RESUMEN
OBJECTIVE. To evaluate the prevalence of dysmenorrhoea, its impact, and management approaches in Hong Kong university students, and to compare between medical and non-medical students for any potential differences in coping strategies. DESIGN. Cross-sectional questionnaire survey. SETTING. The University of Hong Kong, Hong Kong. PARTICIPANTS. A total of 240 undergraduate (128 medical and 112 non-medical) students. MAIN OUTCOME MEASURES. Data on the presence and severity of dysmenorrhoea, its impact on daily life, management approaches, specific strategies, and their self-perceived effectiveness were obtained and analysed. RESULTS. In these subjects, the prevalence of dysmenorrhoea was 80% (95% confidence interval, 75-85%) with a mean (standard deviation) pain score of 5.0 (1.7). The most common impacts on daily life included reduced ability to concentrate and/or disturbance with study (75%) and changes in normal physical activity (60%). Only 6% sought medical advice, while 70% practised self-management. Pain scores and pain affecting normal physical activities were important predictive factors for self-management and for management based on pharmacological or non-pharmacological means. The commonest specific strategies used were a warm beverage (62%), paracetamol (57%), and sleeping (45%), while the most effective strategies were non-steroidal anti-inflammatory drugs (100%), traditional Chinese medicine (93%), and dietary/nutritional supplements (92%). Regarding the comparison of medical and non-medical students, the former used fewer pharmacological strategies among the various management approaches investigated. CONCLUSION. With data showing dysmenorrhoea as a very common condition having a significant impact in the Hong Kong community, primary care doctors should reassure young women with dysmenorrhoea that it is a common experience in the same age-group. Health education on the existence of effective treatment from medical practitioners could help women whose dysmenorrhoea was not controlled by self-management.
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Dismenorrea/epidemiología , Educación en Salud , Autocuidado/métodos , Estudiantes/estadística & datos numéricos , Estudios Transversales , Dismenorrea/terapia , Femenino , Hong Kong/epidemiología , Humanos , Prevalencia , Índice de Severidad de la Enfermedad , Estudiantes de Medicina/estadística & datos numéricos , Encuestas y Cuestionarios , Universidades , Adulto JovenRESUMEN
BACKGROUND: Retinoic acid-regulated nuclear matrix-associated protein (RAMP) is a WD40 repeat-containing protein that is involved in various biological functions, but little is known about its role in human cancer. This study aims to delineate the oncogenic role of RAMP in gastric carcinogenesis. METHODS: RAMP expression was examined by real-time quantitative RT-PCR, immunohistochemistry and western blotting. Inhibition of RAMP expression was performed by siRNA-mediated knockdown. The functional effects of RAMP on cell kinetics were measured by cell viability assay, colony formation assay and flow cytometry. Cell lines stably expressing RAMP were established to investigate the oncogenic effects of RAMP in vitro. RESULTS: Ramp was readily expressed in all seven gastric cancer cell lines and was significantly increased in human gastric cancer tissues when compared with their adjacent non-cancerous tissues (P<0.001). In keeping with this, expression of RAMP protein was higher in gastric cancer tissues compared with their adjacent non-cancerous tissues, whereas moderate protein expression were noted in intestinal metaplasia. Knockdown of RAMP in gastric cancer cells significantly reduced cell proliferation (P<0.01) and soft agar colony formation (P<0.001), but induced apoptosis and G(2)/M arrest. In additional, knockdown RAMP induced cell apoptosis is dependent on functional accumulation of p53 and p21 and induction of cleaved caspases-9, caspases-3 and PARP. Strikingly, overexpression of RAMP promoted anchorage-independent cell growth in soft agar. CONCLUSION: Our findings demonstrate that RAMP plays an oncogenic role in gastric carcinogenesis. Inhibition of RAMP may be a promising approach for gastric cancer therapy.
