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1.
Curr Oncol ; 29(8): 5748-5762, 2022 08 15.
Article in English | MEDLINE | ID: mdl-36005191

ABSTRACT

CCS often wish to have biological children yet harbour concerns about fertility impairment, pregnancy risks and the general health risks of prospective offspring. To clarify these concerns, health outcomes in survivor offspring born following ART (n = 74, 4.5%) or after spontaneous conception (n = 1585) were assessed in our European offspring study by descriptive and bivariate analysis. Outcomes were compared to a sibling offspring cohort (n = 387) in a 4:1 matched-pair analysis (n = 1681). (i) Survivors were more likely to employ ART than their siblings (4.5% vs. 3.7%, p = 0.501). Successful pregnancies were achieved after a median of one cycle with, most commonly, intracytoplasmic sperm injection (ICSI) using non-cryopreserved oocytes/sperm. (ii) Multiple-sibling births (p < 0.001, 29.7% vs. 2.5%), low birth weight (p < 0.001; OR = 3.035, 95%-CI = 1.615−5.706), and preterm birth (p < 0.001; OR = 2.499, 95%-CI = 1.401−4.459) occurred significantly more often in survivor offspring following ART utilisation than in spontaneously conceived children. ART did not increase the prevalence of childhood cancer, congenital malformations or heart defects. (iii) These outcomes had similar prevalences in the sibling population. In our explorative study, we could not detect an influence on health outcomes when known confounders, such as multiple births, were taken into account.


Subject(s)
Cancer Survivors , Neoplasms , Premature Birth , Child , Female , Humans , Infant, Newborn , Male , Neoplasms/epidemiology , Pregnancy , Pregnancy Outcome , Premature Birth/epidemiology , Prospective Studies , Reproductive Techniques, Assisted/adverse effects , Semen
2.
BMJ Open ; 12(8): e062398, 2022 08 23.
Article in English | MEDLINE | ID: mdl-35998971

ABSTRACT

INTRODUCTION: The perioperative period is high risk for older adults. Depression and anxiety are common perioperative problems, frequently coexisting with cognitive impairment. Older patients with these conditions are more likely than younger patients to experience postoperative delirium, long hospital stays, poor quality of life and rehospitalisation. These experiences can, in turn, exacerbate anxiety and depressive symptoms. Despite these risks, little is known about how to treat perioperative anxiety and depression among older adults. METHODS AND ANALYSIS: We designed a feasibility study of a perioperative mental health intervention bundle to improve perioperative mental health, specifically depression and anxiety. The overarching goals of this study are twofold: first, to adapt and refine an intervention bundle comprised of behavioural activation and medication optimisation to meet the needs of older adults within three surgical patient populations (ie, orthopaedic, oncological and cardiac); and second, to test the feasibility of study procedures and intervention bundle implementation. Quantitative data on clinical outcomes such as depression, anxiety, quality of life, delirium, falls, length of stay, hospitalisation and pain will be collected and tabulated for descriptive purposes. A hybrid inductive-deductive thematic approach will be employed to analyse qualitative feedback from key stakeholders. ETHICS AND DISSEMINATION: The study received approval from the Washington University Institutional Review Board. Results of this study will be presented in peer-reviewed journals, at professional conferences, and to our perioperative mental health advisory board. TRIAL REGISTRATION NUMBER: NCT05110690.


Subject(s)
Delirium , Mental Health , Aged , Anxiety/psychology , Feasibility Studies , Humans , Quality of Life
3.
Methods Mol Biol ; 2224: 29-45, 2021.
Article in English | MEDLINE | ID: mdl-33606204

ABSTRACT

Reporter mice transgenically expressing the bacterial (E. coli) lacZ gene encoding ß-galactosidase (ß-gal, EC 3.2.1.23) are a versatile and extensively used tool to study gene expression and cell lineage patterns. Enzymatic activity of the ß-gal reporter can be effectively visualized at cellular resolution either histochemically using 5-bromo-4-chloro-3-indolyl-ß-D-galactopyranoside (X-gal) or by immunofluorescent detection using a ß-gal-specific antibody. Here, we summarize protocols for the localization of ß-gal expressing cells in whole embryos or organs as well as in histological tissue sections of lacZ reporter mice and discuss their limitations and common pitfalls.


