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OBJECTIVES: To describe the change in the caudal vena cava to aorta ratio (CVC:Ao) ratio during fluid resuscitation of circulatory shock in dogs and compare these results with those of the physical examination and blood lactate. MATERIALS AND METHODS: Perfusion parameters and blood lactate were recorded at admission. An abdominal point-of-care ultrasound protocol was performed, during which the caudal vena cava to aorta ratio was measured on the spleno-renal view. Measurements were performed within 5 minutes before and after a 10 mL/kg crystalloid fluid bolus. Investigators were not blinded to therapeutic interventions. RESULTS: Twenty-nine dogs with physical signs of circulatory shock were enrolled. Caudal vena cava to aorta ratios were below reference interval in 28 of 29 dogs. After bolus administration, median caudal vena cava diameter increased by 0.14 cm (0.69 to 0.83 cm) and median aorta diameter increased by 0.03 cm (0.87 to 0.90 cm) and caudal vena cava to aorta ratio returned to within reference range in 65% of dogs (13/29). Bolus administration was associated with an increase in median caudal vena cava to aorta ratio of 0.10 (95% CI:0.05 to 0.16, P=0.0005). Blood lactate did not change significantly. Heart rate and capillary refill time decreased significantly after fluid bolus (heart rate: estimate=-19 bpm, 95% CI:-30 to -8, P=0.002; capillary refill time: estimate=-1.0 s, 95% CI:-1.3 to -0.7, P < 0.0001). CLINICAL SIGNIFICANCE: In this population of dogs with circulatory shock, the caudal vena cava to aorta ratio significantly increased after a fluid bolus. Future studies that implement blinding of the outcome assessors are warranted to confirm these findings.
Assuntos
Aorta , Hidratação , Cães , Animais , Aorta/diagnóstico por imagem , Hidratação/veterinária , Veia Cava Inferior/diagnóstico por imagem , Ultrassonografia/veterinária , LactatosRESUMO
PURPOSE: To report the results in a series of Peters Anomaly cases, and propose management and treatment approaches according to the alterations associated with each case. MATERIAL AND METHODS: A retrospective analysis was performed on the records of 27 patients (32 eyes) clinically diagnosed with Peters Anomaly. Each patient was subjected to different treatment modalities according to the type of Peters Anomaly, anywhere from medical follow-up clinics to a Penetrating Keratoplasty procedure (PKP). RESULTS: Of the 27 patients (32 eyes), 74% were male and 26% female, with 18.5% (5) being bilateral and 81.5% (22) unilateral. The mean number of years of follow-up was 10.2 years (Range: 3.5 to 18 years). The results of long-term VA correlate directly with the type of Peters Anomaly. For the total number of patients, the VA results were LogMAR 1.71⯱â¯1.04. The results by groups were: Type I with only medical monitoring LogMAR 0.3⯱â¯0, Type I with only Optical Iridectomy (OI) LogMAR 0.97⯱â¯0.78, Type I with PKP LogMAR 1.22⯱â¯0.97, Type II without a compromised posterior pole with PKP LogMAR 2.41⯱â¯0.80, and Type II with a compromised posterior pole with PKP LogMAR 2.56⯱â¯0.48. CONCLUSIONS: The result of VA and long-term corneal failure is directly related to the type of Peters Anomaly. Patients with Type I who only required medical follow-ups had the most favourable prognosis. Patients who underwent Peripheral Iridectomy followed and patients in which PKP was performed had an inferior prognosis.
