RESUMO
The KIF5A gene (OMIM 602821) encodes a neuron-specific kinesin heavy chain involved in intracellular transport of mitochondria and other cargoes. KIF5A protein comprises the N terminal motor domain, the stalk domain and the C-terminal cargo binding domain. The binding between KIF5A and its cargoes is mediated by kinesin adaptor proteins such as TRAK1 and TRAK2. Numerous missense KIF5A mutations in the motor and stalk domains cause spastic paraplegia type 10 (SPG10, OMIM 604187). Conversely, the role of loss-of-function mutations, especially those affecting the cargo binding domain, is unclear. We describe a novel de novo KIF5A p.Ser974fs/c.2921delC mutation found by whole exome sequencing in a patient with a congenital severe disease characterized by myoclonic seizures and progressive leukoencephalopathy. Since this phenotype differs considerably from the KIF5A/SPG10 disease spectrum we propose that the KIF5A p.Ser974fs and possibly other mutations which lead to truncation of the C-terminal tail of the protein cause a novel disorder. We speculate that the unique effect of the C-terminal truncating KIF5A mutations may result from the previously described complex role of this protein domain in binding of the TRAK2 and possibly other kinesin adaptor protein(s).
Assuntos
Epilepsias Mioclônicas/genética , Mutação da Fase de Leitura , Cinesinas/genética , Leucoencefalopatias/genética , Idade de Início , Proteínas de Transporte/metabolismo , Humanos , Recém-Nascido , Peptídeos e Proteínas de Sinalização Intracelular , Cinesinas/metabolismo , Leucoencefalopatias/diagnóstico por imagem , Imageamento por Ressonância Magnética , Masculino , Proteínas do Tecido Nervoso/metabolismoRESUMO
In 1999, based on a single family, spondyloepimetaphyseal dysplasia (SEMD) with mental retardation (MR) was described as a novel syndrome with probably X-linked recessive inheritance and unknown molecular defect (MIM 300232). Our purpose was to search for the causative defect in the originally described family and in an independently ascertained second family. All patients had slowly progressive neurodegeneration with central and peripheral involvement and identical skeletal dysplasia. Whole exome sequencing performed in two subjects showed a single plausible candidate - the p.Asp237Gly variant in AIFM1 (chr. Xq26.1). The p.Asp237Gly segregated with disease as indicated by linkage analysis [maximum logarithm of odds score (LOD) score at theta 0 for the two families was 3.359]. This variant had not been previously reported and it was predicted to be pathogenic by Polyphen2, SIFT, MutationTaster and Mutation Assessor. AIFM1 encodes mitochondria associated apoptosis-inducing factor. The AIFM1 gene has been linked with COXPD6 encephalomyopathy (MIM 300816), Cowchock syndrome (MIM 310490) and X-linked deafness with neuropathy (DFNX5, MIM 300614), none of which are similar to SEMD-MR. Our results place SEMD as the third instance of a skeletal phenotype associated with a mitochondrial disease (the others being EVEN-PLUS syndrome caused by mutations of HSPA9 and CODAS syndrome due to LONP1 mutations).
Assuntos
Fator de Indução de Apoptose/genética , Predisposição Genética para Doença/genética , Doenças Mitocondriais/genética , Mutação , Doenças Neurodegenerativas/genética , Osteocondrodisplasias/genética , Sequência de Aminoácidos , Sequência de Bases , Exoma/genética , Saúde da Família , Feminino , Doenças Genéticas Ligadas ao Cromossomo X/diagnóstico , Doenças Genéticas Ligadas ao Cromossomo X/genética , Humanos , Masculino , Doenças Mitocondriais/diagnóstico , Doenças Neurodegenerativas/diagnóstico , Osteocondrodisplasias/diagnóstico , Linhagem , Fenótipo , Análise de Sequência de DNA/métodos , Homologia de Sequência de Aminoácidos , SíndromeRESUMO
BACKGROUND: Thoracic aortic aneurysms and dissections (TAAD) are silent but possibly lethal condition with up to 40 % of cases being hereditary. Genetic background is heterogeneous. Recently next-generation sequencing enabled efficient and cost-effective examination of gene panels. Aim of the study was to define the diagnostic yield of NGS in the 51 TAAD patients and to look for genotype-phenotype correlations within families of the patients with TAAD. METHODS: 51 unrelated TAAD patients were examined by either whole exome sequencing or TruSight One sequencing panel. We analyzed rare variants in 10 established thoracic aortic aneurysms-associated genes. Whenever possible, we looked for co-segregation in the families. Kaplan-Meier survival curve was constructed to compare the event-free survival depending on genotype. Aortic events were defined as acute aortic dissection or first planned aortic surgery. RESULTS AND DISCUSSION: In 21 TAAD patients we found 22 rare variants, 6 (27.3 %) of these were previously reported, and 16 (73.7 %) were novel. Based on segregation data, functional analysis and software estimations we assumed that three of novel variants were causative, nine likely causative. Remaining four were classified as of unknown significance (2) and likely benign (2). In all, 9 (17.6 %) of 51 probands had a positive result when considering variants classified as causative only and 18 (35.3 %) if likely causative were also included. Genotype-positive probands (n = 18) showed shorter mean event free survival (41 years, CI 35-46) than reference group, i.e. those (n = 29) without any plausible variant identified (51 years, CI 45-57, p = 0.0083). This effect was also found when the 'genotype-positive' group was restricted to probands with 'likely causative' variants (p = 0.0092) which further supports pathogenicity of these variants. The mean event free survival was particularly low (37 years, CI 27-47) among the probands with defects in the TGF beta signaling (p = 0.0033 vs. the reference group). CONCLUSIONS: This study broadens the spectrum of genetic background of thoracic aneurysms and dissections and supports its potential role as a prognostic factor in the patients with the disease.