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Biomarcadores de Tumor/análisis , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/metabolismo , Apoptosis/fisiología , Western Blotting , Línea Celular Tumoral , Citometría de Flujo , Expresión Génica , Humanos , Inmunohistoquímica , ARN Mensajero/análisis , ARN Interferente Pequeño , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Matrices Tisulares , Ubiquitina-Proteína LigasasRESUMEN
Treatment of human immunodeficiency virus type 1 with protease inhibitors (PIs) is associated with the emergence of resistance-associated mutations. Treatment-characterized datasets have been used to identify novel treatment-associated protease mutations. In this study, we utilized two large reference laboratory databases (>115,000 viral sequences) to identify non-established resistance-associated protease mutations. We found 20 non-established protease mutations occurring in 82% of viruses with a PI resistance score of 4-7, 62% of viruses with a resistance score of 1-3, and 35% of viruses with no predicted PI resistance. We correlated mutational prevalence to treatment duration in a treatment-characterized dataset of 2161 patients undergoing non-suppressive PI therapy. In the non-suppressed dataset, 24 mutations became more prevalent and three mutations became less prevalent after more than 48 months of non-suppressive PI-therapy. Longer durations of non-suppressive treatment correlated with higher PI resistance scores. Mutations at eight non-established positions that were more common in viruses with the longest duration of non-suppressive therapy were also more common in viruses with the highest PI resistance score. Covariation analysis of 3036 protease amino acid substitutions identified 75 positive and nine negative correlations between resistance associated positions. Our findings support the utility of reference laboratory datasets for surveillance of mutation prevalence and covariation.
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Infecciones por VIH/tratamiento farmacológico , Inhibidores de la Proteasa del VIH/farmacología , Proteasa del VIH/genética , VIH-1/enzimología , VIH-1/genética , Secuencia de Aminoácidos , Bases de Datos como Asunto , Farmacorresistencia Viral/genética , Infecciones por VIH/virología , VIH-1/fisiología , Humanos , Mutación Puntual/genética , Prevalencia , ARN Viral/química , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Estadísticas no Paramétricas , Replicación Viral/efectos de los fármacosRESUMEN
The incidence of bacteremia related to transesophageal echocardiography was studied in 140 consecutive patients (71 women and 69 men with a mean age of 53.7 +/- 15 years). Thirty-four patients had one or more prosthetic heart valves. Blood cultures were obtained from each patient through separate venipuncture sites immediately before and after transesophageal echocardiography. An additional late blood culture was obtained in 114 patients 1 h later. The skin was cleaned with povidone-iodine and venipunctures were performed with separate butterfly needles with use of sterile gloves and drapes. Blood samples were drawn into separate syringes, transferred to aerobic and anaerobic culture bottles and processed with use of a semiautomated system. The overall incidence of blood cultures positive for bacteremia was 2% (8 of 394 bottles) and all positive cultures grew in a single blood culture bottle. Positive cultures occurred in 4 (1.4%) of 280 bottles before the procedure, in 2 (0.7%) of 280 bottles immediately after the procedure and in 2 (0.9%) of 228 late (1-h) blood culture bottles. Bacterial isolates were coagulase-negative staphylococci (n = 5), Propionibacterium (n = 2) and Moraxella (n = 1). All were considered contaminants. Mean endoscopic time in these patients was not significantly different from that in the other patients. Follow-up of patients with a blood culture positive for bacteremia revealed no clinical evidence of systemic infection. It is concluded that 1) the incidence of bacteremia related to transesophageal echocardiography is very low, and 2) the incidence of blood cultures positive for bacteremia after transesophageal echocardiography is indistinguishable from the anticipated contamination rate.
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Bacteriemia/epidemiología , Ecocardiografía/efectos adversos , Esófago/diagnóstico por imagen , Adulto , Anciano , Anciano de 80 o más Años , Bacteriemia/sangre , Bacteriemia/microbiología , Recolección de Muestras de Sangre/métodos , Recolección de Muestras de Sangre/normas , Ecocardiografía/métodos , Reacciones Falso Positivas , Femenino , Prótesis Valvulares Cardíacas , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Ontario/epidemiología , Faringe/microbiología , Estudios Prospectivos , Factores de Riesgo , Factores de TiempoRESUMEN
OBJECTIVES: Angiotensin II (AngII) generation in response to vascular injury has long been assumed to influence neointimal proliferation contributing to restenosis. This concept has been supported by evidence that ACE inhibition and AT1 receptor blockade effectively limits restenosis in the rat. On the other hand, ACE inhibition has proven ineffective in clinical trails. The present study examines the response of the porcine coronary artery after angioplasty in vitro and compares the actions of an ACE inhibitor to AngII receptor antagonists. METHODS AND RESULTS: Captopril, an ACE inhibitor, and the AngII receptor antagonists, losartan and PD123319, were evaluated for their ability to attenuate neointimal proliferation in a porcine organ culture model of coronary restenosis. The neointima was significantly increased by 300% after angioplasty compared to non-angioplasty controls. The AT1 receptor antagonist, losartan, produced a significant reduction in neointimal index at 10(-5) mol/l, while its in vivo metabolite, EXP3174, reduced neointimal proliferation at 10(-6) mol/l. PD123319, a selective antagonist of the AT2 receptor, also restricted neointimal proliferation at 10(-5) mol/l. Treatment with captopril (10(-6) mol/l) increased the neointimal proliferation by approximately 200% after angioplasty. CONCLUSIONS: Direct blockade of AngII receptors effectively inhibits cell proliferation and restenosis post-angioplasty in vitro. ACE inhibition, exclusive of flow, does not attenuate proliferative restenosis. These data suggest that AngII contributes to neointimal proliferation and validates the concept that receptor antagonists could contribute to the therapeutic management of restenosis.
