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BACKGROUND: The outcome of anti-tuberculosis treatment varies according to patient factors. OBJECTIVE: To retrospectively identify risks related to the extension of time to negative sputum culture (Tn) and to determine their clinical significance. DESIGN: Patients with bacilli susceptible to isoniazid and rifampicin who received initial standard treatment without cessation were recruited into the study. A total of 630 consecutive in-patients were included in the risk development analysis (development cohort) and another 611 consecutive in-patients in the risk validation analysis (validation cohort). RESULTS: Univariate analysis showed that Tn was related to sex, body mass index (BMI), white blood cell count (WBC), serum albumin, fasting blood sugar, haemoglobin A1c, C-reactive protein and total cholesterol levels and sputum smear positivity (SSP). Multivariate analysis showed that BMI, WBC and SSP were significant risk factors related to extended Tn. Optimal cut-offs of BMI and WBC for predicting good (Tn < 46 days) and poor responders (Tn ⩾ 46 days) according to each risk were determined by receiver operating characteristics analysis. Risks were verified with the validation cohort. Tn increased according to the number of risks; the median Tn for patients with three risks was 21 days longer than that of patients with none. CONCLUSION: The nutritional state of a TB patient can be used to predict Tn.
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Antituberculosos/uso terapéutico , Mycobacterium tuberculosis/aislamiento & purificación , Estado Nutricional , Esputo/microbiología , Tuberculosis/tratamiento farmacológico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/análisis , Índice de Masa Corporal , Proteína C-Reactiva/análisis , Femenino , Intolerancia a la Glucosa , Humanos , Isoniazida/uso terapéutico , Japón , Leucocitos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Estudios Retrospectivos , Rifampin/uso terapéutico , Factores de Riesgo , Adulto JovenRESUMEN
Epithelial cell scattering recapitulates the first steps of carcinoma invasion/metastasis. While the balance between cell-cell adhesive activity and cell motility ultimately determines this process, its molecular mechanisms remain unclear. Adherence junctions and tight junctions (TJs) are primarily responsible for cell-cell adhesive activity and subjected to dynamic remodeling. We previously showed that Rab13 and its effector protein JRAB/MICAL-L2 mediate the endocytic recycling of the integral TJ protein occludin and the assembly of functional TJs. In this study, we examined the role of Rab13 and JRAB/MICAL-L2 in the scattering of Madin-Darby canine kidney (MDCK) cells in response to 12-O-tetradecanoylphorbol-13-acetate (TPA). Knockdown of Rab13 in canine MDCK cells suppressed the TPA-induced scattering, and this phenotype was restored by re-expression of human Rab13. During TPA-induced MDCK cell scattering, Rab13 was transiently activated and returned to its basal level, and both Rab13 and JRAB/MICAL-L2 were colocalized with F-actin at cell-cell contact sites and then accumulated at emerging lamellipodial structures. TPA-induced MDCK cell scattering was also inhibited by knockdown of canine JRAB/MICAL-L2 and rescued by re-expression of mouse JRAB/MICAL-L2. These results indicate that Rab13 and JRAB/MICAL-L2 are involved in epithelial cell scattering.
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Proteínas del Citoesqueleto/fisiología , Células Epiteliales/metabolismo , Proteínas de Microfilamentos/fisiología , Proteínas de Unión al GTP rab/fisiología , Animales , Carcinógenos/farmacología , Adhesión Celular/efectos de los fármacos , Adhesión Celular/fisiología , Línea Celular , Movimiento Celular/efectos de los fármacos , Movimiento Celular/fisiología , Cricetinae , Perros , Células Epiteliales/efectos de los fármacos , Humanos , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
A local Schrödinger equation (LSE) method is proposed for solving the Schrödinger equation (SE) of general atoms and molecules without doing analytic integrations over the complement functions of the free ICI (iterative-complement-interaction) wave functions. Since the free ICI wave function is potentially exact, we can assume a flatness of its local energy. The variational principle is not applicable because the analytic integrations over the free ICI complement functions are very difficult for general atoms and molecules. The LSE method is applied to several 2 to 5 electron atoms and molecules, giving an accuracy of 10(-5) Hartree in total energy. The potential energy curves of H2 and LiH molecules are calculated precisely with the free ICI LSE method. The results show the high potentiality of the free ICI LSE method for developing accurate predictive quantum chemistry with the solutions of the SE.
