self-report questionnaires about COVID-19 diagnosis/quarantine (being diagnosed with and/or quarantined because of COVID-19) and personal exposure to bushfire harm (being physically injured, evacuated from home and/or having possessions destroyed). Validated standardised scales were used to assess depression, psychological distress, anxiety, insomnia, and suicidal ideation. Trauma related to COVID-19 and the bushfires was also assessed. The survey was completed in two large school-based cohorts between October 2020 and November 2021. RESULTS: Exposure to COVID-19 diagnosis/quarantine was associated with increased probability of elevated trauma. Exposure to personal harm by the bushfires was associated with increased probability of elevated insomnia, suicidal ideation, and trauma. There were no interactive effects between disasters on adolescent mental health. Effects between personal risk factors and disasters were generally additive or sub-additive. CONCLUSIONS: Adolescent mental health responses to community-level disasters are multi-faceted. Complex psychosocial factors associated with mental ill health may be relevant irrespective of disaster. Future research is needed to investigate synergistic effects of disasters on young mental health.
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OBJECTIVES: The Future Proofing Study (FPS) was established to examine factors associated with the onset and course of mental health conditions during adolescence. This paper describes the design, methods, and baseline characteristics of the FPS cohort. METHODS: The FPS is an Australian school-based prospective cohort study with an embedded cluster-randomized controlled trial examining the effects of digital prevention programs on mental health. Data sources include self-report questionnaires, cognitive functioning, linkage to health and education records, and smartphone sensor data. Participants are assessed annually for 5 years. RESULTS: The baseline cohort (N = 6388, M = 13.9 years) is broadly representative of the Australian adolescent population. The clinical profile of participants is comparable to other population estimates. Overall, 15.1% of the cohort met the clinical threshold for depression, 18.6% for anxiety, 31.6% for psychological distress, and 4.9% for suicidal ideation. These rates were significantly higher in adolescents who identified as female, gender diverse, sexuality diverse, or Aboriginal and/or Torres Strait Islander (all ps < 0.05). CONCLUSIONS: This paper provides current and comprehensive data about the status of adolescent mental health in Australia. The FPS cohort is expected to provide significant insights into the risk, protective, and mediating factors associated with development of mental health conditions during adolescence.
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Saúde Mental , Humanos , Adolescente , Feminino , Austrália/epidemiologia , Estudos Prospectivos , Inquéritos e QuestionáriosRESUMO
Objective As direct jejunal feeding often causes great fluctuation in glucose levels, continuous or slow infusion is recommended for jejunal tube-fed patients. However, continuous feeding results in prolonged immobility and the loss of activities of daily living. We investigated whether or not intermittent feeding of a low-carbohydrate high-monounsaturated fatty acid (LC/HM) nutrient formula reduces glucose fluctuation in patients who have undergone jejunotomy. Methods Ten bed-ridden non-diabetic patients receiving enteral feeding via a jejunostomy tube were enrolled in this study. LC/HM formula and standard control formula were infused in cross-over order for each patient at a speed of 160 kcal/h. Blood glucose levels were monitored by a continuous glucose monitoring system during the investigation period. Results The mean and standard deviation of the glucose concentrations and mean amplitude of glucose excursion (MAGE) were markedly lower while receiving LC/HM formula than while receiving control standard formula (104 vs. 136 mg/dL, 18.1 vs. 58.1 mg/dL, 50.8 vs. 160 mg/dL, respectively). The post-infusion hyperglycemia [area under the curve (AUC) >140 mg/dL] and peak value of the glucose level were also significantly lower in patients fed LC/HM than the control (25.7 vs. 880 mgã»h/dL and 153 vs. 272 mg/dL, respectively). Reactive hypoglycemia (AUC <70 mg/dL) was also significantly lower (0.63 vs. 16.7 mgã»h/dL) and the minimum value of the glucose level higher (78.4 vs. 61.8 mg/dL) in patients fed LC/HM than the control. Conclusion The LC/HM formula is considered to markedly inhibit glycemic spikes and prevent rebound hypoglycemia in patients who receive enteral feeding after jejunostomy.