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Angiotensina II , Antagonistas de Receptores de Angiotensina , Enfermedad Coronaria/prevención & control , Imidazoles/uso terapéutico , Losartán/uso terapéutico , Piridinas/uso terapéutico , Análisis de Varianza , Angioplastia Coronaria con Balón , Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Animales , Antihipertensivos/uso terapéutico , Captopril/uso terapéutico , División Celular/efectos de los fármacos , Enfermedad Coronaria/patología , Enfermedad Coronaria/terapia , Vasos Coronarios , Técnicas de Cultivo , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/patología , Recurrencia , Porcinos , Tetrazoles/uso terapéuticoRESUMEN
Large plastic (>5mm) and microplastic (0.315-5mm) debris were collected from 25 beaches along the Hong Kong coastline. More than 90% consisted of microplastics. Among the three groups of microplastic debris, expanded polystyrene (EPS) represented 92%, fragments represented 5%, and pellets represented 3%. The mean microplastic abundance for Hong Kong was 5595items/m(2). This number is higher than international averages, indicating that Hong Kong is a hotspot of marine plastic pollution. Microplastic abundance was significantly higher on the west coast than on the east coast, indicating that the Pearl River, which is west of Hong Kong, may be a potential source of plastic debris. The amounts of large plastic and microplastic debris of the same types (EPS and fragments) were positively correlated, suggesting that the fragmentation of large plastic material may increase the quantity of beach microplastic debris.
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Monitoreo del Ambiente/métodos , Plásticos/análisis , Residuos , Contaminantes Químicos del Agua/análisis , Estuarios , Hong Kong , Poliestirenos/análisis , Ríos , Residuos/análisisRESUMEN
Thrombolytic therapy with tissue plasminogen activator (tPA) for acute myocardial infarction may result in major bleeding complications such as gastrointestinal or intracranial bleeding. A case is described of severe splenic hemorrhage and rupture which developed 3 h after completion of tPA infusion for suspected acute myocardial infarction. The patient developed hypovolemic shock with abdominal pain and distension and further evidence of myocardial necrosis. A computed tomography scan of the abdomen was helpful in elucidating the diagnosis, and surgical splenectomy resulted in a good patient outcome, though the period of hypotension had increased the extent of myocardial necrosis.
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Hemorragia/inducido químicamente , Rotura del Bazo/inducido químicamente , Activador de Tejido Plasminógeno/efectos adversos , Anciano , Femenino , Humanos , Hipotensión/etiología , Infarto del Miocardio/tratamiento farmacológico , Activador de Tejido Plasminógeno/uso terapéuticoRESUMEN
To examine physician bias in reporting percutaneous transluminal coronary angioplasty (PTCA) results and analyze this potential source of errors, and to examine the ability to estimate absolute lumen diameters visually, the authors reviewed 56 successful PTCAs from their institution. Pre- and postprocedural cineangiograms were blindly reviewed by an experienced consensus panel (three members) and compared with the interventional cardiologist's reported outcome (percentage diameter stenosis) and quantitative coronary angiography (QCA) using the Cardiac Measurement System. Staff cardiologists significantly overestimated pre-PTCA stenosis severity (staff 83.7 versus panel 75.2%, P < 0.05) while underestimating residual narrowing (staff 18.4 versus panel 22.8%, P < 0.05), thus exaggerating overall angioplasty benefit (staff -65.3 versus panel -52.4%, P < 0.05). The cumulative error varied greatly among individual staff members (3.4 to 18.0%). Despite these findings, the consensus panel accurately identified pre-PTCA minimum lumen diameter, as measured by quantitative angiography (panel 0.66 versus QCA 0.67 mm, not significant), although they tended to overestimate absolute postprocedural luminal dimensions (panel 2.28 versus QCA 2.00 mm, P < 0.05) and thereby ultimate changes in minimum lumen diameter (panel 1.62 versus QCA 1.33 mm, P < 0.05). Therefore, substantial bias exists in the reporting of PTCA outcomes, which tends to magnify the perceived benefits of the procedure. Well-trained observers can accurately estimate pre-PTCA absolute lumen diameters, although difficulties occur in evaluating residual dimensions.