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The photoelectron shake-up satellite spectra that accompany the C1s and O1s main lines of carbon monoxide have been studied by a combination of high-resolution x-ray photoelectron spectroscopy and accurate ab initio calculations. The symmetry-adapted cluster-expansion configuration-interaction general-R method satisfactorily reproduces the satellite spectra over a wide energy region, and the quantitative assignments are proposed for the 16 and 12 satellite bands for C1s and O1s spectra, respectively. Satellite peaks above the pi(-1)pi(*) transitions are mainly assigned to the Rydberg excitations accompanying the inner-shell ionization. Many shake-up states, which interact strongly with three-electron processes such as pi(-2)pi(*2) and n(-2)pi(*2), are calculated in the low-energy region, while the continuous Rydberg excitations are obtained with small intensities in the higher-energy region. The vibrational structures of low-lying shake-up states have been examined for both C1s and O1s ionizations. The vibrational structures appear in the low-lying C1s satellite states, and the symmetry-dependent angular distributions for the satellite emission have enabled the Sigma and Pi symmetries to be resolved. On the other hand, the potential curves of the low-lying O1s shake-up states are predicted to be weakly bound or repulsive.
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We have measured the vibrational structures of the N 1s photoelectron mainline and satellites of the gaseous N2 molecule with the resolution better than 75 meV. The gerade and ungerade symmetries of the core-ionized (mainline) states are resolved energetically, and symmetry-dependent angular distributions for the satellite emission allow us to resolve the Sigma and Pi symmetries of the shake-up (satellite) states. Symmetry-adapted cluster-expansion configuration-interaction calculations of the potential energy curves for the mainline and satellite states along with a Franck-Condon analysis well reproduce the observed vibrational excitation of the bands, illustrating that the theoretical calculations well predict the symmetry-dependent geometry relaxation effects. The energies of both mainline states and satellite states, as well as the splitting between the mainline gerade and ungerade states, are also well reproduced by the calculation: the splitting between the satellite gerade and ungerade states is calculated to be smaller than the experimental detection limit.
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A theoretical method for calculating magnetic circular dichroism (MCD) of molecules is presented. We examined the numerical accuracy and the stability of the finite perturbation (FP) method and the sum-over-state (SOS) perturbation method. The relativistic effects are shown to be important for the MCD spectra of molecules containing heavy elements. Calculations using the FP and the SOS methods were carried out for ethylene, para- and ortho-benzoquinone, showing that the FP method is superior to the SOS method, as expected. The relativistic effect was examined using the second-order Douglas-Kroll Hamiltonians for the halogen molecules F2, Cl2, Br2, and I2. The Faraday terms of I2 and Br2 were strongly affected by the relativistic effects, while the effect was negligible for Cl2 and F2.
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OBJECTIVE: We evaluated the efficacy of betamethasone ointment in the treatment of childhood phimosis and determined the most effective concentration of betamethasone. MATERIAL AND METHODS: Between August 1996 and May 2000, 69 patients (median age 3.0, range 0-12) presented to our department with severe phimosis (grade 4, Tsugaya's classification), and were enrolled in this study and treated with betamethasone ointment for 4 weeks. The ointment was applied on the narrowed preputial skin twice a day by parents. During the first week, parents were asked not to retract the foreskin. After this period, parents were encouraged to retract the foreskin gently and without intense tension. The concentration of betamethasone were 0.12%, 0.05% and 0.025%. If the patients become to be able to retract foreskin satisfactory, we consider the state as complete response. If he become to be able to retract to allow partial exposure of the glans, we consider the state as partial response. We compared the efficacy among each of the concentrations, and at 3 months after the end of the treatment, we also assessed the adverse effects and recurrence. RESULTS: The overall success rate was 85.5%. Success rates with 0.12%, 0.05% and 0.025% ointment were 96.8%, 82.8% and 55.6%, respectively (p = 0.0001). The 0.12% ointment was most effective. Recurrence was observed in 3 patients (4.3%). No adverse effects were observed in any of the patients. CONCLUSIONS: Treatment with betamethasone ointment is very effective, easy and safe. We recommend betamethasone ointment as the first treatment of choice for childhood phimosis.
