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1.
Springerplus ; 5(1): 2077, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-28018785

RESUMEN

BACKGROUND: Establishing a stand-alone cryogenic test stand is of vital importance to ensure the highly reliable and available operation of superconducting radio-frequency module in a synchrotron light source. Operating a cryogenic test stand relies strongly on a capability to deliver two-phase helium along long cryogenic transfer lines. A newly constructed cryogenic test stand with flexible cryogenic transfer lines of length 220 m at National Synchrotron Radiation Research Center is required to support a superconducting radio-frequency module operated at 126.0 kPa with a 40-W dynamic load for a long-term reliability test over weeks. It is designed based on a simple analytical approach with the introduction of a so-called tolerance factor that serves to estimate the pressure drops in transferring a two-phase helium flow with a substantial transfer cryogenic heat load. Tolerance factor 1.5 is adopted based on safety factor 1.5 commonly applied in cryogenic designs to estimate the total mass flow rate of liquid helium demanded. A maximum 60-W dynamic load is verified with experiment measured with heater power 60 W instead after the cryogenic test stand has been installed. RESULTS: Aligning the modeled cryogenic accumulated static heat load with the results measured in situ, actual tolerance factor 1.287 is obtained. The feasibility and validity of our simple analytical approach with actual tolerance factor 1.287 have been scrutinized by using five test cases with varied operating conditions. Calculated results show the discrepancies of the pressure drops between the estimated and measured values for both liquid helium and cold gaseous helium transfer lines have an underestimate 0.11 kPa and an overestimate 0.09 kPa, respectively. A discrepancy is foreseen, but remains acceptable for engineering applications from a practical point of view. CONCLUSIONS: The simple analytical approach with the introduction of a tolerance factor can provide not only insight into optimizing the choice of each lossy cryogenic piping element of the transfer lines in the design phase but also firm guidance for upgrading the present cryogenic transfer lines for its subsequent application.

2.
Biochim Biophys Acta ; 1494(1-2): 144-8, 2000 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-11072077

RESUMEN

The CDC37 gene was isolated from a round-spotted pufferfish genomic library and characterized. This gene is composed of nine exons spanning 3.5 kb. Exon 1 contains the 5'-untranslated region and exon 2 contains the putative translation initiation site. By 5'-RACE (rapid amplication of cDNA ends) and sequence analysis, we deduced the promoter region for the CDC37 gene and found that it does not contain typical TATA or CCAAT box. The 1.8 kb DNA fragment upstream of the putative transcription initiation site contains numerous potential binding sites for transcription factors including CREB, E2A, Ets-1, GATA, NF-IL6 and PEA3. When this DNA fragment was placed upstream of the chloramphenicol acetyltransferase (CAT) reporter gene and transfected into a carp CF cell line, it could drive the synthesis of CAT enzyme four times more efficiently than the promoterless pCAT-Basic did. In addition, the CDC37 gene is linked to the TYK2 gene in a tail-to-head manner with a small intergenic region of 292 bp.


Asunto(s)
Proteínas de Ciclo Celular/genética , Proteínas de Drosophila , Exones/genética , Peces/genética , Intrones/genética , Chaperonas Moleculares , Regiones Promotoras Genéticas/genética , Proteínas Tirosina Quinasas , Regiones no Traducidas 5'/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Carpas , Proteínas de Ciclo Celular/química , Línea Celular , Clonación Molecular , Codón Iniciador/genética , Regulación de la Expresión Génica , Orden Génico , Genes Reporteros/genética , Genoma , Datos de Secuencia Molecular , Proteínas/genética , Elementos de Respuesta/genética , Alineación de Secuencia , Transfección
3.
Comp Biochem Physiol B Biochem Mol Biol ; 121(2): 169-76, 1998 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-9972292

