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1.
Child Psychiatry Hum Dev ; 51(5): 853, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32797336

RESUMO

The original version of this article were unfortunately published with an error in "Methods" section. This has been corrected by publishing this correction article.

2.
Transfus Med ; 26(6): 432-439, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27726216

RESUMO

BACKGROUND: Restrictive red blood cell (RBC) transfusion has been widely described in transfusion guidelines. However, compliance with these guidelines is often poor. Therefore, we developed a care bundle for the transfusion of RBCs in intensive care. We investigated the effect of the application of the transfusion bundle on transfusion practice, hypothesising that the implementation of the transfusion bundle would lead to a reduction of inappropriate RBC transfusions. STUDY DESIGN AND METHODS: We conducted a before and after study between January and December 2014 in a medical-surgical intensive care unit (ICU) of a university hospital in Amsterdam, the Netherlands. The primary outcome was the percentage of appropriate transfusions, referring to those transfusions that were in accordance to the patients' individual preset haemoglobin threshold. RESULTS: The mean pre-transfusion haemoglobin level was 7·3 g dL-1 [standard deviation (SD) = 1·15] during baseline and significantly decreased to 7·1 g dL-1 (SD = 1·04) after transfusion bundle implementation; 95% confidence interval (CI): 0·009-0·308, P-value = 0·04. The number of inappropriate transfusions significantly decreased from 25% (111/439) during baseline to 15% (42/280) during implementation, a difference of 10%; 95% CI: -0·164 to -0·0416, P-value 0·001. This further decreased to 12% (45/370) in the post-implementation phase. A logistic regression analysis showed that the chance to find an appropriate transfusion is approximately twice as high after transfusion bundle implementation. CONCLUSIONS: Introduction of a transfusion bundle results in a significant reduction of the number of inappropriate RBC transfusions in the medical-surgical ICU. Our results show that the introduction of a transfusion care bundle helps to improve compliance with transfusion guidelines in daily practice.


Assuntos
Cuidados Críticos/métodos , Transfusão de Eritrócitos/métodos , Fidelidade a Diretrizes , Hemoglobinas/metabolismo , Feminino , Humanos , Masculino , Países Baixos
3.
Neuroimage ; 119: 252-261, 2015 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-26093329

RESUMO

While being in the center of attention and exposed to other's evaluations humans are prone to experience embarrassment. To characterize the neural underpinnings of such aversive moments, we induced genuine experiences of embarrassment during person-group interactions in a functional neuroimaging study. Using a mock-up scenario with three confederates, we examined how the presence of an audience affected physiological and neural responses and the reported emotional experiences of failures and achievements. The results indicated that publicity induced activations in mentalizing areas and failures led to activations in arousal processing systems. Mentalizing activity as well as attention towards the audience were increased in socially anxious participants. The converging integration of information from mentalizing areas and arousal processing systems within the ventral anterior insula and amygdala forms the neural pathways of embarrassment. Targeting these neural markers of embarrassment in the (para-)limbic system provides new perspectives for developing treatment strategies for social anxiety disorders.


Assuntos
Ansiedade/fisiopatologia , Emoções/fisiologia , Comportamento Social , Estresse Psicológico/fisiopatologia , Adolescente , Adulto , Mapeamento Encefálico , Feminino , Fixação Ocular , Humanos , Imageamento por Ressonância Magnética , Masculino , Pupila/fisiologia , Adulto Jovem
4.
Child Psychiatry Hum Dev ; 44(1): 152-65, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22798203

RESUMO

A behaviorally inhibited temperament in early childhood has been identified as a potential risk factor for anxiety disorders in children and adolescents. The purpose of our investigation was the development and evaluation of the factor structure, reliability and validity of the first retrospective parent report measure to assess behavioral inhibition in infants and toddlers. Principal Component Analysis of the Retrospective Infant Behavioral Inhibition Scale (RIBI) supported a three factor solution of the core features of BI in two unselected samples. Internal consistency and inter-rater agreement of both parent judgments were >.90 and >.70. Scores of the RIBI were positively correlated with the parent report temperament questionnaire IBQ and a laboratory-based test at age 14 months with the child.