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Angioplastia Coronaria con Balón/normas , Enfermedad Coronaria/terapia , Adulto , Anciano , Anciano de 80 o más Años , Angioplastia Coronaria con Balón/estadística & datos numéricos , Sesgo , Canadá , Angiografía Coronaria , Enfermedad Coronaria/diagnóstico por imagen , Enfermedad Coronaria/patología , Vasos Coronarios/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Variaciones Dependientes del Observador , Recurrencia , Reproducibilidad de los Resultados , Índice de Severidad de la Enfermedad , Factores de Tiempo , Resultado del TratamientoRESUMEN
Ecto ATP-diphosphohydrolase (apyrase) activity of human endothelial cells following aspirin treatment has been studied in-vitro. It was shown by HPLC analysis of supernatant samples that pre-incubation of the cultures with aspirin resulted in a significantly increased turnover of supplemented ATP into its degradation products (ADP and AMP). Enhanced expression of ectoenzyme after aspirin treatment could be observed as demonstrated by immunofluorescence-staining with monoclonal anti-apyrase antibodies. This suggests enhancement of endothelial ATP-diphosphohydrolase activity induced by aspirin. The present data obtained in human vascular cells in-vitro are in line with results from previous animal studies in-vivo, suggesting a novel cyclooxygenase-independent antithrombotic activity of aspirin.
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Apirasa/metabolismo , Aspirina/farmacología , Endotelio Vascular/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Adenosina Difosfato/metabolismo , Adenosina Monofosfato/metabolismo , Adenosina Trifosfato/metabolismo , Células Cultivadas , Cromatografía Líquida de Alta Presión , Endotelio Vascular/citología , Endotelio Vascular/enzimología , Fluoresceína-5-Isotiocianato/química , Humanos , Espectrometría de Fluorescencia , Venas Umbilicales/efectos de los fármacos , Venas Umbilicales/metabolismoAsunto(s)
Ecocardiografía Doppler , Electrocardiografía , Infarto del Miocardio/diagnóstico , Infarto del Miocardio/etiología , Mordeduras de Serpientes/complicaciones , Adulto , Angiografía Coronaria , Trombosis Coronaria/diagnóstico por imagen , Trombosis Coronaria/etiología , Ecocardiografía , Humanos , Masculino , Infarto del Miocardio/fisiopatologíaRESUMEN
The drug-eluting stent (DES) era has seen an increasing number of stent fractures, which is considered one of the mechanisms of restenosis in DES. This increase in recognition could be due to various factors such as increased diagnosis compared to the bare-metal stent era, platform design and strut thickness, higher inflation pressures for DES deployment, and use of DES in more complex lesions (e.g., angulated, diffuse, calcified, bifurcated). The angiographic presentation of DES fracture has been reported as in-stent restenosis (either symptomatic or asymptomatic) in all published cases. We discuss a case of DES fracture presented as an ST-elevation myocardial infarction that was associated with a large coronary artery aneurysm.
