RESUMEN
BACKGROUND: Several methods are used for reconstruction in colon cancer surgery, including hand-sewn or stapled anastomosis. However, few reports have compared short-term outcomes among reconstruction methods. This study compared short-term outcomes between delta-shaped anastomosis (Delta) and functional end-to-end anastomosis (FEEA). METHODS: We retrospectively reviewed 1314 consecutive patients who underwent colorectal surgery with FEEA or Delta reconstruction between January 2016 and December 2023. Patients were divided into two groups according to reconstruction by FEEA (F group; n = 1242) or Delta (D group; n = 72). Propensity score matching was applied to minimize the possibility of selection bias and to balance covariates that could affect postoperative complications. Short-term outcomes were compared between groups. RESULTS: Postoperative complications occurred in 215 patients (17.3%) in F group and 8 patients (11.1%) in D group. Before matching, transverse colon cancer was more frequent (p = 0.002), clinical N-positive status was less frequent (44.1% versus 16.7%, p < 0.001), distant metastasis was less frequent (11.7% versus 1.4%, p = 0.003), and laparoscopic approach was more frequent (87.8% versus 100%, p < 0.001) in D group. After matching, no differences in any clinical factor were evident between groups. Blood loss was significantly lower (28 mL versus 10 mL, p = 0.002) in D group, but operation time and postoperative complication rates were similar between groups. CONCLUSIONS: Delta and FEEA were both considered safe as reconstruction methods. Further studies are needed to clarify appropriate case selection for Delta and FEEA.
Asunto(s)
Anastomosis Quirúrgica , Neoplasias del Colon , Complicaciones Posoperatorias , Puntaje de Propensión , Humanos , Anastomosis Quirúrgica/métodos , Anastomosis Quirúrgica/efectos adversos , Femenino , Masculino , Estudios Retrospectivos , Persona de Mediana Edad , Neoplasias del Colon/cirugía , Anciano , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/epidemiología , Resultado del Tratamiento , Engrapadoras Quirúrgicas , Grapado Quirúrgico/métodos , Colon/cirugía , Colectomía/métodos , Colectomía/efectos adversos , Tempo Operativo , Laparoscopía/métodos , Laparoscopía/efectos adversos , Laparoscopía/estadística & datos numéricosRESUMEN
It remains poorly understood how symptoms in allergic rhinitis are most severe during overnight or early in the morning. The circadian clock consisting of a network of several 'clock genes' including Clock drives daily rhythms in physiology. This study showed that allergen-induced surface CD203c expression on basophils in seasonal allergic rhinitis caused by Japanese cedar pollen exhibited a time-of-day-dependent variation associated with temporal variations in canonical circadian clock gene expression. We also found that bone-marrow-derived basophils (BM basophils) generated from wild-type mice exhibited a time-of-day-dependent variation in IgE-mediated IL-4 and histamine production, which was not observed in BM basophils generated from Clock-mutated mice. Therefore, allergen-specific basophil reactivity shows daily variations depending on the circadian clock activity in basophils, which could partly explain temporal symptomatic variations in allergic rhinitis. Additionally, circadian variations in CD203c expression should be considered for interpretation of this biomarker in clinical research.