Assuntos
Aneurisma da Aorta Torácica/diagnóstico , Aneurisma da Aorta Torácica/genética , Dissecção Aórtica/diagnóstico , Dissecção Aórtica/genética , Estudos de Associação Genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Mutação/genética , Adulto , Análise Mutacional de DNA , Diagnóstico por Imagem , Feminino , Heterozigoto , Humanos , Estimativa de Kaplan-Meier , Masculino , LinhagemRESUMO
AIM: Type of delivery onset is not currently evaluated for its predictive impact. This study explored whether the type of preterm delivery onset was an antenatal predictor for post-natal mortality in preterm infants <30 weeks' gestation and should be included in antenatal counselling. METHODS: This retrospective cohort study included 1117 preterm infants <30 weeks' gestation born between 1999 and 2008 in a tertiary perinatal referral centre. Study patients were classified into spontaneous or iatrogenic preterm deliveries. Spontaneous deliveries included deliveries after preterm premature rupture of membranes (PPROM) and preterm labour. The study outcome was infant mortality before discharge from hospital. RESULTS: We included 499 patients born after PPROM (44.7%) and 247 born after preterm labour (22.1%). Iatrogenic preterm birth was noted in 282 patients (25.2%) and 89 patients fulfilled both criteria for spontaneous and iatrogenic preterm delivery (8.0%). Babies born after iatrogenic preterm delivery in gestational weeks 25-29 had significantly higher mortality rates. Logistic regression revealed that type of preterm delivery onset was an independent antenatal predictor for post-natal mortality. CONCLUSION: Type of preterm delivery onset had a significant impact on post-natal mortality in preterm infants <30 weeks' gestation, with a higher mortality rate after iatrogenic preterm delivery.
Assuntos
Parto Obstétrico/efeitos adversos , Mortalidade Infantil , Recém-Nascido Prematuro , Nascimento Prematuro/etiologia , Áustria/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Estudos RetrospectivosRESUMO
BACKGROUND: Blood transfusions are required by most extremely low birth weight (ELBW) infants, but sometimes an adequate peripheral venous access cannot be achieved. Under these circumstances, we used 27 Gauge (G) peripherally inserted central catheter (PICC) lines that are routinely inserted on the second day of life. Due to their narrow lumen, hemolysis of transfused erythrocytes was a major concern. We therefore performed a retrospective study in ELBW infants to analyze the incidence, safety and feasibility of PRBC transfusions via 27 G PICC lines. METHODS: ELBW infants admitted from 08/2011-07/2012 were screened for packed red blood cell (PRBC) transfusions. Those applied via 27 G PICC lines were identified. For analysis of transfusion safety (hemolysis), hemoglobin and potassium levels as well as cardiovascular variables (invasive mean arterial blood pressure and heart rate) were evaluated before and after transfusion. For analysis of transfusion feasibility, catheter removal after transfusion and the reason for removal were recorded. RESULTS: A total of 648 transfusions were applied in 110 ELBW infants. 27 infants (24%) received no transfusion. In 12/83 (14.5%) infants who received PRBCs, transfusions were applied using a 27 G PICC line (38/648, 5.9%). Patients who received PRBCs via the PICC line were smaller at birth (582 g [range 380-752 g] vs. 710 g [430-972 g]; 23+6 [23+1-27+6] vs. 26+0 [23+1-31+4]) and required a higher number of PRBC transfusions (n=13 vs. n=5) overall. Transfusion analysis showed an appropriate increase of blood hemoglobin levels and stable potassium levels as well as cardiovascular parameters. 4/38 of PICC lines were removed within 24 h after transfusion, one due to occlusion (15 h after transfusion). CONCLUSIONS: We conclude that PRBC transfusions via 27 G PICC lines were feasible and performed without signs of hemolysis in ELBW infants. Our findings may help clinicians in the management of ELBW infants requiring transfusions if a peripheral venous access is not achievable.