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Opacidade da Córnea , Segmento Anterior do Olho/anormalidades , Anormalidades do Olho , Feminino , Seguimentos , Humanos , Masculino , Estudos Retrospectivos , Acuidade VisualRESUMO
ObjetivoComunicar los resultados de una serie de casos con anomalía de Peters, y proponer el manejo y sugerencias terapéuticas según alteraciones asociadas.Material y métodosSe analizaron las historias clínicas de 27 pacientes (32 ojos) con diagnóstico clínico de anomalía de Peters, los cuales fueron sometidos a diferentes tratamientos. Distintos parámetros y condiciones asociadas fueron evaluadas: presión intraocular (PIO), agudeza visual (AV), técnica quirúrgica y complicaciones postoperatorias.ResultadosDe los 27 pacientes (32 ojos), 74% sexo masculino y 26% femenino. El promedio de años de seguimiento fue de 10,2 años. La PIO preoperatoria fue de 23 ± 9,21 mmHg y al último control fue de 18,81 ± 7,45 mmHg. El resultado de AV a largo plazo se correlaciona directamente con el tipo de anomalía de Peters. Para el total los resultados de AV fueron LogMAR 1,71 ± 1,04 y por grupos: Tipo I sólo con seguimiento médico LogMAR 0,3 ± 0, Tipo I sólo con iridectomía periférica LogMAR 0,97 ± 0,78, Tipo I con queratoplastia penetrante (QPP) LogMAR 1,22 ± 0,97, Tipo II sin compromiso de polo posterior LogMAR 2,41 ± 0,80 y Tipo II con compromiso de polo posterior LogMAR 2,56 ± 0,48.ConclusionesEl resultado de AV y fracaso del injerto corneal a largo plazo se correlaciona directamente al tipo de anomalía de Peters, con mejor pronóstico la Tipo I en que se realizó sólo seguimiento médico, luego en los que se practicó iridectomía periférica, en los que se realizó QPP los que presentan peor pronóstico visual son con anomalía de Peters Tipo II con compromiso de polo posterior (p = 0.0087).
PurposeTo report the results in a series of Peters Anomaly cases, and propose management and treatment approaches according to the alterations associated with each case.Material and methodsA retrospective analysis was performed on the records of 27 patients (32 eyes) clinically diagnosed with Peters Anomaly. Each patient was subjected to different treatment modalities according to the type of Peters Anomaly, anywhere from medical follow-up clinics to a Penetrating Keratoplasty procedure (PKP).ResultsOf the 27 patients (32 eyes), 74% were male and 26% female, with 18.5% (5) being bilateral and 81.5% (22) unilateral. The mean number of years of follow-up was 10.2 years (Range: 3.5 to 18 years). The results of long-term VA correlate directly with the type of Peters Anomaly. For the total number of patients, the VA results were LogMAR 1.71 ± 1.04. The results by groups were: Type I with only medical monitoring LogMAR 0.3 ± 0, Type I with only Optical Iridectomy (OI) LogMAR 0.97 ± 0.78, Type I with PKP LogMAR 1.22 ± 0.97, Type II without a compromised posterior pole with PKP LogMAR 2.41 ± 0.80, and Type II with a compromised posterior pole with PKP LogMAR 2.56 ± 0.48.ConclusionsThe result of VA and long-term corneal failure is directly related to the type of Peters Anomaly. Patients with Type I who only required medical follow-ups had the most favourable prognosis. Patients who underwent Peripheral Iridectomy followed and patients in which PKP was performed had an inferior prognosis.
Assuntos
Humanos , Ciências da Saúde , Oftalmologia , Anormalidades do Olho , Iridectomia , Ceratoplastia Penetrante , Opacidade da CórneaRESUMO
OBJECTIVES: To prospectively describe the impact of gas flow rate and temperature on dog's tolerance of high-flow nasal oxygen therapy during recovery from anaesthesia, hypothesizing that higher flow rates and temperatures will decrease tolerance. MATERIALS AND METHODS: Twelve non-dyspnoeic client-owned dogs recovering from general anaesthesia were included in this study. After extubation, a nasal cannula was positioned and high-flow nasal oxygen therapy was initiated. Two flow rates (two or four time the theoretical minute ventilation: HF2 and HF4), each of them combined with two temperatures (31 and 37°C: T31 and T37), were randomly applied (four conditions per dog). For each condition, cardiovascular and respiratory parameters (heart rate, respiratory rate, systolic arterial blood pressure and pulse oximeter oxygen saturation), sedation score and tolerance score were recorded at initiation (T0 ) and after 10 minutes of accommodation (T10 ). RESULTS: Sedation scores were not significantly different between the four conditions. Cardiovascular and respiratory parameters were not significantly different between any condition at both T0 and T10 . Tolerance scores were good and not significantly different between any flow rate or temperature (HF2-T31: 4 (2-4), HF4-T31: 4 (2-4), HF2-T37: 4 (2-4), HF4-T37: 4 (1-4)). CLINICAL SIGNIFICANCE: The gas flow rates and temperatures studied have no impact on tolerance during the recovery period of non-dyspnoeic dogs, and high-flow nasal cannula is well tolerated. Further studies are required to confirm these results in dyspnoeic dogs.
