RESUMEN
High Throughput Screening (HTS) now plays an important role in the discovery of new lead compounds for novel therapeutic targets. The advantage of HTS over the conventional method, now termed as Low Throughput Screening (LTS), is that valuable compounds can be selected rapidly from a large number of samples with minimal human involvement. In spite of the growing awareness of HTS, the importance of the LTS in the drug discovery and development is still not changed. Advances in pharmacogenomics will also provide us many pharmacological targets, and thus increase the number of compounds that should be assayed by HTS and LTS. In this review, we will first describe the outline of HTS. We will next describe new approaches to develop and brush up the LTS: 1) screening method of drugs acting on ion channels by voltage-sensitive fluorescent dye, 2) functional assay method using reconstituted smooth muscle fiber, and 3) organ culture method as a useful model of vascular proliferative disease. These approaches, which work cooperatively with HTS, will contribute greatly to the development of new drugs.
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Química Farmacéutica/métodos , Diseño de Fármacos , Evaluación Preclínica de Medicamentos/métodos , Tecnología Farmacéutica/métodos , Animales , Autoanálisis , Genómica , Humanos , Canales Iónicos , Músculo Liso , Análisis de Secuencia por Matrices de Oligonucleótidos , Técnicas de Cultivo de Órganos , Farmacogenética , RobóticaRESUMEN
A hospital-based prospective study of 99 patients with community-acquired pneumonia (CAP) was carried out in Kampala, Uganda. We evaluated microbiological etiologies, clinical features and effectiveness of short-term parenteral ampicillin followed by oral amoxicillin for these patients in relation to HIV-status. We demonstrated a very high prevalence (75%) of HIV-1 infection. No significant difference was observed with respect to age, gender, prior antibiotic usage, symptoms, laboratory data or bacterial etiology between HIV-1-infected and HIV-uninfected CAP patients. Most strains of Streptococcus pneumoniae (n = 19) and Haemophilus influenzae (n = 8) isolated from HIV-1-infected patients were penicillin-resistant (95%) and beta-lactamase producing (75%) strains, respectively. A high percentage of good clinical response was found in both HIV-1-infected (81%) and HIV-uninfected (86%) among 39 patients with CAP due to a defined bacterial pathogen. These data support the use of short-term parenteral ampicillin for patients with bacterial CAP irrespective of HIV-status.
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Ampicilina/administración & dosificación , Infecciones Comunitarias Adquiridas/tratamiento farmacológico , Infecciones por VIH/epidemiología , VIH-1/aislamiento & purificación , Penicilinas/administración & dosificación , Neumonía Bacteriana/tratamiento farmacológico , Administración Oral , Adulto , Amoxicilina/administración & dosificación , Infecciones Comunitarias Adquiridas/complicaciones , Esquema de Medicación , Farmacorresistencia Microbiana , Femenino , Infecciones por VIH/complicaciones , Haemophilus influenzae/efectos de los fármacos , Humanos , Infusiones Intravenosas , Masculino , Pruebas de Sensibilidad Microbiana , Neumonía Bacteriana/complicaciones , Prevalencia , Estudios Prospectivos , Streptococcus pneumoniae/efectos de los fármacos , Resultado del Tratamiento , Uganda/epidemiologíaRESUMEN
Recently, our group and others cloned the TRMA disease gene, SLC19A2, which encodes a thiamin transporter. Here, we report the cloning and characterization of the full-length cDNA and genomic sequences of mouse Slc19a2. The Slc19a2 cDNA contained a 1494-bp open-reading frame, and had 5'- and 3'-untranslated regions of 189 and 1857 bp, respectively. A putative GC-rich, TATA-less promoter was identified in genomic sequence directly upstream of the identified 5' end. The Slc19a2 gene spanned 16.3 kb and was organized into six exons, a gene structure conserved with the human orthologue. The predicted Slc19a2 protein, like SLC19A2, was predicted to have 12 transmembrane domains and shared a number of other conserved sequence motifs with the human orthologue, including one potential N-glycosylation site (N(63)) and several potential phosphorylation sites. Comparison of the Slc19a2 amino acid sequence with those of the other known SLC19A solute carriers highlighted interesting patterns of conservation and divergence in various domains, allowing insight into potential structure-function relationships. The identification of the mouse Slc19a2 cDNA and genomic sequences will facilitate the generation of an animal model of TRMA, permitting future studies of disease pathogenesis.