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Carboidratos da Dieta/administração & dosagem , Nutrição Enteral/métodos , Hiperglicemia/dietoterapia , Hipoglicemia/dietoterapia , Atividades Cotidianas , Idoso , Idoso de 80 Anos ou mais , Glicemia , Estudos Cross-Over , Feminino , Humanos , Intubação Gastrointestinal/métodos , Jejunostomia/métodos , Masculino , Monitorização Fisiológica , Estado NutricionalRESUMO
BACKGROUND: Appropriate pain management is essential to improve the postoperative recovery after total hip arthroplasty (THA). Various case reports have indicated that anterior quadratus lumborum block (QLB) provides effective postoperative analgesia in lower limb surgeries. However, few randomized controlled trials have confirmed the efficacy of anterior QLB for lower limb surgeries. The aim of this single-center, double-blind, randomized controlled trial is to confirm the efficacy of anterior QLB for postoperative recovery after THA. METHODS: The participants will be randomly assigned to either the anterior QLB or placebo groups, using a set of random numbers for the allocation sequence. Only pharmacists will be aware of the allocations; other investigators will be blinded until study completion. After induction of general anesthesia, anterior QLB will be performed by using 0.25% levobupivacaine or normal saline. Fentanyl will be administered according to blood pressure change during the surgery. The primary outcome will be the quality of recovery 40 score (QoR-40). Secondary outcomes will include the visual analog scale score of pain intensity at rest and movement, intraoperative and postoperative doses of fentanyl, and incidence of postoperative nausea and vomiting. Statistical analysis will be performed by using the Student's t test, Mann-Whitney U test, and Fisher's exact test as appropriate. A P value of less than 0.05 will be considered statistically significant. DISCUSSION: The results of our study will reveal whether anterior QLB is effective for postoperative recovery after THA. TRIAL REGISTRATION: UMIN Clinical Trials Registry, UMIN000032255. Registered on 15 April 2018.
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Artroplastia de Quadril/efeitos adversos , Bloqueio Nervoso/métodos , Manejo da Dor/métodos , Dor Pós-Operatória/diagnóstico , Náusea e Vômito Pós-Operatórios/epidemiologia , Músculos Abdominais/inervação , Adulto , Analgésicos Opioides/administração & dosagem , Anestésicos Locais/administração & dosagem , Método Duplo-Cego , Feminino , Humanos , Incidência , Masculino , Medição da Dor , Dor Pós-Operatória/epidemiologia , Dor Pós-Operatória/etiologia , Náusea e Vômito Pós-Operatórios/etiologia , Náusea e Vômito Pós-Operatórios/prevenção & controle , Período Pós-Operatório , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
Bilateral quadratus lumborum blockade (QLB) using ultrasound guidance has been introduced as an abdominal truncal block to improve postoperative analgesia and quality of recovery (QoR) after abdominal surgery, but efficacy remains controversial. The primary aim of this study was to evaluate the efficacy of posterior QLB on the postoperative QoR, and secondarily to evaluate postoperative pain after gynaecological laparoscopic surgery (LS). This study was a single-centre randomized controlled trial. QLB group patients underwent bilateral posterior quadratus lumborum injections with 25-30 mL of 0.25% levobupivacaine after induction of general anaesthesia; the control group underwent no block. Both groups were administered fentanyl-based intravenous patient-controlled analgesia postoperatively. The postoperative QoR was measured using the Quality of Recovery 40 (QoR-40) questionnaire score; postoperative pain was evaluated using the visual analogue scale (VAS) and the cumulative postoperative fentanyl dose. Thirty-one and 29 patients were randomised to the QLB and control groups, respectively. The intraoperative remifentanil dosage was significantly less in the QLB group. The median (interquartile range) for the QoR-40 score was not different between the groups: 154 (133-168) in the QLB group and 158 (144-172) in the control group. There were no statistically significant differences in secondary outcome variables. Single-shot QLB did not improve the QoR or postoperative pain in patients managed by multimodal analgesia after gynaecological LS.