Assuntos
Transtornos de Ansiedade/psicologia , Ansiedade/psicologia , Comportamento Infantil/psicologia , Inibição Psicológica , Pais , Temperamento , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Psicometria , Estudos Retrospectivos , Fatores de Risco , Inquéritos e Questionários
5.
Pulmonology ; 2022 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-36274046

RESUMO

OBJECTIVE: We determined the prevalences of hyperoxemia and excessive oxygen use, and the epidemiology, ventilation characteristics and outcomes associated with hyperoxemia in invasively ventilated patients with coronavirus disease 2019 (COVID-19). METHODS: Post hoc analysis of a national, multicentre, observational study in 22 ICUs. Patients were classified in the first two days of invasive ventilation as 'hyperoxemic' or 'normoxemic'. The co-primary endpoints were prevalence of hyperoxemia (PaO2 > 90 mmHg) and prevalence of excessive oxygen use (FiO2 ≥ 60% while PaO2 > 90 mmHg or SpO2 > 92%). Secondary endpoints included ventilator settings and ventilation parameters, duration of ventilation, length of stay (LOS) in ICU and hospital, and mortality in ICU, hospital, and at day 28 and 90. We used propensity matching to control for observed confounding factors that may influence endpoints. RESULTS: Of 851 COVID-19 patients, 225 (26.4%) were classified as hyperoxemic. Excessive oxygen use occurred in 385 (45.2%) patients. Acute respiratory distress syndrome (ARDS) severity was lowest in hyperoxemic patients. Hyperoxemic patients were ventilated with higher positive end-expiratory pressure (PEEP), while rescue therapies for hypoxemia were applied more often in normoxemic patients. Neither in the unmatched nor in the matched analysis were there differences between hyperoxemic and normoxemic patients with regard to any of the clinical outcomes. CONCLUSION: In this cohort of invasively ventilated COVID-19 patients, hyperoxemia occurred often and so did excessive oxygen use. The main differences between hyperoxemic and normoxemic patients were ARDS severity and use of PEEP. Clinical outcomes were not different between hyperoxemic and normoxemic patients.

6.
Pulmonology ; 28(1): 18-27, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34836830

RESUMO

OBJECTIVE: Invasively ventilated patients with acute respiratory failure related to coronavirus disease 2019 (COVID-19) potentially benefit from tracheostomy. The aim of this study was to determine the practice of tracheostomy during the first wave of the pandemic in 2020 in the Netherlands, to ascertain whether timing of tracheostomy had an association with outcome, and to identify factors that had an association with timing. METHODS: Secondary analysis of the 'PRactice of VENTilation in COVID-19' (PRoVENT-COVID) study, a multicenter observational study, conducted from March 1, 2020 through June 1, 2020 in 22 Dutch intensive care units (ICU) in the Netherlands. The primary endpoint was the proportion of patients receiving tracheostomy; secondary endpoints were timing of tracheostomy, duration of ventilation, length of stay in ICU and hospital, mortality, and factors associated with timing. RESULTS: Of 1023 patients, 189 patients (18.5%) received a tracheostomy at median 21 [17 to 28] days from start of ventilation. Timing was similar before and after online publication of an amendment to the Dutch national guidelines on tracheostomy focusing on COVID-19 patients (21 [17-28] vs. 21 [17-26] days). Tracheostomy performed ≤ 21 days was independently associated with shorter duration of ventilation (median 26 [21 to 32] vs. 40 [34 to 47] days) and higher mortality in ICU (22.1% vs. 10.2%), hospital (26.1% vs. 11.9%) and at day 90 (27.6% vs. 14.6%). There were no patient demographics or ventilation characteristics that had an association with timing of tracheostomy. CONCLUSIONS: Tracheostomy was performed late in COVID-19 patients during the first wave of the pandemic in the Netherlands and timing of tracheostomy possibly had an association with outcome. However, prospective studies are needed to further explore these associations. It remains unknown which factors influenced timing of tracheostomy in COVID-19 patients.