Asunto(s)
Proteínas Portadoras/genética , Proteínas de Transporte de Membrana , Secuencia de Aminoácidos , Anemia Megaloblástica/tratamiento farmacológico , Anemia Megaloblástica/genética , Animales , Secuencia de Bases , Northern Blotting , Clonación Molecular , ADN/química , ADN/genética , ADN Complementario/química , ADN Complementario/genética , ADN Complementario/aislamiento & purificación , Exones , Expresión Génica , Genes/genética , Intrones , Masculino , Ratones , Datos de Secuencia Molecular , ARN Mensajero/genética , ARN Mensajero/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Tiamina/uso terapéutico , Distribución TisularRESUMEN
It has been reported that activation of autonomic effectors during mental simulation of voluntary motor actions (motor imagery: MI) may be explained by two different factors, i.e., functions of preparation or anticipation of actual exercise (motor anticipation) and the central motor programming/planning which acts during actual motor action (motor programming). This study was designed to clarify how these factors participate during MI, utilizing two mental tasks with high mental stress, i.e., MI and mental arithmetic (MA). Several autonomic effectors' responses were compared between MI of a 500 m speed skating sprint and MA. Subjects were eight 18 to 25 year old young male speed skate athletes, all of them could easily and vividly imagine a 500 m speed skating sprint. Duration of the MI ranged from 35 to 38 sec and these were very close to each subject's actual best record (means of absolute differences were less than 0.6 sec, i.e., less than 1.7% relatively). A significant decrease of skin resistance (SR), increases of heart rate (HR) and respiration rate were observed in both MI and MA when compared to each control resting level (excluding one subject for respiration rate during MI). SR decreased during MI (mean and SD of 8 subjects: 45.9 +/- 17.7%) and MA (39.7 +/- 16.8%), with no significant differences between MI and MA (t = 1.29, by paired t-test). HR increased significantly above control values in MA (10.3 +/- 4.3%) and MI (44.3 +/- 18.8%). However, the increase during MA was significantly smaller (t = 4.99, p < 0.001) than in MI. Respiratory rate increased significantly in both MI (46.5 +/- 30.9%) and MA (27.7 +/- 14.6%), with no significant difference between MI and MA (t = 1.82) due to the large individual variation in MI. The frequency of respiration was fairly regular during MA, but quite irregular during MI (similar to those during actual motor actions). The central nervous system which acts in MI may possess the function of activation of target effectors which play an important role in actual exercise, on the basis of incremental vigilance level induced by the function of motor anticipation.
Asunto(s)
Sistema Nervioso Autónomo/fisiología , Imágenes en Psicoterapia , Adolescente , Adulto , Respuesta Galvánica de la Piel/fisiología , Frecuencia Cardíaca/fisiología , Humanos , Pruebas de Inteligencia , Masculino , Respiración , Análisis y Desempeño de TareasRESUMEN
The cDNA clone encoding a novel isoform of protein kinase PKN, termed PKNbeta, was isolated from a HeLa cDNA library. PKNbeta had high sequence homology with PKNalpha, originally isolated PKN, especially in the repeats of charged amino acid-rich region with leucine-zipper like sequences (CZ region/HR1), in the carboxyl-terminal catalytic domain, and in approximately 130 amino acid stretch (D region/HR2), located between CZ region/HR1 and the catalytic domain. However, the amino acid sequence of PKNbeta differed from that of PKNalpha in the region immediately amino-terminal to the catalytic domain, which contained two distinct proline-rich sequences consistent with the class II consensus sequence, PXXPXR, for binding to SH3 domain. Distribution of PKNbeta differed from that of PKNalpha in the following two respects: (1) Northern blotting indicated that PKNbeta mRNA could not be detected in human adult tissues, but was expressed abundantly in human cancer cell lines; (2) immunochemical analysis indicated that PKNbeta localized in nucleus and perinuclear Golgi apparatus, and was almost absent in cytoplasmic region in NIH3T3 cells. Recombinant PKNbeta expressed in COS7 cells displayed autophosphorylation and peptide kinase activity, but was found to be significantly less responsive to arachidonic acid than PKNalpha. The identification of this novel isoform underscores the diversity of PKN signaling pathway.
