RESUMO
When divorce or separation of a couple occurs, children will no longer live with both parents at the same time. There may be multiple causes for divorce, and both literature and our own experien ce as child mental health providers, report some short- and long-term consequences for children, especially where the divorce has been conflictive. In these cases, increased risk of developing be havioral disorders, poor school performance, and substance abuse has been documented as well as consequences in adult life with higher risk of psychiatric pathologies or difficulties in interpersonal relationships, if an intervention that addresses some potentially traumatic situations for children is not done. Pediatricians are in a privileged relationship with children and their families in order to detect signs of parental discord and altered mental health in children. Children behaviors as a result of divorce and parental conflict will depend on the age of the child and stage of development, and it is important to recognize them in order to intervene properly. This article proposes some guidelines for parents. Good management of high conflict situations related to divorce may prevent some of the consequences that these can have on children.
Assuntos
Comportamento Infantil , Divórcio/psicologia , Conflito Familiar/psicologia , Transtornos Mentais/prevenção & controle , Pediatria , Papel do Médico , Relações Profissional-Família , Criança , Humanos , Transtornos Mentais/diagnóstico , Transtornos Mentais/etiologia , Pais/psicologia , Psicologia da CriançaRESUMO
BACKGROUND AND PURPOSE: The Rio score (RS) and the modified Rio score (MRS) are two scoring systems that can identify the early predictive factors of disability progression in relapsing-remitting multiple sclerosis (RRMS) patients treated with interferon-ß (IFN-ß). The objective of the study was to validate the usefulness of the RS and MRS in a large cohort of multiple sclerosis patients treated with IFN-ß in daily clinical practice. METHODS: The analysis included a cohort of RRMS patients treated with different formulations of IFN-ß for at least 1 year. The RS and MRS were used to classify the patients after 1 year of treatment. Multivariate analysis was performed to identify predictive variables of suboptimal response at 5 years, defined as Expanded Disability Status Scale confirmed progression or switching to a second-line therapy. RESULTS: Sixty-nine of 416 included patients were considered as suboptimal responders at 5-year evaluation. The possible score range was 0-3. A higher risk of suboptimal response was found for RS and MRS in the presence of ≥2 scores (hazard ratio 3.0, P = 0.002, and hazard ratio 5.0, P < 0.0001, respectively). CONCLUSIONS: Our study confirmed, in a daily clinical setting, that MRS had a better specificity and accuracy than RS in identifying the patients who will have a poor response to long-term IFN-ß treatment.
Assuntos
Fatores Imunológicos/farmacologia , Interferon beta/farmacologia , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Avaliação de Resultados em Cuidados de Saúde/métodos , Índice de Gravidade de Doença , Adulto , Progressão da Doença , Feminino , Seguimentos , Humanos , Masculino , PrognósticoRESUMO
The objective of this work is to compare the effectiveness and the side effects of two different drugs, amoxicillin and clavulanic acid vs ceftazidime, used as antibiotic prophylaxis in the surgical extraction of third molars and to demonstrate that the use of second choice antibiotic has no significant advantages in comparison with a first choice antibiotic. One hundred and seven patients with impacted third molar were selected and divided into two groups: amoxicillin and clavulanic acid were administered to group 1 and ceftazidime to group 2 for five days after surgery and we observed the postoperative period. The statistical analysis showed no differences between the two groups which lead to the conclusion that there is no indication to routinely administrate intramuscular second-choice antibiotic prophylatic therapy (ceftazidime) in case of surgical extraction of the third molar.