Assuntos
Cateterismo Periférico/instrumentação , Transfusão de Eritrócitos/instrumentação , Recém-Nascido de Peso Extremamente Baixo ao Nascer , Doenças do Prematuro/terapia , Peso ao Nascer , Pressão Sanguínea/fisiologia , Remoção de Dispositivo , Desenho de Equipamento , Segurança de Equipamentos , Estudos de Viabilidade , Frequência Cardíaca/fisiologia , Hemoglobinometria , Humanos , Recém-Nascido , Doenças do Prematuro/sangue , Poliuretanos , Potássio/sangue , Estudos RetrospectivosRESUMO
Blood transfusions are required by the majority of extremely premature infants. Packed red blood cells (PRBCs) are usually applied via simple peripheral cannulas. In situations where no peripheral venous access is achievable, 27 Gauge (G) neonatal PICC lines - that are ideally exclusively dedicated to application of parenteral nutrition - may represent a useful alternative access for PRBC transfusions. However, transfusion via small scaled catheters may damage PRBCs and lead to hemolysis. We here evaluate whether transfusion of irradiated PRBCs via 27 G PICC lines leads to hemolysis in vitro.Experimental transfusions of gamma-irradiated PRBCs were performed at increasing velocities (2.5, 3.7, 5 ml/h; full force manual push approximating 30 ml/h) via 27 G PICC lines of 20 and 30 cm length. Parameters of hemolysis (lactate dehydrogenase, potassium and free hemoglobin) were measured from the supernatants of transfused PRBCs and the percentage of hemolysis was calculated.Potassium and lactate dehydrogenase after transfusion at increasing velocities did not differ significantly from negative controls. Free hemoglobin levels showed a small but significant increase at the slowest transfusion speed (2.5 ml/h) using the 30 cm 27 G PICC line, with a relative hemolysis of only 0.13%. A manual push (approximating 30 ml/h) showed no significant changes of parameters from baseline.We conclude that transfusion of gamma-irradiated PRBCs using a 27 G neonatal PICC line does not cause clinically relevant hemolysis in vitro. Clinical studies are needed to confirm the feasibility and safety of the approach in vivo.
Assuntos
Segurança do Sangue , Cateterismo Venoso Central/instrumentação , Transfusão de Eritrócitos/instrumentação , Hemólise , Recém-Nascido de Peso Extremamente Baixo ao Nascer , Doenças do Prematuro/terapia , Velocidade do Fluxo Sanguíneo , Feminino , Hemoglobinometria , Humanos , Técnicas In Vitro , Recém-Nascido , Doenças do Prematuro/sangue , L-Lactato Desidrogenase/sangue , Masculino , Potássio/sangueRESUMO
PURPOSE: Despite a decreasing incidence, intraventricular hemorrhage (IVH) remains a point of major concern in neonatology due to its association to adverse neurodevelopmental outcome (NDO). Aim of this study was to compare outcome of preterm infants with different grades of IVH born below 32 weeks of gestational age (GA) with outcome of controls without IVH and to especially evaluate the influence of low grade IVH on NDO. METHODS: Four hundred seventy-one preterm infants with a GA below 32 weeks were admitted to our neonatal intensive care unit between 1994 and 2005 and included into analysis. RESULTS: IVH patients showed significantly lower mean psychomotor and mental developmental indices and a significantly higher percentage of cerebral palsy and visual impairment. Results of IVH patients born below 28 weeks of GA were significantly worse than results of IVH patients born at or above 28 weeks of GA. In all parameters, an increase of abnormal results with increasing grade of IVH could be observed; even patients with low-grade IVH (grades I and II) showed higher percentages of impairment compared to controls without any IVH. CONCLUSION: Even low-grade IVH has an significant impact on neurodevelopmental outcome of preterm patients and gestational age influences the impact of intraventricular hemorrhage on neurodevelopmental outcome.
