RESUMO
Variants in CLCN4, which encodes the chloride/hydrogen ion exchanger CIC-4 prominently expressed in brain, were recently described to cause X-linked intellectual disability and epilepsy. We present detailed phenotypic information on 52 individuals from 16 families with CLCN4-related disorder: 5 affected females and 2 affected males with a de novo variant in CLCN4 (6 individuals previously unreported) and 27 affected males, 3 affected females and 15 asymptomatic female carriers from 9 families with inherited CLCN4 variants (4 families previously unreported). Intellectual disability ranged from borderline to profound. Behavioral and psychiatric disorders were common in both child- and adulthood, and included autistic features, mood disorders, obsessive-compulsive behaviors and hetero- and autoaggression. Epilepsy was common, with severity ranging from epileptic encephalopathy to well-controlled seizures. Several affected individuals showed white matter changes on cerebral neuroimaging and progressive neurological symptoms, including movement disorders and spasticity. Heterozygous females can be as severely affected as males. The variability of symptoms in females is not correlated with the X inactivation pattern studied in their blood. The mutation spectrum includes frameshift, missense and splice site variants and one single-exon deletion. All missense variants were predicted to affect CLCN4's function based on in silico tools and either segregated with the phenotype in the family or were de novo. Pathogenicity of all previously unreported missense variants was further supported by electrophysiological studies in Xenopus laevis oocytes. We compare CLCN4-related disorder with conditions related to dysfunction of other members of the CLC family.
Assuntos
Canais de Cloreto/genética , Síndromes Epilépticas/genética , Deficiência Intelectual/genética , Adolescente , Adulto , Idoso , Animais , Criança , Pré-Escolar , Canais de Cloreto/metabolismo , Epilepsia/genética , Síndromes Epilépticas/fisiopatologia , Família , Feminino , Genes Ligados ao Cromossomo X , Doenças Genéticas Ligadas ao Cromossomo X/genética , Mutação em Linhagem Germinativa , Humanos , Deficiência Intelectual/metabolismo , Masculino , Pessoa de Meia-Idade , Mutação , Oócitos , Linhagem , Fenótipo , Síndrome , Substância Branca/fisiopatologia , Xenopus laevisRESUMO
INTRODUCTION: Most preclinical trials are designed to identify potential torsadogenicity test only for surrogates of torsade de pointes, most commonly prolongation of the heart rate corrected QT interval (QTc). This study was conducted to determine which correction method best accounts for the effects of changes in the RR interval on the QT interval of conscious rabbits. This study was also conducted to validate the use of conscious, sling-trained rabbits to assess the QTc interval, and to evaluate the reliability and accuracy of this preparation in predicting drug-induced QTc prolongation in humans. METHODS: ECGs were recorded via bipolar transthoracic ECG leads in 7 conscious rabbits previously trained to rest quietly in slings. The heart rate was slowed with 2.0 mg/kg zatebradine to assess the effects of heart rate on the QT interval. The same ECG and sling preparation was used to evaluate the effects in of three drugs known to be torsadogenic in humans (cisapride, dofetilide and haloperidol), two drugs known to be non-torsadogenic in humans (propranolol and enalaprilat) and a control article (vehicle). All of the test articles were administered intravenously to 4 rabbits, and both RR and QT intervals were measured and the corrected QT values were calculated by an investigator blinded to the test article, utilizing our own algorithm (QTc=QT/(RR)(0.72)) which permitted the least dependency of QTc on RR interval. RESULTS: The following regression equations were obtained relating QT to RR: QT=2.4RR(0.72), r(2)=0.79, with RR intervals varying between 210 and 350 ms. QTc lengthened significantly in all conscious rabbits given intravenous cisapride, dofetilide and haloperidol (p<0.05), and QTc did not change with DMSO (vehicle control), propranolol or enalaprilat. DISCUSSION: Results indicate that a bipolar transthoracic ECG recorded in conscious, sling-trained rabbits may provide an easy and economical methodology useful in predicting QTc lengthening of novel pharmacological entities.