Subject(s)
Gene Expression/genetics , Genes, Reporter/genetics , Lac Operon/genetics , Animals , Cell Lineage/genetics , Embryo, Mammalian/metabolism , Escherichia coli/genetics , Galactosides/genetics , Indoles , Mice , Staining and Labeling/methods
4.
BMJ Open ; 10(12): e041987, 2020 12 16.
Article in English | MEDLINE | ID: mdl-33328261

ABSTRACT

INTRODUCTION: Sedation is a cornerstone therapy in the management of patients receiving mechanical ventilation and is highly influential on outcome. Early sedation depth appears especially influential, as early deep sedation is associated with worse outcome when compared with light sedation. Our research group has shown that patients receiving mechanical ventilation in the emergency department (ED) are exposed to deep sedation commonly, and ED sedation depth is impactful on intensive care unit (ICU) care and clinical outcomes. While extensive investigation has occurred for patients in the ICU, comparatively little data exist from the ED. Given the influence that ED sedation seems to carry, as well as a lack of ED-based sedation trials, there is significant rationale to investigate ED-based sedation as a means to improve outcome. METHODS AND ANALYSIS: This is a multicentre (n=3) prospective, before-and-after pilot trial examining the feasibility of implementing targeted sedation in the immediate postintubation period in the ED. A cohort of 344 patients receiving mechanical ventilation in ED will be included. Feasibility outcomes include: (1) participant recruitment; (2) proportion of Richmond Agitation-Sedation Scale (RASS) scores in the deep sedation range; (3) reliability (agreement) of RASS measurements performed by bedside ED nurses; and (4) adverse events. The proportion of deep sedation measurements before and after the intervention will be compared using the χ2 test. Logistic regression will be used to compare before-and-after differences, adjusting for potential confounders. The inter-rater correlation coefficient will be used to assess paired observations between a study team member and bedside ED nurses, and to describe reliability of RASS measurements. ETHICS AND DISSEMINATION: The Human Research Protection Office at Washington University in St. Louis School of Medicine has approved the study. The publication of peer-reviewed manuscripts and the presentation of abstracts at scientific meetings will be used to disseminate the work. REGISTRATION: ClinicalTrials.gov identifier NCT04410783; Pre-results.


Subject(s)
Intensive Care Units , Respiration, Artificial , Emergency Service, Hospital , Feasibility Studies , Humans , Hypnotics and Sedatives/therapeutic use , Pilot Projects , Prospective Studies , Reproducibility of Results , Washington
5.
BMJ Open ; 9(10): e033379, 2019 10 07.
Article in English | MEDLINE | ID: mdl-31594905

ABSTRACT

INTRODUCTION: Awareness with paralysis is a complication with potentially devastating psychological consequences for mechanically ventilated patients. While rigorous investigation into awareness has occurred for operating room patients, little attention has been paid outside of this domain. Mechanically ventilated patients in the emergency department (ED) have been historically managed in a way that predisposes them to awareness events: high incidence of neuromuscular blockade use, underdosing of analgesia and sedation, delayed administration of analgesia and sedation after intubation, and a lack of monitoring of sedation targets and depth. These practice patterns are discordant to recommendations for reducing the incidence of awareness, suggesting there is significant rationale to examine awareness in the ED population. METHODS AND ANALYSIS: This is a single centre, prospective cohort study examining the incidence of awareness in mechanically ventilated ED patients. A cohort of 383 mechanically ventilated ED patients will be included. The primary outcome is awareness with paralysis. Qualitative reports of all awareness events will be provided. Recognising the potential problem with conventional multivariable analysis arising from a small number of events (expected less than 10-phenomenon of separation), Firth penalised method, exact logistic regression model or penalised maximum likelihood estimation shrinkage (Ridge, LASSO) will be used to assess for predictors of awareness. ETHICS AND DISSEMINATION: Approval of the study by the Human Research Protection Office has been obtained. This work will be disseminated by publication of peer-reviewed manuscripts, presentation in abstract form at scientific meetings and data sharing with other investigators through academically established means.