Assuntos
COVID-19/complicações , Síndrome do Desconforto Respiratório/terapia , Insuficiência Respiratória/terapia , Traqueostomia/métodos , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Países Baixos , Respiração Artificial , Insuficiência Respiratória/etiologia , SARS-CoV-2 , Resultado do Tratamento , Ventilação
7.
J Crit Care ; 67: 118-125, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34749051

RESUMO

INTRODUCTION: Hypotension in the ICU is common, yet management is challenging and variable. Insight in management by ICU physicians and nurses may improve patient care and guide future hypotension treatment trials and guidelines. METHODS: We conducted an international survey among ICU personnel to provide insight in monitoring, management, and perceived consequences of hypotension. RESULTS: Out of 1464 respondents, 1197 (81.7%) were included (928 physicians (77.5%) and 269 nurses (22.5%)). The majority indicated that hypotension is underdiagnosed (55.4%) and largely preventable (58.8%). Nurses are primarily in charge of monitoring changes in blood pressure, physicians are in charge of hypotension treatment. Balanced crystalloids, dobutamine, norepinephrine, and Trendelenburg position were the most frequently reported fluid, inotrope, vasopressor, and positional maneuver used to treat hypotension. Reported complications believed to be related to hypotension were AKI and myocardial injury. Most ICUs do not have a specific hypotension treatment guideline or protocol (70.6%), but the majority would like to have one in the future (58.1%). CONCLUSIONS: Both physicians and nurses report that hypotension in ICU patients is underdiagnosed, preventable, and believe that hypotension influences morbidity. Hypotension management is generally not protocolized, but the majority of respondents would like to have a specific hypotension management protocol.


Assuntos
Hipotensão , Médicos , Cuidados Críticos , Humanos , Hipotensão/terapia , Unidades de Terapia Intensiva , Inquéritos e Questionários
8.
J Am Heart Assoc ; 11(4): e022833, 2022 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-35112889

RESUMO

Background Natriuretic peptides are routinely quantified to diagnose heart failure (HF). Their concentrations are also elevated in atrial fibrillation (AF). To clarify their value in predicting future cardiovascular events, we measured natriuretic peptides in unselected patients with cardiovascular conditions and related their concentrations to AF and HF status and outcomes. Methods and Results Consecutive patients with cardiovascular conditions presenting to a large teaching hospital underwent clinical assessment, 7-day ECG monitoring, and echocardiography to diagnose AF and HF. NT-proBNP (N-terminal pro-B-type natriuretic peptide) was centrally quantified. Based on a literature review, four NT-proBNP groups were defined (<300, 300-999, 1000-1999, and ≥2000 pg/mL). Clinical characteristics and NT-proBNP concentrations were related to HF hospitalization or cardiovascular death. Follow-up data were available in 1616 of 1621 patients (99.7%) and analysis performed at 2.5 years (median age, 70 [interquartile range, 60-78] years; 40% women). HF hospitalization or cardiovascular death increased from 36 of 488 (3.2/100 person-years) in patients with neither AF nor HF, to 55 of 354 (7.1/100 person-years) in patients with AF only, 92 of 369 (12.1/100 person-years) in patients with HF only, and 128 of 405 (17.7/100 person-years) in patients with AF plus HF (P<0.001). Higher NT-proBNP concentrations predicted the outcome in patients with AF only (C-statistic, 0.82; 95% CI, 0.77-0.86; P <0.001) and in other phenotype groups (C-statistic in AF plus HF, 0.66; [95% CI, 0.61-0.70]; P <0.001). Conclusions Elevated NT-proBNP concentrations predict future HF events in patients with AF irrespective of the presence of HF, encouraging routine quantification of NT-proBNP in the assessment of patients with AF.


Assuntos
Fibrilação Atrial , Insuficiência Cardíaca , Fibrilação Atrial/complicações , Fibrilação Atrial/diagnóstico , Biomarcadores , Ecocardiografia , Feminino , Insuficiência Cardíaca/diagnóstico , Hospitalização , Humanos , Masculino , Peptídeo Natriurético Encefálico , Fragmentos de Peptídeos , Prognóstico , Vasodilatadores
9.
J Crit Care ; 65: 142-148, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34148010