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Músculos Abdominais , Procedimentos Cirúrgicos em Ginecologia , Laparoscopia , Bloqueio Nervoso , Anestésicos Locais , Feminino , Humanos , Dor Pós-OperatóriaRESUMO
OBJECTIVE: Examine the feasibility and acceptability of an electrocardiogram (ECG) attached to a mobile phone (iECG) screening device for atrial fibrillation (AF) in Aboriginal Controlled Community Health Services (ACCHS) and other community settings. METHODS: Semi-structured interviews were conducted with ACCHS staff in urban, rural and remote communities in three Australian states/territories. Quantitative and qualitative questions identified the enabling factors and barriers for staff and Aboriginal patients' receptiveness to the device. Mean quantitative scores and their standard deviation were calculated in Microsoft Excel and qualitative questions were thematically analysed. RESULTS: Eighteen interviews were conducted with 23 staff across 11 ACCHS. Quantitative data found staff were confident in providing iECG screening and managing the referral pathway, and thought the process was beneficial for patients. Qualitative data highlighted the usefulness of the device to undertake opportunistic screening and acceptability in routine practice, and provided opportunities to engage patients in education around AF. CONCLUSION: The iECG device was well accepted within ACCHSs and was feasible to use to screen for AF among Aboriginal patients. Implications for public health: The device can be used in clinical and community settings to screen Aboriginal people for atrial fibrillation to help reduce rates of stroke and other cardiovascular diseases.
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Fibrilação Atrial/diagnóstico , Telefone Celular , Eletrocardiografia/instrumentação , Programas de Rastreamento/métodos , Aplicativos Móveis , Havaiano Nativo ou Outro Ilhéu do Pacífico , Aceitação pelo Paciente de Cuidados de Saúde , Adulto , Austrália , Eletrocardiografia/métodos , Estudos de Viabilidade , Feminino , Humanos , Entrevistas como Assunto , Masculino , Atenção Primária à Saúde , Pesquisa Qualitativa , Saúde da População Rural , População Rural , População UrbanaRESUMO
OBJECTIVE: Aspiration of subglottic secretion is a widely used intervention to prevent ventilator-associated pneumonia (VAP). This study aimed to compare the efficacy of continuous and intermittent subglottic secretion drainage (SSD) in preventing VAP. METHODS: A single-center randomized controlled trial was conducted on adult postoperative patients who were expected to undergo mechanical ventilation for more than 48 hours. Primary outcome measure was incidence of VAP and secondary outcome measures were length of mechanical ventilation and intensive-care unit (ICU) stay. RESULTS: Fifty-nine patients received continuous SSD, while 60 patients received intermittent SSD. Of these 119 patients, 88 (74%) were excluded and 15 and 16 patients were allocated to receive continuous and intermittent SSD, respectively. VAP was detected in 4 (26.7%) and 7 (43.8%) patients in the continuous and intermittent groups, respectively, (p=0.320). The length of mechanical ventilation was significantly shorter (p=0.034) in the continuous group (99.5±47.1 h) than in the intermittent group (159.9±94.5 h). The length of ICU stay was also shorter (p=0.0097) in the continuous group (6.3±2.1 days) than the intermittent group (9.8±4.8 days). CONCLUSIONS: Although continuous SSD did not reduce the incidence of VAP, it reduced the length of mechanical ventilation and ICU stay when compared to intermittent SSD.
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Previous screening of a single-gene knockout library consisting of 3,908 disrupted-mutant strains allowed us to identify 51 thermotolerant genes that are essential for survival at a critical high temperature (CHT) in Escherichia coli [Murata M, Fujimoto H, Nishimura K, Charoensuk K, Nagamitsu H, Raina S, Kosaka T, Oshima T, Ogasawara N, Yamada M (2011) PLoS ONE 6: e20063]. In this study, we identified another 21 thermotolerant genes. E. coli thus has 72 thermotolerant genes in total. The genes are classified into 8 groups: genes for energy metabolism, outer membrane organization, DNA double-strand break repair, tRNA modification, protein quality control, translation control, cell division and transporters. This classification and physiological analysis indicate the existence of fundamental strategies for survival at a CHT, which seems to exclude most of the heat shock responses.