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Angioplastia Coronaria con Balón , Angiografía Coronaria , Enfermedad de la Arteria Coronaria/terapia , Reestenosis Coronaria/diagnóstico por imagen , Stents Liberadores de Fármacos/efectos adversos , Anciano , Aneurisma Coronario/diagnóstico por imagen , Aneurisma Coronario/etiología , Aneurisma Coronario/cirugía , Puente de Arteria Coronaria , Reestenosis Coronaria/etiología , Reestenosis Coronaria/cirugía , Humanos , Masculino , Falla de PrótesisRESUMEN
A mutant strain of Escherichia coli suppressed a frameshift and some UAG, UAA, and UGA mutants of bacteriophage T4 at 37 degrees C but not at 31 degrees C. This suppression was inhibited by the addition of thymine or thymidine to the medium used to test phage growth. Furthermore, the suppressor strain required thymine or thymidine for growth on minimal medium at 43 degrees C and if this auxotrophy was removed by reversion or recombination the strain no longer suppressed. These results suggest a link between thymidine nucleotide biosynthesis and suppression.
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Escherichia coli/genética , ARN de Transferencia/genética , Supresión Genética , Timina/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Genes , Genes Bacterianos , ARN Bacteriano/genéticaRESUMEN
Early constrictive pericarditis and anaemia developed in a 52 year old man after he had an inferior wall myocardial infarction complicated by Dressler's syndrome. Total pericardiectomy at the time of coronary artery bypass surgery resulted in complete resolution of signs and symptoms.
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Anemia/etiología , Infarto del Miocardio/complicaciones , Pericarditis Constrictiva/etiología , Anemia/patología , Puente de Arteria Coronaria , Humanos , Masculino , Persona de Mediana Edad , Infarto del Miocardio/patología , Infarto del Miocardio/cirugía , Pericardiectomía , Pericarditis Constrictiva/patología , Pericarditis Constrictiva/cirugía , Pericardio/patología , SíndromeRESUMEN
Circulating factors, including the plasma protease (100 KF) described previously, have been suspected to play a role in the pathogenesis of minimal change disease (MCD) for several decades. This factor was able to induce MCD-like alterations in kidney tissue in vitro, i.e. impairment of glomerular polyanion (GPA), as well as glomerular ecto-ATPase. We conducted permeability studies using alternate perfusion of the rat kidney ex vivo according to standard techniques. Either native 100 KF (n = 7) or control factor (n = 7) perfusion, followed by perfusion with diluted rat serum was carried out, while urine samples were collected by ureter cannulation. Total urinary protein (by spectrophotometry) as well as IgG (by ELISA) and albumin (by rocket electrophoresis) were measured. Sections of perfused kidneys were stained (immuno-) histochemically for GPA and glomerular ecto-ATPase, and the stainability was quantified using image analysis and expressed as arbitrary units. The results show significantly increased protein leakage after perfusion of 100 KF versus control factor (150.0 +/- 48.9 vs. 33.2 +/- 7.7 micrograms/min, p < or = 0.01), while the IgG/albumin ratio has decreased (12.0 +/- 9.4 vs 26.9 +/- 14.4%, p < or = 0.01). Plasma protein leakage after 100KF perfusion is associated with a significant loss of GPA (57.3 +/- 27.5 vs. 98.4 +/-12.0, p < or = 0.01) and significant decrease of glomerular ecto-ATPase expression (28.7 +/- 11.5 vs. 79.5 +/- 15.0, p < or = 0.001). The capability of 100KF to induce MCD-like glomerular lesions, in association with selectively increased permeability for plasma proteins, suggests that this human plasma constituent may be important in the pathogenesis of MCD.