Assuntos
Ventrículos Cerebrais , Desenvolvimento Infantil/fisiologia , Recém-Nascido Prematuro/fisiologia , Hemorragias Intracranianas/psicologia , Envelhecimento/fisiologia , Peso ao Nascer , Paralisia Cerebral/etiologia , Estudos de Coortes , Feminino , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Hemorragias Intracranianas/complicações , Masculino , Doenças do Sistema Nervoso/epidemiologia , Testes Neuropsicológicos , Transtornos da Visão/etiologiaRESUMO
BACKGROUND: The use of recombinant lactic acid bacteria (LAB) as vehicles for mucosal delivery of recombinant allergens is an attractive concept for antigen-defined allergy prevention/treatment. Interventions with LAB are of increasing interest early in life when immune programming is initiated. Here, we investigated the effect of neonatal colonization with a recombinant LAB producing the major birch pollen allergen Bet v 1 in a murine model of type I allergy. METHODS: We constructed a recombinant Lactobacillus (L.) plantarum NCIMB8826 strain constitutively producing Bet v 1 to be used for natural mother-to-offspring mono-colonization of germ-free BALB/c mice. Allergen-specific immunomodulatory effects of the colonization on humoral and cellular immune responses were investigated prior and after sensitization to Bet v 1. RESULTS: Mono-colonization with the Bet v 1 producing L. plantarum induced a Th1-biased immune response at the cellular level, evident in IFN-γ production of splenocytes upon stimulation with Bet v 1. After sensitization with Bet v 1 these mice displayed suppressed IL-4 and IL-5 production in spleen and mesenteric lymph node cell cultures as well as decreased allergen-specific antibody responses (IgG1, IgG2a, and IgE) in sera. This suppression was associated with a significant up-regulation of the regulatory marker Foxp3 at the mRNA level in the spleen cells. CONCLUSION: Intervention at birth with a live recombinant L. plantarum producing a clinically relevant allergen reduces experimental allergy and might therefore become an effective strategy for early intervention against the onset of allergic diseases.
Assuntos
Antígenos de Plantas/imunologia , Imunização , Lactobacillus plantarum/genética , Lactobacillus plantarum/imunologia , Linfócitos T Reguladores/imunologia , Células Th1/imunologia , Alérgenos/administração & dosagem , Alérgenos/genética , Alérgenos/imunologia , Animais , Animais Recém-Nascidos , Antígenos de Plantas/genética , Betula/genética , Betula/imunologia , Citocinas/biossíntese , Citocinas/imunologia , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Hipersensibilidade Imediata , Isotipos de Imunoglobulinas/sangue , Isotipos de Imunoglobulinas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Pólen/genética , Pólen/imunologia , Baço/citologia , Baço/imunologia , Células Th2/imunologiaRESUMO
HL-60 promyelocytic leukemia cells differentiated to eosinophils and eosinophilic precursors when cultured under mildly alkaline conditions (pH 7.6-7.8) for 7 d without refeeding. New cytoplasmic granules appeared blue in the least mature cells and red in the most mature cells when stained with Wright-Giemsa. The granules also stained with Luxol-fast-blue, a characteristic of eosinophil granules. Furthermore, most cells contained the eosinophil major basic protein (MBP); the Charcot-Leyden Crystal (CLC) protein (lysophospholipase), eosinophil peroxidase, acid phosphatase, and arylsulfatase were also detected in a portion of these cells. The eosinophil major basic protein was found in a high proportion of undifferentiated cells, and thus may be constituitively produced. By examining finely banded chromosomes, translocation break points were demonstrated at q22 on one chromosome 16 and at q23 on the other homologue; abnormalities in this region of the long arm of 16 are a characteristic finding in the recently described syndrome of acute myelomonocytic leukemia (AMMoL) with abnormal bone marrow eosinophils. In common with the bone marrow eosinophils in these patients, the HL-60 eosinophil granules contained chloroacetate esterase and periodic-acid Schiff (PAS) reactive material; crystalloid inclusions were rare. Therefore, the HL-60 cell line appears to be an in vitro model for eosinophilopoiesis and may be specially suited for the study of the abnormal eosinophils seen in certain malignant conditions.
Assuntos
Transformação Celular Neoplásica/patologia , Eosinófilos/patologia , Leucemia Mieloide Aguda/patologia , Ribonucleases , Proteínas Sanguíneas/análise , Linhagem Celular , Transformação Celular Neoplásica/metabolismo , Transformação Celular Neoplásica/ultraestrutura , Cromossomos Humanos 16-18 , Grânulos Citoplasmáticos/enzimologia , Grânulos Citoplasmáticos/ultraestrutura , Proteínas Granulares de Eosinófilos , Eosinófilos/enzimologia , Eosinófilos/ultraestrutura , Células-Tronco Hematopoéticas/patologia , Histocitoquímica , Humanos , Concentração de Íons de Hidrogênio , Cariotipagem , Leucemia Mieloide Aguda/metabolismoRESUMO
The purpose of the study was to assess the haemodynamic (blood pressure and heart rate) changes and stress responses (serum cortisol and serum amyloid A [SAA] concentrations) to surgery in piglets during total intravenous anaesthesia (TIVA) with propofol and fentanyl. After preanaesthetic medication with intramuscular midazolam (0.5 mg/kg body mass), ketamine (10 mg/kg) and butorphanol (0.5 mg/kg) anaesthesia was induced in five piglets, with intravenous propofol (1 mg/kg) followed by tracheal intubation and mechanical lung ventilation. Soft tissue surgery was performed in the jugular and inguinal regions during TIVA with propofol (8 mg/kg/h) and fentanyl (35 microg/kg/h). Anaesthesia was maintained for 300 min after surgery as the piglets were the control group of a project involving extracorporeal membrane oxygenation. Mean plasma cortisol concentration decreased significantly (P<0.05) from 59+/-39.9 nmol/L (mean+/-1 SD) before surgery to 7.5+/-2.5 nmol/L 300 min after end of surgical procedure. The mean SAA concentrations increased over the same period from 1.6+/-2.3 microg/mL to 4.2+/-5.6 microg/mL without statistical significance. The baseline (presurgery) mean arterial pressure (MAP) was 72+/-9 mmHg compared with 72+/-11 mmHg 300 min after end of surgery. Neither heart rate nor lactate concentrations changed significantly over the same time points: heart rate was 104+/-11 and 103+/-15 beats/min whereas mean lactate concentrations were reduced from 1.14+/-0.45 mmol/L to 0.90+/-0.22 mmol/L. Haemodynamic stability, a decrease in serum cortisol and a non-statistically significant rise in mean SAA concentrations suggest that the anaesthetic described suppresses the stress response of piglets to surgery without adverse cardiovascular effects. Therefore, it may prove useful in cardiovascular research.