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Doenças do Cão , Oxigenoterapia , Animais , Cânula , Doenças do Cão/terapia , Cães , Dispneia/veterinária , Oxigênio , Oxigenoterapia/veterinária , TemperaturaRESUMO
BACKGROUND: Leptospirosis in dogs is occasionally associated with a hemorrhagic syndrome, the pathophysiology of which is not fully understood. HYPOTHESIS/OBJECTIVES: To characterize hematologic, hemostatic, and thromboelastometric abnormalities in dogs with leptospirosis and to study their association with hemorrhagic diatheses and outcomes. ANIMALS: Thirty-five client-owned dogs. METHODS: A prospective observational single cohort study was conducted. Results from the CBC, coagulation tests (prothrombin, activated partial thromboplastin and thrombin times, fibrinogen, fibrin(ogen) degradation products, and D-dimer concentrations), rotational thromboelastometry (TEM), signalment, hemorrhagic diatheses, occurrence of disseminated intravascular coagulation (DIC) at admission, and survival to discharge were recorded. RESULTS: The most common hematologic and hemostatic abnormalities were anemia (30/35), thrombocytopenia (21/35), and hyperfibrinogenemia (15/35). Eight dogs were diagnosed with DIC. A normal TEM profile was found in 14 dogs, a hypercoagulable profile in 14 dogs, and a hypocoagulable profile in 7 dogs. The 8 dogs with hemorrhagic diatheses at admission had significantly decreased platelet counts (P = .037) and increased D-dimer concentrations (P = .015) compared with other dogs. Dogs with a hypocoagulable profile exhibited more hemorrhagic diatheses compared with the dogs that had normal and hypercoagulable profiles (P = .049). The mortality rate was lower in dogs with a hypercoagulable profile than in those with a hypocoagulable profile (21% vs 57%; P = .043). Disseminated intravascular coagulation was not a significant prognostic factor. CONCLUSIONS AND CLINICAL IMPORTANCE: Thromboelastometric parameters were altered in dogs with both hypercoagulable and hypocoagulable profiles. A hypocoagulable profile was significantly correlated with hemorrhagic diathesis and higher mortality rate.
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Coagulação Intravascular Disseminada/veterinária , Doenças do Cão/mortalidade , Leptospirose/veterinária , Animais , Estudos de Coortes , Coagulação Intravascular Disseminada/sangue , Coagulação Intravascular Disseminada/complicações , Coagulação Intravascular Disseminada/mortalidade , Doenças do Cão/sangue , Cães , Feminino , Produtos de Degradação da Fibrina e do Fibrinogênio/metabolismo , França , Hemostasia , Leptospirose/sangue , Leptospirose/complicações , Leptospirose/mortalidade , Masculino , Tempo de Tromboplastina Parcial/veterinária , Estudos Prospectivos , Análise de SobrevidaRESUMO
In human medicine, age is a risk factor for thromboembolic diseases associated with hypercoagulable and antifibrinolytic states, but information in veterinary medicine is limited. This study compared the thromboelastometric (TEM) profiles of two groups of dogs of distinct ages. Ten healthy old (>10 years) Beagles and 10 healthy young (<3 years) Beagles were recruited. White blood cell counts and haematocrit were significantly lower in the old group compared to the young group, and fibrinogen, total proteins, globulins and monocyte chemoattractant protein-1 plasma concentrations were significantly higher in the old group. Comparisons of the TEM profiles indicated a hypercoagulable profile and a decrease in fibrinolytic activity in all old Beagles. The findings support the need to consider age as a possible risk factor for thrombosis in dogs.