RESUMEN

A clottable protein was purified from the hemolymph of tiger shrimp (Penaeus monodon) by sequential DEAE anion-exchange chromatography. The protein formed stable clots in the presence of Ca2+ and the transglutaminase in hemocyte lysate. It is thermostable at temperatures up to 66 degrees C. The molecular mass of the clottable protein was determined to be 380 kDa by SDS-PAGE and MALDI-TOF mass spectrometry, and the protein exists as disulfide-linked homodimers and oligomers. The size and amino acid composition of the clottable protein are similar to those of several other shrimps, prawns, lobster and crayfish, and their N-terminal amino acid sequences are 60-80% identical. Monosaccharide analysis of the clottable protein revealed the presence of mannose, glucosamine or N-acetylglucosamine and possibly glucose in this glycoprotein of about 5% sugar content. Lipid in the protein upon electrophoresis was hardly detectable with the Oil Red O staining method. In immunodiffusion and immunoblotting analyses, the anti-clottable protein antibodies reacted with the clottable proteins from the penaeid shrimps but not with those from other crustaceans.


Asunto(s)
Hemolinfa/química , Penaeidae/química , Penaeidae/genética , Proteínas/química , Proteínas/genética , Secuencia de Aminoácidos , Aminoácidos/análisis , Animales , Dimerización , Inmunoquímica , Datos de Secuencia Molecular , Peso Molecular , Polisacáridos/análisis , Conformación Proteica , Proteínas/aislamiento & purificación , Homología de Secuencia de Aminoácido
4.
Br Dent J ; 191(6): 325-9, 2001 Sep 22.
Artículo en Inglés | MEDLINE | ID: mdl-11587504

RESUMEN

AIM: To examine the relationship between the Index of Complexity, Outcome and Need (ICON) and the subjective opinions of patients attending for routine dental care. MATERIALS AND METHODS: This study was undertaken at two general dental practices in Cardiff and Bedford. 50 patients aged between 11-14 years and 50 patients aged 30-40 years presenting for routine dental treatment were selected in each. The subjects were assessed objectively using the ICON guidelines by two examiners trained and calibrated in the use of this index. The scores were recorded directly from the patient. Subjective assessments were obtained from the patients by means of a questionnaire consisting of four simple questions addressing aesthetics, function, speech and treatment need using a five point Likert scale. RESULTS: The mean ICON scores for the different genders and age groups participating in this study were; 11-14 year old males 58.4 (SE 3.17); 11-14 year old females 51.8 (SE 3.51); 30-40 year old males 51.2 (SE 2.70); 30-40 year old females 45.3 (SE 2.56). There were statistically significant differences in ICON scores between the younger and older groups (P = 0.024) and females and males (P = 0.04). Adult patients were more likely to reject treatment than younger patients. Analysis of the professional scores in relation to subjective assessments, using the Spearman rank correlation coefficient, for 11-14 and 30-40 year olds, and for the male and female genders, revealed that the ICON has a significant correlation with patients' perceptions of aesthetics, function, speech and treatment need (r2 = 0.01 to 0.28). The only exceptions were patients' perceptions of speech in the 30-40 year old group, and function in the female gender, which did not show a statistically significant correlation to the professional assessments. CONCLUSION: In this study, the ICON was found to correlate with patients' opinions of aesthetics, function, speech and treatment need. The strength of association, however, was low. It can be concluded that the ICON alone is not necessarily a suitable predictor for appearance, function, speech or treatment need for those individuals attending general dental practice for routine dental care. In combination with a simple question to assess the patients desire for treatment, the shared decision for any particular individual to enter the treatment process can be determined.


Asunto(s)
Odontología General/estadística & datos numéricos , Maloclusión/terapia , Evaluación de Necesidades/estadística & datos numéricos , Ortodoncia Correctiva/estadística & datos numéricos , Evaluación de Resultado en la Atención de Salud/métodos , Satisfacción del Paciente , Adolescente , Adulto , Factores de Edad , Niño , Encuestas de Salud Bucal , Estética Dental , Estudios de Factibilidad , Femenino , Humanos , Masculino , Maloclusión/psicología , Masticación , Ortodoncia Correctiva/psicología , Aceptación de la Atención de Salud , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Factores Sexuales , Habla , Estadísticas no Paramétricas , Encuestas y Cuestionarios
5.
Nanoscale Res Lett ; 5(12): 1982-91, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21170405

