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tRNAHis guanylyltransferase (Thg1) has unique reverse (3'-5') polymerase activity occurring in all three domains of life. Most eukaryotic Thg1 homologs are essential genes involved in tRNAHis maturation. These enzymes normally catalyze a single 5' guanylation of tRNAHis lacking the essential G-1 identity element required for aminoacylation. Recent studies suggest that archaeal type Thg1, which includes most archaeal and bacterial Thg1 enzymes is phylogenetically distant from eukaryotic Thg1. Thg1 is evolutionarily related to canonical 5'-3' forward polymerases but catalyzes reverse 3'-5'polymerization. Similar to its forward polymerase counterparts, Thg1 encodes the conserved catalytic palm domain and fingers domain. Here we investigate the minimal requirements for reverse polymerization. We show that the naturally occurring minimal Thg1 enzyme from Ignicoccus hospitalis (IhThg1), which lacks parts of the conserved fingers domain, is catalytically active. And adds all four natural nucleotides to RNA substrates, we further show that the entire fingers domain of Methanosarcina acetivorans Thg1 and Pyrobaculum aerophilum Thg1 (PaThg1) is dispensable for enzymatic activity. In addition, we identified residues in yeast Thg1 that play a part in preventing extended polymerization. Mutation of these residues with alanine resulted in extended reverse polymerization. PaThg1 was found to catalyze extended, template dependent tRNA repair, adding up to 13 nucleotides to a truncated tRNAHis substrate. Sequencing results suggest that PaThg1 fully restored the near correct sequence of the D- and acceptor stem, but also produced incompletely and incorrectly repaired tRNA products. This research forms the basis for future engineering efforts towards a high fidelity, template dependent reverse polymerase.
Assuntos
Desulfurococcaceae/enzimologia , Methanosarcina/enzimologia , Nucleotidiltransferases/metabolismo , Pyrobaculum/enzimologia , RNA de Transferência de Histidina/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/enzimologia , Sequência de Aminoácidos , Sítios de Ligação , Domínio Catalítico , Sequência Conservada , Desulfurococcaceae/genética , Expressão Gênica , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/metabolismo , Cinética , Methanosarcina/genética , Modelos Moleculares , Mutação , Nucleotidiltransferases/química , Nucleotidiltransferases/genética , Polimerização , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Engenharia de Proteínas/métodos , Pyrobaculum/genética , RNA de Transferência de Histidina/química , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
STUDY OBJECTIVE: The authors present the first ever laparoscopic-assisted uterus retrieval in a live donor for uterus transplant. DESIGN: A step-by-step surgical demonstration. SETTING: Galaxy CARE Laparoscopy Institute, Pune, India. PATIENTS: Two patients, ages 21 and 26 years, with Mayer-Rokitansky-Küster-Hauser syndrome and Asherman syndrome, respectively, with their mothers as donors. INTERVENTIONS: A 12-member team was formed. After a review of the available literature on uterine transplant, a protocol was formulated and submitted to the Institutional Review Board (IRB). Approval from the Institutional Review Board was obtained. Thorough screening of the candidates was done. Two consecutive uterine transplants were done on 2 successive days. Vessels were harvested laparoscopically in both donors. Uterus was retrieved through a small abdominal incision, to prevent any injury to the uterus and harvested vessels. Uterus was transplanted in the recipients by end-to-side anastomosis of the harvested vessels to the external iliac vessels, followed by anchoring of supports of the donor uterus to those of the recipients. MEASUREMENTS AND MAIN RESULTS: Surgical intra- and postoperative parameters, postoperative investigations, and follow-up data of 4 months. The operative time for laparoscopic donor surgery was 4 hours. Bench surgery took 45 minutes. The recipient surgery was completed in 4 hours. There were no intraoperative or immediate postoperative complications. Both recipients started menstruating after 34 days and 48 days, respectively, and have had 3 cycles of menses at regular intervals to date. After discharge, follow-up cervical biopsies at 3 weekly intervals showed no signs of rejection. Uterine artery Doppler ultrasound showed good flow in both patients. CONCLUSION: Laparoscopic-assisted donor retrieval is feasible and affords all advantages of a minimally invasive technique. It helps in better dissection of vessels, shortens the operative time, and helps minimize tissue handling, thereby reducing the morbidity of the procedure.