Assuntos
Avaliação Pré-Clínica de Medicamentos/métodos , Eletrocardiografia/métodos , Síndrome do QT Longo/induzido quimicamente , Testes de Toxicidade , Animais , Cisaprida/efeitos adversos , Estado de Consciência , Eletrocardiografia/instrumentação , Enalaprilato/farmacologia , Feminino , Haloperidol/efeitos adversos , Injeções Intravenosas , Síndrome do QT Longo/fisiopatologia , Masculino , Fenetilaminas/efeitos adversos , Propranolol/farmacologia , Coelhos , Reprodutibilidade dos Testes , Sulfonamidas/efeitos adversosRESUMO
This 10-day (D) study was conducted to evaluate changes in traditional and newer kidney safety biomarker expression levels in dogs. Animals received cisplatin (CDDP, 0.75 mg per kg per day) or 0.9% Saline (vehicle) for 5 days. Serum/urine samples were collected at various time points. Cage-side observations included emesis (D1-2/D4-D5/D7-9), absence of stool (D5-9/D11), soft stool (D4-7/D12), excessive salivation (D1/D3/D5-6), decreased food consumption (D5-8), decreased activity (D7-8) and/or dehydration (D7). Animals were necropsied when serum creatinine (sCr) levels measured at ≥1.9 mg dL-1, indicating significant loss of renal function; or at the end of the study (D11). When compared to controls, increases in BUN/sCr were detected on D3, D5 and/or D8. Increases in urinary total protein (Ur TP) were noted on D6. The moribund dog that was euthanized early on D7 showed insignificant increases in urinary osteopontin (Ur OPN), urinary neutrophil gelatinase-associated lipocalin (Ur NGAL), urinary clusterin (Ur CLU), sCr, serum cystatin C (sCYS C) and urinary cystatin C (Ur CYS C) on D5 when compared to controls. Insignificant increases in urinary albumin (Ur ALB) were observed from an animal that was euthanized on D7 and 1 : 2 surviving animals on D8 relative to baseline. From three dogs that were euthanized on D9, increases in Ur CLU, and/or sCYS C were noted on D8 relative to baseline. The two surviving dogs showed elevated Ur CLU and 1 : 2 surviving dogs showed elevated Ur CYS C. Decreased urinary kidney injury molecule 1 (Ur KIM-1) on D3/D5 was evident (versus baseline and controls). CDDP-induced cortico-medullary lesions were characterized as minimal to mild tubule degeneration/necrosis, dilatation, regeneration, cell alteration, intratubular casts, interstitial inflammation and vacuolization. Increased Ur OPN and Ur CLU correlated with enhanced OPN and CLU immunopositive staining in damaged cortical epithelium in the proximal tubules. Enhanced KIM-1 staining in damaged cortico-medullary tubular epithelium appeared in the absence of rises in Ur KIM-1. This study showed changes in kidney safety protein biomarkers associated with CDDP nephrotoxicity in dogs and possibly in humans.
RESUMO
Polyclonal antithymocyte globulin (ATG)/antilymphocyte and antilymphoblast globulins (ALG) antibodies have been used successfully in transplantation, aplastic anemia and graft-versus-host disease. Flow cytometry has been used to analyze peripheral blood lymphocyte populations in transplant patients receiving polyclonal ATG/ALG preparations for immunosuppression. Recent reports have indicated clinical dose adjustment based on levels of patient's cells expressing various CD antigens. In vitro analysis of individual polyclonal ATG/ALG CD antigen specificity could identify appropriate antigens for clinical monitoring as well as provide useful in vitro activity data. Therefore, a flow cytometry based assay to characterize and compare activities to specific CD antigens found on the surface of peripheral blood lymphocytes has been developed. Activities found in four lots each of horse ATG (ATGAM, Upjohn), rabbit and horse ATG (thymoglobulin and lymphoglobulin, Merieux), horse ALG (Minnesota), and rabbit ATG (Fresenius) have been compared for CD2, CD3, CD4, CD5, CD7, CD8, CD11a, CD18, CD28, CD44, CD45, and TCR-alpha/beta antigens. Quantitation is achieved by measuring the concentration of ATG/ALG required to give 50% inhibition of antigen specific fluorescent-labeled monoclonal antibody relative to buffer controls. The three horse products tested have similar activity to most antigens tested. However, Fresenius rabbit ATG has the lowest activity for almost all antigens tested whereas the Merieux rabbit ATG has activities closer to the horse products. This technique allows for rapid in vitro comparison of reactivities to individual lymphocyte antigens as well as in vitro analysis of consistency.