Assuntos
Combinação Amoxicilina e Clavulanato de Potássio/uso terapêutico , Antibacterianos/uso terapêutico , Antibioticoprofilaxia , Ceftazidima/uso terapêutico , Dente Serotino/cirurgia , Infecção da Ferida Cirúrgica/prevenção & controle , Extração Dentária , Dente Impactado/cirurgia , Administração Oral , Adolescente , Adulto , Combinação Amoxicilina e Clavulanato de Potássio/administração & dosagem , Combinação Amoxicilina e Clavulanato de Potássio/efeitos adversos , Antibacterianos/administração & dosagem , Antibacterianos/efeitos adversos , Ceftazidima/administração & dosagem , Ceftazidima/efeitos adversos , Esquema de Medicação , Feminino , Humanos , Injeções Intramusculares , Itália , Masculino , Infecção da Ferida Cirúrgica/microbiologia , Fatores de Tempo , Extração Dentária/efeitos adversos , Resultado do Tratamento , Adulto JovemRESUMO
The meta-analysis aimed to investigate the prevalence of gallstones (GS) in Inflammatory bowel disease (IBD), especially ulcerative colitis (UC). A systematic and thorough search was conducted on online electronic databases (PubMed/Medline, Cochrane Library, and Google Scholar) from the databases' inception to April 30th, 2022. Review Manager 5.4.1 (The Nordic Cochrane Centre, The Cochrane Collaboration, Copenhagen) was used for all statistical analyses and forest plots. Only studies that met inclusion criteria were selected. The selected studies were pooled using a random-effect model and the results were reported in the odds ratio (OR) with their corresponding 95% confidence interval (CI). Ten studies met the final inclusion criteria and were analyzed. Patients with UC had significantly higher prevalence of GS than those in the control group (OR=1.67 [1.32-2.11]; p < 0.0001; I2=58%). There was also significant prevalence of GS in Crohn's disease (CD) than those in control group (OR=2.22 [1.82, 2.69]; p < 0.00001; I2=31%). Analysis also showed the prevalence of GS in studies conducted in Asia (OR=2.00 [1.48, 2.70]; p < 0.00001; I2=80%) and Europe (OR= 1.84 [1.32, 2.55]; p = 0.0003; I2=45%) compared to the control group. This study provided a conclusive answer to whether GS is significant in UC or not. Our meta-analysis provides a well-powered estimate that there is a prevalence of GS in UC. CD is also significantly associated with GS.
RESUMO
Isotretinoin is an analog of vitamin A, used to treat severe, recalcitrant nodular acne, psoriasis and disorders of keratinization. However, its benefits come with a broad range of serious side effects, including ocular adverse effects. We report and describe a rare case of bilateral myopia in a female patient treated with oral isotretinoin for acne vulgaris. A female patient, 29-year old, presented to the office for blurry vision. The patient was treated with oral isotretinoin for four weeks. She had a past medical history of myopia precisely corrected with laser-assisted in situ keratomileusis without any residual refractive error. After a thorough examination and laboratory workup, isotretinoin was thought to be the causative agent of her current complaint, so it was immediately stopped. The follow-up showed no further decrement of visual acuity in the patient. However, the vision loss acquired as an adverse effect of oral isotretinoin persisted.
RESUMO
Coronavirus disease 2019 (COVID-19) is an infectious respiratory disease caused by the novel coronavirus severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Initially, it was reported in December 2019 and became a global pandemic in March 2020, with many presentations and after-effects. We report the case of a 68-year-old female patient who presented to the emergency room with the chief complaint of a skin rash and itching all over her body, developing within a few minutes of insulin injection. The patient had tested positive for COVID-19 almost eight days ago and was self-quarantined. She was a known diabetic for the past 28 years. Her blood glucose levels were maintained within the normal range by a combination regimen of oral anti-diabetic drugs and subcutaneous humulin 70/30 (70% neutral protamine Hagedorn (NPH) insulin and 30% regular human insulin) injections. After careful examination and thorough history taking, a newly acquired insulin allergy was diagnosed in the patient, attributed to her disrupted immune system due to the recent COVID-19 infection.
RESUMO
Supraventricular tachycardia (SVT) refers to the narrow complex tachycardia originating at or above the bundle of His. Several risk factors are associated with the development and recurrence of SVT, but its association with gastric problems, especially dyspepsia, is relatively rare. We report the case of a 54-year-old female who presented to the emergency room (ER) with palpitations, which were diagnosed as an episode of paroxysmal supraventricular tachycardia (PSVT). She had a history of PSVT in the past, along with hypertension and dyspepsia. After thorough history and examination, dyspepsia was identified as the common trigger of her PSVT episodes, pointing towards the likelihood of gastrocardiac symptoms. Therefore, an appropriate regimen of beta-blockers, proton pump inhibitors (PPIs), and anti-foaming agents (simethicone) was prescribed to manage her symptoms with the plan to perform a catheter ablation later.