Assuntos
Laparoscopia/métodos , Doadores Vivos , Coleta de Tecidos e Órgãos/métodos , Útero/transplante , Transtornos 46, XX do Desenvolvimento Sexual/cirurgia , Adulto , Anormalidades Congênitas/cirurgia , Dissecação/métodos , Estudos de Viabilidade , Feminino , Ginatresia/cirurgia , Humanos , Índia , Mães , Ductos Paramesonéfricos/anormalidades , Ductos Paramesonéfricos/cirurgia , Duração da Cirurgia , Adulto JovemRESUMO
STUDY OBJECTIVE: To report the first ever laparoscopic-assisted live donor uterus retrieval in 2 patients for uterus transplant. DESIGN: Case study (Canadian Task Force classification III). SETTING: Galaxy CARE Laparoscopy Institute, Pune, India. PATIENTS: Two patients with absolute uterine factor infertility with their mothers as donors. INTERVENTIONS: In vitro fertilization and uterine transplant. MEASUREMENTS AND MAIN RESULTS: A 12-member team was formed, and approval for transplant was obtained from the institutional review board. Pretransplant, in vitro fertilization for both patients was done. Two consecutive uterine transplants were done on 2 successive days. Vessels were harvested laparoscopically in both donors. Uterus and harvested vessels were retrieved by a small abdominal incision to prevent injury and infection. The uterus was transplanted in the recipients by end to side anastomosis of the harvested vessels to external iliac vessels, followed by anchoring of supports of the donor uterus to those of the recipients. Surgical intra- and postoperative parameters, postoperative investigations, and follow-up data of 6 months were measured. Operative time for laparoscopic donor surgery was 4 hours. Bench surgery took 45 minutes. Recipient surgery time was 4 hours. There were no intraoperative or immediate postoperative complications. Both the recipients started menstruating after 34 days and 48 days, respectively, and have had 6 cycles of menses at regular intervals. Uterine artery Doppler showed good flow in both patients. Hysteroscopy-guided cervical biopsies were used as a method of surveillance of graft rejection after uterine transplant. Office hysteroscopy was done after 2 months in both patients, and hysteroscopy-guided endometrial and cervical biopsies were taken. Minimal slough was seen on the endometrium in the patient with Mayer-Rokitansky-Küster-Hauser syndrome, which was removed. Repeat hysteroscopy after 10 days showed a healthy endometrium. CONCLUSIONS: Laparoscopic-assisted uterus donor retrieval is feasible and affords all the advantages of a minimally invasive technique, thereby reducing the morbidity of the procedure. It helps in better dissection of the vessels, shortens the operative time, and helps to minimize tissue handling of the harvested uterus and vessels.
Assuntos
Doação Dirigida de Tecido , Infertilidade Feminina/cirurgia , Laparoscopia , Doadores Vivos , Útero/transplante , Adulto , Feminino , Fertilização in vitro , Humanos , Histeroscopia , Índia , Pessoa de Meia-Idade , Gravidez , Adulto JovemRESUMO
The pelvic anatomy is constant, with few variations. It has a distinct appearance when observed using the 2-dimensional laparoscope. Thus it is important to master the laparoscopic anatomy and use this knowledge to perform better surgery. The laparoscope offers better vision in a narrow space and thus helps better understanding of the anatomy than what can be seen during open surgery. The objectives of this video are to enable the observer to become familiar with the surgical anatomy, to apply anatomical knowledge to develop fine surgical skills, and to address the myths of open surgical anatomy. The lucid graphics, images, and commentary will enable easy understanding of the pelvic anatomy.