Assuntos
Anestesia/veterinária , Anestésicos Intravenosos/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Fentanila/farmacologia , Frequência Cardíaca/efeitos dos fármacos , Propofol/farmacologia , Estresse Psicológico/induzido quimicamente , Adjuvantes Anestésicos/administração & dosagem , Amiloide/sangue , Analgésicos Opioides/administração & dosagem , Anestesia/métodos , Animais , Butorfanol/administração & dosagem , Combinação de Medicamentos , Oxigenação por Membrana Extracorpórea , Feminino , Hidrocortisona/sangue , Injeções Intramusculares , Ketamina/administração & dosagem , Midazolam/administração & dosagem , Complicações Pós-Operatórias/etiologia , Complicações Pós-Operatórias/veterinária , Estresse Psicológico/sangue , Cirurgia Veterinária/métodos , SuínosRESUMO
Hippocampal functions vary across the estrous cycle but metabolic changes at the protein level have not been systematically studied so far. It was therefore the aim of the study to screen expression of metabolic proteins mainly represented by metabolic enzymes in the hippocampus over the estrous cycle and in males. Female and male OFA Sprague-Dawley rats were used and female estrous phases were determined by vaginal smears, according to which females were separated into groups of proestrous, estrous, early and late metestrous and diestrous. Proteins were extracted from hippocampal tissue and separated on two-dimensional gel electrophoresis followed by identification with mass spectrometry methods (MALDI-TOF-TOF and nano-LC-ESI-MS/MS). Comparative analysis of protein levels was carried out by quantifying protein spot volumes by means of specific software. Levels of one expression form of gamma-enolase were different between diestrous and early metestrous; C-1-tetrahydrofolate synthase levels were elevated in proestrous as compared with estrous and serotransferrin levels were increased in diestrous as compared with proestrous, estrous, metestrous and in males. The outcome of estrous cycle- and gender-dependent protein fluctuations is relevant for the interpretation of previous and future work as well as for the design of further studies at the protein level in the hippocampus.
Assuntos
Aminoidrolases/metabolismo , Ciclo Estral/metabolismo , Ciclo Estral/fisiologia , Formiato-Tetra-Hidrofolato Ligase/metabolismo , Hipocampo/metabolismo , Hipocampo/fisiologia , Metilenotetra-Hidrofolato Desidrogenase (NADP)/metabolismo , Complexos Multienzimáticos/metabolismo , Fosfopiruvato Hidratase/metabolismo , Transferrina/metabolismo , Animais , Interpretação Estatística de Dados , Eletroforese em Gel Bidimensional , Feminino , Hidrólise , Masculino , Espectrometria de Massas , Proteômica , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Anterior gradient protein 2 homolog is a metastasis-inducing protein in a rat model of rat breast cancer and prognostic for outcome in hormonally treated breast cancer patients. Carrying out protein profiling in several mammalian cells and tissues, we detected this protein (synonym: secreted cement gland protein XAG-2 homolog) that was originally described in toad skin, in human bronchial epithelia. Tissues obtained from biopsies were homogenised and extracted proteins were run on two-dimensional gel electrophoresis. Following in-gel digestion with proteases trypsin, AspN, LysC and chymotrypsin, mass spectrometrical analysis was carried out by MALDI-TOF/TOF. The use of MS following multi-enzyme digestion of the protein resulted into 100% sequence coverage. MS/MS analysis enabled sequencing of 87% of the protein structure. This percentage does not include the signal peptide that was not observed in our protein due to processing. No posttranslational modifications were detectable and no sequence conflicts were observed. Complete analysis, unambiguous identification and characterisation of this biologically important protein could be shown, which is relevant for the definition of a marker protein that has been described so far by immunochemical methods only. Complete analysis is of importance as it forms the basis for all future work on this protein and, moreover, may serve as an analytical tool for further studies.