RESUMO
BACKGROUND AND PURPOSE: The effects of bergamot essential oil (BEO; Citrus bergamia, Risso) on excitotoxic neuronal damage was investigated in vitro. EXPERIMENTAL APPROACH: The study was performed in human SH-SY5Y neuroblastoma cells exposed to N-methyl-D-aspartate (NMDA). Cell viability was measured by dye exclusion. Reactive oxygen species (ROS) and caspase-3 activity were measured fluorimetrically. Calpain I activity and the activation (phosphorylation) of Akt and glycogen synthase kinase-3beta (GSK-3beta) were assayed by Western blotting. KEY RESULTS: NMDA induced concentration-dependent, receptor-mediated, death of SH-SY5Y cells, ranging from 11 to 25% (0.25-5 mM). Cell death induced by 1 mM NMDA (21%) was preceded by a significant accumulation of intracellular ROS and by a rapid activation of the calcium-activated protease calpain I. In addition, NMDA caused a rapid deactivation of Akt kinase and this preceded the detrimental activation of the downstream kinase, GSK-3beta. BEO (0.0005-0.01%) concentration dependently reduced death of SH-SY5Y cells caused by 1 mM NMDA. In addition to preventing ROS accumulation and activation of calpain, BEO (0.01%) counteracted the deactivation of Akt and the consequent activation of GSK-3beta, induced by NMDA. Results obtained by using specific fractions of BEO, suggested that monoterpene hydrocarbons were responsible for neuroprotection afforded by BEO against NMDA-induced cell death. CONCLUSIONS AND IMPLICATIONS: Our data demonstrate that BEO reduces neuronal damage caused in vitro by excitotoxic stimuli and that this neuroprotection was associated with prevention of injury-induced engagement of critical death pathways.
Assuntos
N-Metilaspartato/toxicidade , Fármacos Neuroprotetores/farmacologia , Óleos Voláteis/farmacologia , Óleos de Plantas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Cálcio/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Humanos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fosfatidilinositol 3-Quinases/fisiologia , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismoRESUMO
OBJECTIVE: To determine the outcomes of patients with chest pain (CP) and prior history of coronary artery disease (CAD) managed with observation followed by outpatient stress myocardial perfusion imaging (MPI). METHODS: Retrospective analysis of patients with CP managed with observation followed by outpatient stress MPI, comparing cardiovascular (CV) event rates stratified by CAD history. RESULTS: 375 patients were included: 111 with and 264 without a CAD history. All patients underwent outpatient stress MPI within 72 h of observation. MPI identified patients at risk for CV events. However, while patients with negative MPI and without a CAD history had very low rates of short- and long-term CAD events (0.8%, 0.8%, and 1.3% at 30 days, 1 year, and 3 years, respectively), event rates of those with a negative test but a CAD history were significantly higher (2.6%, 5.3%, and 6.6% at 30 days, 1 year and 3 years, respectively; p = 0.044 and p = 0.034 compared to CAD- patients at 1 year and 3 years, respectively). In a multivariable logistic regression model, a positive MPI proved to be an independent predictor of long-term CV events in patients with CP and prior CAD. CONCLUSION: Observation followed by stress MPI can effectively risk stratify CP patients with prior CAD for CV risk. These patients are at increased risk of CV events even after a low-risk stress MPI study. Patients presenting with CP and managed with a strategy of observation followed by a negative stress MPI warrant close short- and long-term monitoring for recurrent events.
RESUMO
To evaluate rejection episodes in lung-transplanted patients, we analyzed 31 bronchoalveolar lavage specimens for lymphocyte levels and lymphocyte expression of two intracytoplasmic activation markers, perforin, the pore-forming lytic protein, and granzyme B, a member of the serine esterase family. Using anti-human granzyme B and perforin mAbs, we show that their expression in alveolar lymphocytes is correlated with the severity of rejection as assessed by histological parameters and the patients' clinical status. The presence of these molecules may provide a prognostic parameter that will facilitate the patients' monitoring, particularly in cases with minimal acute lung rejection susceptible to rapid progression to severe rejection.