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Adaptação Fisiológica , Escherichia coli/genética , Escherichia coli/fisiologia , Genes Bacterianos , Temperatura Alta , Teste de Complementação GenéticaRESUMO
STUDY OBJECTIVE: The glass vial of acetaminophen as an intravenous preparation (Acelio®, Terumo, Japan) has a strong internal negative pressure. The aim of our study was to determine if this negative pressure could result in medication administration errors if not released prior to connecting to the IV set. DESIGN: Questionnaire survey and simulation study. SETTING: University hospital and its affiliated hospitals. SUBJECTS: Fifty-two anesthesiologists in 6 different hospitals in Yokohama. MEASUREMENTS: A questionnaire on current practice was sent to the subjects. The authors then first calculated the internal pressure of the Acelio® vial followed by a simulation set-up. This set-up measured the amount of saline that could be aspirated from a syringe loaded on a syringe pump connected via a secondary IV line when the Acelio® vial was attached to the primary line without prior release of the internal pressure. The volume of aspiration was tested with two syringe sizes and with a fully open IV clamp vs partially closed. MAIN RESULTS: Twenty-nine (56.9%) of 51 anesthesiologists who responded to the survey had connected the Acelio® vial at least once without releasing the internal negative pressure, and 21 experienced consequences such as backflow of the venous blood. The pressure inside the Acelio® vial was 81.8±19.6Torr. With the clamp of the simulated IV line fully open, the amount of saline aspirated before the alarm of the syringe pump went off was 1.5±0.1ml and 3.2±0.3ml when 20ml and 50ml syringes were used, respectively. When the clamp was partially closed to allow 2 drops per second, this value was 1.3±0.1ml and 2.3±0.1ml, respectively. After removing the plunger from the holder of the syringe pump, an additional 7ml (clamp partially closed) or 15-18ml (clamp fully open) was aspirated in the subsequent 1min. CONCLUSIONS: A considerable number of anesthesiologists experienced consequences caused by the negative pressure inside the Acelio® vial. This can also cause aspiration of the contents of the syringe pump.
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Acetaminofen/administração & dosagem , Administração Intravenosa/efeitos adversos , Bombas de Infusão/efeitos adversos , Erros de Medicação/prevenção & controle , Pressão/efeitos adversos , Administração Intravenosa/instrumentação , Anestesiologistas/educação , Falha de Equipamento , Humanos , Treinamento por Simulação , Inquéritos e Questionários , SeringasRESUMO
Regional anesthesia, especially epidural anesthesia, rarely causes involuntary movement Here we present a case of a patient who demonstrated myoclonus-like involuntary movement of the lower limbs during continuous infusion of ropivacaine, fentanyl, and droperidol through the thoracic epidural catheter. This movement disappeared when the epidural infusion was stopped, but reappeared when the epidural infusion was restarted. Naloxone did not eliminate the movement The patient was thereafter discharged uneventfully. This case and other reports in the literature suggest that involuntary movement associated with regional anesthesia is rare and self-limiting. However, careful consideration should be given to exclude other, potentially dangerous complications.
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Amidas/efeitos adversos , Anestesia Epidural/efeitos adversos , Droperidol/efeitos adversos , Discinesia Induzida por Medicamentos/fisiopatologia , Discinesias/fisiopatologia , Fentanila/efeitos adversos , Extremidade Inferior/fisiopatologia , Combinação de Medicamentos , Feminino , Humanos , Neoplasias Pulmonares/cirurgia , Pessoa de Meia-Idade , RopivacainaRESUMO
Candida glabrata was continuously isolated in cultured urine samples from a subject with thrombotic thrombocytopenic purpura. Yeast-like fungal phagocytosis found in gram staining led to agents being tested for antifungal susceptibility, revealing hyposensitivity to micafungin (MCFG) of MIC < 2 mg/mL. MCFG administered for 10 days failed to cure C. glabrata infection. To clarify why hyposensitivity occurred, we analyzed the FKS gene sequence using the PCR, finding a deficit of 3 bases coding phenylalanine at FKS2 gene amino acid 659. MCFG hyposensitivity may thus occur in long-term candin-class anti-fungal agent treatment. Candin-class agents have potent anti-fungal activity with fewer adverse effects and are widely used clinically. Hyposensitivity due to resistant C. glabrata species showed thus be considered in fungal infection treatment.