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Betametasona/uso terapéutico , Fimosis/tratamiento farmacológico , Niño , Preescolar , Humanos , Lactante , Recién Nacido , Masculino , PomadasRESUMEN
Erection is a hemodynamic event and accordingly, erectile dysfunction (ED) is closely related with ischemic heart disease. We should confirm that the cardiac condition of the ED patient is safe enough to perform sexual intercourse prior to beginning treatment for ED. Asymptomatic ischemic heart disease cannot be diagnosed only in an interview, but it's difficult to perform cardiac exercise tests on all patients complaining of ED. Therefore, screening methods to evaluate patients who should undergo exercise tests are needed. Sixty patients with erectile dysfunction participated in this study. Physical examinations, interviews, and color Doppler examinations were conducted. Chest X-rays and electrocardiograms of all patients in the resting position were obtained, as were electrocardiograms following exercise. Echocardiograms, treadmill test results, thallium exercise scintigrams, and coronary angiograms were obtained as required for diagnosis. Two patients were excluded because they had obvious arteriogenic ED due to perineal injury. Fifty-eight patients underwent Doppler evaluations of their cavernous arteries and heart exercise tests. Fourteen patients (24.1%) were diagnosed with ischemic heart disease. Although six of them had already been diagnosed with ischemic heart disease, eight were newly diagnosed by the exercise tests. Cardiovascular risk factors such as advanced age, hyperlipidemia, diabetes mellitus, hypertension, smoking, and obesity were not sufficient predictive factors. The mean peak systolic velocity of the patients without ischemic heart disease was 34.6 cm/s vs 22.0 cm/s in those with ischemic heart disease. Only 3.7% of patients whose peak systolic velocity in the cavernous artery was equal to or exceeded 35 cm/s had ischemic heart disease. On the other hand, 41.9% of patients with peak systolic velocity of less than 35 cm/s had ischemic heart disease. The sensitivity of peak systolic velocity against ischemic heart disease was 92.9%, and specificity was 59.1%. In ED patients, incidences of complications involving symptomatic or asymptomatic ischemic heart disease were found to be high. The peak systolic velocity in the cavernous artery is thought to be a useful predictive factor of ischemic heart disease in ED patients. When a patient reveals a peak systolic velocity of less than 35 cm/s, he should undergo heart exercise tests prior to treatment of ED.
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Disfunción Eréctil/complicaciones , Isquemia Miocárdica/complicaciones , Isquemia Miocárdica/diagnóstico , Pene/irrigación sanguínea , Adulto , Anciano , Arterias/diagnóstico por imagen , Arterias/fisiopatología , Velocidad del Flujo Sanguíneo , Prueba de Esfuerzo , Humanos , Masculino , Persona de Mediana Edad , Flujo Sanguíneo Regional , Sensibilidad y Especificidad , Sístole , Ultrasonografía Doppler en ColorRESUMEN
BACKGROUND: CT angiography reconstructed by a multidetector-row helical CT scanner is a newly developed form of imaging. We compared CT angiography and ultrasonic Doppler examination with digital subtraction angiography (DSA) in the diagnosis of arterial lesion. METHOD: Eighteen patients with arteriogenic erectile dysfunction (ED) underwent color Doppler study, DSA, and CT angiography after providing informed consent. The CT angiography images were obtained by a multidetector-row helical CT scanner, Asteion TSX021A (TOSHIBA). We injected prostaglandin E1 into the penile cavernous body, and then rapidly infused nonionic contrast medium into the antecubital vein. DSA and CT angiography images of the bilateral internal pudendal arteries and cavernous arteries were examined for stenotic lesions or occlusion. We also compared the peak systolic blood flow velocity in the cavernous artery measured by color Doppler ultrasound with CT angiography and DSA. RESULTS: The CT angiography and color Doppler studies were performed on an outpatient basis, but DSA required hospitalization. In the 36 internal pudendal arteries, DSA represented 22 normal arteries and 14 stenosis or occlusions. CT angiography showed 15 normal arteries and 21 occlusions. For the diagnosis of stenosis or occlusion in the internal pudendal artery, the CT angiography image had a good agreement, with a sensitivity of 1.00, specificity of 0.68, and accuracy of 0.81. For diagnosis in the cavernous artery, CT angiography image also showed a good agreement with DSA; however, the quality of the images of fine arteries was better in the DSA images. The inferior view and internal view of the pelvis in CT angiography were helpful for visualizing the internal pudendal artery, especially at the pubic bone. There was insufficient correlation between peak systolic blood flow velocity and DSA findings. There were no serious complications involved in either examination. CONCLUSIONS: CT angiography has not yet reached the same level as DSA in the evaluation of fine arteries. However, CT angiography can produced images sufficient for the diagnosis of arteriogenic ED with some advantages. We believe that with improvement, CT angiography will become an adequate replacement for DSA in the diagnosis of penile arterisl lesion.