Assuntos
Rejeição de Enxerto/fisiopatologia , Transplante de Pulmão/imunologia , Linfócitos/química , Glicoproteínas de Membrana/fisiologia , Serina Endopeptidases/fisiologia , Biópsia , Brônquios/patologia , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Contagem de Células , Granzimas , Humanos , Imuno-Histoquímica , Linfócitos/enzimologia , Perforina , Fenótipo , Proteínas Citotóxicas Formadoras de Poros , Alvéolos Pulmonares/citologiaRESUMO
El divorcio o separación de una pareja con hijos pone fin a la convivencia de ambos padres en con junto con sus hijos. Puede haber múltiples causas para un divorcio y tanto la literatura como la ex periencia de los autores trabajando con familias en esta situación, dan cuenta de algunas consecuen cias a corto y largo plazo para los niños, especialmente en aquellos casos en que el divorcio ha sido conflictivo. En estos casos, se ha observado mayor riesgo de presentar trastornos de conducta, bajo rendimiento escolar y abuso de sustancias. Por otra parte puede tener consecuencias en la vida adulta, presentando mayor riesgo de patologías psiquiátricas o dificultades en las relaciones interpersonales si no se realiza una intervención que aborde algunas situaciones potencialmente traumáticas para los niños. Los pediatras se encuentran en una relación privilegiada con respecto a los niños y sus familias para detectar signos de discordia parental y alteración de la salud mental infantil. Las conductas que se manifiesten en los niños a consecuencia del divorcio y el conflicto parental van a depender de la edad y etapa de desarrollo del niño, siendo importante reconocerlas para poder intervenir de forma adecuada. El presente artículo propone directrices para guiar a los padres, ya que con un adecuado manejo de las situaciones de conflicto que ocurren alrededor del divorcio es posible prevenir algunas de las consecuencias que éste puede tener en los hijos.
When divorce or separation of a couple occurs, children will no longer live with both parents at the same time. There may be multiple causes for divorce, and both literature and our own experien ce as child mental health providers, report some short- and long-term consequences for children, especially where the divorce has been conflictive. In these cases, increased risk of developing be havioral disorders, poor school performance, and substance abuse has been documented as well as consequences in adult life with higher risk of psychiatric pathologies or difficulties in interpersonal relationships, if an intervention that addresses some potentially traumatic situations for children is not done. Pediatricians are in a privileged relationship with children and their families in order to detect signs of parental discord and altered mental health in children. Children behaviors as a result of divorce and parental conflict will depend on the age of the child and stage of development, and it is important to recognize them in order to intervene properly. This article proposes some guidelines for parents. Good management of high conflict situations related to divorce may prevent some of the consequences that these can have on children.
Assuntos
Humanos , Criança , Pediatria , Papel do Médico , Relações Profissional-Família , Divórcio/psicologia , Comportamento Infantil , Conflito Familiar/psicologia , Transtornos Mentais/prevenção & controle , Pais/psicologia , Psicologia da Criança , Transtornos Mentais/diagnóstico , Transtornos Mentais/etiologiaRESUMO
Modern drug discovery for Central Nervous System pathologies has recently focused its attention to in vitro neuronal networks as models for the study of neuronal activities. Micro Electrode Arrays (MEAs), a widely recognized tool for pharmacological investigations, enable the simultaneous study of the spiking activity of discrete regions of a neuronal culture, providing an insight into the dynamics of networks. Taking advantage of MEAs features and making the most of the cross-correlation analysis to assess internal parameters of a neuronal system, we provide an efficient method for the evaluation of comprehensive neuronal network activity. We developed an intra network burst correlation algorithm, we evaluated its sensitivity and we explored its potential use in pharmacological studies. Our results demonstrate the high sensitivity of this algorithm and the efficacy of this methodology in pharmacological dose-response studies, with the advantage of analyzing the effect of drugs on the comprehensive correlative properties of integrated neuronal networks.
Assuntos
Algoritmos , Córtex Cerebral/citologia , Córtex Cerebral/fisiologia , Eletrofisiologia/métodos , Modelos Neurológicos , Rede Nervosa/citologia , Rede Nervosa/fisiologia , Neurofarmacologia/métodos , Potenciais de Ação/efeitos dos fármacos , Potenciais de Ação/fisiologia , Animais , Células Cultivadas , Córtex Cerebral/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos , Rede Nervosa/efeitos dos fármacos , Processamento de Sinais Assistido por ComputadorRESUMO
Cell death requires coordinated intracellular signalling before disassembly of cell architecture by degradative enzymes. Although the death signalling cascades that involve the mitochondria, the ER and the plasma membrane have been extensively characterized, only a handful of studies have examined the functional and structural alterations of the nuclear pore complex (NPC) during neuronal death. Here, we show that during excitotoxic neuronal degeneration calpains redistributed across the nuclear envelope and mediated the degradation of NPC components causing altered permeability of the nuclear membrane. In primary dissociated neurons, simultaneous recording of cytosolic [Ca(2+)] and localization of fluorescent proteins showed that the onset of Ca(2+) overload signalled a progressive increase in the diffusion of small reporter molecules across the nuclear envelope. Later, calpain-mediated changes in nuclear pore permeability allowed accumulation of large proteins in the nucleus. Further, in a model of excitotoxic neuronal degeneration in Caenorhabditis elegans, we found similar nuclear changes and redistribution of fluorescent probes across the nuclear membrane in dying neurons. Our findings strongly suggest that increased leakiness of the nuclear barrier affects nucleocytoplasmic transport, alters the localization of proteins across the nuclear envelope and it is likely to be involved in Ca(2+)-dependent cell death, including ischemic neuronal demise.