Assuntos
Biomarcadores Tumorais/análise , Proteínas/análise , Sequência de Aminoácidos , Asparagina/química , Quimotripsina/química , Eletroforese em Gel Bidimensional , Humanos , Lisina/química , Espectrometria de Massas/métodos , Mucoproteínas , Proteínas Oncogênicas , Sensibilidade e Especificidade , Análise de Sequência de Proteína , Homologia de Sequência de Aminoácidos , Tripsina/químicaRESUMO
Handling and detoxification of metals by enzymes is a major issue that is not in the focus of current biomedical research concepts. The finding of the presence of arsenic (+3 Oxidation State) methyltransferase (AS3MT) in neuroblastoma cells NE-115 as a high abundance protein made us investigate primary structure of AS3MT reflecting an example of metal-handling in eucaryotes. Proteins extracted from NE-115 cells were run on 2-DE followed by two different mass spectrometrical methods. High sequence coverage was obtained by multiple protease digestion and a sequence conflict was solved at arginine 335. These findings are important when future studies on this enzyme are designed at the protein level and in particular, when antibodies against this protein will be generated.
Assuntos
Arsenitos/química , Metiltransferases/química , Neuroblastoma/enzimologia , Animais , Cromatografia Líquida/métodos , Eletroforese em Gel Bidimensional , Espectrometria de Massas/métodos , Metiltransferases/isolamento & purificação , Camundongos , Células Tumorais CultivadasRESUMO
Hereditary fructose intolerance (HFI) is caused by a deficiency of aldolase B due to mutations of the ALDOB gene. The disease poses diagnostic problems because of unspecific clinical manifestations. We report three cases of HFI all of whom had a chronic disease with neurological, nephrological or gastroenterological symptoms, whereas nutritional fructose intolerance, the pathognomonic sign of HFI, was apparent only in retrospect. In all patients a hypoglycosylated pattern of transferrin isoforms was found but was misinterpreted as a sign of CDG Ix. The correct diagnosis was achieved with marked delay (26, 36 and 24 months, respectively) by sequencing of the ALDOB gene two common mutations were identified on both alleles or on one (A150P/A175D, A150P/-, and A150P/A175D). The diagnosis was further supported by normalization of transferrin isoforms on a fructose-free diet. Data available in two patients showed that following the fructose restriction the type I pattern of carbohydrate-deficient transferrin detectable on fructose-containing diet disappeared after 3-4 weeks. These cases illustrate that in the first years of life HFI may show misleading variability in clinical presentation and that protein glycosylation analysis such as transferrin isofocusing may give important diagnostic clues. However, care should be taken not to misinterpret the abnormal results as CDG Ix as well as to remember that a normal profile does not exclude HFI due to the possibility of spontaneous fructose restriction in the diet. The presented data also emphasize the usefulness of ALDOB mutation screening for diagnosis of HFI.
Assuntos
Intolerância à Frutose/diagnóstico , Intolerância à Frutose/genética , Frutose-Bifosfato Aldolase/genética , Transferrina/metabolismo , Diagnóstico Diferencial , Glicosilação , Humanos , MutaçãoRESUMO
OBJECTIVE: To report a case of recurrent tako-tsubo syndrome that developed despite treatment with calcium channel antagonists. CASE SUMMARY: A 76-year-old woman with past medical history of ischemic heart disease and mild chronic asthma presented in 2001 with clinical characteristics and laboratory markers consistent with myocardial ischemia. Coronary angiogram was done with successful balloon angioplasty to LAD stenosis. Ventriculogram and echocardiography demonstrated apical ballooning believed to represent aneurysm formation. Several months later, a follow-up echocardiogram (ECG) revealed normal LV size and function with no wall motion abnormalities. ECG was unremarkable. In 2004, the patient was admitted with dyspnea, chest pain and ST elevation in ECG with normal troponin. Coronary angiogram demonstrated patent coronary tree. Left ventriculogram revealed apical ballooning sparing the base of the heart. Medically controlling the asthma attack led to clinical, echocardiographic and remarkable electrocardiographic normalization within days. Rest thallium perfusion scan done within 48 h demonstrated isolated fully reversible defect in the apex after 24 h suggesting a microvessel etiology. CONCLUSION: Tako-tsubo cardiomyopathy is an increasingly recognized condition. We report here the first case of tako-tsubo recurrence despite treatment with verapamil, and suggest a microvessel pathophysiology supported by rest thallium scan.