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Antifúngicos/farmacologia , Candida glabrata/efeitos dos fármacos , Candida glabrata/genética , Farmacorresistência Fúngica , Equinocandinas/farmacologia , Genes Fúngicos/genética , Lipopeptídeos/farmacologia , Mutação , Idoso , Feminino , Proteínas Fúngicas/genética , Glucosiltransferases/genética , Humanos , Proteínas de Membrana/genética , MicafunginaRESUMO
Two α-1,3-glucanase isozymes, designated as α-1,3-glucanase 1 (Agl-FH1) and α-1,3-glucanase 2 (Agl-FH2), were purified from the culture medium of Paenibacillus glycanilyticus FH11. Agl-FH1 and Agl-FH2 exhibited similar characteristics such as optimal pH, pH stability, optimal temperature, thermostability, and molecular masses on SDS-PAGE. However, their hydrolysis products of α-1,3-glucan varied somewhat. Agl-FH1 hydrolyzed α-1,3-glucan into a mixture of maltotriose and maltotetraose, and maltotetraose was the major hydrolysis product of Agl-FH2. N-terminal amino acid sequence analysis and LC-MS/MS analysis of trypsin digested fragments revealed several differences between the amino acid sequences of Agl-FH1 and Agl-FH2. Genes of Agl-FH1 and Agl-FH2 were subcloned into an expression plasmid, and both enzymes were successfully expressed in Escherichia coli. The recombinant Agl-FH1 and Agl-FH2 exhibited the same enzymatic properties as those of each wild-type enzyme, and both of the recombinants showed the activity on the protoplast formation of Schizophyllum commune mycelia. A great diversity was detected in the C-terminal region of family 87 α-1,3-glucanases. Compared with Agl-FH2 which is highly sequence-related to the known α-1,3-glucanases, the C-terminal region of Agl-FH1 has only slight similarity to them (approximately 20% identity). Our analysis revealed that Agl-FH1 was the first member of a new subgroup of family 87 α-1,3-glucanases.
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Proteínas de Bactérias/química , Glucanos/química , Glicosídeo Hidrolases/química , Paenibacillus/química , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/farmacologia , Estabilidade Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Glucanos/metabolismo , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Glicosídeo Hidrolases/farmacologia , Concentração de Íons de Hidrogênio , Hidrólise , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Isoenzimas/farmacologia , Maltose/análogos & derivados , Maltose/química , Maltose/metabolismo , Dados de Sequência Molecular , Micélio/efeitos dos fármacos , Micélio/crescimento & desenvolvimento , Paenibacillus/enzimologia , Estrutura Terciária de Proteína , Protoplastos/efeitos dos fármacos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Schizophyllum/efeitos dos fármacos , Schizophyllum/crescimento & desenvolvimento , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , TemperaturaRESUMO
Appropriate parental care by the father can greatly facilitate healthy human family life. Much less is known from animal studies about the factors leading to paternal parental care than those favoring maternal parent care. Recently, we have reported that sires of the ICR strain of laboratory mice can express maternal-like retrieval behavior when separated from their pups through ultrasound and pheromonal signals from the dam, i.e. mate-dependent parental care. The sire's retrieval behavior was inhibited by prior treatment of scopolamine, a muscarinic cholinergic inhibitor, and recovered by physostigmine. KCNQ K(+)-channel blocking and enhancing drugs, linopiridine and retigabine, were also examined. Linopiridine alone did not enhance care after pairing with the dam, nor change scopolamine-induced inhibition of care. Retigabine totally suppressed parental care, and this effect was partially rescued by co-administration of linopiridine. These results indicate the involvement of cholinergic cellular signaling in the central nervous system in the maternal induction of paternal parental behavior in ICR mice.
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Comportamento Apetitivo/fisiologia , Neurônios Colinérgicos/fisiologia , Comportamento Materno/fisiologia , Comportamento Paterno/fisiologia , Escopolamina/administração & dosagem , Animais , Comportamento Apetitivo/efeitos dos fármacos , Antagonistas Colinérgicos/administração & dosagem , Neurônios Colinérgicos/efeitos dos fármacos , Feminino , Masculino , Comportamento Materno/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos ICR , Comportamento Paterno/efeitos dos fármacosRESUMO
Compared with the knowledge of maternal care, much less is known about the factors required for paternal parental care. Here we report that new sires of laboratory mice, though not spontaneously parental, can be induced to show maternal-like parental care (pup retrieval) using signals from dams separated from their pups. During this interaction, the maternal mates emit 38-kHz ultrasonic vocalizations to their male partners, which are equivalent to vocalizations that occur following pheromone stimulation. Without these signals or in the absence of maternal mates, the sires do not retrieve their pups within 5 min. These results show that, in mice, the maternal parent communicates to the paternal parent to encourage pup care. This new paradigm may be useful in the analysis of the parental brain during paternal care induced by interactive communication.