Subject(s)
Awareness , Consciousness , Emergency Service, Hospital , Paralysis/therapy , Respiration, Artificial/psychology , Adolescent , Adult , Aged , Aged, 80 and over , Clinical Protocols , Female , Hospitalization , Humans , Male , Middle Aged , Paralysis/psychology , Prospective Studies , Young Adult
6.
BMJ Open ; 9(8): e026649, 2019 08 26.
Article in English | MEDLINE | ID: mdl-31455698

ABSTRACT

INTRODUCTION: Obstructive sleep apnoea (OSA) is common among older surgical patients, and delirium is a frequent and serious postoperative complication. Emerging evidence suggests that OSA increases the risk for postoperative delirium. We hypothesise that OSA is an independent risk factor for postoperative delirium, and that in patients with OSA, perioperative adherence to positive airway pressure (PAP) therapy decreases the incidence of postoperative delirium and its sequelae. The proposed retrospective cohort analysis study will use existing datasets to: (i) describe and compare the incidence of postoperative delirium in surgical patients based on OSA diagnosis and treatment with PAP; (ii) assess whether preoperatively untreated OSA is independently associated with postoperative delirium; and (iii) explore whether preoperatively untreated OSA is independently associated with worse postoperative quality of life (QoL). The findings of this study will inform on the potential utility and approach of an interventional trial aimed at preventing postoperative delirium in patients with diagnosed and undiagnosed OSA. METHODS AND ANALYSIS: Observational data from existing electronic databases will be used, including over 100 000 surgical patients and ~10 000 intensive care unit (ICU) admissions. We will obtain the incidence of postoperative delirium in adults admitted postoperatively to the ICU who underwent structured preoperative assessment, including OSA diagnosis and screening. We will use doubly robust propensity score methods to assess whether untreated OSA independently predicts postoperative delirium. Using similar methodology, we will assess if untreated OSA independently predicts worse postoperative QoL. ETHICS AND DISSEMINATION: This study has been approved by the Human Research Protection Office at Washington University School of Medicine. We will publish the results in a peer-reviewed venue. Because the data are secondary and high risk for reidentification, we will not publicly share the data. Data will be destroyed after 1 year of completion of active Institutional Review Board (IRB) approved projects.


Subject(s)
Delirium , Postoperative Complications , Sleep Apnea, Obstructive , Age Factors , Aged, 80 and over , Delirium/diagnosis , Delirium/etiology , Delirium/prevention & control , Female , Humans , Incidence , Male , Observational Studies as Topic , Perioperative Period , Positive-Pressure Respiration/methods , Postoperative Complications/diagnosis , Postoperative Complications/physiopathology , Postoperative Complications/prevention & control , Prognosis , Risk Factors , Sleep Apnea, Obstructive/diagnosis , Sleep Apnea, Obstructive/epidemiology
7.
BMJ Open ; 8(11): e023137, 2018 11 21.
Article in English | MEDLINE | ID: mdl-30467132

ABSTRACT

INTRODUCTION: Delirium is a common, serious postoperative complication. For clinical studies to generate valid findings, delirium assessments must be standardised and administered accurately by independent researchers. The Confusion Assessment Method (CAM) is a widely used delirium assessment tool. The objective was to determine whether implementing a standardised CAM training protocol for researchers at multiple international sites yields reliable inter-rater assessment and accurate delirium diagnosis. METHODS: Patients consented to video recordings of CAM delirium assessments for research purposes. Raters underwent structured training in CAM administration. Training entailed didactic education, role-playing with intensive feedback, apprenticeship with experienced researchers and group discussions of complex cases. Raters independently viewed and scored nine video-recorded CAM interviews. Inter-rater reliability was determined using Fleiss kappa. Accuracy was judged by comparing raters' scores with those of an expert delirium researcher. RESULTS: Twenty-seven raters from eight international research centres completed the study and achieved almost perfect agreement for overall delirium diagnosis, kappa=0.88 (95% CI 0.85 to 0.92). Agreement of the four core CAM features ranged from fair to substantial. The sensitivity and specificity for identifying delirium were 72% (95% CI 60% to 81%) and 99% (95% CI 96% to 100%), considering an expert rater's scores as the reference standard (delirious, n=3; non-delirious, n=6). Delirium severity ratings were tightly clustered, with most scores within 5% of the median. CONCLUSION: Our results demonstrate that, with appropriate training and ongoing scoring discussions, researchers at multiple sites can reliably detect delirium in postsurgical patients. These results support the premise that methodologically rigorous multi-centre studies can yield standardised and accurate determinations of delirium.