Assuntos
Bloqueadores dos Canais de Cálcio/uso terapêutico , Cardiomiopatias/diagnóstico , Infarto do Miocárdio/diagnóstico , Disfunção Ventricular Esquerda/diagnóstico , Verapamil/uso terapêutico , Idoso , Cardiomiopatias/etiologia , Angiografia Coronária , Diagnóstico Diferencial , Eletrocardiografia , Feminino , Aneurisma Cardíaco/etiologia , Aneurisma Cardíaco/prevenção & controle , Sistema de Condução Cardíaco , Humanos , Infarto do Miocárdio/complicações , Isquemia Miocárdica/diagnóstico , Isquemia Miocárdica/tratamento farmacológico , RecidivaRESUMO
OBJECTIVE: Ultra-long gap (> 3.5 cm) esophageal atresias (ULGEA) are very rare congenital disorders usually treated by esophageal substitution. Since the introduction of Foker's elongation technique for primary anastomosis, there is much controversy over whether the Foker method or esophageal substitution results in a better outcome. Until now, there was only one series, which had been reported by Foker himself. We therefore present the outcome 2.5 years after one of the first children was operated on with the Foker method in Europe (2003). PATIENT AND METHOD: A premature child (1820 g, 33 + 4 gestational week) born with isolated ULGEA (6-cm gap length/30.5-cm body length) was treated with the Foker technique. We present a critical review of the course together with any complications that can help in the clinical evaluation of the new technique. RESULTS AND CONCLUSION: Three thoracotomies and 5 dilating procedures over a period of 3 months were necessary to achieve primary anastomosis. Two major complications occurred (anastomotic leak and subsequent stricture). The boy was under mechanical ventilation for a total of 15 days. The Foker method seems to be quite feasible and quickly results in the expected primary anastomosis. The result in this single case is excellent.
Assuntos
Procedimentos Cirúrgicos do Sistema Digestório/métodos , Atresia Esofágica/cirurgia , Esôfago/cirurgia , Técnicas de Sutura/instrumentação , Anastomose Cirúrgica/métodos , Atresia Esofágica/diagnóstico por imagem , Fluoroscopia , Seguimentos , Humanos , Recém-Nascido , Masculino , Radiografia TorácicaRESUMO
Previous studies have reported antitumor activity and reduced cardiotoxicity for a putative 3:1 complex of iron:Adriamycin (ADR). We have studied the tissue distribution and metabolism of a wide variety of freshly prepared and lyophilized iron:ADR preparations after administration to BALB/c mice (ADR, 16 mg/kg i.v.). Tissue concentrations of ADR given without iron were initially highest in kidney and liver, and ADR fluorescence was lost from all tissues except the spleen with a t1/2 of 15 to 18 hr. ADR remained the major fluorescent species in liver and kidney from 0.5 to 72 hr after treatment. Freshly prepared iron:ADR (1:1) behaved similarly to ADR except for a slightly longer tissue t1/2 in heart, liver, and kidney. The tissue distribution of freshly prepared 2:1 and 3:1 iron:ADR was very different from that of ADR without iron; lung containing the highest concentrations of ADR fluorescence. Administration of freshly prepared 1:1, 2:1, and 3:1 iron:ADR resulted in some increase in adriamycinol in the liver, but ADR was always the major fluorescent species present. The tissue distribution of 1:1 iron:ADR that had been aged for 48 or 96 hr was similar to that of fresh 2:1 and 3:1 iron:ADR rather than ADR or fresh 1:1 iron:ADR. When lyophilized iron:ADR preparations were reconstituted and administered, the 0.5-hr tissue distribution of 0.1:1, 0.2:1, 0.25:1, and 0.33:1 iron:ADR was the same as ADR alone, but 0.5:1, 1:1, 2:1, and 3:1 iron:ADR were all accumulated primarily in the lung. Physicochemical studies confirm the production of microaggregated iron:ADR complexes and light microscopy allows visualization and sizing of these aggregates. We feel that trapping of these iron:ADR aggregates in the pulmonary vascular bed accounts for the observed dramatic alteration in tissue distribution. Light and electron microscopic studies confirm the intravascular sequestration of iron in pulmonary capillaries.
Assuntos
Doxorrubicina/análogos & derivados , Animais , Doxorrubicina/análise , Doxorrubicina/metabolismo , Fluorescência , Histocitoquímica , Injeções Intravenosas , Ferro/análise , Pulmão/análise , Pulmão/irrigação sanguínea , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Coloração e Rotulagem , Fatores de Tempo , Distribuição TecidualRESUMO
To study the nature of virus-cell interaction in persistently infected cells we have examined production of infectious virus, synthesis of viral DNA and DNA polymerase in a human leukemic cell line K562. It was found that only one of three K562 cell lines was permissive for limited growth of HSV-2 and infectious virus was released in a cyclical fashion. Intranuclear inclusions with electron-dense fibrils and particles resembling viral structures were observed in the virus-infected but not control K562 cells. Viral DNA synthesis could not be detected by centrifugation in CsCl density gradients; but was readily identified by Southern blot hydridization of virus-infected intracellular DNA with purified viral DNA. Viral DNa polymerase was synthesized by infected cells during active infectious virus production. In one of the two K562 cell lines that did not produce infectious virus, a few DNA fragments from infected cells were found to hybridize with purified viral DNA. These results suggest that variable lengths of HSV-2 genome can be harbored and propagated by different human leukemic K562 cells.
