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SETTING: Four tuberculosis (TB) diagnostic health facilities of the Chongwe District, Zambia. OBJECTIVE: To determine the frequency of bacteriologically confirmed TB patients lost to follow-up (LTFU) before treatment from January to December 2017. DESIGN: This is a retrospective cohort study involving the review of TB registers. Information on presumptive TB patients who tested positive either by smear microscopy or Xpert® MTB/RIF assay was extracted from the laboratory TB registers of the TB diagnostic facilities and cross-matched with the TB treatment registers of TB treatment facilities. RESULTS: Two hundred and seventeen bacteriologically confirmed TB patients were found in the laboratory TB registers. Of these, 145 (67%) were males and seven (3%) were children; 177 (81%) patients were diagnosed using Xpert, while the remaining 40 (19%) were diagnosed using sputum smear microscopy. A total of 71 (33%) were not linked to treatment. Those diagnosed using smear microscopy were 2.5 times (95% CI 1.1-5.3) more likely to be LTFU before treatment than those diagnosed using Xpert. CONCLUSION: About one third of TB patients who were not linked to treatment could potentially extend the duration of bacilli transmission in their communities. National TB control programmes should consider including LTFU patients before treatment in routine monitoring and evaluation.
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Facial nerve paralysis (FNP) after super-selective intra-arterial chemotherapy (SSIAC) is a relatively rare local side effect of SSIAC to the maxillary artery (MA) or the middle meningeal artery (MMA). The incidence and prognosis of FNP after SSIAC in 381 patients with oral cancer (133 with catheterization of the MA, 248 without) was investigated retrospectively. Only three patients (two male and one female) had FNP, for an incidence of 0.8%. All patients with FNP had undergone catheterization of the MA, and the incidence of FNP in this group was 2.3% (3/133). One of the three patients with FNP had paralysis of the third branch of the trigeminal nerve. FNP occurred a mean of 8.7 days (range 5-11 days) after initial SSIAC, and the mean total dose of cisplatin was 55.8mg (range 42.5-67.2mg) and of docetaxel was 25.4mg (range 17.0-33.6mg). FNP resolved completely a mean of 12.7 months (range 6-19 months) after onset. Because the administration of anticancer agents via the MA or MMA carries a risk of FNP, this information will be useful when obtaining informed consent from patients before treatment.
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Antineoplásicos/administración & dosificación , Antineoplásicos/efectos adversos , Cateterismo/efectos adversos , Parálisis Facial/inducido químicamente , Neoplasias de la Boca/tratamiento farmacológico , Anciano , Parálisis Facial/epidemiología , Femenino , Humanos , Incidencia , Masculino , Arteria Maxilar , Arterias Meníngeas , Persona de Mediana Edad , Neoplasias de la Boca/radioterapia , Pronóstico , Estudios RetrospectivosRESUMEN
We developed a cryogenic system on a rotating table that achieves sub-Kelvin conditions. The cryogenic system consists of a helium sorption cooler and a pulse tube cooler in a cryostat mounted on a rotating table. Two rotary-joint connectors for electricity and helium gas circulation enable the coolers to be operated and maintained with ease. We performed cool-down tests under a condition of continuous rotation at 20 rpm. We obtained a temperature of 0.23 K with a holding time of more than 24 h, thus complying with catalog specifications. We monitored the system's performance for four weeks; two weeks with and without rotation. A few-percent difference in conditions was observed between these two states. Most applications can tolerate such a slight difference. The technology developed is useful for various scientific applications requiring sub-Kelvin conditions on rotating platforms.