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Isoenzimas/genética , Isoenzimas/metabolismo , Proteína Quinasa C/genética , Proteína Quinasa C/metabolismo , Adenosina Trifosfato/metabolismo , Secuencia de Aminoácidos , Animales , Ácido Araquidónico/farmacología , Secuencia de Bases , Sitios de Unión , Células COS , Núcleo Celular/enzimología , Secuencia de Consenso , ADN Complementario/aislamiento & purificación , Células HeLa , Humanos , Isoenzimas/química , Leucina Zippers , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Proteína Quinasa C/química , ARN Mensajero/análisis , Homología de Secuencia , Células Tumorales CultivadasRESUMEN
Thiamine-responsive megaloblastic anaemia syndrome (TRMA; MIM 249270) is an autosomal recessive disorder with features that include megaloblastic anaemia, mild thrombocytopenia and leucopenia, sensorineural deafness and diabetes mellitus. Treatment with pharmacologic doses of thiamine ameliorates the megaloblastic anaemia and diabetes mellitus. A defect in the plasma membrane transport of thiamine has been demonstrated in erythrocytes and cultured skin fibroblasts from TRMA patients. The gene causing TRMA was assigned to 1q23.2-q23.3 by linkage analysis. Here we report the cloning of a new gene, SLC19A2, identified from high-through-put genomic sequences due to homology with SLC19A1, encoding reduced folate carrier 1 (refs 8-10). We cloned the entire coding region by screening a human fetal brain cDNA library. SLC19A2 encodes a protein (of 497 aa) predicted to have 12 transmembrane domains. We identified 2 frameshift mutations in exon 2. a 1-bp insertion and a 2-bp deletion, among four Iranian families with TRMA. The sequence homology and predicted structure of SLC19A2, as well as its role in TRMA, suggest that its gene product is a thiamine carrier, the first to be identified in complex eukaryotes.
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Anemia Megaloblástica/genética , Proteínas Portadoras/genética , Proteínas de Transporte de Membrana , Mutación , Tiamina/metabolismo , Tiamina/uso terapéutico , Secuencia de Aminoácidos , Anemia Megaloblástica/complicaciones , Anemia Megaloblástica/tratamiento farmacológico , Secuencia de Bases , Clonación Molecular , ADN Complementario/genética , Sordera/complicaciones , Sordera/genética , Complicaciones de la Diabetes , Diabetes Mellitus/genética , Femenino , Humanos , Masculino , Datos de Secuencia Molecular , Linaje , Homología de Secuencia de Aminoácido , SíndromeRESUMEN
A 41-year-old male received acupuncture in the right shoulder for the sake of arthralgia. Three days after acupuncture he was admitted due to severe epigastralgia. Erythematous change and swelling were observed around the right shoulder. A study by magnetic resonance showed an increased signal intensity in a portion of the right subscapular muscle. Four hours after admission he became hypotensive. The erythematous and necrotic change in the right shoulder skin rapidly spread. Excisional debridement in the right lateral chest wall was immediately done. However, the patient died one day after admission despite administration of a high-dose ampicillin and other supportive therapies. Bacteriological and histological examinations confirmed severe streptococcal myositis. This is a case report of toxic shock-like syndrome probably caused by acupuncture.
Asunto(s)
Terapia por Acupuntura/efectos adversos , Choque Séptico/transmisión , Infecciones Estreptocócicas/transmisión , Streptococcus pyogenes , Adulto , Artralgia/terapia , Resultado Fatal , Humanos , MasculinoRESUMEN
1. cDNA encoding a functional inwardly rectifying K+ (IRK1) channel was transfected into COS-1 cells (a Green Monkey kidney cell line) using the liposome method, and voltage clamp experiments were done after 48-72 h. 2. Transfected cells showed inward rectification under whole-cell recording. The unitary current-voltage relationships in the inside-out configuration were almost linear in the absence of internal Mg2+ and polyamines, and the channel conductance averaged 34.1 +/- 2.0 pS (n = 15) at 23-26 degrees C. 3. Internal Mg2+ (2-10 microM) induced sublevels in the outward current with one-third and two-thirds of the unitary amplitude as in native channels. 4. To determine the subunit stoichiometry, we constructed tandem multimeric cDNAs consisting of the coding sequences of the IRK1 gene linked in a head-to-tail fashion. Cells transfected with tandem homomultimers up to octamers showed similar inwardly rectifying K+ channels. 5. A mutation (E138Q) eliminated the ionic conductance of the channel. Channels expressed by dimeric constructs containing a single mutant have a conductance ranging between 5 and 35 pS. 6. The E138Q mutant cotransfected with a wild-type dimeric, trimeric or tetrameric construct did not alter the channel conductance. The results do not support the notion that IRK1 channel proteins consist of four subunits.