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Histerectomia Vaginal/métodos , Laparoscopia/métodos , Diafragma da Pelve/anatomia & histologia , Útero/anatomia & histologia , Feminino , Humanos , Útero/irrigação sanguínea , Gravação em VídeoRESUMO
Surfactant replacement therapy is currently approved by the United States Food and Drug Administration (FDA) for premature infants with respiratory distress syndrome (RDS) caused by surfactant deficiency due to immaturity. There is strong evidence that surfactant decreases mortality and air leak syndromes in premature infants with RDS. However, surfactant is also used "off-label" for respiratory failure beyond classic RDS. This review discusses current evidence for the use of off-label surfactant therapy for (1) term infants with lung disease such as meconium aspiration syndrome (MAS), pneumonia/sepsis, and congenital diaphragmatic hernia (2) premature infants after 72 h for acute respiratory failure, and (3) the use of surfactant lavage. At last, we briefly describe the use of surfactants for drug delivery and the current evidence on evaluating infants for surfactant deficiency.
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Síndrome de Aspiração de Mecônio , Surfactantes Pulmonares , Síndrome do Desconforto Respiratório do Recém-Nascido , Recém-Nascido , Humanos , Feminino , Tensoativos/uso terapêutico , Síndrome de Aspiração de Mecônio/tratamento farmacológico , Síndrome do Desconforto Respiratório do Recém-Nascido/tratamento farmacológico , Surfactantes Pulmonares/uso terapêutico , Recém-Nascido PrematuroRESUMO
Background: Gender disparities in emergency medicine (EM) persist, with women underrepresented in leadership positions and faced with unique challenges, such as gender discrimination and harassment. To address these issues, professional development programs for women have been recommended. Objectives: The purpose of this scoping review was to examine current women's professional development programs for EM and develop a collection of program characteristics, meeting topics, and tips for success that can be useful to new or existing women's professional development programs. Methods: The authors systematically searched research databases for literature detailing current women's professional development programs for EM physicians. Studies detailing professional development programs for female physicians in EM were included. Results: After 149 unique articles were screened, 11 studies met inclusion criteria, describing 10 professional development programs for women in EM. The most commonly cited program objectives included providing mentors and role models (n = 9, 90%), offering career advice and promoting professional advancement and leadership skills (n = 5, 50%), increasing academic recognition for women (n = 4, 40%), and promoting work-life balance and integration (n = 2, 20%). The most common topics covered in program sessions included mentorship and coaching, compensation and/or negotiation, leadership skills, and career advancement and promotion. Challenges and barriers to the success of these programs included a lack of funding and support, difficulty in recruiting participants, lack of institutional recognition and support, lack of time, and difficulty in sustaining the program over time. Conclusions: The study's findings can inform the development of programs that promote gender equity and support the advancement of women in EM.
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OBJECTIVE: To describe the population to which we administered recombinant erythropoietin and to determine the effectiveness of this treatment as quantified by the change in hematocrit. STUDY DESIGN: This retrospective chart review study included infants who received erythropoietin for the treatment of anemia of prematurity. RESULTS: There were 132 infants representing 162 unique treatment courses included in the study. The average duration of therapy was 9 days (±7) and 6 doses (±2). The average change in hematocrit (Hct) was 6.2% (SD 3.9%, p < 0.001). Rise in Hct was associated with a higher number of rEPO doses (p < 0.001) and higher postmenstrual age (p < 0.001). In our small cohort we did not find an association between the number of rEPO doses and retinopathy of prematurity (ROP) requiring treatment. CONCLUSION: Erythropoietin is safe and effective at treating anemia of prematurity as evidenced by a clinically and statistically significant increase in Hct from baseline.