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PURPOSE: The aim of this study was to review the efficacy and toxicity of radiation therapy with concurrent retrograde superselective intra-arterial chemotherapy in the treatment of gingival carcinoma. METHODS AND MATERIALS: In all, 34 patients (21 men and 13 women) with squamous cell carcinoma of the gingiva underwent radiation therapy with concurrent retrograde superselective intra-arterial chemotherapy. Treatment consisted of daily external irradiation and concurrent retrograde superselective intra-arterial infusion with cisplatin and docetaxel. A median total dose of 60 Gy in 30 fractions was delivered to tumors. RESULTS: Of the 34 patients, 29 (85 %) achieved a complete response (CR) and 5 had residual tumors. Of the 29 patients with a CR, 2 had local recurrences and 1 had distant metastasis 1-15 months after treatment. Twenty-six of the 36 patients had survived at a median follow-up time of 36 months (range 12-79 months); 4 died of cancer and 4 died of non-cancer-related causes. At both 3 and 5 years after treatment, the overall survival rates were 79 % and the cause-specific survival rates were 85 %. Osteoradionecrosis of the mandibular bone only developed in 1 patient after treatment. CONCLUSION: Radiation therapy with concurrent retrograde superselective intra-arterial chemotherapy was effective and safe in the treatment of gingival carcinoma. This treatment may be a promising curative and organ-preserving treatment option for gingival carcinoma.
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Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Carcinoma de Células Escamosas/terapia , Quimioradioterapia , Neoplasias Gingivales/terapia , Infusiones Intraarteriales , Anciano , Anciano de 80 o más Años , Carcinoma de Células Escamosas/mortalidad , Carcinoma de Células Escamosas/patología , Cisplatino/administración & dosificación , Terapia Combinada , Progresión de la Enfermedad , Docetaxel , Femenino , Neoplasias Gingivales/mortalidad , Neoplasias Gingivales/patología , Humanos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Tratamientos Conservadores del Órgano , Tasa de Supervivencia , Taxoides/administración & dosificaciónRESUMEN
In the field of radiowave detection, enlarging the receiver aperture to enhance the amount of light detected is essential for greater scientific achievements. One challenge in using radio transmittable apertures is keeping the detectors cool. This is because transparency to thermal radiation above the radio frequency range increases the thermal load. In shielding from thermal radiation, a general strategy is to install thermal filters in the light path between aperture and detectors. However, there is difficulty in fabricating metal mesh filters of large diameters. It is also difficult to maintain large diameter absorptive-type filters in cold because of their limited thermal conductance. A technology that maintains cold conditions while allowing larger apertures has been long-awaited. We propose radio-transparent multi-layer insulation (RT-MLI) composed from a set of stacked insulating layers. The insulator is transparent to radio frequencies, but not transparent to infrared radiation. The basic idea for cooling is similar to conventional multi-layer insulation. It leads to a reduction in thermal radiation while maintaining a uniform surface temperature. The advantage of this technique over other filter types is that no thermal links are required. As insulator material, we used foamed polystyrene; its low index of refraction makes an anti-reflection coating unnecessary. We measured the basic performance of RT-MLI to confirm that thermal loads are lowered with more layers. We also confirmed that our RT-MLI has high transmittance to radiowaves, but blocks infrared radiation. For example, RT-MLI with 12 layers has a transmittance greater than 95% (lower than 1%) below 200 GHz (above 4 THz). We demonstrated its effects in a system with absorptive-type filters, where aperture diameters were 200 mm. Low temperatures were successfully maintained for the filters. We conclude that this technology significantly enhances the cooling of radiowave receivers, and is particularly suitable for large-aperture systems. This technology is expected to be applicable to various fields, including radio astronomy, geo-environmental assessment, and radar systems.
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We developed a system that continuously maintains a cryocooler for long periods on a rotating table. A cryostat that holds the cryocooler is set on the table. A compressor is located on the ground and supplies high-purity (>99.999%) and high-pressure (1.7 MPa) helium gas and electricity to the cryocooler. The operation of the cryocooler and other instruments requires the development of interface components between the ground and rotating table. A combination of access holes at the center of the table and two rotary joints allows simultaneous circulation of electricity and helium gas. The developed system provides two innovative functions under the rotating condition, cooling from room temperature and the maintenance of a cold condition for long periods. We have confirmed these abilities as well as temperature stability under a condition of continuous rotation at 20 rpm. The developed system can be applied in various fields, e.g., in tests of Lorentz invariance, searches for axion, radio astronomy, and cosmology, and application of radar systems. In particular, there is a plan to use this system for a radio telescope observing cosmic microwave background radiation.