Assuntos
Apoptose , Cálcio/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Poro Nuclear/fisiologia , Animais , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/metabolismo , Calpaína/metabolismo , Células Cultivadas , Ácido Glutâmico/farmacologia , Gliceraldeído 3-Fosfato Desidrogenase (NADP+)/metabolismo , Células HeLa , Humanos , Ratos , Receptores Nicotínicos/metabolismo , Transdução de SinaisAssuntos
Antígenos CD8/análise , Linfócitos T CD8-Positivos/imunologia , Sobrevivência de Enxerto/imunologia , Transplante de Pulmão/imunologia , Glicoproteínas de Membrana/análise , Subpopulações de Linfócitos T/imunologia , Adulto , Antígenos CD/análise , Complexo CD3/análise , Quimioterapia Combinada , Citometria de Fluxo , Seguimentos , Rejeição de Enxerto/epidemiologia , Rejeição de Enxerto/imunologia , Transplante de Coração-Pulmão/imunologia , Humanos , Imunossupressores/uso terapêutico , Células Matadoras Naturais/imunologia , Seleção de Pacientes , Perforina , Proteínas Citotóxicas Formadoras de Poros , Valores de Referência , Linfócitos T Citotóxicos/imunologiaRESUMO
Parental Alienation Syndrome is characterized by the presence of a campaign of denigration towards a previously loved parent, which is done by instigating unjustified fear and hatred in the course of a divorce custody battle. In this campaign one parent instills resentment in the child, and is successful when the child itself directly shuns the accused parent. The resentment in the child extends towards the family of the accused parent. The syndrome arises in the absence of actual abuse, which would justify the child's rejection. Pediatricians have access to their patients' family crisis and are in a special stance to acknowledge it. The parental alienation syndrome is a form of emotional child abuse and both psychological assessment and multidisciplinary intervention are required to stop it. In this article we provide an update on parental alienation syndrome and was motivated by the authors' intervention in clinical cases where this diagnosis was posed.
El Síndrome de Alienación Parental se caracteriza por la presencia de una campaña de denigración hacia un progenitor previamente querido por el niño, la que se inicia instigando temor y animadversión injustificadas y que suele producirse durante el litigio por la custodia del niño en un proceso de divorcio. En esta campaña habitualmente participa un progenitor que instiga el resentimiento y que culmina cuando el niño rechaza abiertamente al progenitor acusado, haciendo suyas las descalificaciones esgrimidas en su contra. Este síndrome se produce en ausencia de maltrato físico o abuso, en cuyo caso el rechazo del niño es justificado. Los pediatras tienen acceso a las crisis por las que atraviesan las familias de sus pacientes y están en condiciones de detectar y reconocer a tiempo una situación de esta naturaleza. El síndrome de alienación parental es una forma grave de maltrato psicológico y requiere de una intervención multidisciplinaria para detenerlo. En el presente artículo se presenta una actualización del síndrome de alienación parental basado en la experiencia de los autores en casos clínicos en los cuales se propuso este diagnóstico.
Assuntos
Humanos , Feminino , Criança , Maus-Tratos Infantis , Divórcio/psicologia , Relações Pais-Filho , Pais/psicologia , Alienação Social , SíndromeRESUMO
BACKGROUND: Even though studying Medicine and perceiving abuse seem to be two opposite situations, recent investigations in Chile and abroad find that this is a frequent and pervasive combination. These studies also report the negative effects in the lives of students as well as the impact on the profession as a whole. AIM: To ascertain the perception of abusive situations in medical students during training at the University of Chile. MATERIAL AND METHODS: Descriptive and cross sectional study in which a questionnaire was applied to all the students enrolled in 2nd, 3rd, 5th and 7th year during 2001 and 2002. Using short vignettes, they were asked if they had experienced verbal, psychological, physical and sexual abuse, at least once, during their training, by whom and the eventual effects derived from it. RESULTS: We obtained 757 questionnaires. Of the surveyed students, 91% reported having perceived at least one abusive episode during training. Teachers and fellow students were identified as the main offenders. Among the effects of such behavior, 32% mentioned that they considered dropping out of the career as a consequence of this experience. DISCUSSION: The perception of abuse in medical students is common and has adverse effects. Efforts should be made to draw attention to this problem to prevent it.