Assuntos
Envelhecimento , Coagulação Sanguínea , Citocinas/sangue , Tromboelastografia/veterinária , Trombose/veterinária , Animais , Cães , Feminino , Masculino , Fatores de Risco , Trombose/etiologiaRESUMO
Reactive oxygen species fulfill key roles in development and signaling, but lead at high concentration to damage in macromolecules. In proteins, methionine (Met) is particularly prone to oxidative modification and can be oxidized into Met sulfoxide (MetO). MetO reduction is catalyzed by specialized enzymes, termed methionine sulfoxide reductases (MSRs), involved in senescence and protection against diseases and environmental constraints. The precise physiological functions of MSRs remain often elusive because of very poor knowledge of their substrates. In this study, affinity chromatography was used to isolate partners of Arabidopsis thaliana plastidial methionine sulfoxide reductase B1 (MSRB1). Twenty-four proteins involved in photosynthesis, translation, and protection against oxidative stress, as well as in metabolism of sugars and amino acids, were identified. Statistical analysis shows that the abundance of MSRB1 partners in chromatography affinity samples is proportional to Met content. All proteins, for which structural modeling was feasible, display surface-exposed Met and are thus potentially susceptible to oxidation. Biochemical analyses demonstrated that H(2)O(2) treatment actually converts several MSRB1-interacting proteins into MSRB substrates. In consequence, we propose that affinity chromatography constitutes an efficient tool to isolate physiological targets of MSRs.
Assuntos
Proteínas de Arabidopsis/isolamento & purificação , Proteínas de Arabidopsis/metabolismo , Cromatografia de Afinidade/métodos , Metionina Sulfóxido Redutases/metabolismo , Arabidopsis/metabolismo , Proteínas de Cloroplastos/metabolismo , Oxirredução , Fator Tu de Elongação de Peptídeos/metabolismo , Ligação ProteicaRESUMO
Cytogenetic stratification remains insufficient for almost half of the acute myeloblastic leukemia (AML) cases, with AML patients requiring subsequent molecular investigation. In our study, we used mass spectrometry (MS)-based proteomic approaches to characterize de novo AML. Fifty-four samples (mononuclear cells from bone marrow or peripheral blood mononuclear cells collected and frozen before treatment) from two independent cohorts of newly diagnosed AML patients were analyzed. We showed that the protein signature of leukemic cells defined two clusters that displayed significant variation for overall and disease-free survival (P=0.001 and 0.0004, respectively). This proteomic classification refines the cytogenetic classes. AML patients with intermediate and unfavorable cytogenetic classifications could be subdivided according to their protein profiles into subgroups with significantly different survival rates. Among the proteins expressed by leukemic cells, we isolated a 10,800-Da marker that retained the highest discriminative value between living and deceased patients. The 10,800-Da marker was identified by MS peptide sequencing as S100A8 (also designated MRP8 or calgranulin A). Western blot analysis confirmed its expression mainly in AML patients with the worst prognosis, arguing for a selective deregulation associated with poor prognosis. These results suggest that the expression of S100A8 in leukemic cells is a predictor of low survival.
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Calgranulina A/sangue , Leucemia Mieloide Aguda/mortalidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/sangue , Western Blotting , Proteínas Estimuladoras de Ligação a CCAAT/genética , Feminino , Humanos , Leucemia Mieloide Aguda/sangue , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteômica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
Research of new diagnosis or prognosis biomarkers is a major challenge for the management of patients with complex pathologies like cancer. Clinical proteomics is one of the recent approaches to identify these biomarkers in biological fluids. Over the last five years, many problems related to the variability and the quality control of these analyses have been observed. This was notably related to the different preanalytical status of each sample. A strong need for standardization of the critical preanalytical phases (collection, transport, processing, storage...) has been therefore recognized. With this goal in mind, working groups of the "Institut national du cancer" (INCa) and the "Société française de biologie clinique" (SFBC) proposed here preanalytical proteomics guidelines for the most common biological fluids: plasma, serum, urine and cerebrospinal fluid. To goal is to provide the basis for the harmonization of the procedures in clinical laboratories and biobanks to allow an optimal use of biological collections.