RESUMO

INTRODUCTION: Although hypotension in ICU patients is associated with adverse outcome, currently used definitions are unknown and no universally accepted definition exists. METHODS: We conducted an international, peer-reviewed survey among ICU physicians and nurses to provide insight in currently used definitions, estimations of incidence, and duration of hypotension. RESULTS: Out of 1394 respondents (1055 physicians (76%) and 339 nurses (24%)), 1207 (82%) completed the questionnaire. In all patient categories, hypotension definitions were predominantly based on an absolute MAP of 65 mmHg, except for the neuro(trauma) category (75 mmHg, p < 0.001), without differences between answers from physicians and nurses. Hypotension incidence was estimated at 55%, and time per day spent in hypotension at 15%, both with nurses reporting higher percentages than physicians (estimated mean difference 5%, p = 0.01; and 4%, p < 0.001). CONCLUSIONS: An absolute MAP threshold of 65 mmHg is most frequently used to define hypotension in ICU patients. In neuro(trauma) patients a higher threshold was reported. The majority of ICU patients are estimated to endure hypotension during their ICU admission for a considerable amount of time, with nurses reporting a higher estimated incidence and time spent in hypotension than physicians.


Assuntos
Hipotensão , Unidades de Terapia Intensiva , Cuidados Críticos , Humanos , Hipotensão/epidemiologia , Incidência , Inquéritos e Questionários
10.
Expert Rev Respir Med ; 15(11): 1403-1413, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34047244

RESUMO

Introduction: INTELLiVENT-Adaptive Support Ventilation (INTELLiVENT-ASV), an advanced closed-loop ventilation mode for use in intensive care unit (ICU) patients, is equipped with algorithms that automatically adjust settings on the basis of physiologic signals and patient's activity. Here we describe its effectiveness, safety, and efficacy in various types of ICU patients.Areas covered: A systematic search conducted in MEDLINE, EMBASE, the Cochrane Central register of Controlled Trials (CENTRAL), and in Google Scholar identified 10 randomized clinical trials.Expert opinion: Studies suggest INTELLiVENT-ASV to be an effective automated mode with regard to the titrations of tidal volume, airway pressure, and oxygen. INTELLiVENT-ASV is as safe as conventional modes. However, thus far studies have not shown INTELLiVENT-ASV to be superior to conventional modes with regard to duration of ventilation and other patient-centered outcomes. Future studies are needed to test its efficacy.


Assuntos
Unidades de Terapia Intensiva , Respiração Artificial , Cuidados Críticos , Humanos , Pulmão , Volume de Ventilação Pulmonar
11.
Oral Oncol ; 44(4): 335-44, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17689286

RESUMO

Two mechanisms of radiotherapy resistance of major importance in head and neck cancer are tumour cell repopulation and hypoxia. Hypoxic tumour cells that retain their clonogenic potential can survive radiation treatment and lead to local recurrences. The aim of this study was to quantify this cellular population in a cohort of human head and neck carcinomas and to investigate the prognostic significance. The proliferation marker iododeoxyuridine (IdUrd) and the hypoxia marker pimonidazole were administered intravenously prior to biopsy taking in patients with stage II-IV squamous cell carcinoma of the head and neck. Triple immunohistochemical staining of blood vessels, IdUrd and pimonidazole was performed and co-localization of IdUrd and pimonidazole was quantitatively assessed by computerized image analysis. The results were related with treatment outcome. Thirty-nine biopsies were analyzed. Tumours exhibited different patterns of proliferation and hypoxia but generally the IdUrd signal was found in proximity to blood vessels whereas pimonidazole binding was predominantly at a distance from vessels. Overall, no correlations were found between proliferative activity and oxygenation status. The fraction of IdUrd-labelled cells positive for pimonidazole ranged from 0% to 16.7% with a mean of 2.4% indicating that proliferative activity was low in hypoxic areas and occurring mainly in the well-oxygenated tumour compartments. IdUrd positive cells in hypoxic areas made up only 0.09% of the total viable tumour cell mass. There were no associations between the magnitude of this cell population and local tumour control or survival. Co-localization between proliferating cells and hypoxia in head and neck carcinomas was quantified using an immunohistochemical triple staining technique combined with a computerized simultaneous analysis of multiple parameters. The proportion of cells proliferating under hypoxic conditions was small and no correlation with treatment outcome could be found.