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Alérgenos/inmunología , Basófilos/inmunología , Basófilos/metabolismo , Relojes Circadianos/genética , Rinitis Alérgica Estacional/genética , Rinitis Alérgica Estacional/inmunología , Adulto , Animales , Proteínas CLOCK/genética , Proteínas CLOCK/metabolismo , Regulación de la Expresión Génica , Humanos , Masculino , Ratones , Persona de Mediana Edad , Mutación , Proteínas Circadianas Period/genética , Proteínas Circadianas Period/metabolismo , Hidrolasas Diéster Fosfóricas/genética , Hidrolasas Diéster Fosfóricas/metabolismo , Polen/inmunología , Pirofosfatasas/genética , Pirofosfatasas/metabolismo , Factores de Tiempo , Adulto JovenRESUMEN
We have revealed that about one and a half thousand tiny clusters, filled with one thousand closely packed lymphocytes, can be found throughout the murine small and large intestinal mucosa. They are located in crypt lamina propria (cryptopatches; CP) and can be first detected at 14-17 d after birth. A large fraction of lymphocytes in CP expresses c-kit, IL-7R, Thy1 and a lymphocyte function-associated antigen, LFA-1, whereas most of them remain CD3-, TCR alpha beta-, TCR gamma delta-, sIgM-, and B220-. The population size of IL-2R alpha+, HSA+ and Pgp-1+ subsets is variable (20-50%) and the composition of CD8+, Ly-1+, and CD4+ subsets is smaller but also variable (3-20%). In the small intestine, CP do not contain cells undergoing apoptosis nor cells bearing RAG-1 molecules, but do contain dendritic stromal cells bearing CD11c/CD18 molecules. The frequency of DNA replicating cells in CP is higher than that in Peyer's patches (PP), is lower than that in the thymic cortex and is almost comparable with that in the thymic medulla. The numbers of CP remain the same in aged mice (> 114 wk) but double after estrogen treatment even though the thymi are attenuated sharply in both conditions. Thus, with respect to histogenesis, lymphocyte composition and tissue level of cellular behavior, neither PP, isolated lymphoid follicles, peripheral LNs, nor thymus are identical with CP. Finally, CP are virtually absent in lamina propria of IL-7R-deficient mice that display a profound reduction in thymic and peripheral lymphoid cellularity. By contrast, CP are present in germ-free mice and in athymic (nu/nu), SCID, TCR beta x delta-/-, RAG-2-/-, PP-deficient (aly/aly), stem cell factor (Sl/Sld) and c-kit (W/Wv) mutant mice. Taking all of these results together, CP are the first identification of gut-associated murine lymphoid tissues where the generation of IL-7-dependent lympho-hematopoietic progenitors for T and/or B cell descendants may start to take place at the age of commencement of weaning.
Asunto(s)
Antígenos de Diferenciación , Células Madre Hematopoyéticas/citología , Mucosa Intestinal/citología , Tejido Linfoide/citología , Animales , Antígenos CD/aislamiento & purificación , Colon/citología , Vida Libre de Gérmenes , Íleon/citología , Yeyuno/citología , Antígeno-1 Asociado a Función de Linfocito/aislamiento & purificación , Ratones , Ratones Endogámicos , Ratones Mutantes , Proteínas Proto-Oncogénicas c-kit/aislamiento & purificación , Receptores de Interleucina/aislamiento & purificación , Receptores de Interleucina-7 , Antígenos Thy-1/aislamiento & purificación , DesteteRESUMEN
The composition of asteroids and their connection to meteorites provide insight into geologic processes that occurred in the early Solar System. We present spectra of the Nightingale crater region on near-Earth asteroid Bennu with a distinct infrared absorption around 3.4 micrometers. Corresponding images of boulders show centimeters-thick, roughly meter-long bright veins. We interpret the veins as being composed of carbonates, similar to those found in aqueously altered carbonaceous chondrite meteorites. If the veins on Bennu are carbonates, fluid flow and hydrothermal deposition on Bennu's parent body would have occurred on kilometer scales for thousands to millions of years. This suggests large-scale, open-system hydrothermal alteration of carbonaceous asteroids in the early Solar System.
RESUMEN
We assessed the effect of reconstructing the pulmonary artery during arterial switch surgery for transposition of the great arteries on late pulmonary stenosis. Sixty-five patients who underwent Lecompte procedure between September 1991 and December 2006 were divided, by the procedure used chronologically to reconstruct the pulmonary artery, into group XP (single pantaloon patch with equine pericardium, n = 11), group P (direct reconstruction, n = 47), and group AP (single pantaloon patch with fresh autopericardium, n = 7). Outcome and pulmonary stenosis on the most recent ultrasound cardiography (UCG) were compared in the 3 groups. The median follow-up was 13, 7.5, and 1.3 years, respectively. Both early and late mortalities were 1.5% (1/65). Although percutaneous trans-pulmonary angioplasty was necessary in 1, 13, and 3 patients, there was 1, 1, and 0 reoperation for pulmonary stenosis in the 3 groups, respectively. Pulmonary stenosis (pulmonary arterial maximum flow velocity > 3 m/sec on UCG) was present in 4 (40%). 14 (30%). and 3 patients (43%). Although there was no significant difference among the 3 procedures in preventing pulmonary stenosis 10 years after arterial switch surgery, direct reconstruction of the pulmonary artery may show a superior outcome, in particular, over 10 years after arterial switch surgery.
Asunto(s)
Procedimientos Quirúrgicos Cardíacos/métodos , Arteria Pulmonar/cirugía , Transposición de los Grandes Vasos/cirugía , Estudios de Seguimiento , Humanos , Lactante , Recién Nacido , Estenosis de la Válvula Pulmonar/prevención & controlRESUMEN
Two types of cDNAs encoding novel human FGF receptors were isolated. These two cDNAs were found to be closely related to the oncogene bek. Products from these genes were membrane-bound when their cDNAs were transiently expressed in COS cells, whereas products from the regions coding extracellular domains were free of membrane attachment and found in the culture medium.