Assuntos
Líquidos Corporais/fisiologia , Técnicas de Laboratório Clínico/normas , Técnicas e Procedimentos Diagnósticos/normas , Guias de Prática Clínica como Assunto , Proteômica/métodos , Análise Química do Sangue/normas , Humanos , Prognóstico , Proteinúria/diagnóstico , Proteômica/normas , Urina/químicaRESUMO
OBJECTIVES: The use of biologicals such as infliximab has dramatically improved the treatment of rheumatoid arthritis (RA). However, factors predictive of therapeutic response need to be identified. A proteomic study was performed prior to infliximab therapy to identify a panel of candidate protein biomarkers of RA predictive of treatment response. METHODS: Plasma profiles of 60 patients with RA (28 non-responders (as defined by the American College of Rheumatology 20% improvement criteria (ACR20)) negative and 32 responders (ACR70 positive) to infliximab) were studied by surface enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF MS) technology on two types of arrays, an anion exchange array (SAX2) and a nickel affinity array (IMAC3-Ni). Biomarker characterisation was carried out using classical biochemical methods (purification by ammonium sulfate precipitation or metal affinity chromatography) and identification by matrix assisted laser desorption/ionisation time-of-flight (MALDI-TOF) MS analysis. RESULTS: Two distinct protein profiles were observed on both arrays and several proteins were differentially expressed in both patient populations. Five proteins at 3.86, 7.77, 7.97, 8.14 and 74.07 kDa were overexpressed in the non-responder group, whereas one at 28 kDa was increased in the responder population (sensitivity>56%, specificity>77.5%). Moreover, combination of several biomarkers improved the sensitivity and specificity of the detection of patient response to over 97%. The 28 kDa protein was characterised as apolipoprotein A-I and the 7.77 kDa biomarker was identified as platelet factor 4. CONCLUSIONS: Six plasma biomarkers are characterised, enabling the detection of patient response to infliximab with high sensitivity and specificity. Apolipoprotein A-1 was predictive of a good response to infliximab, whereas platelet factor 4 was associated with non-responders.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Antirreumáticos/uso terapêutico , Apolipoproteína A-I/sangue , Artrite Reumatoide/tratamento farmacológico , Fator Plaquetário 4/sangue , Adulto , Idoso , Artrite Reumatoide/sangue , Biomarcadores/sangue , Monitoramento de Medicamentos/métodos , Feminino , Humanos , Infliximab , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteômica/métodos , Sensibilidade e Especificidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Resultado do TratamentoRESUMO
Viruses require viral and cellular chaperones during their life cycle and interactions of these molecules with the immune system are probable during the infection. Thus, an anti-chaperone antibody response has been firstly investigated in hepatitis C patients in this paper. A HepG2-lysate antigen (90, 79, 72, 70, 62, 54 and 48 kDa) was assayed in sera from 59 (19F/40M) chronic hepatitis C patients without cirrhosis before therapy. Forty of them were positive for anti-HepG2 lysate antigen antibodies and this test may evaluate biological autoimmunity. Hsp70.1, Hsp90 and calreticulin levels were significantly higher in this antigen than in a control HepG2 antigen. Secondly, Hsp70.1 was identified as Hsp 70 kDa protein-1 by proteomic analysis and studied as a possible antibody target. Fourteen out of 59 patients were positive for anti-Hsp70.1 antibodies that were inversely correlated with alanine aminotransferase levels, the Metavir activity index and viraemia. Finally, for comparative purposes, 50 sera from systemic lupus erythematosus (SLE) patients have been tested: eight and 41 of them were positive for anti-Hsp70.1 and anti-HepG2 lysate antigen antibodies, respectively. Therefore, anti-Hsp70.1 autoantibodies may be produced and can partially lead to biological autoimmunity in chronic hepatitis C patients.
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Autoanticorpos/imunologia , Autoimunidade , Transportador de Glucose Tipo 1/imunologia , Proteínas de Choque Térmico HSP70/imunologia , Hepatite C Crônica/imunologia , Adolescente , Adulto , Idoso , Autoanticorpos/sangue , Linhagem Celular , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Chaperonas Moleculares/imunologia , Curva ROC , Sensibilidade e Especificidade , Estatísticas não ParamétricasRESUMO
We present a case, pathologically proven, of a patient with multiple papillary renal cell carcinoma (PRCC) with bilateral and synchronous affectation. CT showed fatty tissue inside one of the lesions and numerous calcified lesions. The study with MR demonstrated multiple and hypointense lesions in T2 and contrast enhancement in T1. Our observations confirm that the presence of multiple lesions with fat and calcified deposits and poor contrast enhancement should be diagnosed as PRCC, rather than renal clear cell carcinoma (RCCC) or renal angiomyolipoma.