Assuntos
Antígenos CD/imunologia , Soro Antilinfocitário , Linfócitos/imunologia , Animais , Anticorpos Monoclonais , Soro Antilinfocitário/uso terapêutico , Citometria de Fluxo/métodos , Cavalos , Humanos , Terapia de Imunossupressão/métodos , Coelhos , Timo/imunologiaRESUMO
The high affinity of biotin for streptavidin has made this pair of molecules very useful for in vivo applications. To optimize reagents for one potential in vivo application, antibody-based pretargeting of cancer, we have prepared a number of new biotin and streptavidin derivatives. The derivatives developed include new radiolabeled biotin reagents, new protein biotinylation reagents, and new biotin multimers for cross-linking and/or polymerization of streptavidin. We have also modified streptavidin by site-directed mutation and chemical modification to improve its in vivo characteristics, and have developed new reagents for cross-linking antibody fragments with streptavidin. A brief overview of these new reagents is provided.
Assuntos
Biotina , Estreptavidina , Marcadores de Afinidade , Anticorpos Monoclonais , Biotina/química , Reagentes de Ligações Cruzadas , Humanos , Indicadores e Reagentes , Radioisótopos do Iodo , Mutagênese Sítio-Dirigida , Neoplasias/radioterapia , Engenharia de Proteínas , Estrutura Quaternária de Proteína , Radioimunoterapia , Estreptavidina/química , Estreptavidina/genéticaRESUMO
We have examined the reproducibility, sensitivity, and specificity of detecting Plasmodium falciparum using the polymerase chain reaction (PCR) and the species-specific probe pPF14 under field conditions in the Venezuelan Amazon. Up to eight samples were field collected from each of 48 consenting Amerindians presenting with symptoms of malaria. Sample processing and analysis was performed at the Centro Amazonico para la Investigacion y Control de Enfermedades Tropicales Simon Bolivar. A total of 229 samples from 48 patients were analyzed by PCR methods using four different P. falciparum-specific probes. One P. vivax-specific probe and by conventional microscopy. Samples in which results from PCR and microscopy differed were reanalyzed at a higher sensitivity by microscopy. Results suggest that microscopy-negative, PCR-positive samples are true positives, and that microscopy-positive and PCR-negative samples are true negatives. The sensitivity of the DNA probe/PCR method was 78% and its specificity was 97%. The positive predictive value of the PCR method was 88%, and the negative predictive value was 95%. Through the analysis of multiple blood samples from each individual, the DNA probe/PCR methodology was found to have an inherent reproducibility that was highly statistically significant.
Assuntos
DNA de Protozoário/sangue , Malária Falciparum/diagnóstico , Plasmodium falciparum/isolamento & purificação , Reação em Cadeia da Polimerase , Animais , Sequência de Bases , Primers do DNA/química , Sondas de DNA , DNA de Protozoário/química , Humanos , Indígenas Sul-Americanos , Malária Falciparum/sangue , Malária Vivax/sangue , Malária Vivax/diagnóstico , Dados de Sequência Molecular , Plasmodium falciparum/genética , Plasmodium vivax/genética , Plasmodium vivax/isolamento & purificação , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Especificidade da Espécie , VenezuelaRESUMO
In the course of computed tomographic (CT) evaluations involving 200 patients with suspected pelvic disease we have found that scan quality is often suboptimal. A preliminary report employing a low density bladder opacification method is presented whereby an indwelling catheter is inserted to control bladder volume and facilitate the instillation of low density iodinated contrast agent (0.6% Renografin-60). The patient remains supine throughout the twenty-five-minute procedure. Observations during the CT staging and follow-up of a controlled group of 8 patients undergoing immunotherapy and/or radiation therapy for bladder carcinoma indicate that this is a simple, safe, and effective staging procedure. We have obtained reliable clinical-radiologic-pathologic corrlation as a result of careful surgical staging and biopsy, followed by open surgery and full pathologic examination. It is hoped that this information will help initiate other similar studies to determine the diagnostic accuracy of this method and thus its use in pretherapeutic evaluation of the bladder lesion and its subsequent response to treatment.