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ADN/genética , Proteínas Tirosina Quinasas Receptoras , Receptores de Superficie Celular/genética , Secuencia de Aminoácidos , Secuencia de Bases , Western Blotting , Línea Celular , Humanos , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Proteínas Tirosina Quinasas/genética , Receptor Tipo 2 de Factor de Crecimiento de Fibroblastos , Receptores de Factores de Crecimiento de Fibroblastos , Homología de Secuencia de Ácido NucleicoRESUMEN
A DNA fragment that carries the gene coding for poly(3-hydroxybutyrate) (PHB) depolymerase was cloned from the chromosomal DNA of Alcaligenes faecalis AE122 isolated from seawater. The open reading frame encoding the precursor of the PHB depolymerase was 1905 base pairs (bp) long, corresponding to a protein of 635 amino acid residues (M(r) = 65,208). The promoter site, which could be recognized by Escherichia coli RNA polymerase, was upstream from the gene, and the sequence adhering to the ribosome-binding sequence was found in front of the gene. The deduced amino acid sequence agreed with the N-terminal amino acid sequence of the purified PHB depolymerase from amino acid 28 onwards. Analysis of the deduced amino acid sequence revealed the domain structure of the protein; a signal peptide of 27 amino acids long was followed by a catalytic domain of about 400 amino acids, a fibronectin type III module sequence, and a putative substrate binding domain. The molecular mass (62,526) of the mature protein deduced from the nucleotide sequence was significantly lower than the value (95 kDa) estimated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but coincided well with the value (62,426) estimated from matrix-assisted laser desorption ionization mass spectra. By comparison of the primary structure with those of other PHB depolymerases, the substrate binding domain was found to consist of two domains, PHB-specific and poly(3-hydroxyvalerate)-specific ones, connected by a linker region. The PHB depolymerase gene was expressed in Escherichia coli under the control of the tac promoter. The enzyme expressed in E. coli was purified from culture broth and showed the same catalytic properties as the enzyme from A. faecalis.
Asunto(s)
Alcaligenes/genética , Hidrolasas de Éster Carboxílico/genética , Secuencia de Aminoácidos , Secuencia de Bases , Hidrolasas de Éster Carboxílico/química , Hidrolasas de Éster Carboxílico/metabolismo , Clonación Molecular , ADN/química , Escherichia coli/metabolismo , Expresión Génica , Datos de Secuencia Molecular , Peso Molecular , Alineación de SecuenciaRESUMEN
Two cases of somatostatin (SRIF)-producing medullary carcinoma of the thyroid are presented. The plasma levels of SRIF and calcitonin changed in accordance with the clinical course before and after resection of the tumor. On chromatographic analysis of tumor extracts derived from primary and metastatic tumors of medullary thyroid carcinoma, at least two components of immunoreactive SRIF (IR-SRIF) of large molecular size were found, besides a component corresponding to the tetradecapeptide (SRIF 14); one of these seemed to have almost the same molecular weight as the octacosapeptide (SRIF 28). Most of the IR-SRIF in one patient's plasma was eluted in the same position as tetradecapeptide SRIF. The biological activity of SRIF recovered from the tumor extract was assessed by measuring activity for inhibiting GH release using dispersed rat anterior pituitary cells. GH release from the cells into the incubation medium tended to decrease at a concentration of 1 x 10(-9) M IR-SRIF derived from the tumor (3000 +/- 482 ng/10(5) cells . 3 h) compared with the control value (4288 +/- 567 ng/10(5) cells . 3 h). A significant decrease was observed at concentrations of 1 x 10(-8) M (1880 +/- 403 ng/10(5) cells . 3 h) and 5 x 10(-8) M (938 +/- 262 ng/10(5) cells . 3 h). Responses to equivalent amounts of synthetic SRIF 14 were similar. This suggests that IR-SRIF in the tumor has almost the same biological activity as synthetic tetradecapeptide SRIF.