Subject(s)
Delirium/diagnosis , Health Personnel/education , Postoperative Complications/diagnosis , Research Personnel/education , Aged , Female , Humans , Male , Middle Aged , Observer Variation , Reproducibility of Results , Severity of Illness Index , Video Recording
8.
BMJ Open ; 8(4): e020124, 2018 04 10.
Article in English | MEDLINE | ID: mdl-29643160

ABSTRACT

INTRODUCTION: Mortality and morbidity following surgery are pressing public health concerns in the USA. Traditional prediction models for postoperative adverse outcomes demonstrate good discrimination at the population level, but the ability to forecast an individual patient's trajectory in real time remains poor. We propose to apply machine learning techniques to perioperative time-series data to develop algorithms for predicting adverse perioperative outcomes. METHODS AND ANALYSIS: This study will include all adult patients who had surgery at our tertiary care hospital over a 4-year period. Patient history, laboratory values, minute-by-minute intraoperative vital signs and medications administered will be extracted from the electronic medical record. Outcomes will include in-hospital mortality, postoperative acute kidney injury and postoperative respiratory failure. Forecasting algorithms for each of these outcomes will be constructed using density-based logistic regression after employing a Nadaraya-Watson kernel density estimator. Time-series variables will be analysed using first and second-order feature extraction, shapelet methods and convolutional neural networks. The algorithms will be validated through measurement of precision and recall. ETHICS AND DISSEMINATION: This study has been approved by the Human Research Protection Office at Washington University in St Louis. The successful development of these forecasting algorithms will allow perioperative healthcare clinicians to predict more accurately an individual patient's risk for specific adverse perioperative outcomes in real time. Knowledge of a patient's dynamic risk profile may allow clinicians to make targeted changes in the care plan that will alter the patient's outcome trajectory. This hypothesis will be tested in a future randomised controlled trial.


Subject(s)
Algorithms , Machine Learning , Postoperative Complications , Adult , Humans , Missouri , Retrospective Studies
9.
BMJ Open ; 8(3): e017079, 2018 03 17.
Article in English | MEDLINE | ID: mdl-29550773

ABSTRACT

INTRODUCTION: Postoperative delirium (POD) is a common complication in elderly patients, characterised by a fluctuating course of altered consciousness, disordered thinking and inattention. Preliminary research has linked POD with persistent cognitive impairment and decreased quality of life. However, these findings maybe confounded by patient comorbidities, postoperative complications and frailty. Our objective is to determine whether POD is an independent risk factor for persistent impairments in attention and executive function after elective surgery. Our central hypothesis is that patients with POD are more likely to have declines in cognition and quality of life 1 year after surgery compared with patients without POD. We aim to clarify whether these associations are independent of potentially confounding factors. We will also explore the association between POD and incident dementia. METHODS AND ANALYSIS: This study will recruit 200 patients from the ongoing Electroencephalography Guidance of Anesthesia to Alleviate Geriatric Syndromes (ENGAGES) study. Patients who live ≤45 miles from the study centre or have a planned visit to the centre 10-16 months postoperatively will be eligible. Patients with POD, measured by the Confusion Assessment Method, will be compared with patients without delirium. The primary outcome of cognitive function and secondary outcomes of quality of life and incident dementia will be compared between cohorts. Cognition will be measured by Trails A and B and Stroop Color and Word Test, quality of life with Veteran's RAND 12-item Health Survey and incident dementia with the Short Blessed Test. Multivariable regression analyses and a Cox proportional hazards analysis will be performed. All results will be reported with 95% CIs and α=0.05. ETHICS AND DISSEMINATION: The study has been approved by the Washington University in St. Louis Institutional Review Board (IRB no 201601099). Plans for dissemination include scientific publications and presentations at scientific conferences. TRIAL REGISTRATION NUMBER: NCT02241655.