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OBJECTIVES: We examined gender and age differences in the impact of multiple role occupancy on health-related behaviours and health status among working age Japanese adults. METHODS: We analysed the individually linked, nationally representative data of 5693 respondents aged 20-59, who completed the Comprehensive Survey of the Living Conditions of People on Health and Welfare and the National Nutrition Survey, conducted by the Japanese government in 1995. RESULTS: Younger women benefited from multiple roles (less smoking), while younger men demonstrated more high-risk behaviours (more smoking, heavier drinking). By contrast, middle-aged men benefited from multiple roles (less smoking, fewer health problems), while middle-aged women reported lower health maintenance behaviours (less exercise, fewer health check-ups). CONCLUSIONS: Japanese society appears to be undergoing a transition in gender roles, as reflected by age and gender differences in the impact of multiple roles on health and health-related behaviours. Middle-aged males benefit from multiple roles (being the primary bread-winner and being married), while middle-aged women do not. This pattern seems to break down for younger Japanese men and women.
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Composición Familiar/etnología , Identidad de Género , Conductas Relacionadas con la Salud/etnología , Psicología Social , Adulto , Factores de Edad , Cuidadores , Empleo , Femenino , Promoción de la Salud , Humanos , Japón , Masculino , Persona de Mediana Edad , Morbilidad , Responsabilidad Parental , Medición de Riesgo , Factores de Riesgo , Factores Sexuales , Estrés Psicológico , Mujeres Trabajadoras/psicologíaRESUMEN
PURPOSE: To investigate the relationship between skipping breakfast and cardiovascular disease risk factors such as blood pressure, serum lipids, smoking, and lock of exercise. MATERIALS AND METHODS: Data from the National Nutrition survey conducted by the Ministry of Health and Welfare, from 1995 through 1997 were employed for the analysis. Data were used only if subjects were between 20 and 59 years old and had participated in the nutrition survey, and values for height and weight, blood pressure, serum lipids, blood sugar, number of steps per day, and smoking, drinking, and exercise habits were available. The number of subjects with data available for all of the above was 11,778 (4,438 men and 7,340 women). Analyses were performed separately by sex. The relationship between skipping breakfast and continuous variables such as nutrient intake ratio (nutrient intake divided by recommended dietary allowances), body mass index, blood pressure, serum lipids, blood sugar, and the number of steps per day was examined controlling for age using analysis of covariance (ANCOVA). The relationship between skipping breakfast and categorical variables such as smoking, drinking, and exercise habits was assessed controlling for age using Cochran-Mantel-Haenszel statistics. RESULTS: The younger the generation, the higher the rate of skipping breakfast. The rate of skipping breakfast in men was twice as high as in women. The breakfast skippers tended to intake less energy and calcium, both in men and women, that non-skippers. Female skippers tended to intake less iron and vitamin D. The number of steps per day was smaller in breakfast skippers than that in non-skippers both in men and women. Male skippers tended to have higher blood pressure than non-skippers, and female skippers tended to have a higher serum total cholesterol level than non-skippers. The breakfast skippers tended to smoke more than non-skippers, both in men and women, and female skippers tended to drink more alcohol and take less exercise. CONCLUSIONS: Skipping breakfast is related to cardiovascular risk factors such as lack of exercise, smoking, high blood pressure, and high serum total cholesterol.