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Angiografía de Substracción Digital , Impotencia Vasculogénica/diagnóstico , Pene/irrigación sanguínea , Tomografía Computarizada por Rayos X/métodos , Ultrasonografía Doppler , Adulto , Angiografía , Arterias/patología , Humanos , Masculino , Persona de Mediana Edad , Pene/diagnóstico por imagenRESUMEN
BACKGROUND: We report the results of microscopic penile revascularization in patients with arteriogenic erectile dysfunction. METHODS: One patient with localized obstruction of the common penile artery underwent the Michal II penile revascularization, 13 patients underwent the Furlow-Fisher procedure, and 5 patients underwent the Hauri procedure. The mean age was 33.0 years and the mean follow-up period was 32 months (4-80 months). Eight patients were tobacco smokers, one patient was over 50 years old. Surgery was considered successful when the patients had a permeable anastomosis and were able to achieve satisfactory erection resulting in normal sexual intercourse. RESULTS: All surgery was successful except for one patient who had undergone the Furlow-Fisher procedure. In Spite of antithrombotic therapy, graft occlusion occurred in two patients. Post operative glans hypervascularity occurred in two patients. CONCLUSION: Penile revascularization surgery is a highly effective treatment for selected patients. There is a need for further study of graft occlusion and glans hypervascularity.
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Implantación de Prótesis Vascular/métodos , Disfunción Eréctil/cirugía , Microcirugia/métodos , Pene/irrigación sanguínea , Adulto , Anastomosis Quirúrgica , Arteriopatías Oclusivas/complicaciones , Arterias/cirugía , Disfunción Eréctil/etiología , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Factores de Tiempo , Resultado del TratamientoRESUMEN
BACKGROUND: Erection is an hemodynamic event and accordingly, erectile dysfunction is closely related with ischemic heart diseases. Sufficient study has yet to be made of this relation. METHOD: Fifty-eight patients with erectile dysfunction participated in the present study. Intracavernous injection tests and color Doppler examination were conducted for assessment of erectile function. For patients with history of ischemic heart disease, we referred to their doctor for data from their examinations. For patients otherwise, chest X-rays and electrocardiograms in the resting position were obtained and also electrocardiograms following exercise. Echocardiograms, treadmill test results, thallium exercise scintigrams, and coronary angiograms were obtained as requires for diagnosis. RESULTS: Eighteen patients (31.0%) were diagnosed as ischemic heart disease or at high risk. Although 6 of them had been already diagnosed with ischemic heart disease, 12 were newly diagnosed by the exercise tests. Among these 18 patients 16 had one or more significant risk factors for ischemic heart disease such as age, hyperlipidemia, and diabetes mellitus. 3.7% of patients whose peak systolic velocity in the cavernous artery was equal to or exceeded 35 cm/sec had ischemic heart disease or were at high risk. 54.8% of patients with peak systolic velocity of less than 35 cm/sec had ischemic heart disease or were at high risk. CONCLUSIONS: In erectile dysfunctional patients, incidence of complications with symptomatic or asymptomatic ischemic heart disease was found to be high. Therefore, in patients with risk factors or low peak systolic velocity in the cavernous artery, exercise tests should be implemented prior to treatment of erectile dysfunction.
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Disfunción Eréctil/complicaciones , Isquemia Miocárdica/complicaciones , Anciano , Angiografía Coronaria , Disfunción Eréctil/fisiopatología , Prueba de Esfuerzo , Hemodinámica , Humanos , Masculino , Persona de Mediana Edad , Isquemia Miocárdica/diagnóstico , Isquemia Miocárdica/fisiopatología , Pene/irrigación sanguíneaRESUMEN
We report here three cases of bilateral synchronous renal cell carcinoma. One of the 3 patients underwent bilateral partial nephrectomy, while the other 2 underwent combined partial nephrectomy and radical nephrectomy. All patients received adjuvant therapy of interferon-alpha and tegafur uracil. In the management of synchronous bilateral renal cell carcinoma, we discussed the selection of surgical procedure for primary lesions, i.e., based on the renal function of both sides, and the necessity of adjuvant therapy in such cases.