Asunto(s)
Carcinoma/metabolismo , Somatostatina/metabolismo , Neoplasias de la Tiroides/metabolismo , Adulto , Anciano , Animales , Bioensayo , Femenino , Hormona del Crecimiento/metabolismo , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/secundario , Masculino , Peso Molecular , Adenohipófisis/efectos de los fármacos , Adenohipófisis/metabolismo , Radioinmunoensayo , Ratas , Ratas EndogámicasRESUMEN
The hst-1 transforming gene encodes a protein which belongs to the FGF family of growth factors. We showed previously that a human hst-1 protein produced in silkworm cells has in vitro mitogenic activity to vascular endothelial cells. Here we report effective synthesis of an unfused human hst-1 protein in E. coli and a potent in vivo angiogenic activity of this hst-1 protein by two in vivo assays for angiogenesis, chick chorioallantoic membrane assay and rat cornea assay. The NIH3T3 transformant transfected with the hst-1 gene appeared to develop a highly-vascularized tumor on nude mice. These data showed that the hst-1 protein has an angiogenic activity in vivo as well as in vitro.
Asunto(s)
Factores de Crecimiento de Fibroblastos/farmacología , Neovascularización Patológica , Proteínas Proto-Oncogénicas/farmacología , Animales , Embrión de Pollo , Córnea/irrigación sanguínea , Factor 4 de Crecimiento de Fibroblastos , Técnicas In Vitro , Ratones , Ratones Desnudos , Neoplasias Experimentales/irrigación sanguínea , Ratas , Proteínas RecombinantesRESUMEN
Phospholipase A2 from the venom of Trimeresurus flavoviridis (Habu snake) on treatment with cyanogen bromide is split into less than Glu-Gly-Leu-Trp-Gln-Phe-Asn-Hse greater than, which is derived from the N-terminal moiety and is designated as S-peptide, and the remaining large peptide, which is designated as L-peptide. It should be stressed that the N-terminal residue is pyroglutamyl, unlike other phospholipases A2. The L-peptide alone was about 6% as active as the parent molecule. It occurs in dimeric form, like the parent molecule. When L-peptide was mixed with increasing amounts of S-peptide, the activity increased in a hyperbolic manner, indicating the formation of an ordered complex between L-peptide and S-peptide. The dissociation constant of the complex was 2.1 x 10(-7) M and its specific activity was 2.8 times that of L-peptide.
Asunto(s)
Venenos de Crotálidos/análisis , Bromuro de Cianógeno , Fragmentos de Péptidos/análisis , Fosfolipasas A/análisis , Fosfolipasas/análisis , Secuencia de Aminoácidos , Aminoácidos/análisis , Cromatografía en Gel , Sustancias Macromoleculares , Fosfolipasas A2 , Ácido Pirrolidona Carboxílico/análisisRESUMEN
The products derived from the degradation of the sixteen possible diribonucleoside monophosphates (NpN') by Fusarium phosphodiesterase-phosphomonoesterase were analyzed by means of thin layer chromatography. The analysis showed that NpN' was first cleaved into nucleoside N and 5'-nucleotide pN', which was then dephosphorylated to yield nucleoside N'. The dephosphorylation was fast when N' was adenosine or cytidine but slow when N' was guanosine or uridine. The cleavage reaction was followed by measuring the increase of absorbance due to hyperchromicity, and the kinetic constants, Km and kcat, were determined for the sixteen dinucleoside phosphates. The Km value was higher, for a given N, when N' was a pyrimidine nucleoside than when N' was a purine nucleoside. For a given N', uridine as N gave the highest Km value and adenosine gave the lowest one. The kcat value was the highest, for a given N, when N' was cytidine. For a given N', uridine as N gave by far the lowest kcat value. These results can be interpreted in terms of two binding sites on the enzyme with different base preferences. Comparison of kcat/Km values suggested that the base of nucleoside N plays an important role in determining whether a dinucleoside phosphate is a good substrate of the enzyme. The dinucleoside phosphates with uridine as N were found to be particularly poor substrates of the enzyme.
Asunto(s)
Fusarium/enzimología , Oligonucleótidos/metabolismo , Hidrolasas Diéster Fosfóricas/metabolismo , Monoéster Fosfórico Hidrolasas/metabolismo , Cromatografía en Capa Delgada , Fosfatos de Dinucleósidos , Proteínas Fúngicas , Hidrólisis , Cinética , Nucleósidos/metabolismoRESUMEN
Phospholipase A was purified from venom of Trimeresurus flavoviridis (Habu snake) via three steps consisting of Sephadez G-100, CM-cellulose, and DEAE-cellulose column chromatographies. The apparent molecular weights determined by gel filtration on Sephadex G-75 and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were 28,000 and 14,000, respectively, suggesting that the enzyme is composed of two identical subunits. The isoelectric point was 7.9. The enzyme is characterized by high contents of aspartic acid, glycine, tyrosine, and lysine and contains one residue each of histidine and methionine, three or four tryptophans, and eight disulfide bonds per subunit. The enzyme was inactivated by reaction with p-bromophenacyl bromide following pseudo first order kinetics. The loss of activity was accompanied by loss of histidine, indicating that a single histidine residue is essential for activity. Oxidation with N-bromosuccinimide decreased the enzymic activity. Tryptophan residues appear to play some role in catalysis.