RESUMEN

Powdery calcium carbonates, predominantly calcite and aragonite, with planar defects and cation-anion mixed surfaces as deposited on low-carbon steel by magnetic water treatment (MWT) were characterized by X-ray diffraction, electron microscopy, and vibration spectroscopy. Calcite were found to form faceted nanoparticles having 3x (01̄14) commensurate superstructure and with well-developed {112̄0} and {101̄4} surfaces to exhibit preferred orientations. Aragonite occurred as laths having 3x (01̄1) commensurate superstructure and with well-developed (01̄1) surface extending along [100] direction up to micrometers in length. The (hkil)-specific coalescence of calcite and rapid lath growth of aragonite under the combined effects of Lorentz force and a precondensation event account for a beneficial larger particulate/colony size for the removal of the carbonate scale from the steel substrate. The coexisting magnetite particles have well-developed {011} surfaces regardless of MWT.

6.
Biochem Biophys Res Commun ; 199(2): 969-76, 1994 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-7510963

RESUMEN

Polymerase chain reaction (PCR) was employed to amplify cDNAs constructed from the poly(A)+RNA of venom glands in Taiwan cobras to facilitate the cloning and sequencing of phospholipase A2 (PLA2) gene. The PCR product was then subcloned into pUC18 vector and transformed in E. coli strain JM109. Plasmids purified from the positive clones were prepared for nucleotide sequencing by dideoxynucleotide chain-termination method. Sequencing several clones containing about 0.5 kb DNA inserts constructed a complete and unambiguous full-length reading frame of 468 base pairs covering a precursor for phospholipase A2 with a deduced mature protein sequence of 119 amino acids and a 27 amino-acid segment of signal peptide. The sequenced major PLA2 with pI 4.991 shows a high degree of sequence homology to those PLA2 of the same or closely-related genus. The deduced protein sequence allows us to correct and resolve some discrepancy between the sequences determined by conventional protein sequencing (Toxicon, 19, 141(1981)) and X-ray crystallography (Science, 250, 1560(1990)). Expression of PLA2 in E. coli vector generated a polypeptide which can cross-react with the antiserum against the native and purified PLA2 from the same cobra venom albeit with a much lower activity.


Asunto(s)
Venenos Elapídicos/metabolismo , Fosfolipasas A/biosíntesis , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bungarotoxinas/metabolismo , Bovinos , Clonación Molecular , Cartilla de ADN , Elapidae/genética , Datos de Secuencia Molecular , Páncreas/enzimología , Fosfolipasas A/química , Fosfolipasas A/aislamiento & purificación , Fosfolipasas A2 , Plásmidos , Poli A/aislamiento & purificación , Poli A/metabolismo , Reacción en Cadena de la Polimerasa , Señales de Clasificación de Proteína/biosíntesis , Señales de Clasificación de Proteína/química , ARN/aislamiento & purificación , ARN/metabolismo , ARN Mensajero , Mapeo Restrictivo , Homología de Secuencia de Aminoácido , Taiwán
7.
J Biol Chem ; 276(13): 9631-9, 2001 Mar 30.
Artículo en Inglés | MEDLINE | ID: mdl-11133989

RESUMEN

A Sepharose CL-4B-binding protein, Tachypleus plasma lectin 1 (TPL-1), and a lipopolysaccharide (LPS)-binding protein, Tachypleus plasma lectin-2 (TPL-2), have been isolated from the plasma of Tachypleus tridentatus and biochemically characterized. Each protein is coded by a homologous family of multigenes. TPL-1 binds to Sepharose CL-4B and was eluted with buffer containing 0.4 m GlcNAc. The deduced amino acid sequence of TPL-1 consisted of 232 amino acids with an N-glycosylation site, Asn-Gly-Ser at residues 74-76. It shares a 65% sequence identity and similar internal repeats of about 20 amino acid motifs with tachylectin-1. Tachylectin-1 was identified as a lipopolysaccharide-agarose binding nonglycosylated protein from the amebocytes of T. tridentatus. TPL-2 was eluted from the LPS-Sepharose CL-4B affinity column in buffer containing 0.4 m GlcNAc and 2 m KCl. The deduced amino acid sequence of TPL-2 consisted of 128 amino acids with an N-glycosylation site, Asn-Cys-Thr, at positions 3-5. It shares an 80% sequence identity with tachylectin-3, isolated from the amebocytes of T. tridentatus. TPL-2 purified by LPS-affinity column from the plasma predominantly exists as a dimer of a glycoprotein with an apparent molecular mass of 36 kDa. Tachylectin-3 is an intracellular nonglycosylated protein that also exists as a dimer in solution with an apparent molecular mass of 29 kDa. It recognizes Gram-negative bacteria through the 0-antigen of LPS. Western blot analyses showed that, in the plasma, TPL-1 and TPL-2 exist predominantly as oligomers with molecular masses above 60 kDa. They both bind to Gram-positive and Gram-negative bacteria, and this binding is inhibited by GlcNAc. Possible binding site of TPL-1 and TPL-2 to the bacteria could be at the NAc moiety of GlcNAc-MurNAc of the peptidoglycan. The physiological function of TPL-1 and TPL-2 is most likely related to their ability to form a cluster of interlocking molecules to immobilize and entrap invading organisms.