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Carcinoma de Células Renales/cirugía , Neoplasias Renales/cirugía , Anciano , Antineoplásicos/uso terapéutico , Carcinoma de Células Renales/tratamiento farmacológico , Quimioterapia Adyuvante , Femenino , Humanos , Interferón-alfa/uso terapéutico , Neoplasias Renales/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Nefrectomía/métodos , Tegafur/uso terapéutico , Resultado del TratamientoRESUMEN
Forty-five patients with muscle-invasive bladder cancer treated with intra-arterial doxorubicin chemotherapy plus low-dose radiotherapy between September 1979 and March 1990 were retrospectively studied. Twenty-eight (62%) patients achieved a complete response (CR) and in all of them, a functional bladder could be preserved. The 10-year cause-specific survival rate of patients with CR was 95.5%, but that of patients not achieving a CR was 39%. These results demonstrate that in patients who achieve a CR with this treatment, we may be able to preserve a functional bladder. In a prospective study, we designed a new intra-arterial chemotherapy regimen in order to achieve a higher degree of effectiveness and to preserve a functional bladder. Twenty-three patients were treated with concurrent pirarubicin/cisplatin intra-arterial chemotherapy and radiotherapy after complete transurethral resection. Twenty-one (91%) patients achieved CR. One of these patients had relapse with lung metastases and was treated surgically. Two patients who did not achieve a CR died of cancer, and 21 patients are alive with preservation of functional bladder. For treatment of prostate cancer, we now administer only adjuvant intra-arterial chemotherapy plus irradiation for patients after radical prostatectomy.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/radioterapia , Neoplasias de la Vejiga Urinaria/tratamiento farmacológico , Neoplasias de la Vejiga Urinaria/radioterapia , Adulto , Anciano , Anciano de 80 o más Años , Cisplatino/administración & dosificación , Terapia Combinada , Doxorrubicina/administración & dosificación , Doxorrubicina/análogos & derivados , Esquema de Medicación , Femenino , Humanos , Infusiones Intraarteriales , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Estudios RetrospectivosRESUMEN
Eight patients with locally advanced bladder cancer who were not candidates for radical cystectomy or concurrent intra-arterial chemotherapy and radiotherapy were treated with combined platinum-based chemotherapy and radiation therapy. Six of the eight patients (75%) achieved a clinical complete response (CR). The effect of therapy in four patients whose histopathological responses were evaluated was effect grade 3. One of the eight patients died of treatment-related myelosuppression. The other two patients died of intercurrent disease, while the remaining five patients are alive with preservation of a functional bladder. The 2-year overall survival rate was 87.5%. Adverse events due to chemotherapy were mainly bone marrow suppression. Those caused by radiation therapy were rectal irritability. We considered concurrent platinum-based chemotherapy and radiotherapy useful for the treatment of locally advanced bladder cancer.
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Antineoplásicos/uso terapéutico , Compuestos Organoplatinos/uso terapéutico , Neoplasias de la Vejiga Urinaria/terapia , Anciano , Anciano de 80 o más Años , Carboplatino/uso terapéutico , Cisplatino/uso terapéutico , Terapia Combinada , Humanos , Terapia de Inmunosupresión/efectos adversos , Tasa de Supervivencia , Neoplasias de la Vejiga Urinaria/mortalidadRESUMEN
A 57-year-old man was admitted with the chief complaint of macrohematuria. Digital rectal examination showed a slightly enlarged, irregular prostate with stony consistency. Serum levels of prostate specific antigen (PSA), neuron-specific enolase (NSE) and progastrin-releasing peptide (ProGRP) were elevated. Transurethral resection (TUR)-biopsy of the prostate revealed small cell carcinoma with poorly differentiated adenocarcinoma. Various radiological examinations revealed metastases to pelvic lymph nodes and liver. He was treated with chemoendocrine therapy consisting of cisplatin, etoposide, flutamide and luleinizing hormone-releasing hormone (LH-RH) agonist. The primary tumor and metastatic lesion decreased and serum PSA, NSE and ProGRP levels were decreased to normal ranges after 5 cycles of chemotherapy. After the 5-cycle chemotherapy, TUR-biopsy proved viable tumor cells. During the additional chemotherapy, tumor markers increased and 4 months later liver metastasis progressed. He died 13 months after diagnosis of small cell carcinoma of the prostate.