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Fosfolipasas A/aislamiento & purificación , Fosfolipasas/aislamiento & purificación , Aminoácidos/análisis , Animales , Sitios de Unión , Bromosuccinimida/farmacología , Disulfuros/análisis , Cinética , Sustancias Macromoleculares , Peso Molecular , Fosfolipasas A/metabolismo , Venenos de SerpienteRESUMEN
Two proteins that act as alpha-amylase inhibitors, Haim I and Paim I, were crystallized and preliminary X-ray diffraction studies on them were carried out. We also sequenced Haim I prepared from Streptomyces griseosporeus YM-25 and confirmed that it is composed of 78 amino acid residues. Crystals of Haim I were grown from ammonium sulfate solution mixed with ethanol by the vapor diffusion technique. The crystals grew as hexagonal bipyramids and diffracted X-rays beyond 2.0 A resolution. They belong to the space group P6(1)22 (or P6(5)22) with unit cell dimensions of a = b = 36.7 A, c = 192.4 A, and contain one molecule per asymmetric unit. Paim I, a protein of 39 amino acid residues produced by Streptomyces corchorusii, was crystallized under similar conditions to Haim I. The crystals diffracted X-rays beyond 2.5 A. They belong to the space group P4(1)2(1)2 (or P4(3)2(1)2) with unit cell dimensions of a = b = 65.4 A, c = 96.1 A, and contain three molecules per asymmetric unit.
Asunto(s)
Proteínas Bacterianas/química , Péptidos/química , alfa-Amilasas/antagonistas & inhibidores , Secuencia de Aminoácidos , Cristalización , Cristalografía por Rayos X , Datos de Secuencia Molecular , Streptomyces/químicaRESUMEN
Immunohistochemical localization of voltage-gated potassium channel Kv3.1b subunit was studied in the cochlea. Intense Kv3.1b-like immunoreactivity was present in the type I, type III, type IV and suprastrial fibrocytes of the cochlear lateral wall. Immunostaining was also found in the interdental cells and the fibrocytes of the spiral limbus and in the supralimbal dark cells. K+ ions, which play a pivotal role in the mechanosensory transduction process in the inner ear, are recycled via gap junctional networks in the cochlea. These results suggest that the voltage-gated potassium channel, containing Kv3.1b, in the cochlear lateral wall fibrocytes may control the intracellular potential and play an important role in regulating the potassium ion recycling mechanism via gap junctions in the inner ear.
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Cóclea/metabolismo , Neuropéptidos/metabolismo , Canales de Potasio con Entrada de Voltaje , Canales de Potasio/metabolismo , Subunidades de Proteína , Ácido Acético , Animales , Cóclea/citología , Formaldehído , Uniones Comunicantes/metabolismo , Cobayas , Inmunohistoquímica , Masculino , Potasio/metabolismo , Canales de Potasio Shaw , Fijación del TejidoRESUMEN
Two phenanthropyran derivatives, 3-methoxy-2,7-dihydroxy-5H-phenanthro[4,5-bcd]pyran and 2,3,7-trihydroxy-5H-phenanthro[4,5-bcd]pyran were isolated from the orchid Phalaenopsis equestri. Their chemical structures were elucidated from spectroscopic (NMR, MS etc.) analyses.
Asunto(s)
Orchidaceae/química , Fenantrenos/aislamiento & purificación , Piranos/aislamiento & purificación , Espectroscopía de Resonancia Magnética , Estructura Molecular , Fenantrenos/química , Piranos/química , Espectrometría de Masa Bombardeada por Átomos VelocesRESUMEN
Two new flavone glucosides, 5,2',6'-trihydroxy-6,7,8-trimethoxyflavone 2'-O-glucoside and 5,2',6'-trihydroxy-6,7-dimethoxyflavone 2'-O-glucoside were isolated from the aqueous methanol extract of the roots of Scutellaria baicalensis. From the extract, seven phenolics, 5,7,2',6'-terahydroxyflavone, 5,7,2',5'-tetrahydroxy-8,6'-dimethoxyflavone, skullcapflavone II, baicalin, baicalin methyl ester, wogonin 7-glucuronide and 3,5,7,2',6'-pentahydroxyflavanone were also isolated.