Asunto(s)
ADN Complementario/metabolismo , Cangrejos Herradura/química , Cangrejos Herradura/metabolismo , Lectinas/genética , Lectinas/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Biotinilación , Western Blotting , Cromatografía de Afinidad , Cromatografía Líquida de Alta Presión , Clonación Molecular , Relación Dosis-Respuesta a Droga , Electroforesis en Gel de Poliacrilamida , Escherichia coli/metabolismo , Glicosilación , Hemolinfa/metabolismo , Cinética , Lectinas/química , Datos de Secuencia Molecular , Antígenos O/metabolismo , Peptidoglicano/metabolismo , Reacción en Cadena de la Polimerasa , Pruebas de Precipitina , Unión Proteica , Sefarosa/metabolismo , Homología de Secuencia de Aminoácido , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción
8.
Eur J Biochem ; 266(2): 624-33, 1999 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-10561606

RESUMEN

To investigate the coagulation system in crustacean decapoda, a homodimeric glycoprotein of 380 kDa was purified from the hemolymph of tiger shrimp (Penaeus monodon) by sequential DEAE anion exchange chromatography. The purified protein was coagulated by the shrimp hemocyte transglutaminase in the presence of Ca2+. The clottable protein contains 44% alpha helices and 26% beta sheets as determined by circular dichroism spectra. Its conformation is stable in buffer of pH 4-9. To solve its primary structure, partial sequences of the purified polypeptides from cyanogen bromide cleavage and endopeptidase digestion were also determined. A shrimp cDNA expression library was constructed. By combination with antibody screening, reverse transcriptase PCR using degenerate primers from determined amino acid sequences and cDNA library screening with digoxigenin-labeled DNA probes, the entire cDNA of 6124 bp was obtained. This cDNA encodes a protein of 1670 amino acids, including a 14-amino acid signal peptide. With four potential N-glycosylation sites, the clottable protein was found to contain 3.8% high-mannose glycan; and Man8GlcNAc and Man9GlcNAc were released upon endo-beta-N-acetylglucosaminidase hydrolysis. Upon conducting a protein sequence database survey, the shrimp clottable protein shows 36% identities to the crayfish clotting protein and lower similarities to members of insect vitellogenins, apolipoprotein B and mammalian von Willebrand factor. Notably, a region rich in Gln residues, a polyGln motif and five Ser-Lys-Thr-Ser repeats are present in the shrimp protein, suggesting this protein might be a transglutaminase substrate. Northern blot analysis revealed that the clottable protein is expressed in most of the shrimp tissues but not in the mature hemocytes.


Asunto(s)
Hemolinfa/química , Secuencia de Aminoácidos , Animales , Biotinilación , Coagulación Sanguínea , Northern Blotting , Carbohidratos/química , Cromatografía por Intercambio Iónico , Dicroismo Circular , Clonación Molecular , Fragmentación del ADN , ADN Complementario/metabolismo , Biblioteca de Genes , Hemolinfa/metabolismo , Concentración de Iones de Hidrógeno , Modelos Genéticos , Datos de Secuencia Molecular , Oligosacáridos/química , Penaeidae , Reacción en Cadena de la Polimerasa , Polisacáridos/química , Homología de Secuencia de Aminoácido , Factores de Tiempo , Distribución Tisular
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