Asunto(s)
Canales de Potasio de Rectificación Interna , Canales de Potasio/biosíntesis , Transfección , Animales , Células COS , ADN Complementario/química , Desoxirribonucleasa EcoRI/metabolismo , Magnesio/metabolismo , Mutagénesis , Canales de Potasio/genética , Conformación ProteicaRESUMEN
Transgenic plants have significant potential in the bioproduction of complex human therapeutic proteins due to ease of genetic manipulation, lack of potential contamination with human pathogens, conservation of eukaryotic cell machinery mediating protein modification, and low cost of biomass production. Tobacco has been used as our initial transgenic system because Agrobacterium-mediated transformation is highly efficient, prolific seed production greatly facilitates biomass scale-up, and development of new "health-positive" uses for tobacco has significant regional support. We have targeted bioproduction of complex recombinant human proteins with commercial potential as human pharmaceuticals. Human protein C (hPC), a highly processed serum protease of the coagulation/anticoagulation cascade, was produced at low levels in transgenic tobacco leaves. Analogous to its processing in mammalian systems, tobacco-synthesized hPC appears to undergo multiple proteolytic cleavages, disulfide bond formation, and N-linked glycosylation. Although tobacco-derived hPC has not yet been tested for all posttranslational modifications or for enzymatic (anticlotting) activity, these results are promising and suggest considerable conservation of protein processing machinery between plants and animals. CropTech researchers have also produced the human lysosomal enzyme glucocerebrosidase (hGC) in transgenic tobacco. This glycoprotein has significant commercial potential as replacement therapy in patients with Gaucher's disease. Regular intravenous administration of modified glucocerebrosidase, derived from human placentae or CHO cells, has proven highly effective in reducing disease manifestations in patients with Gaucher's disease. However, the enzyme is expensive (dubbed the "world's most expensive drug" by the media), making it a dramatic model for evaluating the potential of plants to provide a safe, low-cost source of bioactive human enzymes. Transgenic tobacco plants were generated that contained the human glucocerebrosidase cDNA under the control of an inducible plant promoter. hGC expression was demonstrated in plant extracts by enzyme activity assay and immunologic cross-reactivity with anti-hGC antibodies. Tobacco-synthesized hGC comigrates with human placental-derived hGC during electrophoretic separations, is glycosylated, and, most significantly, is enzymatically active. Although expression levels vary depending on transformant and induction protocol, hGC production of > 1 mg/g fresh weight of leaf tissue has been attained in crude extracts. Our studies provide strong support for the utilization of tobacco for high-level production of active hGC for purification and eventual therapeutic use at potentially much reduced costs. Furthermore, this technology should be directly adaptable to the production of a variety of other complex human proteins of biologic and pharmaceutical interest.
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Enzimas/biosíntesis , Nicotiana/genética , Plantas Tóxicas , Proteínas Recombinantes/biosíntesis , Glucosilceramidasa/biosíntesis , Humanos , Plantas Modificadas Genéticamente , Proteína C/biosíntesisRESUMEN
Nitrogen microwave-induced plasma mass spectrometry was studied for its applicability to the isotope dilution analysis of selenium in biological samples. Spectroscopic interference by calcium, which is present in high concentrations in biological samples, was investigated. No detectable background spectrum was observed for the major selenium isotopes of 78Se and 80Se. No detectable interferences by sodium, potassium, calcium and phosphorus on the isotope ratio 80Se/78Se were observed up to concentration of 200 mg/ml. The method was applied to the analysis of selenium in biological reference materials of marine organisms. The results showed good agreement between the certified and found values.