Assuntos
Anemia Neonatal , Eritropoetina , Recém-Nascido Prematuro , Unidades de Terapia Intensiva Neonatal , Proteínas Recombinantes , Humanos , Estudos Retrospectivos , Recém-Nascido , Eritropoetina/uso terapêutico , Eritropoetina/administração & dosagem , Feminino , Masculino , Proteínas Recombinantes/uso terapêutico , Proteínas Recombinantes/administração & dosagem , Anemia Neonatal/tratamento farmacológico , Hematócrito , Retinopatia da Prematuridade/tratamento farmacológico , Resultado do Tratamento , Idade Gestacional , Anemia/tratamento farmacológicoRESUMO
OBJECTIVE: To systematically evaluate the measurement properties of 12 patient-reported outcome measures (PROMs) used to measure depression symptom severity in adolescents with depression. Depression symptom severity was chosen as the outcome of focus given its importance as an outcome to measure in adolescents with depression across clinical trials and/or care. METHOD: MEDLINE, PsycInfo, Scopus, CINAHL, and Web of Science were searched from year of inception up to December 7th, 2023. Study appraisal (i.e., risk of bias), evaluation of measurement properties, and evidence synthesis followed the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) guidelines. Included studies evaluated at least one of nine measurement properties as detailed in the COSMIN taxonomy within a reported sample or subgroup of youth 12-24 years, with at least 40% meeting criteria for any depressive disorder. RESULTS: Of the 15,560 records identified, 31 studies for seven PROMs were included in the COSMIN appraisal. Although several PROMs have the potential to accurately measure depression symptom severity in adolescents with depression, at this time none of the PROMs can be recommended for use without further evaluative work. High-quality evidence was generally lacking, largely due to few or inconsistent findings, small sample sizes, and other methodological concerns. CONCLUSION: This systematic review of the measurement properties of 12 PROMs used to measure depression symptom severity in adolescents with depression found that none of the PROMs can be recommended for use until further evaluative work is conducted. Clinicians and researchers should proceed with caution when using these PROMs.
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Laparoendoscopic single-site surgery (LESS) and natural orifice translumenal endoscopic surgery are now being used in gynecologic oncologic procedures. We used our expertise with LESS to perform nerve-sparing laparoscopic radical hysterectomy. A 45-year-old woman with stage IA2 cervical cancer was referred to us. The biopsy specimen showed grade II invasive keratinizing squamous cell carcinoma. We duplicated the steps of our laparoscopic nerve-sparing radical hysterectomy procedure to perform a nerve-sparing radical hysterectomy via LESS using conventional ports and instruments. Oncologic clearance was comparable to that in conventional laparoscopic radical hysterectomy. Bladder function recovered completely after removal of the Foley catheter. Nerve-sparing laparoscopic radical hysterectomy using fewer ports is technically feasible. The oncologic clearance and functional results are comparable to those in the multiport variant.
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Carcinoma de Células Escamosas/cirurgia , Histerectomia/métodos , Laparoscopia/métodos , Neoplasias do Colo do Útero/cirurgia , Carcinoma de Células Escamosas/patologia , Feminino , Humanos , Pessoa de Meia-Idade , Resultado do Tratamento , Neoplasias do Colo do Útero/patologiaRESUMO
STUDY OBJECTIVE: To access the technical feasibility of performing laparoscopic repair of vesicovaginal fistula. MATERIAL AND METHODS: We attempted a laparoscopic repair of vesicovaginal fistula in five women with a history of urinary leakage via the vagina after vaginal hysterectomy. Five pelvic ports were used. The surgical procedure was performed using the same principles as for open surgery, i.e., separation of the vaginal wall from the bladder wall, repair of the fistula, and interposition of the omentum. RESULTS: There was early recovery of the patients in terms of continence, with less chance of recurrence. Results were comparable to those with the vaginal approach. CONCLUSION: The use of minimally invasive surgery for vesicovaginal fistula repair helps to ease the suturing deep in the pelvis, and the magnification facilitates good identification of tissues planes and thus better mobilization of the vaginal and bladder walls and decreases postoperative morbidity.
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Laparoscopia/métodos , Fístula Vesicovaginal/cirurgia , Feminino , Humanos , Cirurgia Endoscópica por Orifício Natural , Bexiga UrináriaRESUMO
The value of involving people with lived experience in the research process (ie, patient engagement) is increasingly being recognised within youth mental health research. The wide-reaching benefits of patient engagement have been documented in the literature, including the empowerment of patients, improvement of research outcomes, and increased relevance of research findings. Although various models exist to guide patient engagement, there are key concepts that deserve exploration to ensure the authentic implementation of these models and development of patient roles. Our Personal View aims to: identify and discuss barriers to patient engagement roles in the context of youth mental health research; consider how key concepts of relational empowerment, fluidity, and flexibility can address some of these barriers; and provide tangible recommendations for implementing authentic patient engagement throughout the research process.