Assuntos
Carcinoma de Células Escamosas/patologia , Hipóxia Celular , Neoplasias de Cabeça e Pescoço/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Carcinoma de Células Escamosas/irrigação sanguínea , Carcinoma de Células Escamosas/radioterapia , Proliferação de Células , Feminino , Neoplasias de Cabeça e Pescoço/irrigação sanguínea , Neoplasias de Cabeça e Pescoço/radioterapia , Humanos , Idoxuridina , Processamento de Imagem Assistida por Computador/métodos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neovascularização Patológica/metabolismo , Nitroimidazóis , Inibidores da Síntese de Ácido Nucleico , Radiossensibilizantes , Resultado do Tratamento
12.
Int J Radiat Oncol Biol Phys ; 62(4): 1157-68, 2005 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-15913908

RESUMO

PURPOSE: Most solid tumors contain hypoxic cells, and the amount of tumor hypoxia has been shown to have a negative impact on the outcome of radiotherapy. The efficacy of combined modality treatments depends both on the sequence and timing of the treatments. Hypoxic cell turnover in tumors may be important for optimal scheduling of combined modality treatments, especially when hypoxic cell targeting is involved. METHODS AND MATERIALS: Previously we have shown that a double bioreductive hypoxic marker assay could be used to detect changes of tumor hypoxia in relation to the tumor vasculature after carbogen and hydralazine treatments. This assay was used in the current study to establish the turnover rate of hypoxic cells in three different tumor models. The first hypoxic marker, pimonidazole, was administered at variable times before tumor harvest, and the second hypoxic marker, CCI-103F, was injected at a fixed time before harvest. Hypoxic cell turnover was defined as loss of pimonidazole (first marker) relative to CCI-103F (second marker). RESULTS: The half-life of hypoxic cell turnover was 17 h in the murine C38 colon carcinoma line, 23 h and 49 h in the human xenograft lines MEC82 and SCCNij3, respectively. Within 24 h, loss of pimonidazole-stained areas in C38 and MEC82 occurred concurrent with the appearance of pimonidazole positive cell debris in necrotic regions. In C38 and MEC82, most of the hypoxic cells had disappeared after 48 h, whereas in SCCNij3, viable cells that had been labeled with pimonidazole were still observed after 5 days. CONCLUSIONS: The present study demonstrates that the double hypoxia marker assay can be used to study changes in both the proportion of hypoxic tumor cells and their lifespan at the same time. The present study shows that large differences in hypoxic cell turnover rates may exist among tumor lines, with half-lives ranging from 17-49 h.


Assuntos
Hipóxia Celular , Corantes/análise , Neoplasias/patologia , Nitroimidazóis/análise , Animais , Benzimidazóis/administração & dosagem , Benzimidazóis/análise , Linhagem Celular Tumoral , Sobrevivência Celular , Corantes/administração & dosagem , Meia-Vida , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias/irrigação sanguínea , Nitroimidazóis/administração & dosagem , Fatores de Tempo
13.
J Mol Biol ; 245(1): 22-33, 1995 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-7823317

RESUMO

In the protist Euglena gracilis, the small subunit of the chloroplast enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase is encoded by nuclear rbcS genes and synthesized as a polyprotein precursor containing eight mature small subunit molecules. This large precursor is encoded by at least eight different nuclear genes as ascertained by transcript analysis. The structure of three rbcS genes was established and the coding sequences were found to be interrupted by many intervening sequences (IVS). Apart from the first 5' intron involved in trans-splicing, none of these IVSs obeys the GT-AG rule characteristic of introns in higher eukaryote genes. Surprisingly, these IVSs are located at identical positions within the three genes studied. Moreover, extensive sequence homologies were found between IVSs located in the same gene as well as in different genes. The sequences of these homologous IVSs differ only by inserted and/or deleted sequences. The striking conservation of the 5' and 3' regions of these IVSs is correlated to their potential secondary structures. These structures, which bring the IVS extremities together with the exon boundaries, could be involved in a novel splicing process. The second 5' IVS is shown to be excised before the addition of the spliced leader sequence to the pre-mRNA. Similarly, two 3' IVSs are excised before the polyadenylation step. These results suggest that IVS splicing is faster than eukaryotic genomic cis-splicing and involves components other than those of the classical spliceosomes.