Assuntos
Carcinoma/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Neoplasias da Bexiga Urinária/diagnóstico por imagem , Bexiga Urinária/diagnóstico por imagem , Idoso , Carcinoma de Células Escamosas/diagnóstico por imagem , Carcinoma de Células Escamosas/patologia , Carcinoma de Células de Transição/diagnóstico por imagem , Carcinoma de Células de Transição/patologia , Diatrizoato de Meglumina , Feminino , Humanos , Masculino , Estadiamento de Neoplasias , Intensificação de Imagem Radiográfica , Neoplasias da Bexiga Urinária/patologiaRESUMO
Thirty-four patients wih infiltrative bladder carcinoma, Stage B2C or higher were treated with immunotherapy and irradiation. Seventeen patients are alive, and 17 have succumbed to their disease. Eight patients underwent cystectomy after immunotherapy and irradiation; 6 of 8 are alive and well at the present time. The technique of immunotherapy is outlined. New methodology for sequential CT scans and scheduled bladder biopsies is mentioned. The 17 patients have survived twelve to sixty-nine months after immunotherapy and irradiation. Downstaging is demonstrated based on sequential CT scans of the bony pelvis and histologic biopsy. The biopsies reveal eosinophilia and multinucleated giant cells, a specific response to immunotherapy. A prospective randomized study will be initiated.
Assuntos
Carcinoma de Células de Transição/terapia , Imunoterapia , Linfonodos/citologia , Neoplasias da Bexiga Urinária/terapia , Adulto , Idoso , Animais , Carcinoma de Células de Transição/mortalidade , Feminino , Humanos , Linfonodos/imunologia , Masculino , Pessoa de Meia-Idade , Dosagem Radioterapêutica , Suínos/imunologia , Tomografia Computadorizada por Raios X , Neoplasias da Bexiga Urinária/mortalidadeRESUMO
Computed tomography (CT) was utilized as part of the surgical-pathologic-radiologic evaluation of 21 patients who were treated for bladder carcinoma with attenuated irradiation and immunotherapy. Fifteen patients had moderately infiltrative (Stage B2-C or less) disease, and it was found that a routine high resolution CT technique using a modern fast scanner delineated the tumor in most cases. More accurate assessment of tumor response to therapy and evaluation of tumor progression were facilitated using a gas insufflation technique combined with intravenous contrast fusion. This was followed in selected cases by quantitative measurements of CT attenuation values using a recently introduced CT software program. Using this program, individual pixel values were obtained in selected areas and evaluation of the resulting numerical data and pixel histograms aided in the differentiation of tumor tissue from adjacent bladder wall and mapped out areas of tumor necrosis. Our preliminary observations suggest that quantitative CT studies incorporating assessment of printouts of attenuation values of adjacent pixels within a region of interest will improve the delineation of smaller (B1/B2) lesions and will aid objective characterization of tumor tissue during and following therapy.