Subject(s)
Cognition/physiology , Cognitive Dysfunction/physiopathology , Delirium/physiopathology , Executive Function/physiology , Aged , Attention/physiology , Delirium/epidemiology , Dementia/epidemiology , Dementia/physiopathology , Elective Surgical Procedures/adverse effects , Female , Humans , Incidence , Male , Middle Aged , Postoperative Complications , Predictive Value of Tests , Prospective Studies , Quality of Life , Regression Analysis , Risk Factors
10.
BMJ Open ; 7(9): e017389, 2017 Sep 27.
Article in English | MEDLINE | ID: mdl-28963302

ABSTRACT

INTRODUCTION: The importance of effective communication, a key component of teamwork, is well recognised in the healthcare setting. Establishing a culture that encourages and empowers team members to speak openly in the cardiothoracic (CT) operating room (OR) is necessary to improve patient safety in this high-risk environment. METHODS AND ANALYSIS: This study will take place at Barnes-Jewish Hospital, an academic hospital in affiliation with Washington University School of Medicine located in the USA. All team members participating in cardiac and thoracic OR cases during this 17-month study period will be identified by the primary surgical staff attending on the OR schedule.TeamSTEPPS (Team Strategies and Tools to Enhance Performance and Patient Safety) training course will be taught to all CT OR staff. Before TeamSTEPPS training, staff will respond to a 39-item questionnaire that includes constructs from the Agency for Healthcare Research and Quality Hospital Survey on Patient Safety Culture, Edmondson's 'Measure of psychological safety' questionnaire, and questionnaires on turnover intentions, job satisfaction and 'burnout'. The questionnaires will be readministered at 6 and 12 months.The primary outcomes to be assessed include the perceived psychological safety of CT OR team members, the overall effect of TeamSTEPPS on burnout and job satisfaction, and observed turnover rate among the OR nurses. As secondary outcomes, we will be assessing self-reported rates of medical error and near misses in the ORs with a questionnaire at the end of each case. ETHICS AND DISSEMINATION: Ethics approval is not indicated as this project does not meet the federal definitions of research requiring the oversight of the Institutional Review Board (IRB). Patient health information (PHI) will not be generated during the implementation of this project. Results of the trial will be made accessible to the public when published in a peer-reviewed journal following the completion of the study.


Subject(s)
Inservice Training/organization & administration , Operating Rooms/standards , Patient Safety , Safety Management/organization & administration , Humans , Models, Organizational , Patient Care Team/standards , Program Evaluation , Research Design
11.
Am J Ophthalmol ; 176: 134-140, 2017 Apr.
Article in English | MEDLINE | ID: mdl-27993591

ABSTRACT

PURPOSE: To characterize the expression of vascular endothelial growth factor (VEGF) in a patient with retinopathy of prematurity (ROP) treated with ranibizumab (Case 1) and compare it with a case of ROP without treatment (Case 2), a case of a premature baby without ROP (Case 3), and a case of a baby without history of ROP or prematurity (Case 4). DESIGN: Observational case series. METHODS: The eyes of the deceased babies were removed postmortem and were sent to the Florida Lions Ocular Pathology Laboratory, where they were processed. The specimens were immunostained using an antibody against VEGF. RESULTS: All eyes except for the eyes in Case 4 disclosed positive VEGF staining. Positive staining was present within the nerve fiber layer, inner plexiform layer, and inner and outer nuclear layers and within the spindle-shaped cell population in the vanguard in Case 1. In the posterior pole, positive staining was only observed at the level of the nerve fiber layer. This case also demonstrated less positive staining when compared with Case 2, where positive staining was found within all layers of the retina. CONCLUSION: Less VEGF staining was observed within the retina of the eyes treated with ranibizumab when compared with the VEGF staining in Case 2. This supports the idea that anti-VEGF agents are effective in reducing the amount of VEGF present in the retina. Furthermore, the fact that some expression of VEGF remains in the immature retina after injection supports the idea that anti-VEGF agents can suppress uncontrolled neovascularization without completely blocking the vascular drive for the vascularization of the immature retina.