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Enfermedades Cardiovasculares/epidemiología , Conducta Alimentaria , Encuestas Nutricionales , Adulto , Factores de Edad , Consumo de Bebidas Alcohólicas , Presión Sanguínea , Enfermedades Cardiovasculares/etiología , Colesterol/sangre , Ejercicio Físico , Femenino , Humanos , Japón/epidemiología , Masculino , Persona de Mediana Edad , Factores de Riesgo , Factores Sexuales , FumarRESUMEN
OBJECTIVES: The purpose of the present study was to examine the long-term effect of pumping injection of sodium hyaluronate into the TMJ in patients with non-reducing disc displacement. PATIENTS: Sixty patients with non-reducing disc displacement underwent pumping injection of sodium hyaluronate (pumping group). Seventy-six patients with non-reducing disc displacements were observed without any active treatment (observation group). STUDY DESIGN: In both patient groups clinical signs and symptoms were observed periodically for 2 years. Variables such as age, range of maximum mouth opening, angle of posterior slope of the articular eminence and degenerative bony changes of the condyle at the initial visit were also examined. Cox hazards analysis was applied to examine the clinical outcome for such variables in addition to the results of pumping injection of sodium hyaluronate. RESULTS: Pumping injection of sodium hyaluronate seemed to have a favourable effect when compared with the control group (untreated) (p = 0.0002). However, the four background variables mentioned could not be explained as predictors of outcome. CONCLUSION: Pumping injection of sodium hyaluronate seems to be effective for non-reducing disc displacement of the temporomandibular joint.
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Ácido Hialurónico/uso terapéutico , Luxaciones Articulares/tratamiento farmacológico , Disco de la Articulación Temporomandibular/efectos de los fármacos , Adolescente , Adulto , Factores de Edad , Anciano , Niño , Estudios de Cohortes , Femenino , Estudios de Seguimiento , Humanos , Ácido Hialurónico/administración & dosificación , Inyecciones Intraarticulares , Luxaciones Articulares/fisiopatología , Estudios Longitudinales , Masculino , Cóndilo Mandibular/patología , Persona de Mediana Edad , Osteoartritis/clasificación , Modelos de Riesgos Proporcionales , Rango del Movimiento Articular/fisiología , Estudios Retrospectivos , Factores Sexuales , Tensoactivos/administración & dosificación , Tensoactivos/uso terapéutico , Hueso Temporal/patología , Disco de la Articulación Temporomandibular/fisiopatología , Trastornos de la Articulación Temporomandibular/clasificación , Resultado del TratamientoRESUMEN
In the early stage of ripening of cherry-tomato fruits (Lycopersicon esculentum var. cherry), the lectin activity increased logarithmically and reached a plateau at day 10 after flowering. During purification of lectin from ripe and unripe fruits, a 42-kDa protein was found abundantly in unripe fruits. The protein cross-reacted with anti-cherry-tomato-lectin serum, retained chitin-binding ability, but showed no lectin activity. Comparative studies between the structure of the lectin and the 42-kDa protein were done. N-Terminal amino acid sequences of the lectin, peptides derived from the S-pyridylethylated lectin, and fragments generated by limited proteolysis of the native lectin showed that the lectin was comprised of three domains, Hyp-rich, Cys-rich, and Gln-rich, and the alignment of them was as this order from the N-terminus. Studies on the 42-kDa protein showed that it contained two of the three domains, Cys-rich and Gln-rich, but the amino acid sequence analysis showed that the protein should be a product of another gene.
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Lectinas/química , Proteínas de Plantas/química , Estructura Terciaria de Proteína , Solanum lycopersicum/química , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Aminoácidos/química , Quitina/metabolismo , Frutas/química , Immunoblotting , Lectinas/genética , Lectinas/aislamiento & purificación , Lectinas/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/fisiología , Datos de Secuencia Molecular , Lectinas de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
Methods of electromigration in laboratory apparatus of small-bore size have recently undergone development at a remarkably rapid pace, leading to a variety of new analytical techniques. One such technique is called "capillary electrophoresis" (CE), which is further classified on the basis of electromigration mode, viz., "capillary zone electrophoresis" (CZE), which, in turn, has several variations. This review aims to give a short overview of the various electromigration methods for amino compounds by using CE. Firstly, this review briefly summarizes the detection methods employed for detection of monoamines and polyamines by CE for both native and derivative forms. Next, current CE methods are described, and their applications to detection of amino acids, biogenic amines, aromatic amines, including heteroaromatic amines and their enantiomers, are introduced from representative papers. Finally, new methods for single-cell analysis and microchip CE techniques are focused on.