Assuntos
Estágio Clínico/estatística & dados numéricos , Docentes de Medicina , Percepção , Estudantes de Medicina/psicologia , Violência/psicologia , Adulto , Chile , Feminino , Humanos , Internato e Residência , Relações Interprofissionais , Masculino , Cultura Organizacional , Fatores Sexuais , Estudantes de Medicina/estatística & dados numéricos , Inquéritos e Questionários , Violência/estatística & dados numéricosRESUMO
A combinatorial approach towards identifying inhibitors of VALA-4 was investigated. A library of piperazine-peptoid-bisarylureas was assembled in solid phase and screening in the novel v-well assay enabled the identification of active compounds.
Assuntos
Técnicas de Química Combinatória , Integrinas/antagonistas & inibidores , Integrinas/química , Peptídeos/química , Piperazinas/farmacologia , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Receptores de Retorno de Linfócitos/química , Cromatografia Líquida de Alta Pressão , Bases de Dados como Assunto , Dimetil Sulfóxido/química , Dimetilformamida/química , Integrina alfa4beta1 , Espectrometria de Massas , Modelos Químicos , Biblioteca de Peptídeos , Peptídeos/síntese química , Peptoides , Piperazina , Software , Triazóis/químicaRESUMO
The TU25 mutant strain of the basidiomycete Coprinus cinereus grows well at 28 degrees C but not at 37 degrees C. Microscopic examination revealed that TU25 exhibited swelling at hyphal apices after a shift-up from 28 degrees C to 37 degrees C. The temperature sensitivity and hyphal swelling co-segregated through meiosis as expected for a single Mendelian factor, designated hyt1 (hyphal tip). Both defects could be suppressed by the presence of osmotic stabilizers in the medium, suggesting that the hyt1 gene product is required for proper cell-wall function. Chemical analysis of the cell walls, however, failed to detect any clear difference in the composition of the cell-wall polysaccharides between the wild-type and TU25. A DNA fragment which complements the hyt1-1 mutation was cloned and sequenced. The predicted protein in the ORF essential for complementation of hyt1-1 is a novel protein.
Assuntos
Coprinus/genética , Proteínas Fúngicas/genética , Genes Fúngicos , Mutação , Sequência de Aminoácidos , Proteínas Fúngicas/química , Dados de Sequência Molecular , TemperaturaRESUMO
The synthesis and identification of a novel series of inhibitors of integrin VLA-4 are described. Their in vitro activity and selectivity against closely related integrins are also presented.
Assuntos
Antialérgicos/antagonistas & inibidores , Integrinas/antagonistas & inibidores , Peptídeos/síntese química , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Receptores de Antígeno muito Tardio/antagonistas & inibidores , Desenho de Fármacos , Integrina alfa4beta1 , Peptídeos/química , Peptídeos/farmacologia , Relação Estrutura-Atividade , Ureia/químicaRESUMO
We have developed a homogeneous high-capacity assay format for measuring integrin- and selectin-dependent cell binding to immobilized ligand using V-well microtiter plates. 2',7'-Bis(2-carboxyethyl)-5-(and-6)-carboxylfluorescence, acetoxymethylester-labeled cells are added to ligand-coated V-shaped microtiter wells. Bound cells are separated from free cells using centrifugal force to produce shear stress. Nonadherent cells accumulate in the nadir of the well and are measured using a fluorescence plate reader. Antibody or low-molecular-weight inhibitors of either the ligand or the cell surface receptor result in less cell binding, more cells in the pellet, and increased signal. The optimization and validation of the very late antigen-4/vascular cell adhesion molecule-1 assay is described in detail. We demonstrate that this assay can be rapidly adapted to measure other integrin- and selectin-mediated interactions. This assay format has several advantages over conventional assays. The centrifugal process is biologically relevant and eliminates the washing steps to remove nonadherent cells that can cause well-to-well and plate-to-plate variation. Because the assay is robust with a high signal-to-noise ratio and low variability, it is ideally suited for studying multiple parameters of cell adhesion and for high capacity screening.