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Proteínas Sanguíneas/toxicidad , Nefrosis Lipoidea/inducido químicamente , Nefrosis Lipoidea/patología , Adenosina Trifosfatasas/metabolismo , Albuminuria/orina , Animales , Proteínas Sanguíneas/metabolismo , Permeabilidad Capilar , Endopeptidasas/sangre , Femenino , Histocitoquímica , Humanos , Inmunoglobulina G/orina , Riñón/patología , Glomérulos Renales/metabolismo , Nefrosis Lipoidea/orina , Perfusión , Proteinuria/orina , Ratas , Ratas WistarRESUMEN
BACKGROUND: The pathogenesis of albuminuria in minimal change disease (MCD) is unknown. A human plasma factor (denoted as 100KF) is able to induce minimal change-like glomerular alterations, i.e. loss of glomerular sialoglycoproteins and decreased expression of glomerular ecto-ATPase, following in vitro incubation with kidney tissue. In addition, increased (selective) permeability for plasma proteins occurs after perfusion of 100KF into the rat kidney ex vivo. As in kidney tissue from subjects with minimal change disease, subendothelial injury has also been observed, i.e. reduced anionic sites in the lamina rara interna, the question was raised whether injury induced by the plasma factor 100KF involves vascular endothelium or subendothelial matrix of the glomerular capillary wall. METHODS: Permeability studies were carried out by using confluent endothelial monolayers (HUVEC) cultured on a standard two-compartment system. The permeability for a macromolecular marker (horse radish peroxidase) was tested following incubation of the monolayers with either the native plasma factor 100KF or the control factor (heat-inactivated 100KF), in combination with histochemical evaluation of the cells for ecto-ATPase expression. Also quantification of glomerular anionic sites at the ultrastructural level was carried out, after ex vivo perfusion of 100KF or control factor into rat kidneys. RESULTS: The plasma factor 100KF is able to increase the permeability of human endothelial monolayers for macromolecules in a dose-dependent manner (relative increase 122.4 +/- 24, 178.4 +/- 34 and 236.1 +/- 58% after preincubation with 0.05; 0.5 and 1.5 mg/ml 100KF respectively), concomitant with induction of minimal change-like histochemical alterations such as reduced expression of ecto-ATPase. The number of anionic sites in the lamina rara interna of the glomerular capillary wall is significantly diminished following perfusion with the plasma factor 100KF versus control factor (7.58 +/- 1.60 versus 12.57 +/- 2.05 per 1000 nm; P < or = 0.02); in contrast to the lamina rara externa (22.71 +/- 3.15 versus 22.27 +/- 2.92 per 1000 nm; statistically not significant). CONCLUSIONS: Endothelial cells and subendothelial matrix along the glomerular filtration barrier may be considered as target structures for the plasma factor 100KF, leading to initial minimal change-like alterations associated with glomerular albumin leakage.
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Proteínas Sanguíneas/farmacología , Endotelio Vascular/efectos de los fármacos , Uniones Intercelulares/efectos de los fármacos , Nefrosis Lipoidea/sangre , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Endotelio Vascular/patología , Femenino , Humanos , Ratas , Ratas WistarRESUMEN
The mechanism by which a human plasma factor associated with proteinuria is able to cause experimental glomerular albumin leakage is unknown. This factor (called 100KF) is able to induce glomerular alterations in the rat kidney, similar to those seen in minimal change disease, including loss of glomerular sialoglycoproteins and decreased expression of glomerular ecto-ATPase. It was previously shown that 100KF is able to stimulate release of reactive oxygen species in inflammatory cells in vitro. This prompted us to test whether 100KF-induced injury is oxygen dependent. The expression of glomerular sialoglycoproteins and ecto-ATPase was evaluated by standard histochemistry and computerized image analysis and expressed in arbitrary units. Rat kidney sections were incubated with or without 100KF under normal or oxygen-poor, i.e., nitrogen, conditions, or with supplementation of superoxide dismutase (SOD, 100 U/ml). The effect of 100KF on glomerular ecto-ATPase was oxygen dependent (32.98+/-2.14 under air vs. 65.20+/-5.53 under nitrogen, P< or =0.01), in contrast to the 100KF-induced loss of glomerular sialoglycoproteins that was not significantly altered under nitrogen (62.67+/-10.08 under air vs. 61.74+/-26.05 under nitrogen). Supplementation of SOD to 100KF solution under normal incubation conditions also suggested oxygen-dependent impairment of glomerular ecto-ATPase. Alternate perfusion ex vivo of the rat kidney with 100KF followed by diluted plasma showed that enhanced leakage of plasma proteins could be inhibited with SOD, indicating oxygen dependency of this 100KF-induced enhanced permeability (60.25+/-19.32 microg urinary albumin/ml after 100KF perfusion vs. 25.23+/-12.05 microg/ml after 100KF plus SOD, P< or =0.01). We conclude that the action of 100KF upon specific glomerular matrix molecules is oxygen dependent, as is the albumin leakage induced by 100KF in the present ex vivo model.