Subject(s)
Infant, Premature , Ranibizumab/administration & dosage , Retina/pathology , Retinopathy of Prematurity/diagnosis , Vascular Endothelial Growth Factor A/biosynthesis , Angiogenesis Inhibitors/administration & dosage , Dose-Response Relationship, Drug , Female , Humans , Infant , Infant, Newborn , Male , Retina/metabolism , Retinopathy of Prematurity/drug therapy , Retinopathy of Prematurity/metabolism
12.
J Health Organ Manag ; 30(4): 565-80, 2016 Jun 20.
Article in English | MEDLINE | ID: mdl-27296879

ABSTRACT

Purpose - The purpose of this paper is to investigate how general practitioners' (GPs) decisions about discontinuation of medication are influenced by their institutional context. Design/methodology/approach - In total, 24 GPs were interviewed, three practices were observed and documents were collected. The Gioia methodology was used to analyse data, drawing on a theoretical framework that integrate the sensemaking perspective and institutional theory. Findings - Most GPs, who actively consider discontinuation, are reluctant to discontinue medication, because the safest course of action for GPs is to continue prescriptions, rather than discontinue them. The authors conclude that this is in part due to the ambiguity about the appropriateness of discontinuing medication, experienced by the GPs, and in part because the clinical guidelines do not encourage discontinuation of medication, as they offer GPs a weak frame for discontinuation. Three reasons for this are identified: the guidelines provide dominating triggers for prescribing, they provide weak priming for discontinuation as an option, and they underscore a cognitive constraint against discontinuation. Originality/value - The analysis offers new insights about decision making when discontinuing medication. It also offers one of the first examinations of how the institutional context embedding GPs influences their decisions about discontinuation. For policymakers interested in the discontinuation of medication, the findings suggest that de-stigmatising discontinuation on an institutional level may be beneficial, allowing GPs to better justify discontinuation in light of the ambiguity they experience.


Subject(s)
Decision Making , General Practitioners/psychology , Withholding Treatment , Behavior Observation Techniques , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/administration & dosage , Interviews as Topic , Qualitative Research
13.
J Chem Phys ; 143(13): 134309, 2015 Oct 07.
Article in English | MEDLINE | ID: mdl-26450316

ABSTRACT

The influence of nuclear dynamics in the electronic ground state on the (e,2e) momentum profiles of dimethyl ether has been analyzed using the harmonic analytical quantum mechanical and Born-Oppenheimer molecular dynamics approaches. In spite of fundamental methodological differences, results obtained with both approaches consistently demonstrate that molecular vibrations in the electronic ground state have a most appreciable influence on the momentum profiles associated to the 2b1, 6a1, 4b2, and 1a2 orbitals. Taking this influence into account considerably improves the agreement between theoretical and newly obtained experimental momentum profiles, with improved statistical accuracy. Both approaches point out in particular the most appreciable role which is played by a few specific molecular vibrations of A1, B1, and B2 symmetries, which correspond to C-H stretching and H-C-H bending modes. In line with the Herzberg-Teller principle, the influence of these molecular vibrations on the computed momentum profiles can be unraveled from considerations on the symmetry characteristics of orbitals and their energy spacing.

14.
Proc Natl Acad Sci U S A ; 105(18): 6702-7, 2008 May 06.
Article in English | MEDLINE | ID: mdl-18448676

ABSTRACT

Hypertension, a major cardiovascular risk factor and cause of mortality worldwide, is thought to arise from primary renal abnormalities. However, the etiology of most cases of hypertension remains unexplained. Vascular tone, an important determinant of blood pressure, is regulated by nitric oxide, which causes vascular relaxation by increasing intracellular cGMP and activating cGMP-dependent protein kinase I (PKGI). Here we show that mice with a selective mutation in the N-terminal protein interaction domain of PKGIalpha display inherited vascular smooth muscle cell abnormalities of contraction, abnormal relaxation of large and resistance blood vessels, and increased systemic blood pressure. Renal function studies and responses to changes in dietary sodium in the PKGIalpha mutant mice are normal. These data reveal that PKGIalpha is required for normal VSMC physiology and support the idea that high blood pressure can arise from a primary abnormality of vascular smooth muscle cell contractile regulation, suggesting a new approach to the diagnosis and therapy of hypertension and cardiovascular diseases.