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Saúde Mental , Participação do Paciente , Humanos , AdolescenteRESUMO
Pediatric anxiety disorders (AD) are prevalent disorders with an impact on all aspects of a child's life and functioning.1 Although evidence supports commonly used treatments, there are notable concerns with the research to date.2 Heterogeneity in outcome selection, measurement, analysis, and reporting is a contributing factor to the hinderance of the translation of research into clinical practice.3 Recognition for outcome standardization in pediatric mental health disorders is evolving and there are several initiatives of importance, including the International Consortium for Health Outcomes Measurement (ICHOM), which has developed standardized outcome sets for use in the routine clinical mental health treatment of children and adolescents.4 Similarly, the International Alliance of Mental Health Research Funders5 advocate for use of 1 specific outcome measurement instrument (OMI) in the youth mental health research that they fund. Development of a Core Outcome Set (COS), a minimal set of outcomes that should be measured and reported in clinical trials, has been a solution in other areas of medicine to address heterogeneity in outcome selection and measurement across trials.6 The Core Outcomes and Measures in Pediatric Anxiety Clinical Trials (COMPACT) Initiative will develop a harmonized, evidence- and consensus-based COS that is meaningful to youth and families for use in future trials in pediatric AD.
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Transtornos de Ansiedade , Projetos de Pesquisa , Adolescente , Humanos , Criança , Técnica Delphi , Determinação de Ponto Final , Transtornos de Ansiedade/terapia , Avaliação de Resultados em Cuidados de Saúde , Resultado do TratamentoRESUMO
Postpartum patients may present to the emergency department with complaints ranging from minor issues, requiring only patient education and reassurance, to severe, life-threatening complications that require prompt diagnosis and multidisciplinary consultation and management. At times, vague presentations or overlapping conditions can make it difficult for the emergency clinician to recognize an emergent condition and initiate proper treatment. This issue reviews the major common emergencies that present in postpartum patients, by chief complaint, including hemorrhage, infection, pre-eclampsia, eclampsia, headache, and cardiopulmonary conditions, and reviews the most recent evidence and guidelines.
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Pré-Eclâmpsia , Transtornos Puerperais , Serviço Hospitalar de Emergência , Feminino , Humanos , Período Pós-Parto , Pré-Eclâmpsia/diagnóstico , Pré-Eclâmpsia/terapia , Gravidez , Transtornos Puerperais/diagnóstico , Transtornos Puerperais/terapia , Encaminhamento e ConsultaRESUMO
OBJECTIVES: To determine if temporomandibular joint (TMJ) compressive stresses during incisor biting (1) differed between growing children over time, and (2) were correlated with Frankfort Horizontal-mandibular plane angle (FHMPA, °) and ramus length (Condylion-Gonion (Co-Go), mm). MATERIALS AND METHODS: Three-dimensional anatomical geometries, FHMPA and Co-Go, were measured longitudinally from lateral and posteroanterior cephalographs1 of children aged 6 (T1), 12 (T2), and 18 (T3) years. Geometries were used in numerical models to estimate subject-specific TMJ eminence shape and forces for incisor bite-forces of 3, 5, and 8 Newtons at T1, T2, and T3, respectively. TMJ compressive stresses were estimated via two steps: First, TMJ force divided by age-dependent mandibular condylar dimensions, and second, modified by loading surfaces' congruency. Analysis of variance and Tukey honest significant difference post-hoc tests, plus repeated measures and mixed effects model analyses were used to evaluate differences in variables between facial groups. Regression analyses tested for correlation between age-dependent compressive stresses, FHMPA, and Co-Go. RESULTS: Sixty-five of 842 potential subjects had T1-T3 cephalographs and were grouped by FHMPA at T3. Dolichofacial (FHMPA ≥ 27°, n = 36) compared to meso-brachyfacial (FHMPA< 27°, n = 29) subjects had significantly larger FHMPA at T1-T3, shorter Co-Go at T2 and T3 (all P < .01), and larger increases in TMJ compressive stresses with age (P < .0001). Higher compressive stresses were correlated with larger FHMPA (all R2 ≥ 0.41) and shorter Co-Go (all R2 ≥ 0.49). CONCLUSIONS: Estimated TMJ compressive stress increases from ages 6 to 18 years were significantly larger in dolichofacial compared to meso-brachyfacial subjects and correlated to FHMPA and Co-Go.