Assuntos
Carcinoma de Células Renais/diagnóstico , Neoplasias Renais/diagnóstico , Tecido Adiposo/diagnóstico por imagem , Angiomiolipoma/diagnóstico , Diagnóstico Diferencial , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Tomografia Computadorizada por Raios XRESUMO
Tissue microarray technology (TMA) is nowadays considered as a powerful tool for the high-throughput analysis of molecular expression pattern of cancer. In this manuscript we show the experience of both groups in the design and building of a TMA for the study of protein expression pattern of prostatecancer as well as a summary of the technical points to analyze the results obtained with this technology. Today, different data generated by the immunostained tissues are studied to achieve a molecular profile in different clinical scenarios.
Assuntos
Imuno-Histoquímica/métodos , Análise em Microsséries/instrumentação , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Desenho de Equipamento , Humanos , MasculinoRESUMO
The incorporation and localisation of 133Cs in a plant cellular model and the metabolic response induced were analysed as a function of external K concentration using a multidisciplinary approach. Sucrose-fed photosynthetic Arabidopsis thaliana suspension cells, grown in a K-containing or K-depleted medium, were submitted to a 1 mM Cs stress. Cell growth, strongly diminished in absence of K, was not influenced by Cs. In contrast, the chlorophyll content, affected by a Cs stress superposed to K depletion, did not vary under the sole K depletion. The uptake of Cs was monitored in vivo using 133Cs NMR spectroscopy while the final K and Cs concentrations were determined using atomic absorption spectrometry. Cs absorption rate and final concentration increased in a K-depleted external medium; in vivo NMR revealed that intracellular Cs was distributed in two kinds of compartment. Synchrotron X-ray fluorescence microscopy indicated that one could be the chloroplasts. In parallel, the cellular response to the Cs stress was analysed using proteomic and metabolic profiling. Proteins up- and down-regulated in response to Cs, in presence of K+ or not, were analysed by 2D gel electrophoresis and identified by mass spectrometry. No salient feature was detected excepting the overexpression of antioxidant enzymes, a common response of Arabidopsis cells stressed whether by Cs or by K-depletion. 13C and 31P NMR analysis of acid extracts showed that the metabolome impact of the Cs stress was also a function of the K nutrition. These analyses suggested that sugar metabolism and glycolytic fluxes were affected in a way depending upon the medium content in K+. Metabolic flux measurements using 13C labelling would be an elegant way to pursue on this line. Using our experimental system, a progressively stronger Cs stress might point out other specific responses elicited by Cs.
Assuntos
Arabidopsis/metabolismo , Radioisótopos de Césio/toxicidade , Césio/toxicidade , Potássio/farmacologia , Proteoma , Arabidopsis/efeitos dos fármacos , Arabidopsis/crescimento & desenvolvimento , Divisão Celular/efeitos dos fármacos , Césio/farmacocinética , Radioisótopos de Césio/farmacocinética , Clorofila/metabolismo , Cinética , Espectroscopia de Ressonância MagnéticaRESUMO
Accumulating evidence shows that several cell types have the capacity to secrete membrane proteins by incorporating them into exosomes, which are small lipid vesicles derived from the intralumenal membranes of multivesicular bodies (MVBs) of the endocytic pathway. Exosomes are expelled in the extracellular space upon fusion of the MVB with the plasma membrane. Exosomal release is a way of secreting membrane proteins meant to be discarded, or to be passed on to other cells. Here, we demonstrate, using primary cortical cultures, that neurones and astrocytes can secrete exosomes. We find that exosomes released by cortical neurones contain the L1 cell adhesion molecule, the GPI-anchored prion protein, and the GluR2/3 but not the NR1 subunits of glutamate receptors. We also show that exosomal release is regulated by depolarisation. Our observation suggests that exosomes may have a regulatory function at synapses and could also allow intercellular exchange of membrane proteins within the brain.