Assuntos
Tomografia Computadorizada por Raios X/métodos , Neoplasias da Bexiga Urinária/terapia , Adulto , Idoso , Biópsia , Meios de Contraste , Humanos , Imunoterapia , Pessoa de Meia-Idade , Neoplasias da Bexiga Urinária/diagnóstico por imagem , Neoplasias da Bexiga Urinária/patologiaRESUMO
INTRODUCTION: The purpose of this study was to determine the sensitivity and specificity for predicting the liability of a compound to lengthen QTc using isolated, perfused guinea pig hearts (Langendorff preparation). METHODS: QTc (Fridericia correction) was calculated from bipolar transventricular electrograms. Hearts were exposed to escalating concentrations of 26 compounds thought to lengthen, and 13 compounds thought not to lengthen, QTc in humans. RESULTS: In this preparation, QTc was found to lengthen in 26 of 26 compounds thought to be positive (sensitivity 1.00) and not to lengthen or to lengthen insignificantly in 13 of 13 compounds thought to be negative (specificity 1.0) in man. Probucol and ontazolast could not be studied because of limited solubility. Successful experiments were conducted on over 98% of guinea pigs anesthetized. DISCUSSION: We believe that the isolated perfused guinea pig heart is an in vitro preparation that could be utilized early in preclinical testing for identifying a liability to lengthen QTc in humans, but we do not believe--as is true also for other in vitro methods--that the concentration at which the liability is demonstrated in vitro necessarily predicts the concentration at which a liability exists in man.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Eletrocardiografia/efeitos dos fármacos , Coração/efeitos dos fármacos , Animais , Relação Dose-Resposta a Droga , Cobaias , Coração/fisiopatologia , Sistema de Condução Cardíaco/efeitos dos fármacos , Sistema de Condução Cardíaco/fisiopatologia , Técnicas In Vitro , Síndrome do QT Longo/induzido quimicamente , Masculino , Modelos Biológicos , Perfusão , Sensibilidade e EspecificidadeRESUMO
The synthesis of a nido-carboranyl congener of misonidazole, 1-(3'-nido-carboranyl-2'-hydroxy)propyl-2-nitroimidazole, has been carried out. Alternative methods of preparations were conducted to optimize the chemical yield, with a five step synthesis giving an overall yield (from 1,2-carborane) of 36%. A diastereomeric pair of nido-carboranyl compounds was obtained. The diastereomeric nido-carboranyl misonidazole congeners were (radio)iodinated to yield (> 90%) a mixture of diastereomeric compounds in which the iodine had bonded to a boron atom on the nido-carborane moiety. These compounds will be investigated for their application to boron neutron capture therapy (BNCT) and hypoxia imaging of cancer.
Assuntos
Compostos de Boro/síntese química , Radioisótopos do Iodo/química , Marcação por Isótopo/métodos , Misonidazol/análogos & derivados , Nitroimidazóis/síntese química , Terapia por Captura de Nêutron de Boro , Cromatografia Líquida de Alta Pressão , EstereoisomerismoRESUMO
Recent community based studies have shown that only a minority of visually impaired people who are eligible to be registered as partially sighted or blind are actually registered as such. To determine how many unregistered but eligible people are attending ophthalmic clinics a prospective study was undertaken of all patients (n = 1543) attending ophthalmic outpatient departments, at a single specialty eye hospital and two district general hospitals over a 1 week period. All patients with visual acuity < or = 6/18 or restricted visual field were interviewed. Registration status and factors affecting this were then determined. Although 95/174 patients interviewed were eligible for registration, 68 as partially sighted and 27 as blind, only 46 (48.4%) of these were registered. Asians and Afro-Caribbeans were under-represented in the group eligible for registration. Active treatment impeded registration. Patients having four or more hospital visits were on average 16 times more likely to be registered as those who had fewer attendances. Disabilities, in addition to visual impairment, were present in 40% (n = 38). This study shows that there is unregistered visual impairment in patients attending ophthalmic departments. As registration triggers multidisciplinary support, ophthalmologists need to be more alert to the benefits and criteria for partial sight and blind registration.
Assuntos
Cegueira/epidemiologia , Sistema de Registros , Baixa Visão/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Inglaterra/epidemiologia , Etnicidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oftalmologia , Estudos ProspectivosRESUMO
CT scans were obtained of 11 normal women and 34 women with gynecologic tumors. Twenty-three of 26 women with known malignant tumors had abnormal CT scans while all control patients had normal. The tumor extent demonstrated by CT correlated with either pathologic or surgical staging in 78%. Contrast enhancement increased the diagnostic yield and accuracy of staging. Attenuation values of the normal uterus and uterine tumors are quite similar prior to contrast medium. After administration of contrast agent the normal myometrium attenuation values increase more than other pelvic tissues. Uterine tumors show substantially less enhancement and show a greater variation of density throughout the tumor than normal myometrium. Density readings are not predictive of histologic type or tumor grade. Tumor was best detected and its extent best seen after high doses of intravenous contrast medium infusion, which should be considered a routine part of the diagnostic evaluation of these patients.