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Espectrometría de Masas/métodos , Nitrógeno/química , Selenio/análisis , Animales , Bivalvos , Peces , Indicadores y Reactivos , Espectrometría de Masas/instrumentación , Microondas , Nephropidae , Técnica de Dilución de Radioisótopos , Estándares de Referencia , Radioisótopos de Selenio/análisisRESUMEN
The safety and efficacy of transurethral microwave thermo-therapy (TUMT) for the treatment of symptomatic benign prostatic hyperplasia (BPH) have been demonstrated in the urological literature. Since a radiofrequency electromagnetic wave has deeper and more even transmission of heat to tissues than a microwave, we have developed a new prototype device for transurethral thermotherapy using a radiofrequency electromagnetic wave. The device consists of a microcomputer-controlled heat generator operating at 8 Mhz, a temperature monitoring system, a urethral cooling system and a urethral applicator and a rectal thermosensor probe. A balloon electrode encased in a specialized Foley catheter is connected parallel to twin plate electrodes on both sides of the pelvic region. An 8-MHz electromagnetic wave is directed to the prostate by means of capacitive coupling. The water coolant continuously perfused through the catheter allows high temperatures within the prostate while preserving the urethral mucosa. Heating experiment using agar phantom showed the hot spots to be distributed at 0.5-3 cm from the catheter surface. Heating experiment using canine prostates demonstrated that an intraprostatic temperature of > 48 degrees C could be achieved while the urethral and rectal temperatures had not exceeded 36 and 40 degrees C respectively. Histological examination immediately after the experiment showed the urethral mucosa to be preserved while coagulation necrosis of the periurethral prostate accompanied with congestion and hemorrhage of small blood vessels were observed at 5-8 mm from the urethra. The bladder and the rectum showed no gross alterations. Histopathological examination 10 days after the experiment revealed the intact urethral mucosa and mild mononuclear infiltration around the destroyed periurethral glands.(ABSTRACT TRUNCATED AT 250 WORDS)
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Hipertermia Inducida/instrumentación , Hiperplasia Prostática/terapia , Terapia por Radiofrecuencia , Animales , Perros , Estudios de Evaluación como Asunto , Humanos , Hipertermia Inducida/efectos adversos , Masculino , Modelos Estructurales , Necrosis , Próstata/patología , Ondas de Radio/efectos adversos , Seguridad , TemperaturaRESUMEN
BACKGROUND: Transurethral microwave thermotherapy (TUMT) is a minimally invasive treatment for benign prostatic hyperplasia (BPH). It has been reported that increased thermal dose and higher intraprostatic temperatures resulted in improved clinical response. Recently we treated BPH patients with the prostatron device using a new version of software (Prostasoft 2.5), which was intended to increase thermal delivery by allowing maximum power up to 70W. The safety and clinical results were compared between the patients treated with Prostasoft 2.5 and those treated with the currently available software (Prostasoft 2; maximum power up to 50W). METHODS: A total of 105 patients were treated successively with two treatment protocols. Sixty-three patients were treated with Prostasoft 2 between September 1992 and July 1993, while 42 were treated with Prostasoft 2.5 between August 1993 and April 1994. Therefore, this investigation was a retrospective nonrandomized study. There was no significant difference in the baseline patient characteristics between the two groups. RESULTS: Total thermal dose delivered to the prostate was significantly higher in the Prostasoft 2.5 group than that in the Prostasoft 2 group (137 kJ versus 116 kJ, P < 0.05). No serious complications were encountered in either group. Six months after TUMT, in both the Prostasoft 2.5 and Prostasoft 2 groups there was an improvement in patient condition as measured by the mean I-PSS, QOL, and peak flow rate values, as well as the overall therapeutic efficacy. The two groups differed in the amount of posttreatment improvement from between 8% and 22%, but this difference was not statistically significant. CONCLUSIONS: Our study suggests that higher thermal dose attained by Prostasoft 2.5 does not necessarily result in more pronounced clinical improvement, although clinical response to TUMT has often been reported to be dependent upon thermal dose.
Asunto(s)
Microondas/uso terapéutico , Hiperplasia Prostática/terapia , Anciano , Estudios de Seguimiento , Humanos , Hipertermia Inducida , Masculino , Estudios Retrospectivos , Programas InformáticosRESUMEN
Since September 1992, 63 patients with symptomatic benign prostatic hyperplasia (BPH) have been treated with transurethral microwave thermotherapy (TUMT) using the Prostatron device. The International Prostate Symptom Score (I-PSS) and quality of life (QOL) score were used to evaluate subjective symptoms. The mean I-PSS (total, irritative and obstructive scores) and QOL scores had decreased by 40, 38, 45 and 40%, respectively, at 12 months (p < 0.0001). While the mean peak flow rate had increased by 72% (p < 0.001). The clinical efficacy at 12 months was 42%, using a modification of the response criteria proposed at the 2nd International Consultation on Benign Prostatic Hyperplasia. There were no significant differences in the baseline and treatment parameters between those who responded favorably to TUMT and those who did not. The total thermal dose delivered to the prostate did not predict clinical response. However, there was a positive correlation between I-PSS or QOL at baseline and % reduction at 3, 6 and 12 months, and a negative correlation between peak flow rate at baseline and % increase at 3 and 6 months. There were no major complications associated with TUMT during the follow-up period. In summary, our 1-year clinical results are compatible with previous reports, suggesting that TUMT is a safe, effective and lasting non-surgical treatment for BPH. However, evaluation of efficacy should be based on uniform criteria to facilitate comparisons of different clinical trials. The most suitable patient profiles for TUMT could not be identified by retrospective analysis.