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Proteínas Sanguíneas/fisiología , Riñón/metabolismo , Nefrosis Lipoidea/metabolismo , Oxígeno/metabolismo , Serina Endopeptidasas/fisiología , Adenosina Trifosfatasas/biosíntesis , Albúminas/metabolismo , Animales , Proteínas Sanguíneas/química , Femenino , Humanos , Inmunohistoquímica , Técnicas In Vitro , Glomérulos Renales/metabolismo , Peso Molecular , Nefrosis Lipoidea/sangre , Ratas , Ratas Wistar , Serina Endopeptidasas/química , Sialoglicoproteínas/biosíntesis , Superóxido Dismutasa/farmacologíaRESUMEN
BACKGROUND: The human plasma constituent hemopexin (Hx), following incubation with renal tissue, is able to induce glomerular alterations in vitro that are similar to those seen in minimal change disease (MCD). Whether this acute phase reactant is also able to induce proteinuria and minimal change-like alterations in vivo is questioned. METHODS: In the first set of experiments, Hx (4.0 mg in 5.0 mL saline) or equal amounts of control fraction, that is, heat-inactivated Hx (HI-Hx), were infused into conscious rats (N = 6) that had been surgically equipped with a cannula inserted into the suprarenal artery (SRA), enabling direct contact of the infusate and the renal microvasculature. Each animal received HI-Hx at day 1 for 15 minutes (flow rate 20.0 mL/h), subsequently followed by saline for seven hours (Flow rate 5.0 mL/h), after which the cannula was disconnected. At day 2, identical infusions in the same rat were carried out, using native Hx. Urine samples collected every 30 minutes during the experiments were monitored for protein content using standard methods. In the second set of experiments, unilateral perfusion was done ex vivo in anesthetized rats with Hx (N = 5) or HI-Hx (N = 3; 1.5 mg/mL; 4.0 mL during 6 min). After reconnection of the circulation, urine samples of both kidneys were collected every 30 minutes during five hours via ureter cannulation. Urinary protein (expressed as the difference in excretion between perfused and nonperfused kidney) was calculated in mg/24 h. In additional experiments, rats were sacrificed two hours after perfusion of Hx or heat-inactivated (control) Hx (first set of experiments) or after five hours (second set of experiments), and kidneys were processed for immunohistochemical and ultrastructural examination. RESULTS: The results of experiment 1 show a significant increase of proteinuria after Hx infusion versus HI-Hx (means +/- SD, 41.91 +/- 16.01 mg/24 h vs. control, 21.22 +/- 5.69 mg/24 h; P = 0.03). Histochemical and electron microscopical examination of kidney tissue fragments taken at the time of proteinuria showed diminished expression of glomerular ecto-ATPase and enhanced effacement of epithelial foot processes at the ultrastructural level. In the second set of experiments, the results show significant urinary protein excretion peaking one hour after perfusion (6.63 +/- 7.06 mg/24 h), exclusively in Hx-perfused animals (analysis of variance, P < or = 0.001). CONCLUSION: It is concluded that Hx is able to induce proteinuria associated with MCD-like alterations in rat kidney in vivo.
Asunto(s)
Hemopexina , Proteinuria/inducido químicamente , Adenosina Trifosfatasas/metabolismo , Animales , Sangre/metabolismo , Hemopexina/metabolismo , Histocitoquímica , Humanos , Inyecciones Intraarteriales , Riñón/enzimología , Riñón/ultraestructura , Masculino , Microscopía Electrónica , Proteinuria/patología , Proteinuria/orina , Ratas , Ratas Wistar , Factores de TiempoRESUMEN
The purpose of this study was to describe the effects of moderate intensity exercise training on the muscle energy utilization, blood flow, and exercise performance of four sedentary older individuals (58 +/- 4 yrs). Subjects trained the dominant forearm each day for 12 weeks. The nondominant arm was not trained and served as a within-subject control. 31P nuclear magnetic resonance spectroscopy (31P NMRS) was used to identify the power output in watts (W) at the onset, or threshold, of intracellular acidosis (IT) in the exercising muscle during progressive exercise tests to fatigue. After 6 weeks of training, power output at the IT increased by 14% (p < 0.05) in the dominant arm; however, an additional 6 weeks of the same exercise program failed to produce a further increase in IT power. IT power of the nondominant forearm was not changed. In the dominant forearm, endurance time for a submaximal wrist flexion test was increased 34% and 58% at 6 and 12 weeks, respectively. Maximal voluntary strength was not affected by training, nor was resting or exercising blood flow. The training program delayed the onset of intracellular acidosis during progressive exercise and increased the capacity for submaximal work. These effects did not appear to depend on an increase in muscle blood flow.