Assuntos
Euglena gracilis/genética , Proteínas de Plantas , Splicing de RNA/genética , Ribulose-Bifosfato Carboxilase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , Núcleo Celular/genética , DNA Complementar , Euglena gracilis/enzimologia , Íntrons/genética , Dados de Sequência Molecular , RNA Mensageiro/análise , RNA Mensageiro/genética , Ribulose-Bifosfato Carboxilase/biossíntese , Análise de Sequência , Transcrição Gênica/genética
14.
Radiother Oncol ; 76(2): 194-9, 2005 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-16024110

RESUMO

PURPOSE: In head and neck cancer, it has been shown that hypoxic tumors respond poorly to therapy. Methods to identify hypoxic tumors are, therefore, of importance to select patients for oxygenation modifying or other intensified treatments. The aim of this study was to compare tumor cell hypoxia assessed by the hypoxic cell marker pimonidazole (PIMO) with expression of the endogenous hypoxia-related marker carbonic anhydrase IX (CAIX) in three human head and neck tumor lines. MATERIAL AND METHODS: Forty-five tumors of three human head and neck tumor lines, SCCNij3, SCCNij59 and MEC82, xenografted in athymic mice, were used. CAIX was quantified by biodistribution (% injected dose/g tumor) after injecting 3-5 microl 111In-labeled G250 mouse antibody 3 days prior to euthanizing. In a tissue section from the same tumor, fractions of tumor area positive for PIMO, CAIX and Hoechst 33342 (perfusion marker) were assessed after immunohistochemical staining, using a digital image analysis system. RESULTS: SCCNij3 and MEC82 were relatively hypoxic tumor lines with fractions of tumor area positive for pimonidazole of 0.16 and 0.15, respectively. SCCNij59 was a better-oxygenated tumor line with a PIMO-fraction of 0.03. The three tumor lines showed different levels and patterns of CAIX immunohistochemical staining, but only in MEC82 there was a good correlation between PIMO-fraction and CAIX-fraction (r2=0.92, P<0.0001). Correlations between 111In-G250 uptake and CAIX-fraction or PIMO-fraction within tumor lines were weak or absent. CONCLUSIONS: Assessment of CAIX expression depends largely on the techniques and tumor lines used. Furthermore, the immunohistochemical staining pattern of CAIX relative to PIMO differs between human tumor lines of similar anatomical origin. Therefore, the use of CAIX as endogenous marker of tumor hypoxia remains questionable.


Assuntos
Antígenos de Neoplasias/metabolismo , Anidrases Carbônicas/metabolismo , Hipóxia Celular , Neoplasias de Cabeça e Pescoço/metabolismo , Animais , Benzimidazóis/farmacocinética , Anidrase Carbônica IX , Linhagem Celular Tumoral , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Nitroimidazóis/farmacocinética
15.
Mol Plant Microbe Interact ; 6(3): 393-402, 1993.
Artigo em Inglês | MEDLINE | ID: mdl-8324252

RESUMO

Agrocinopine synthase genes have so far been detected only on the Agrobacterium tumefaciens nopaline Ti plasmid pTiC58 and on the A. rhizogenes Ri plasmid pRiA4. The TA region of the octopine/cucumopine (o/c) A. vitis Ti plasmid pTiTm4 strongly resembles the TL DNA of biotype I octopine Ti plasmids. In addition, it carries an intact and functional agrocinopine synthase gene close to its left border. TA/TL sequence comparison shows that the biotype I TL region still carries a 5'-deleted acs gene, strongly indicating that this gene was originally part of the TL region and belongs to the "common DNA" region of octopine and nopaline Ti plasmids. Tm4 and C58C1 (pTiTm4) remove agrocinopine A from the medium indicating that pTiTm4 also carries agrocinopine uptake genes. In spite of this, Tm4 and related strains are resistant against agrocin 84. Two o/c Ti plasmids that are closely related to pTiTm4, pTiHm1, and pTiAB3, have a mutated acs gene; whereas Hm1 can still take up agrocinopine A, AB3 cannot. These results yield new insights in the evolution of octopine, nopaline, and o/c strains.