Asunto(s)
Hipertermia Inducida , Microondas/uso terapéutico , Hiperplasia Prostática/terapia , Anciano , Estudios de Seguimiento , Humanos , Masculino , Hiperplasia Prostática/fisiopatología , Calidad de Vida , Estudios Retrospectivos , Uretra , UrodinámicaRESUMEN
Abnormal hypertension sometimes occurs following intravenous administration (i.v.) of pancuronium in patients with brain injury. The present experiment was designed to determine whether brain injury contributes to the hypertensive response of i.v. pancuronium. Forty-six Wister strain rats were studied, of which 39 had induced brain injury at (a) upper pons (b) midbrain (c) thalamus region (excluding hypothalamus) and (d) cerebellum or a combination of these sites. The injury was made by single insertion of a 22 Gauge needle through the skull surface. The quantity of pancuronium solution administered i.v. in each case was 1.0 ml containing either 0.8 mg/kg or 8 mg/kg of pancuronium. Group A (n = 7) had no brain injury and the mean arterial pressure (MAP) did not change following i.v. administration of pancuronium. In Group B (n = 9) (a+b+c, 8.0 mg/kg) MAP rose from 90.9 +/- 15.4 to 102 +/- 22.0 mmHg and in Group C (n = 7) (a+b+c, 0.8 mg/kg) MAP rose from 148.4 +/- 13.3 to 160 +/- 14.4 mmHg. In Group D (n = 5) (b+c, 8.0 mg/kg) MAP remained unchanged. In Group E (n = 5) (a, 8.0 mg/kg) MAP rose from 130.3 +/- 18.7 to 146 +/- 27.6 mmHg and in Group F (n = 6) (a, 0.8 mg/kg) MAP rose from 129.7 +/- 15.6 to 135.8 +/- 13.8 mmHg. In Group G (n = 7) (d, 8.0 mg/kg) MAP remained unchanged. Since the MAP was elevated in only those groups that received injury in the upper pons, we concluded that injury in the upper pons can lead to hypertension following i.v. administration of pancuronium.
Asunto(s)
Lesiones Encefálicas/fisiopatología , Hipertensión/inducido químicamente , Pancuronio , Animales , Presión Sanguínea , Lesiones Encefálicas/patología , Relación Dosis-Respuesta a Droga , Femenino , Hipertensión/fisiopatología , Inyecciones Intravenosas , Masculino , Concentración Osmolar , Ratas , Ratas WistarRESUMEN
Transurethral microwave thermotherapy (TUMT) has been shown to produce a clinical benefit in patients with symptomatic benign prostatic hyperplasia. In order to identify the features of the ideal candidate, a retrospective analysis was conducted in 32 patients who were followed for 2 mo or more. Good responders (GR) were defined as having their Siroky peak flow rate (PFR) standard deviation (SD) increase by < 0.5 or a decrease in the International Prostatic Symptom Score (I-PSS) of > 10 (22 patients). Poor responders (PR) were defined as having their PFR SD increase by < or = 0.5 and their I-PSS decrease by < or = 10 (10 patients). The prostate volume, pre-TUMT I-PSS and intravesical opening pressure were significantly greater in the GR group, while there were no significant differences between the 2 groups for the other baseline patient characteristics: age, prostate length, PFR, PFR SD, post-voiding residual volume and quality of life. Concerning the operational parameters, significantly more total energy was delivered to the prostate in the GR group (mean 131 kJ) than in the PR group (mean 101 kJ). Moreover, the 7 patients with anti-androgen therapy pre-TUMT received less total energy and 5 of the 7 were poor responders. These results suggest that patients with apparent obstructive symptoms and with moderate enlargement of prostate could benefit more from this less invasive therapy. Clinical response seems to be dose-dependent and patients with a history of recent anti-androgen treatment may have a less favorable response.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Hipertermia Inducida , Hiperplasia Prostática/terapia , Alilestrenol/uso terapéutico , Acetato de Clormadinona/uso terapéutico , Relación Dosis-Respuesta en la Radiación , Humanos , Masculino , Microondas/uso terapéutico , Presión , Resultado del Tratamiento , MicciónRESUMEN
The ERM family members, ezrin, radixin, and moesin, localizing just beneath the plasma membranes, are thought to be involved in the actin filament/plasma membrane association. To identify the integral membrane protein directly associated with ERM family members, we performed immunoprecipitation studies using antimoesin mAb and cultured baby hamster kidney (BHK) cells metabolically labeled with [35S]methionine or surface-labeled with biotin. The results indicated that moesin is directly associated with a 140-kD integral membrane protein. Using BHK cells as antigens, we obtained a mAb that recognized the 140-kD membrane protein. We next cloned a cDNA encoding the 140-kD membrane protein and identified it as CD44, a broadly distributed cell surface glycoprotein. Immunoprecipitation with various anti-CD44 mAbs showed that ezrin and radixin, as well as moesin, are associated with CD44, not only in BHK cells, but also in mouse L fibroblasts. Furthermore, immunofluorescence microscopy revealed that in both BHK and L cells, the Triton X-100-insoluble CD44 is precisely colocalized with ERM family members. We concluded that ERM family members work as molecular linkers between the cytoplasmic domain of CD44 and actin-based cytoskeletons.