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Mandíbula , Articulação Temporomandibular , Adolescente , Força de Mordida , Criança , Face , Humanos , Mandíbula/diagnóstico por imagem , Côndilo Mandibular , Articulação Temporomandibular/diagnóstico por imagemRESUMO
AIM: The purpose of the present study was to explore the experiences of a diverse group of mental health clinicians both in hospital and in the community, who were required to rapidly adopt virtual-care practices in the delivery of mental health services to children, adolescents, and their families. METHODS: Mental health clinicians (N = 117) completed the Clinician Virtual-Care Experience Survey assessing the following domains: ease of technology use, client/patient-provider interaction quality, and clinician wellbeing. RESULTS: Although over 70% of clinicians had not used virtual-care to deliver mental health services prior to the Coronavirus Disease 2019 pandemic, more than 80% felt it was easy to operate the virtual platforms. Clinicians were divided in their perceptions of the effectiveness of virtual-care, with only 42% reporting that they felt they were as effective in delivering healthcare services virtually as compared to in-person. Virtual-care was described as being more effective for specific populations, while challenges were described in building rapport and when delivering difficult or unexpected feedback. CONCLUSIONS: Clinicians felt there were some benefits of adopting virtual-care practices, while challenges were also identified. Understanding of the impact of virtual-care on service providers is essential in order to strengthen mental healthcare for children, adolescents, and their families even beyond the pandemic.
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COVID-19 , Serviços de Saúde Mental , Adolescente , Criança , Pessoal de Saúde , Hospitais , Humanos , Saúde Mental , Pandemias , SARS-CoV-2RESUMO
INTRODUCTION: Paediatric anxiety disorders (AD) are prevalent and persistent mental health conditions worldwide affecting between 10% and 20% of children and adolescents. Despite the high prevalence of paediatric AD, there is limited understanding of which treatments work best. Outcome heterogeneity across paediatric mental health trials has been a significant factor in hindering the ability to compare results and assess the efficacy of such trials. This scoping review will help to identify and synthesise the outcomes reported in paediatric AD trials to date. METHODS AND ANALYSIS: Following the Joanna Briggs Institute scoping review methodology, a comprehensive electronic bibliographic database search (MEDLINE, APA PsycINFO, Embase, CINAHL) strategy will be applied to identify articles examining interventions for children diagnosed with an AD. Articles will be eligible for inclusion if they assess at least one AD intervention (eg, psychological), in children 4-18 years of age inclusive. Initial title and abstract screening will be completed by two trained reviewers independently and in duplicate. Full-text screening of each included article will be completed independently and in duplicate by two of three trained reviewers. Identified outcomes will be mapped to a standard outcome taxonomy developed for core outcome sets. Trial and outcome characteristics will be synthesised using quantitative metrics (counts and frequencies). ETHICS AND DISSEMINATION: As this is a scoping review of the literature and patient information or records were not accessed, institutional ethics approval was not required. Results of this scoping review will be disseminated to clinicians, researchers inclusive of trialists and other stakeholders invested in outcome selection, measurement and reporting in paediatric AD trials. In addition, scoping review results will inform the development of a Core Outcome Set for paediatric AD trials-a minimum set of outcomes that should be measured across trials in an area of health, without precluding the inclusion of other outcomes.
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Transtornos de Ansiedade , Ansiedade , Adolescente , Transtornos de Ansiedade/terapia , Criança , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto , Projetos de Pesquisa , Literatura de Revisão como AssuntoRESUMO
Bronchopulmonary dysplasia (BPD) is a chronic lung disease commonly affecting extremely preterm infants. Although mechanical ventilation and oxygen requirements in premature infants are identified as inciting mechanisms for inflammation and the development of BPD over time, data now support an array of perinatal events that may stimulate the inflammatory cascade prior to delivery. Corticosteroids, such as dexamethasone and hydrocortisone, have proven beneficial for the prevention and management of BPD postnatally due to their anti-inflammatory characteristics. This review aims to examine the pharmacologic properties of several corticosteroids, appraise the existing evidence for postnatal corticosteroid use in preterm infants, and assess steroid management strategies to ameliorate BPD. Finally, we aim to provide guidance based on clinical experience for managing adrenal suppression resulting from prolonged steroid exposure since this is an area less well-studied.