Assuntos
Arginina/análogos & derivados , DNA Bacteriano/genética , Proteínas de Membrana Transportadoras/genética , Mutação , Doenças das Plantas/microbiologia , Fosfatos Açúcares/genética , Sequência de Aminoácidos , Arginina/genética , Técnicas de Tipagem Bacteriana , Sequência de Bases , Deleção de Genes , Genes Bacterianos , Proteínas de Membrana Transportadoras/química , Dados de Sequência Molecular , Doenças das Plantas/genética , Plasmídeos , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Fosfatos Açúcares/biossíntese , Fosfatos Açúcares/metabolismo
16.
Mol Plant Microbe Interact ; 4(2): 190-7, 1991.
Artigo em Inglês | MEDLINE | ID: mdl-1657255

RESUMO

Agrobacterium tumefaciens biotype III octopine strains have been isolated from grapevine tumors worldwide. They comprise limited and wide host range (LHR and WHR) strains that carry related tumor-inducing (Ti) plasmids with two T-regions, TA and TB. The WHR TA-region resembles the biotype I octopine region, whereas the LHR TA-region is a recent deletion derivative of the WHR TA-region, which lacks the iaa genes and part of the ipt gene. Sequencing of the TA-region of the ubiquitous LHR strain AB3 showed that the deleted region is replaced by an insertion sequence (IS) element, IS868, which resembles the IS51 element of Pseudomonas syringae subsp. savastanoi. The Ti plasmid of LHR strain Ag57 carries essentially the same iaa gene deletion as pTiAB3, but lacks IS868. We propose that the LHR Ti plasmids arose by the recent insertion of an IS868 element into the TA-region of a WHR-type Ti plasmid, followed by transposition to a nearby site. The deletion was caused during the second transposition or by later recombination between the two IS868 copies. Biotype III octopine strains also carry an IS51-like sequence close to the TB iaa genes. Our results confirm and extend earlier observations indicating that IS51-like elements in Pseudomonas and Agrobacterium are associated with iaa genes and played a major role in Ti plasmid evolution.


Assuntos
Agrobacterium tumefaciens/genética , Elementos de DNA Transponíveis , Plasmídeos , Sequência de Aminoácidos , Sequência de Bases , Evolução Biológica , Clonagem Molecular , DNA Bacteriano , Dados de Sequência Molecular , Mapeamento por Restrição , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie
17.
Mol Plant Microbe Interact ; 4(2): 163-72, 1991.
Artigo em Inglês | MEDLINE | ID: mdl-1932812

RESUMO

The limited host range AB3 strain of Agrobacterium tumefaciens induces tumors by transferring two T-regions, TA and TB. TA is a deleted version of the well-known biotype I octopine TL-region that lacks the iaa and ipt genes, but carries an intact oncogene, gene 6b, and typical left and right border sequences. TB carries two iaa genes that together code for the synthesis of indoleacetic acid. Gene 6b and the iaa gene act synergistically when transferred in a coinoculation experiment. The TA-region of the limited host range isolate Ag57 is related to the TA-region of AB3, but differs from it at several positions. The most significant difference is the absence of the right border region. In spite of this, Ag57 and the exconjugant strain C58C9(pTiAg57) induce normal tumors on Nicotiana rustica and Vitis vinifera. Various experiments indicate that gene 6b of the Ag57 TA-region is active and transferred in spite of the absence of the right border. On N. tabacum, C58C9(pTiAg57) is nononcogenic but becomes oncogenic when the pTiAg57 TA-region is restored by the right TA border sequence of pTiAB3. Thus, the right TA border sequence of the biotype III limited host range strains is required for tumor induction on some hosts, but not on others.