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Córtex Cerebral/citologia , Vesículas Citoplasmáticas/metabolismo , Exocitose/fisiologia , Neurônios/metabolismo , Animais , Biomarcadores/metabolismo , Proteínas de Ligação ao Cálcio/metabolismo , Proteínas de Transporte/metabolismo , Células Cultivadas , Vesículas Citoplasmáticas/química , Vesículas Citoplasmáticas/ultraestrutura , Proteínas Luminescentes/metabolismo , Potenciais da Membrana/fisiologia , Camundongos , Camundongos Transgênicos , Neurônios/citologia , RatosRESUMO
El microarray tisular (TMA) es considerado hoy en día una potente herramienta para el análisis masivo del perfil molecular del cáncer. En este trabajo describimos la experiencia de ambos centros en el diseño y creación de un TMA para el estudio de la expresión molecular del cáncer de próstata así como una revisión de los diferentes aspectos técnicos necesarios para el análisis de los resultados obtenidos con esta técnica. En la actualidad, se están estudiando los datos generados por las distintas técnicas inmunohistoquímicas para la obtención de un patrón molecular en diferentes estadios clínicos
Tissue microarray technology (TMA) is nowadays considered as a powerful tool for the high-throughput analysis of molecular expression pattern of cancer. In this manuscript we show the experience of both groups in the design and building of a TMA for the study of protein expression pattern of prostate cancer as well as a summary of the technical points to analyze the results obtained with this technology. Today, different data generated by the immunostained tissues are studied to achieve a molecular profile in different clinical scenarios
Assuntos
Masculino , Humanos , Biomarcadores Tumorais/análise , Imuno-Histoquímica/métodos , Neoplasias da Próstata/diagnóstico , Hematoxilina , Amarelo de Eosina-(YS)RESUMO
Development proceeds through successive activation of different sets of genes by specific transcription factors as a consequence of cell interactions and signaling. It is thus of primary interest to identify new putative transcriptional regulators. We report here the isolation of chicken clones bearing sequences coding for a chicken zinc finger protein (chZFp) which contains four pairs of zinc fingers of mixed type C2-H-C/C2-H2. At least five chZFp isoforms are produced through differential splicing of four small exons. The amino acid domains encoded by these four exons are highly conserved across species. Northern blot analysis and RNase-protection assays showed that chZFp transcripts are present in brain, heart, skin and liver during chick development. Reverse transcription mediated polymerase chain reaction (RT-PCR) experiments suggested that the relative amount of some chZFp isoforms increases at critical stages of development and skin morphogenesis. Finally, the main chZFp isoforms are able to directly interact in vitro with the scaffold attachment factor-A (SAF-A, also known as heterogenous nuclear ribonucleoprotein U) through both their aminoterminal and carboxyterminal domains.
Assuntos
DNA Complementar/genética , Isoformas de Proteínas/biossíntese , Elementos Reguladores de Transcrição/genética , Dedos de Zinco/genética , Processamento Alternativo/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Encéfalo/embriologia , Núcleo Celular/metabolismo , Embrião de Galinha , Clonagem Molecular , Regulação da Expressão Gênica no Desenvolvimento , Coração/embriologia , Ribonucleoproteínas Nucleares Heterogêneas Grupo U/metabolismo , Fígado/embriologia , Camundongos , Dados de Sequência Molecular , Células NIH 3T3 , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Pele/embriologiaRESUMO
OBJECTIVE: To detect the prevalence of uropathogens in community-acquired urinary tract infection in our environment, and the degree of sensitivity to antibiotics used as empirical treatment. PATIENTS AND METHOD: Retrospective longitudinal study on 16,392 consecutive urine cultures collected in the emergency department of Hospital del Mar, between January 1997 and December 2001. Resistance rates were compared through variance analysis. RESULTS: 8,743 urine cultures with significant count were obtained. 6,062 Escherichia coli (69.3%), 517 Proteus mirabilis (5.9%) and 390 Klebsiella pneumoniae (4.5%) were identified. Escherichia coli showed progressive growth rate and significant resistances to most of antibiotics evaluated, especially to quinolones which came close to 30%. Fosfomycin showed the least resistance rate (0.9%) and remained stable along the years studied. CONCLUSIONS: These results suggest that higher rate of resistance to quinolones does not advise its use as empirical in community-acquired urinary tract infection treatment in our environment. According to our experience, fosfomycin can be an excellent option for cystitis treatment in patients without risk factors, while for the treatment of parenchymatous urinary tract infection, complicated urinary tract infections, and urinary tract infections associated to risk factors, preference could be second or third generation oral cephalosporins, or amoxicillin-clavulanic acid.