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Aminas/análisis , Aminoácidos/análisis , Monoaminas Biogénicas/análisis , Cromatografía Liquida/métodos , Electroforesis Capilar/métodosRESUMEN
We report a case of pneumoperitoneum at the end stage of lung cancer. The patient was an 81-year-old man who experienced only mild abdominal pain and fullness. The pneumoperitoneum spontaneously disappeared. Ruptured viscera and peritonitis were not observed at autopsy. We speculated that air had moved from the lung cancer lesion to the mediastinum, and then through the retroperitoneal space into the peritoneal space. When pneumoperitoneum is observed, lung cancer and other intrathoracic diseases should be suspected as causes. Also, it may be necessary to carefully examine the patient for fever, general appearance, abdominal findings, and laboratory findings, including white blood cell count and C-reactive protein level, and on that basis differentiate from ruptured viscus, which requires an emergency laparotomy.
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Carcinoma de Células Escamosas/complicaciones , Neoplasias Pulmonares/complicaciones , Pulmón/patología , Neumoperitoneo/etiología , Anciano , Anciano de 80 o más Años , Carcinoma de Células Escamosas/patología , Humanos , Neoplasias Pulmonares/patología , MasculinoRESUMEN
At the present time, the origin of protein bound D-amino acid (AA) has been fairly well elucidated, but that of free D-AA is still not well understood. To gain greater understanding of this, intestinal absorption in rats of free D,L-AA enantiomers (arginine, alanine and aspartic acid as models for basic, neutral and acidic AAs, respectively, in this study) and the relationship between age and absorption were investigated. The degree of rat intestinal absorption of free D,L-AAs was evaluated using apparent membrane permeability coefficients (Papp) which were obtained from an in situ intestinal single-pass perfusion method with Krebs-Ringer bicarbonate buffer (pH 7.4) solution containing D,L-AA enantiomers. Determinations of D,L-AA enantiomers in perfusion (in- and outflow) solutions were carried out by the in-capillary derivatization high-performance capillary electrophoretic methods (ICD-HPCE methods) that were previously developed by our group. Collectively, our observations suggest: (1) that the Papp of L-AA is higher than that of the D-isomer; (2) that D-AA can be absorbed as well as L-AA using a sodium ion-dependent transporter that is located on the brush border membrane of rat intestinal epithelial cells; (3) that Papp reached a maximum at 8 weeks of age, but were measured at decreased amounts at 52 and 104 weeks of age. These results suggest that free D-AA in a mammalian body originates from 'exogenous sources'.
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Envejecimiento/metabolismo , Aminoácidos/metabolismo , Absorción Intestinal , Animales , Transporte Biológico , Masculino , Perfusión , Ratas , Ratas Wistar , Sodio/metabolismo , EstereoisomerismoRESUMEN
T2* weighted gradient echo image has heightened diagnostic sensitivity to hemorrhage, which is attributed to magnetic susceptibility-induced static field inhomogeneities arising from paramagnetic blood breakdown products such as hemosiderin, which shorten T2*. We examined 4 cases of head injury in chronic stage by T2* weighted images and assessed the clinical application of this sequence for diagnosis of the intracranial lesion. All patients underwent CT scan on acute stage and long-term follow up was performed. In all cases, abnormally low signals in the brain and subarachnoid space were more conspicuous with the T2* weighted image than with any of the conventional sequences. In 2 cases with diffuse axonal injury, of which lesions were not detected on acute stage CT scan, hypointensity area was clearly demonstrated in the cerebral peduncle and corpus callosum on T2* weighted images. Selection of T2* weighted image into the routine MR examination of patients with chronic stage of the head injury is recommended.