Assuntos
Replicação do DNA , Genes Virais , Leucemia Mieloide Aguda/microbiologia , Simplexvirus/genética , Linhagem Celular , Transformação Celular Viral , DNA de Neoplasias/genética , DNA Viral/genética , DNA Polimerase Dirigida por DNA/metabolismo , Humanos , Cinética , Leucemia Mieloide Aguda/genética , Microscopia Eletrônica , Simplexvirus/ultraestrutura , Replicação ViralRESUMO
Six patients with M4 acute myelomonocytic leukemia ( AMMoL ) were identified who had abnormalities of chromosome 16 in bone marrow cells. Five had a pericentric inversion, inv(16)( p13q22 ), and a sixth patient had a translocation, t(16;16)(p13.1;q22). Each of these six patients had bone marrow eosinophils that were abnormal in morphology on light and/or electron microscopy and by cytochemical stains. The eosinophils constituted 1%-24% of nucleated marrow cells. Of 61 acute nonlymphocytic leukemia (ANLL) patients, all those with AMMoL and abnormal bone marrow eosinophils had an inv(16) or a t(16;16). One other patient in this group had a rearrangement of chromosome 16 (with a break in the short arm at band p13); however, the ANLL type was M1 and no abnormal eosinophils were present. Four patients with ANLL types other than M4 had an increase in marrow eosinophils; three in whom the eosinophils appeared normal and one with ANLL-M2 and bizarre eosinophils morphologically distinct from those seen in AMMoL . Chromosome pair 16 was normal in the latter four patients. AMMoL with dysplastic bone marrow eosinophils appears to represent a unique clinicopathologic entity associated with several related abnormalities affecting 16q . The morphologic features of both blasts and eosinophils may be more important than the absolute number of eosinophils in the marrow in identifying this group of patients. This may have prognostic importance as five of six patients achieved complete remission with standard antileukemic therapy and are still alive.
Assuntos
Medula Óssea/patologia , Aberrações Cromossômicas , Cromossomos Humanos 16-18 , Leucemia Mieloide Aguda/genética , Adulto , Medula Óssea/ultraestrutura , Bandeamento Cromossômico , Inversão Cromossômica , Eosinófilos/patologia , Eosinófilos/ultraestrutura , Feminino , Humanos , Cariotipagem , Leucemia Mieloide Aguda/patologia , Masculino , Pessoa de Meia-Idade , Translocação GenéticaRESUMO
OBJECTIVES: This study determined the effect of sotalol on atrial function after electrical cardioversion of atrial fibrillation. BACKGROUND: After electrical cardioversion of atrial fibrillation, the Doppler mitral A wave is often diminished, representing impaired atrial contractile function. Sotalol is an effective atrial antiarrhythmic drug with class III and beta-adrenergic blocking properties. Although the negative inotropic effect of sotalol on the ventricle is minimal in patients with normal ventricular function, it may manifest negative inotropy when ventricular function is impaired. We postulated that after cardioversion, when intrinsic atrial function is impaired, sotalol may have an adverse effect on the atrium. METHODS: Thirty-seven patients enrolled in a randomized, double-blind study of sotalol for maintenance of sinus rhythm were studied by quantitative Doppler echocardiography within 24 h of electrical cardioversion and, for those still in sinus rhythm, again at 1 month. Doppler variables (E and A wave velocities and integrals) in patients receiving sotalol were compared with those in patients receiving placebo. RESULTS: After electrical cardioversion, peak A wave velocity and A wave time-velocity integral in the 20 patients receiving placebo were reduced compared with normal values. In the 17 patients receiving sotalol (median dose 320 mg twice daily) these variables were further reduced (mean [+/- SD] peak A wave velocity 19.4 +/- 5.5 vs. 38.4 +/- 14.7 cm/s, p < 0.001 and mean A wave time-velocity integral 1.7 +/- 0.6 vs. 3.4 +/- 1.4 cm, p < 0.001, in sotalol- vs. placebo-treated patients, respectively). Early diastolic filling (E wave variables) did not differ between sotalol- and placebo-treated groups. At 1 month, five sotalol- and six placebo-treated patients remained in sinus rhythm, and A wave variables had increased for the whole group, with a greater increase in sotalol-treated patients. CONCLUSIONS: After electrical cardioversion, when atrial stunning is prominent, sotalol has a negative atrial inotropic effect. This effect may be temporary, as suggested by resolution at 1 month. Negative inotropic effects of antiarrhythmic drugs on the atrium should be considered in assessing Doppler variables of left ventricular filling.