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Flavonoides/química , Glucósidos/química , Magnoliopsida , Plantas Medicinales , Flavonoides/aislamiento & purificación , Glucósidos/aislamiento & purificación , Espectroscopía de Resonancia Magnética , Estructura Molecular , Raíces de PlantasRESUMEN
An ellagic acid derivative, 3,3'-di-O-methylellagic acid 4-(5"-acetyl)-alpha-L-arabinofuranoside, and two iridoid glucosides, 6alpha-dihydrocornic acid and 6beta-dihydrocornic acid, were isolated from Cornus capitata adventitious roots cultured in Murashige-Skoog (Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473-487) liquid medium containing 10 microM CuSO(4). Three known related metabolites, i.e. stenophyllin H1, dihydrocornin and cornin were also produced in the root cultures. The chemical structures were characterized by analysis of spectroscopic data.
Asunto(s)
Ácido Elágico/análogos & derivados , Glucósidos/aislamiento & purificación , Plantas/química , Piranos/aislamiento & purificación , Glucósidos/química , Iridoides , Espectroscopía de Resonancia Magnética , Raíces de Plantas/química , Piranos/química , Espectrometría de Masa Bombardeada por Átomos VelocesRESUMEN
A hybridoma clone producing a monoclonal antibody (SC78.H81) against scopolamine was established. The monoclonal antibody was an IgG1 (k) antibody with high affinity (1.6 x 10(9) M-1 for methylscopolamine). The monoclonal antibody was cross-reactive with methylscopolamine and butylscopolamine, and showed weak cross-reactivity with 6 beta- and 7 beta-hydroxyhyoscyamine. The cross-reaction with L-hyoscyamine, atropine, scopine and DL-tropic acid was very weak. A competitive enzyme-linked immunosorbent assay using SC78.H81 was established to quantify scopolamine. The sensitivity of the assay allowed detection of 20 pg assay-1 (0.2 ng ml-1) of scopolamine. The assay was applied to the estimation of scopolamine content in hairy root cultures of a Duboisia hybrid.
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Anticuerpos Monoclonales/biosíntesis , Ensayo de Inmunoadsorción Enzimática , Escopolamina/inmunología , Anticuerpos Monoclonales/inmunología , Cromatografía Líquida de Alta Presión , Reacciones Cruzadas , Plantas/química , Radioinmunoensayo , Escopolamina/análisisRESUMEN
A new substrate, 3-ketobutylidene beta-2-chloro-4-nitrophenylmaltopentaoside (3KB-CNPG5), was used for the determination of alpha-amylase (EC 3.2.1.1) in serum and urine. Under this alpha-amylase assay condition, 3KB-CNPG5 is resistant to glucoamylase and alpha-glucosidase, which are auxiliary enzymes, because the 4- and 6-positions of the non-reducing-end glucose residue are modified by the 3-ketobutylidene group. The assay using 3KB-CNPG5 for alpha-amylase activity is a highly sensitive method that uses 2-chloro-4-nitrophenol (CNP) as an aglycone, and is a stable method for determination of alpha-amylase activity in biological fluids.
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Glucósidos , alfa-Amilasas/análisis , Cromatografía Líquida de Alta Presión , Glucosidasas/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Indicadores y Reactivos , Cinética , Páncreas/enzimología , Saliva/enzimología , Espectrofotometría Ultravioleta , Especificidad por Sustrato , alfa-Amilasas/antagonistas & inhibidoresRESUMEN
Expression of Panicum miliaceum L. (proso millet) mitochondrial and cytosolic aspartate aminotransferase (mAspAT and cAspAT, respectively) genes in transgenic tobacco plants (Nicotiana tabacum) and their influences on protein synthesis were examined. The mAspAT- or cAspAT-transformed plants had about threefold or 3.5-fold higher AspAT activity in the leaf than non-transformed plants, respectively. Interestingly, the leaves of both transformed plants had increased levels of phosphoenolpyruvate carboxylase (PEPC) and transformed plants with cAspAT also had increased levels of mAspAT in the leaf. These results suggest that the increased expression of Panicum cAspAT in transgenic tobacco enhances the expression of its endogenous mAspAT and PEPC, and the increased expression of Panicum mAspAT enhances the expression of its endogenous PEPC.