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Traumatismos Craneocerebrales/diagnóstico , Imagen por Resonancia Magnética/métodos , Adulto , Enfermedad Crónica , Femenino , Humanos , Masculino , Tomografía Computarizada por Rayos XRESUMEN
Structural analysis of the sugar chains of human chorionic gonadotropin (hCG) has revealed that abnormal biantennary structures appear specifically on hCG in the urine of choriocarcinoma patients. However, the enzymatic and molecular mechanisms of the biosynthesis of abnormal biantennary sugar chains have not yet been elucidated. In this report, the enzyme activities and the expression levels of mRNAs of N-acetylglucosaminyltransferases (GnT)-I to -V, beta-1,4-galactosyltransferase, and alpha-mannosidase II in normal human placentae and three human choriocarcinoma cell lines were investigated. GnT-IV activities in choriocarcinoma cell lines were increased from 16- to 66-fold and GnT-III activity was increased from 15- to 25-fold as compared with those in human placentae, whereas other enzyme activities were not increased significantly. The mRNA expression levels generally correlated with their enzyme activities. Among the two GnT-IV genes found in human tissues only GnT-IVa gene was strongly expressed in the cancer cells: from three to seven times as much as in the normal tissue, whereas that of GnT-IVb remained constant. On the basis of these results, we proposed that ectopic expression of GnT-IVa gene should occur along with the malignancy of trophoblastic tissues, and that the increased GnT-IV activity should be the main cause of the formation of abnormal biantennary sugar chains in choriocarcinoma. A possible enzymatic basis of the biosynthesis of abnormal biantennary sugar chains is discussed.
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Metabolismo de los Hidratos de Carbono , Coriocarcinoma/enzimología , N-Acetilglucosaminiltransferasas/biosíntesis , Neoplasias Uterinas/enzimología , Secuencia de Carbohidratos , Carbohidratos/química , Carbohidratos/genética , Femenino , Humanos , Datos de Secuencia Molecular , Placenta/enzimología , Embarazo , Células Tumorales CultivadasRESUMEN
A human cDNA for UDP- N -acetylglucosamine:alpha1,3-d-mannoside beta1,4- N- acetylglucosaminyltransferase (GnT-IV) was isolated from a liver cDNA library using a probe based on a partial cDNA sequence of the bovine GnT-IV. The cDNA encoded a complete sequence of a type II membrane protein of 535 amino acids which is 96% identical to the bovine GnT-IV. Transient expression of the human cDNA in COS7 cells increased total cellular GnT-IV activity 25-fold, demonstrating that this cDNA encodes a functional human GnT-IV. Northern blot analysis of normal tissues indicated that at least five different sizes of mRNA (9.7, 7.6, 5.1, 3.8, and 2.4 kb) forGnT-IV are expressed in vivo. Furthermore, these mRNAs are expressed at different levels between tissues. Large amounts of mRNA were detected in tissues harboring T lineage cells. Also, the promyelocytic leukemia cell line HL-60 and the lymphoblastic leukemia cell line MOLT-4 revealed abundant mRNA. Lastly, the gene was mapped at the locus on human chromosome 2, band q12 by fluorescent in situ hybridization.
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Cromosomas Humanos Par 2/genética , Proteínas de la Membrana/genética , N-Acetilglucosaminiltransferasas/genética , Linfocitos T/enzimología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Northern Blotting , Células COS , Linaje de la Célula , Mapeo Cromosómico , ADN Complementario/genética , Expresión Génica , Humanos , Hibridación Fluorescente in Situ , Leucemia/enzimología , Tejido Linfoide/enzimología , Datos de Secuencia Molecular , Proteínas de Neoplasias/genética , ARN Mensajero/aislamiento & purificación , Proteínas Recombinantes/biosíntesis , Distribución Tisular , Células Tumorales CultivadasRESUMEN
A simple method for the detection of endogenous histamine in rat peritoneal mast cells was evaluated using on-line mode in-capillary derivatization high-performance capillary electrophoretic (ICD-HPCE) techniques, which were previously developed by our group [S. Oguri et al., J. Chromatogr. A, 787 (1997) 253-260]. The method involves a suspension of peritoneal mast cells (1 x 10(6) cells/ml of saline) collected from a male Wistar rat (eight weeks of age), which are directly introduced into the capillary tube from the anodic end by hydrostatic injection (at 25 cm height, for 2-20 s). When a high-voltage potential (25 kV) is applied to the capillary, which is already filled with the run buffer containing both a lysing reagent (SDS, sodium dodecyl sulfate,) and a derivatizing reagent (OPA, o-phthalaldehyde; NAC, N-acetylcysteine), histamine in the mast cells was detected at high-sensitivity level without further procedures. During ICD-HPCE, the mast cells injected in the capillary were lysed with the lysing reagent, free histamine released from the cell was labeled with the derivatizing reagent, and its derivative was electromigrated, separated and detected with a fluorescence detector (excitation wavelength at 340 nm, emission wavelength at 450 nm) in a fused-silica capillary (75 cm x effective length x 50 microm I.D.). The run buffer used was a 20 mM phosphate-borate buffer (pH 10) containing 20 mM SDS, 2 mM OPA and 2 mM NAC. This method was also examined with regard to the possibility of its use for determination of histamine at the single mast cell level.