Subject(s)
Hypertension/physiopathology , Muscle, Smooth, Vascular/physiopathology , Aldosterone/blood , Animals , Cyclic GMP-Dependent Protein Kinase Type I , Cyclic GMP-Dependent Protein Kinases/metabolism , Diet , Hypertension/enzymology , Mice , Muscle, Smooth, Vascular/enzymology , Vascular Resistance , Vasomotor System/enzymology , rhoA GTP-Binding Protein/metabolism
15.
Development ; 131(18): 4593-605, 2004 Sep.
Article in English | MEDLINE | ID: mdl-15342483

ABSTRACT

The TGFbeta superfamily plays diverse and essential roles in kidney development. Gdf11 and Bmp4 are essential for outgrowth and positioning of the ureteric bud, the inducer of metanephric mesenchyme. During nephrogenesis, Bmp7 is required for renewal of the mesenchyme progenitor population. Additionally, in vitro studies demonstrate inhibitory effects of BMPs and TGFbetas on collecting duct branching and growth. Here, we explore the predicted models of TGFbeta superfamily function by cell-specific inactivation of Smad4, a key mediator of TGFbeta signaling. Using a HoxB7cre transgene expressed in ureteric bud and collecting duct, we find that development of the collecting duct is Smad4 independent. By contrast, removal of Smad4 in nephrogenic mesenchyme using the Bmp7(cre/+) allele leads to disorganization of the nephrogenic mesenchyme and impairment of mesenchyme induction. Smad4-deficient metanephric mesenchyme does not display defects in inducibility in LiCl or spinal cord induction assays. However, in situ hybridization and lineage analysis of Smad4 null mesenchyme cells at E11.5 show that the nephrogenic mesenchyme does not aggregate tightly around the ureteric bud tips, but remains loosely associated, embedded within a population of cells expressing markers of both nephrogenic mesenchyme and peripheral stroma. We conclude that the failure of recruitment of nephrogenic mesenchyme leaves a primitive population of mesenchyme at the periphery of the kidney. This population is gradually depleted, and by E16.5 the periphery is composed of cells of stromal phenotype. This study uncovers a novel role for TGFbeta superfamily signaling in the recruitment and/or organization of the nephrogenic mesenchyme at early time-points of kidney development. Additionally, we present conclusive genetic lineage mapping of the collecting duct and nephrogenic mesenchyme.


Subject(s)
Kidney/embryology , Kidney/metabolism , Mesoderm/metabolism , Morphogenesis , Signal Transduction , Stem Cells/metabolism , Transforming Growth Factor beta/metabolism , Animals , Bone Morphogenetic Protein 7 , Bone Morphogenetic Proteins/genetics , Bone Morphogenetic Proteins/metabolism , Cell Division , Cell Lineage , DNA-Binding Proteins/deficiency , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Gene Deletion , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Kidney/pathology , Mesoderm/pathology , Mice , Mice, Transgenic , Multigene Family , Nephrosis/genetics , Nephrosis/metabolism , Nephrosis/pathology , Smad4 Protein , Stem Cells/pathology , Trans-Activators/deficiency , Trans-Activators/genetics , Trans-Activators/metabolism , Transforming Growth Factor beta/classification , Transforming Growth Factor beta/genetics
16.
J Am Chem Soc ; 124(25): 7563-72, 2002 Jun 26.
Article in English | MEDLINE | ID: mdl-12071767

ABSTRACT

The gas-phase internal elimination (E(i)) reaction of the sulfoxide (-SO-CH(3)) precursors of ethylene and model oligomers of PPV and PITN has been investigated by means of Hartree-Fock, Møller-Plesset (second and fourth order), and Density Functional Theory (B3LYP, MPW1K) calculations. Considerable differences between the obtained ground state and transition state geometries and the calculated activation energies are observed from one approach to the other, justifying first a careful calibration against the results of a benchmark CCSD(T) study of the E(i) reaction leading to ethylene. In comparison with the CCSD(T) results, as well as with available experimental data, DFT calculations along with the MPW1K functional are found to be a very appropriate choice for describing the E(i) pathway. The leading conformations of the precursors, the relevant transition state structures, and the energy barriers encountered along the lowest energy path to unsubstituted, alpha and beta chloro-, methoxy-, and cyano-substituted ethylene, styrene, stilbene in its cis and trans forms, and at last trans-biisothianaphthene have therefore been identified and characterized in detail employing DFT (MPW1K). Depending on the substituents attached to the C(alpha) and C(beta) atoms, different reaction mechanisms are observed.