Assuntos
Agrobacterium tumefaciens/genética , DNA Bacteriano/fisiologia , Agrobacterium tumefaciens/fisiologia , Sequência de Bases , Clonagem Molecular , Genes Bacterianos , Dados de Sequência Molecular , Oncogenes , Doenças das Plantas , Plantas Tóxicas , Plasmídeos , Mapeamento por Restrição , Especificidade da Espécie , Nicotiana/microbiologia
18.
Int J Radiat Oncol Biol Phys ; 54(1): 215-28, 2002 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-12182995

RESUMO

PURPOSE: To quantify the physiologic status of human tumor cells in relation to the tumor vasculature. METHODS AND MATERIALS: Fourteen tumors of 11 first-generation xenograft lines of human head-and-neck squamous cell carcinoma were injected with the hypoxic cell marker pimonidazole, the proliferation marker BrdUrd, and the perfusion marker Hoechst 33342. Consecutive tissue sections were processed with immunohistochemical methods and analyzed with image-analysis techniques. RESULTS: Three different hypoxic patterns were found: patchy, ribbon-like, and mixed. An image-analysis method was developed to quantify these, and an elongation index (length/width) was calculated for hypoxia. The mean elongation indices ranged from 2.0 to 28.3 and showed a good correlation with the visual scoring of hypoxic patterns. Comparative analysis of hypoxic and proliferating cells in zones around the tumor vasculature showed the presence of both hypoxic and proliferating cells in all zones up to 250 microm from the vessels. The largest coexistence of hypoxic and proliferating cells seemed to occur at 50-100 microm from the vessels. CONCLUSIONS: The three hypoxic patterns could be quantified by an elongation index, which is an additional parameter that allows distinction of tumors with similar fractions of hypoxic cells. The analysis of hypoxic and proliferating cells as a function of distance from the tumor vasculature indicates that proliferation does occur also at low oxygen tensions.


Assuntos
Carcinoma de Células Escamosas/irrigação sanguínea , Hipóxia Celular , Neoplasias de Cabeça e Pescoço/irrigação sanguínea , Animais , Bromodesoxiuridina/metabolismo , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Divisão Celular , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Transplante Heterólogo
19.
Transfus Clin Biol ; 5(5): 336-45, 1998 Oct.
Artigo em Francês | MEDLINE | ID: mdl-9836394

RESUMO

Studying the osmotic resistance or swelling of platelets has often been suggested as a global test to assess the viability of those cells. A number of authors have also analysed the behaviour of platelets in hypotonic media by a variety of complementary methods (cell count, morphology, determinations of substances released, photometric measurement of aggregation induced by aggregating agents, etc). Most studies are currently based on the so-called "osmotic shock response" test, which measures according to time the light transmitted through platelet-rich plasma (PRP) after dilution in distilled water. In this study, the authors describe a new automated and reproducible test using slow dialysis to assess platelet osmotic resistance. The "Fragilimeter", a device initially described by the authors to characterise RBC fragility, has been adapted to the study of platelet osmotic behaviour. The variations in light transmission through a platelet suspension according to NaCl concentration are linked to the change in cellular volume and lysis and characterise the viability of the cells. The results obtained with normal platelets revealed the good reproducibility of the technique. The osmotic resistance is evaluated for two parameters: anticoagulant (citrate, EDTA) and cellular concentration. The test was applied to quality control of stored platelet concentrates for transfusion, prepared with different cell separators.


Assuntos
Plaquetas/fisiologia , Pressão Osmótica , Sobrevivência Celular , Humanos , Luz , Contagem de Plaquetas , Controle de Qualidade , Reprodutibilidade dos Testes , Espalhamento de Radiação , Sensibilidade e Especificidade
20.
J Mal Vasc ; 26(3): 183-90, 2001 Jun.
Artigo em Francês | MEDLINE | ID: mdl-11431623

RESUMO

Vascular engineering developed through advancing knowledge of the physiological and pathological processes operating in cells and tissues subjected to environmental mechanical stress. We review briefly the more relevant cellular and general mechanism characteristic of vascular tissue examining the performances obtained with prosthetic materials (allografts, synthetic grafts) and current research and development of new vessel substitutes (biohybrids or biovessels).


Assuntos
Prótese Vascular , Engenharia Tecidual , Fenômenos Biomecânicos , Vasos Sanguíneos/fisiologia , Colágeno/fisiologia , Criopreservação , Elastina/fisiologia , Humanos , Músculo Liso/citologia , Desenho de Prótese , Veias/transplante
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