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Electroforesis Capilar/métodos , Histamina/análisis , Mastocitos/química , Cavidad Peritoneal/citología , Animales , Recuento de Células , Indicadores y Reactivos , Masculino , Ratas , Ratas Wistar , Reproducibilidad de los Resultados , Dodecil Sulfato de SodioRESUMEN
A new high-performance capillary electrophoretic (HPCE) method with an on-line mode in-capillary derivatization (ICD) procedure for determinations of some amines using 20 mmol/L sodium dodecyl sulfate (SDS) - 2 mmol/L o-phthalaldehyde (OPA) - 2 mmol/L N-acetylcysteine (NAC) - 20 mmol/L phosphate-borate buffer [9] has previously been shown. Although this technique offers direct fluorescence detection of free amines without any derivatization procedures before or after HPCE separation, the presence of spermidine (Spd) is difficult to detect due to low fluorescence intensity. The purpose of this study is to improve the detection sensitivity of Spd by reoptimizing this method with regard to the run buffer; the reoptimized method was applied to the determination of Spd in human plasma. To enhance the fluorescence intensity of the Spd signal, it is effective to use the run buffer in the presence of both beta-cyclodextrin (beta-CD: 8.8 mmol/L) and NAC at high concentration (16 mmol/L). By contrast, the intensity was remarkably decreased when SDS was used in the presence of beta-CD. After ultrafiltrating (UF) spiked human plasma with Spd, UF plasma was directly analyzed using the reoptimized method. Spd peak was detected and separated from the other peaks of blank plasma. The present method gave good linearity (r = 0.999), reproducibility (3.85% coefficient of variation at 5 micromol/L level; n = 10) and specificity. The detection limit and lower limit of quantitation is for 0.2 micromol/L and 1 micromol/L, respectively.
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Electroforesis Capilar/métodos , Espermidina/sangre , Tampones (Química) , Electroforesis Capilar/instrumentación , HumanosRESUMEN
UDP-N-acetylglucosamine:alpha1,3-D-mannoside beta1, 4-N-acetylglucosaminyltransferase (GnT-IV) is one of the essential enzymes in the production of tri- and tetra-antennary Asn-linked sugar chains. Recently, we have successfully purified GnT-IV from bovine small intestine. Based on the partial amino acid sequence of the purified bovine GnT-IV enzyme, its cDNA has been cloned from bovine small intestine. The open reading frame is 1,605 base pairs long, and this sequence produced GnT-IV activity on transient expression in COS-7 cells. Although the deduced amino acid sequence does not have any significant homology with other known N-acetylglucosaminyltransferases (GnTs), the hydrophobicity profile showed a typical type II transmembrane protein structure, which is common to many glycosyltransferases. N-terminal amino acid sequencing of the purified GnT-IV revealed that 92 amino acids, including a transmembrane region, were truncated during purification. Of the three potential N-glycosylation sites Asn-458 was actually glycosylated in the purified enzyme, although this N-glycosylation site could be abolished without any reduction in GnT-IV activity. Serial deletions at both the N and C termini proved that the catalytic domain of GnT-IV is located in the central region of the enzyme. The GnT-IV mRNA level correlated with enzymatic activity in the various bovine tissues tested.