Assuntos
Comportamento Sexual Animal/fisiologia , Comportamento Social , Vocalização Animal/fisiologia , Animais , Animais Recém-Nascidos , Percepção Auditiva/fisiologia , Sinais (Psicologia) , Feminino , Masculino , Comportamento Materno , Camundongos , Camundongos Endogâmicos ICR , Percepção Olfatória/fisiologia , UltrassomRESUMO
H2AX is an important factor for chromatin remodeling to facilitate accumulation of DNA damage-related proteins at DNA double-strand break (DSB) sites. In order to further understand the role of H2AX in the DNA damage response (DDR), we attempted to identify H2AX-interacting proteins by proteomics analysis. As a result, we identified nucleolin as one of candidates. Here, we show a novel role of a major nucleolar protein, nucleolin, in DDR. Nucleolin interacted with γ-H2AX and accumulated to laser micro-irradiated DSB damage sites. Chromatin Immunoprecipitation assay also displayed the accumulation of nucleolin around DSB sites. Nucleolin-depleted cells exhibited repression of both ATM-dependent phosphorylation following exposure to γ-ray and subsequent cell cycle checkpoint activation. Furthermore, nucleolin-knockdown reduced HR and NHEJ activity and showed decrease in IR-induced chromatin accumulation of HR/NHEJ factors, agreeing with the delayed kinetics of γ-H2AX focus. Moreover, nucleolin-knockdown decreased MDC1-related events such as focus formation of 53 BP1, RNF168, phosphorylated ATM, and H2A ubiquitination. Nucleolin also showed FACT-like activity for DSB damage-induced histone eviction from chromatin. Taken together, nucleolin could promote both ATM-dependent cell cycle checkpoint and DSB repair by functioning in an MDC1-related pathway through its FACT-like function.
Assuntos
Quebras de DNA de Cadeia Dupla , Reparo do DNA/fisiologia , Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Proteínas Nucleares/fisiologia , Fosfoproteínas/fisiologia , Proteínas de Ligação a RNA/fisiologia , Transativadores/fisiologia , Proteínas Adaptadoras de Transdução de Sinal , Animais , Pontos de Checagem do Ciclo Celular , Proteínas de Ciclo Celular , Cromatina/metabolismo , Raios gama , Técnicas de Silenciamento de Genes , Células HeLa , Histonas/metabolismo , Histonas/fisiologia , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Camundongos , Proteínas Nucleares/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Fosforilação , Proteômica , Proteínas de Ligação a RNA/genética , Proteínas de Ligação a RNA/metabolismo , Transativadores/metabolismo , Ubiquitinação , NucleolinaRESUMO
The molecular mechanism supporting survival at a critical high temperature (CHT) in Escherichia coli was investigated. Genome-wide screening with a single-gene knockout library provided a list of genes indispensable for growth at 47°C, called thermotolerant genes. Genes for which expression was affected by exposure to CHT were identified by DNA chip analysis. Unexpectedly, the former contents did not overlap with the latter except for dnaJ and dnaK, indicating that a specific set of non-heat shock genes is required for the organism to survive under such a severe condition. More than half of the mutants of the thermotolerant genes were found to be sensitive to H(2)O(2) at 30°C, suggesting that the mechanism of thermotolerance partially overlaps with that of oxidative stress resistance. Their encoded enzymes or proteins are related to outer membrane organization, DNA double-strand break repair, tRNA modification, protein quality control, translation control or cell division. DNA chip analyses of essential genes suggest that many of the genes encoding ribosomal proteins are down-regulated at CHT. Bioinformatics analysis and comparison with the genomic information of other microbes suggest that E. coli possesses several systems for survival at CHT. This analysis allows us to speculate that a lipopolysaccharide biosynthesis system for outer membrane organization and a sulfur-relay system for tRNA modification have been acquired by horizontal gene transfer.