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Carcinoma de Células Pequeñas/diagnóstico , Neoplasias de la Próstata/diagnóstico , Carcinoma de Células Pequeñas/patología , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Fosfopiruvato Hidratasa/sangre , Antígeno Prostático Específico/sangre , Neoplasias de la Próstata/patologíaRESUMEN
The hepatic synthesis of acute phase proteins in ruminants has been suggested to be regulated by some mechanisms different from those in other species such as rodents and human. To explore possible regulatory factors unique to ruminants, we examined effects of interleukin (IL)-6, IL-1 and tumor necrosis factor (TNF), on haptoglobin (Hp) synthesis using a primary culture system of bovine hepatocytes. After bovine primary cultured hepatocytes were incubated in the presence of various concentrations of the cytokines, the synthesis and mRNA level of haptoglobin and albumin were measured by labeling with [35S]-methionine and immunoprecipitation, and by Northern blot analysis, respectively. Hp synthesis was dose-dependently increased by recombinant human (rh) IL-6, and also by rhTNF-alpha, but to a less extent, while it was not affected by rhIL-1 beta. The stimulatory effect is mainly pretranslational, because mRNA level of Hp changed in parallel with protein synthesis. In contrast, albumin synthesis was suppressed by these three cytokines similarly. These results are inconsistent with the previously proposed view that TNF and IL-1 overlap in their pathways leading to the transcriptional activation of many acute phase protein genes. In conclusion, there is a species-specific unique signaling system, especially for TNF, in transcriptional activation of bovine Hp gene.
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Expresión Génica/efectos de los fármacos , Haptoglobinas/biosíntesis , Interleucina-1/farmacología , Interleucina-6/farmacología , Hígado/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Envejecimiento/metabolismo , Animales , Animales Recién Nacidos , Northern Blotting , Bovinos , Células Cultivadas , Femenino , Haptoglobinas/aislamiento & purificación , Humanos , Hígado/efectos de los fármacos , Hígado/crecimiento & desarrollo , Biosíntesis de Proteínas , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Proteínas Recombinantes/farmacología , Albúmina Sérica/biosíntesisRESUMEN
A special bed and a new software program were developed for noninvasive regional cerebral blood flow (rCBF) measurements by 99mTc hexamethylpropylene amine oxime (99mTc-HMPAO) using only three-head SPECT system, Siemens MULTI-SPECT3. A large field gamma camera with a minimum of 38 cm-diameter is required for simultaneous acquisition of time activity curves of brain and aortic arch in this noninvasive method. A special bed with short width was designed and inserted obliquely into the gantry of the SPECT system to acquire both time activity curves of brain and aortic arch with the use of diagonal length (51 cm) of a gamma camera with 31 x 41 cm width. Moreover a software program was developed for processing a series of the graphical analysis of time activity curves, Lassen's linearization correction of SPECT images, and drawing of a rCBF map. This noninvasive rCBF measurement using only MULTISPECT3 in a short period with high spatial resolution is quite useful for routine clinical studies.
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Encéfalo/diagnóstico por imagen , Circulación Cerebrovascular , Compuestos de Organotecnecio , Oximas , Tomografía Computarizada de Emisión de Fotón Único/métodos , Humanos , Exametazima de Tecnecio Tc 99mRESUMEN
We describe a technique for isolation and primary culture of bovine hepatocytes, and their metabolic characterization. Hepatocytes were isolated from the caudate lobe of bovine liver by perfusion with 0.25 mM ethylene-glycol tetraacetic acid and 0.05% collagenase. The viability and yield of the cells were 70-92% and 0.1-3.6 x 10(7) cells/g liver, respectively. When the isolated hepatocytes were cultured in Williams' medium E, they began to spread in 3 hr and formed monolayers in 24 hr. These monolayers were retained for at least 6 days. To monitor the metabolic activities specific to liver, synthesis and secretion of albumin were measured by labeling with [35S]-methionine and immunoprecipitation. This activity was low in isolated hepatocytes, but increased after culturing 1-3 days, and decreased again after 6 days. Glycogenolytic activity was also assessed by measuring glucose release to the medium by stimulation with epinephrine. The glycogenolytic response to epinephrine was also enhanced by culturing the hepatocytes 1-3 days, but was decreased after 6 days. Since the isolated bovine hepatocytes retained the liver-specific activities of albumin synthesis and glycogenolysis for several days in culture, these cells are useful for cellular and molecular studies on the functions of bovine liver.