Assuntos
Neoplasias dos Genitais Femininos/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Meios de Contraste/administração & dosagem , Feminino , Humanos , Histerossalpingografia , Ácido Iotalâmico/administração & dosagem , Intensificação de Imagem Radiográfica , Vagina/diagnóstico por imagemRESUMO
The value of intravenous contract enhancement in computed tomography (CT) of pelvic malignancies was analyzed in 124 cases. Contrast enhancement of pelvic organs depends on the smooth muscle component and the administered iodine dose. Contrary to parenchymal organs in the abdomen the iodine washout from smooth muscle organs is slow. A rapid (approximately 5 minutes) intravenous infusion of 300 ml diatrizoate 30% followed immediately by scanning of the area of interest was found optimum to improve the CT visualization of tumors in all pelvic organs with the exception of the prostate. Depending on the organ to be investigated the intravenous contrast administration has to be combined with retrograde gas insufflation into bladder and/or rectum with insertion of a tampon into the vagina. The described technique is particularly useful for the assessment of early tumor extension beyond the primary organ.
Assuntos
Neoplasias Pélvicas/diagnóstico por imagem , Intensificação de Imagem Radiográfica/métodos , Tomografia Computadorizada por Raios X/métodos , Meios de Contraste/administração & dosagem , Diatrizoato/administração & dosagem , Feminino , Neoplasias dos Genitais Femininos/diagnóstico por imagem , Humanos , Infusões Parenterais , Linfoma/diagnóstico por imagem , Masculino , Neoplasias da Próstata/diagnóstico por imagem , Neoplasias Retais/diagnóstico por imagem , Neoplasias da Bexiga Urinária/diagnóstico por imagemRESUMO
The radiographic manifestation of histiocytic lymphoma in the skeletal and central nervous system of 112 non-selected patients were reviewed in a retrospective study. Histiocytic lymphoma metastatic to bone and brain are late manifestations of the disease occurring in 15 and 4%, respectively. Involvement of the spinal cord and nerve roots is found in 6% and represents often the initial manifestation of the disease. Histiocytic lymphoma of brain presents angiographically as avascular mass without site predilection and on contrast enhanced computer tomography as hyperdense lesion without necrotic center. On myelography an extradural mass or complete block is found. The most common skeletal locations are spine, pelvis, rib and femur. The ratio of multifocal to unifocal skeletal manifestations is approximately 2 to 1. The radiographic appearance varies from completely lytic, often associated with a soft tissue mass, to osteoblastic, often with Pagetoid features. Early recognition of histiocytic lymphoma manifestations is important to guarantee prompt institution of appropriate therapy since the disease can progress extremely rapidly.
Assuntos
Neoplasias Ósseas/diagnóstico por imagem , Neoplasias Encefálicas/diagnóstico por imagem , Linfoma Difuso de Grandes Células B/diagnóstico por imagem , Adolescente , Adulto , Idoso , Neoplasias Ósseas/patologia , Neoplasias Encefálicas/patologia , Criança , Feminino , Humanos , Linfoma Difuso de Grandes Células B/patologia , Masculino , Pessoa de Meia-Idade , Radiografia , Estudos Retrospectivos , Neoplasias da Medula Espinal/diagnóstico por imagem , Neoplasias da Medula Espinal/patologiaRESUMO
We investigated histone-DNA interactions in chromatin of amplified about 1000 fold dihydrofolate reductase (DHFR) domain in methotrexate resistant Chinese hamster ovary cell line CHOC 400. We explored chromatin structure of DHFR-gene and extended region lying 3'-downstream of the DHFR-gene, which includes two zones of DNA replication initiation, Ori-beta and Ori-gamma, and matrix attachment region (MAR). Using the method of hybridization with "protein images" we found that the yield of both core histones and histone H1 cross-linked with DNA in 5'-region of the DHFR-gene, in Ori-gamma and regions flanked from either side MAR is significantly decreased in comparison with 3'-transcribed region of the DHFR-gene, Ori-beta and MAR. It was shown previously that decreased yield of histones cross-linked with DNA is typical for 5'-region of actively transcribed genes. So, histone-DNA interactions in chromatin of the 5'-region of the house-keeping and moderately transcribed DHFR-gene, Ori-gamma and regions which flank MAR resemble those in 5'-regions of actively transcribed genes. Significant differences in amount of histones cross-linked with DNA in various parts of the DHFR-domain testify to high level of heterogeneity of chromatin structure in the domain.