Asunto(s)
Proteínas Sanguíneas/metabolismo , Proteínas Portadoras/metabolismo , Proteínas del Citoesqueleto , Citoesqueleto/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas de Microfilamentos , Fosfoproteínas/metabolismo , Proteínas/metabolismo , Receptores de Superficie Celular/metabolismo , Receptores Mensajeros de Linfocitos/metabolismo , Actinas/metabolismo , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales , Proteínas Sanguíneas/análisis , Proteínas Portadoras/genética , Proteínas Portadoras/inmunología , Proteínas Portadoras/aislamiento & purificación , Línea Celular , Cricetinae , Citoesqueleto/química , ADN Complementario/análisis , Epítopos/genética , Epítopos/inmunología , Receptores de Hialuranos , Proteínas de la Membrana/análisis , Ratones , Datos de Secuencia Molecular , Fosfoproteínas/análisis , Pruebas de Precipitina , Proteínas/análisis , Virus de la Rabia/química , Virus de la Rabia/inmunología , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/inmunología , Receptores de Superficie Celular/aislamiento & purificación , Receptores Mensajeros de Linfocitos/genética , Receptores Mensajeros de Linfocitos/inmunología , Receptores Mensajeros de Linfocitos/aislamiento & purificación , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/inmunología , Análisis de Secuencia de ADNRESUMEN
In order to investigate the changes of motoneuron excitability during mental simulation of a voluntary movement (motor imagery; MI), the soleus H-reflex and several activities of autonomic effectors were recorded simultaneously when MI of speed skate sprint was performed. The subjects were seven elite speed skate athletes ranging in age from 18 to 24 years old. They were all skilled in MI, and could imagine full vivid skating movement internally as if they were really exercising. The subjects were awake, relaxed and blindfolded. At first, rest (5 min), positive relaxation (4 min) and concentration periods (2.5 min) were set up. Then, after the previous announcement for the start, MI was initiated by the sound of a signal gun which was recorded by a tape recorder. MIs were continued for each subject from about 36 s to 38 s, and these imaging times were very close to the actual personal best records of each subject. The autonomic effectors represented by skin conductance response (SCR), heart rate (HR), and respiration rate (RSR) became remarkably active during MI in all trials. SCR was increased by 51% on the average. Furthermore, HR and RSR were increased by 57% and 76% respectively, compared with those during resting period. These changes in the autonomic effectors were all significant and were those which were activated in actual movements. The amplitudes of H-reflex were almost all constant before MI initiation. However, the reduction of the H-reflex amplitude arose just after the start and lasted to the end of MI.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Nivel de Alerta/fisiología , Atención/fisiología , Reflejo H/fisiología , Imaginación/fisiología , Cinestesia/fisiología , Neuronas Motoras/fisiología , Patinación/fisiología , Adolescente , Adulto , Sistema Nervioso Autónomo/fisiología , Humanos , Masculino , Músculo Esquelético/inervaciónRESUMEN
Between April 1984 and September 1988, preoperative radiotherapy or thermoradiotherapy was administered to 49 patients with bladder cancer (T1-4N0M0; UICC classification, 1987). Twenty-one patients were preoperatively treated by radiotherapy alone, with 4 Gy per fraction and three fractions per week to a total dose of 24 Gy (TDF = 53, group 1). The other 28 patients were treated by the same radiotherapy regimen in combination with hyperthermia (group 2). Regional hyperthermia was administered for 35-60 min immediately after irradiation (two sessions per week to a total of four sessions) using an 8 MHz RF capacitive heating device. Group 2 was divided into group 2 (high), in which the average intravesical temperature (T(av)) was > 41.5 degrees C, which was the mean value, and group 2 (low) with a T(av) < 41 x 5 degrees C. Group 2 (high) showed a significantly higher incidence of down-staging and tumour degeneration than both group 1 and group 2 (low). In addition, the local recurrence rate was lower and survival time was longer in group 2 than in group 1, although not significantly so. In particular, the patients with T3-4 or grade 3 bladder cancer in group 2 had a longer average survival than those in group 1, although the difference was not significant. The toxicity associated with hyperthermia was pain during treatment, and complications were not serious.