Asunto(s)
Músculos/metabolismo , Resistencia Física/fisiología , Acidosis/metabolismo , Acidosis/fisiopatología , Prueba de Esfuerzo , Fatiga/metabolismo , Fatiga/fisiopatología , Femenino , Antebrazo , Humanos , Espectroscopía de Resonancia Magnética , Masculino , Persona de Mediana Edad , Contracción Muscular/fisiología , Músculos/irrigación sanguínea , Músculos/fisiología , Fósforo , Educación y Entrenamiento Físico , Flujo Sanguíneo Regional/fisiologíaRESUMEN
Hereditary multiple exostoses (HME), a dominantly inherited genetic disorder characterized by multiple cartilaginous tumors, is caused by mutations in members of the EXT gene family, EXT1 or EXT2. The corresponding gene products, exostosin-1 (EXT1) and exostosin-2 (EXT2), are type II transmembrane glycoproteins which form a Golgi-localized heterooligomeric complex that catalyzes the polymerization of heparan sulfate (HS). Although the majority of the etiological mutations in EXT are splice-site, frameshift, or nonsense mutations that result in premature termination, 12 missense mutations have also been identified. Furthermore, two of the reported etiological missense mutations (G339D and R340C) have been previously shown to abrogate HS biosynthesis (McCormick et al. 1998). Here, a functional assay that detects HS expression on the cell surface of an EXT1-deficient cell line was used to test the remaining missense mutant exostosin proteins for their ability to rescue HS biosynthesis in vivo. Our results show that EXT1 mutants bearing six of these missense mutations (D164H, R280G/S, and R340S/H/L) are also defective in HS expression, but surprisingly, four (Q27K, N316S, A486V, and P496L) are phenotypically indistinguishable from wild-type EXT1. Three of these four "active" mutations affect amino acids that are not conserved among vertebrates and invertebrates, whereas all of the HS-biosynthesis null mutations affect only conserved amino acids. Further, substitution or deletion of each of these four residues does not abrogate HS biosynthesis. Taken together, these results indicate that several of the reported etiological mutant EXT forms retain the ability to synthesize and express HS on the cell surface. The corresponding missense mutations may therefore represent rare genetic polymorphisms in the EXT1 gene or may interfere with as yet undefined functions of EXT1 that are involved in HME pathogenesis.
Asunto(s)
Exostosis Múltiple Hereditaria/enzimología , Exostosis Múltiple Hereditaria/genética , N-Acetilglucosaminiltransferasas/genética , N-Acetilglucosaminiltransferasas/metabolismo , Mutación Puntual/genética , Secuencia de Aminoácidos , Animales , Células CHO , Cromatografía por Intercambio Iónico , Cricetinae , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Prueba de Complementación Genética , Heparitina Sulfato/metabolismo , Humanos , Datos de Secuencia Molecular , Mutación Missense/genética , N-Acetilglucosaminiltransferasas/biosíntesis , N-Acetilglucosaminiltransferasas/química , Polimorfismo Genético/genética , Alineación de SecuenciaRESUMEN
In the majority of cases, the mechanism underlying the resistance to acyclovir (ACV) of herpes simplex viruses (HSVs) is thymidine kinase (TK) deficiency. Plaque isolates from eight ACV-resistant (ACVr) clinical isolates from AIDS patients, of which five reactivated, were sequenced to determine the genetic lesion within the tk gene conferring resistance and whether this may have correlated with reactivation potential. Mutations were clustered within two homopolymer nucleotide stretches. Three plaque isolates (1737-14, 90-150-3, and 89-650-5) had insertion mutations within a stretch of 7 guanosines, while two isolates (89-063-1 and 89-353-1) had frameshift mutations within a stretch of 6 cytosines (a deletion and an insertion, respectively). Mutations resulted in premature termination codons, and the predicted 28- and 32-kDa truncated TK products were detected by Western blot analysis of virus-infected cell extracts. The repair of one homopolymer frameshift mutation (in isolate 1737-14) restored TK activity, demonstrating that this mutation is the basis of TK deficiency. Of the five reactivated isolates, four were TK deficient and contained frameshift mutations while the fifth retained TK activity because of its altered-TK or Pol- phenotype. These data demonstrate that the majority of ACVr clinical isolates contain frameshift mutations within two long homopolymer nucleotide stretches which function as hot spots within the HSV tk gene and produce nonfunctional, truncated TK proteins.