17.
RBCF, Rev. bras. ciênc. farm. (Impr.) ; 35(1): 127-31, jan.-jun. 1999. ilus, tab
Article in Portuguese | LILACS | ID: lil-263406

ABSTRACT

Cloridrato de azelastina, novo anti-histamínico `H IND. 1ï, foi doseado por titulometria em meio não-aquoso, com metóxido de sódio, e por espectrofotometria na região do ultravioleta. Verificou-se serem ambos os métodos adequados à sua determinação, sendo mais prática a técnica volumétrica em função de sua simplicidade e dispensa de instrumentação


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal , Histamine H1 Antagonists/pharmacokinetics , Hypersensitivity/metabolism , Drug Evaluation , Spectrophotometry, Ultraviolet , Titrimetry
18.
Rev. ciênc. farm ; 19(2): 225-34, 1998. ilus, tab, graf
Article in Portuguese | LILACS | ID: lil-229481

ABSTRACT

Dicloridrato de cetirizina, novo anti-histamínico H1, foi doseado volumetricamente em meio anidro com ácido perclórico e por espectrofotometria, em 232 nm. Verificou-se que ambos os métodos säo viáveis e precisos, sendo o doseamento em meio näo-aquoso mais prático por näo requerer instrumental e ser mais rápido e fácil de executar.


Subject(s)
Drug Evaluation/methods , Cetirizine/chemistry , Acetic Acid/administration & dosage , Hydrochloric Acid/administration & dosage , Acetic Anhydrides/administration & dosage , Laboratory Chemicals , Perchlorates/administration & dosage , Quinaldines/administration & dosage , Spectrophotometry, Ultraviolet
19.
São Paulo; s.n; 1989. 160 p. ilus.
Thesis in Portuguese | LILACS, Coleciona SUS | ID: biblio-938393

ABSTRACT

Sete carboxilatos de ródio- acetato, propionato, butirato, trifluoracetato, benzoato, fenilacetato e hdironamato - foram sintetizados e associados a metronidazol e benznidazol, dois ligantes nitroimidazólicos com atividade antiprotozoária. O complexos foram caracterizados por análise elementar, termogravimetria e espectrofotometria no infravermelho, ultravioleta e visível. Alvuns compostos oram também analisados por espectrometria de massa e ressonância magnética nuclear, bem como por difração de raios x. Caraboxilatos e adutos tiveram suas atividades antichagásicas avaliadas em culturas de Trypanosoma cruzi, cepas Fr e U. Os compostos que manifestarm maior atividade neste ensaio - propionato de ródio e respectivos adutos - foram testados quanto ao seu potencial como agentes profiláticos transfusionais, sua toxidade oral e capacidade de prolongarem a sobrevidade de camundongos infectados, por administração oral e intraperitonial


Subject(s)
Male , Female , Humans , Animals , Guinea Pigs , Mice , Chagas Disease/drug therapy , Nitroimidazoles/therapeutic use , Trypanosoma cruzi
20.
São Paulo; s.n; 1989. 160 p. ilus.
Thesis in Portuguese | LILACS | ID: lil-664709

ABSTRACT

Sete carboxilatos de ródio- acetato, propionato, butirato, trifluoracetato, benzoato, fenilacetato e hdironamato - foram sintetizados e associados a metronidazol e benznidazol, dois ligantes nitroimidazólicos com atividade antiprotozoária. O complexos foram caracterizados por análise elementar, termogravimetria e espectrofotometria no infravermelho, ultravioleta e visível. Alvuns compostos oram também analisados por espectrometria de massa e ressonância magnética nuclear, bem como por difração de raios x. Caraboxilatos e adutos tiveram suas atividades antichagásicas avaliadas em culturas de Trypanosoma cruzi, cepas Fr e U. Os compostos que manifestarm maior atividade neste ensaio - propionato de ródio e respectivos adutos - foram testados quanto ao seu potencial como agentes profiláticos transfusionais, sua toxidade oral e capacidade de prolongarem a sobrevidade de camundongos infectados, por administração oral e intraperitonial


Subject(s)
Humans , Animals , Male , Female , Guinea Pigs , Mice , Chagas Disease/drug therapy , Nitroimidazoles/therapeutic use , Trypanosoma cruzi
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