Asunto(s)
Carcinoma de Células Transicionales/radioterapia , Carcinoma de Células Transicionales/terapia , Hipertermia Inducida , Neoplasias de la Vejiga Urinaria/radioterapia , Neoplasias de la Vejiga Urinaria/terapia , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma de Células Transicionales/cirugía , Terapia Combinada , Femenino , Humanos , Hipertermia Inducida/efectos adversos , Masculino , Persona de Mediana Edad , Dolor/etiología , Pronóstico , Neoplasias de la Vejiga Urinaria/cirugíaRESUMEN
Fifty-seven Thai herbs and spices were examined for their retroviral reverse transcriptase inhibitory activity. All herbs and spices were extracted with hot-water and methanol. Reverse transcriptase inhibitory activity of the extracts was determined by using Moloney Murine Leukemia Virus reverse transcriptase (M-MuLV-RT) reacted with 3H-dTTP and radioactivity measured with a scintillation counter. Eighty-one per cent (46/57) of hot-water extracts and 54% (31/57) of methanol extracts showed inhibitory activities. At a concentration of 125 micrograms/ml, 13% (6/46) of hot-water extracts, namely Eugenia caryophyllus Bullock et Harrison, Phyllanthus urinaria Linn., Terminalia belerica Roxb., Nelumbo nucifera Gaertn., Psidium guajava Linn. and Lawsonia inermis Linn., had a relative inhibitory ratio (IR) over 50%. They showed ratios of 100%, 91%, 75%, 74%, 61% and 60%, respectively. For methanol extracts, only 10% (3/31) had IR values over 50%. They were T. belerica, E. caryophyllus and N. nucifera which exhibited IR values of 83%, 54% and 54%, respectively.
Asunto(s)
Inhibidores Enzimáticos/aislamiento & purificación , Plantas Medicinales/química , Retroviridae/enzimología , Inhibidores de la Transcriptasa Inversa , Especias/análisis , Animales , Virus de la Leucemia Murina/enzimología , Ratones , TailandiaRESUMEN
Forty patients with symptomatic benign prostatic hyperplasia were treated with a single session of transurethral microwave thermotherapy (TUMT) using a Prostatron. The clinical effectiveness was evaluated by analyzing the subjective and objective responses following the treatment. The International Prostate Symptom Score (I-PSS) and quality of life (QOL) scale were used to evaluate the subjective symptoms. At three months after treatment, significant improvements in I-PSS (p < 0.0001). QOL (P < 0.0001), and peak flow rate (Qmax) (p < 0.05) were observed. Improvement of both I-PSS and Qmax was found in 90% (18/20) of the patients at 2 months. Although 15 patients noted transient urinary retention and 15 patients had mild to moderate macroscopic hematuria immediately after TUMT, no severe adverse effects occurred during the follow-up period. A significant correlation was found between I-PSS improvement and the total thermal dose delivered. However, the thermal dose could not be predicted in each case. The preliminary findings suggest that TUMT by Prostatron is safe and effective as a nonsurgical treatment for benign prostatic hyperplasia. The clinical response seems to be thermal dose dependent. I-PSS is clinically sensitive and is useful in practice.