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INTRODUCTION: Chlamydia trachomatis is one of the most common sexually transmitted diseases (STDs) globally. However, data on its prevalence and risk factors in Malaysia is still scarce. OBJECTIVE: We aimed to identify the prevalence and risk factors of genitourinary C.trachomatis infection among patients attending STD clinics in northern Peninsular Malaysia. METHODS: A hospital-based cross-sectional study was conducted in STD clinics of Hospital Pulau Pinang and Hospital Sultanah Bahiyah, Kedah from January to November 2014. Participants were individually interviewed using a structured data collection form followed by a physical examination and laboratory tests. Nucleic Acid Amplification Test (NAAT) was used to detect C.trachomatis infection. Analysis was carried out using SPSS Version 15. RESULTS: Eighty-three sexually active patients were enrolled, consisting of 51 males and 32 females. The median age was 28.0 years. In general, 32.5% patients were asymptomatic, the remaining presented with genital discharge (41.0%), genital warty lesion (25.3%), genital ulcer (13.3%), dysuria (13.3%), dyspareunia (2.4%), urine hesistancy (1.2%) and genital swelling (1.2%). The prevalence of genitourinary C.trachomatis infection was 21.7% in the study population; 17.6% in males and 28.1% in females. Among the infected females, 44.4% were pregnant. Of those infected 56.6% did not show any symptoms of genital infection, and 77.8% were aged between 18 and 30 years, of which most were females. Among newly diagnosed HIV patients, the prevalence was 14.3%. From multivariable logistic regression analysis, age under 28 years, being married and engagement in oral sex had significantly increased odds of C.trachomatis infection. CONCLUSIONS: C.trachomatis infection was common among patients attending STD clinics in northern Penisular Malaysia especially in the younger age groups. Majority of the infected patients were asymptomatic.
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Infecciones por Chlamydia/epidemiología , Infecciones por Chlamydia/etiología , Chlamydia trachomatis/aislamiento & purificación , Adolescente , Adulto , Instituciones de Atención Ambulatoria , Infecciones por Chlamydia/diagnóstico , Estudios Transversales , Femenino , Humanos , Entrevistas como Asunto , Malasia/epidemiología , Masculino , Persona de Mediana Edad , Prevalencia , Investigación Cualitativa , Factores de Riesgo , Enfermedades de Transmisión Sexual/epidemiología , Enfermedades de Transmisión Sexual/etiología , Adulto JovenRESUMEN
AIM: Whether some diseases are related to the occurrence of synchronous colorectal carcinoma (sCRC) is unknown. Investigating the risk factors and presentation of sCRC could aid in the treatment of patients with colorectal cancer (CRC). The prognosis of sCRC compared with that of solitary CRC remains unclear. METHODS: A total of 17 093 CRC patients were recruited between 1st January 1995 and 31th December 2016. The risk factors of sCRC development were assessed using univariate and multivariate logistic regression. The effect of sCRC on survival was analysed using the multivariate Cox regression model. RESULTS: The prevalence of sCRC was 5.6% in this study. The independent risk factors of sCRC development were advanced age (P < 0.001), male sex (P < 0.001), hereditary cancer (P < 0.001), hypertension (P < 0.001) and liver cirrhosis (P = 0.024). Compared with solitary CRC, a higher number of patients with sCRC presented with an abnormal carcinoembryonic antigen (CEA) level (P = 0.011), anaemia (P < 0.001) and hypoalbuminemia (P < 0.001). Multivariate analysis revealed that sCRC was a significant factor for poor survival in patients at TNM Stage I [hazard ratio (HR) = 1.86; P < 0.001], Stage II (HR = 1.65; P < 0.001) and Stage III (HR = 1.40; P < 0.001). CONCLUSIONS: In addition to hypertension and liver cirrhosis, other risk factors for sCRC were identified in this study. The prognosis of patients with sCRC was significantly worse than that of those with solitary CRC through TNM Stages I to III. Anaemia, abnormal CEA and hypoalbuminemia were more commonly seen in patients with sCRC.
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Carcinoma/mortalidad , Neoplasias Colorrectales/mortalidad , Neoplasias Primarias Múltiples/mortalidad , Anciano , Biomarcadores de Tumor/sangre , Antígeno Carcinoembrionario/sangre , Carcinoma/patología , Neoplasias Colorrectales/sangre , Neoplasias Colorrectales/patología , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Estadificación de Neoplasias , Neoplasias Primarias Múltiples/sangre , Neoplasias Primarias Múltiples/patología , Pronóstico , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Estudios Retrospectivos , Factores de Riesgo , Tasa de SupervivenciaAsunto(s)
Absceso/cirugía , Enfermedades del Ano/cirugía , Drenaje/métodos , Anciano , Humanos , Conducto Inguinal/cirugía , MasculinoRESUMEN
BACKGROUND: Primary retroperitoneal mucinous cystadenocarcinoma (PRMC) is a rare disease and mostly occurs in females, and there are only three male cases described in the literatures without long-term follow-up. CASE REPORT: A 59-year-old male presented with a left retroperitoneal cystic mass (7.5 ´ 7 ´ 3 cm) that upwardly displaced the left kidney and caused abdominal discomfort. The tumor was totally excised by the hand-assisted laparoscopic method without complications or recurrence in a follow-up period of 79 months. The etiology from coelomic metaplasia of peritoneal epithelium was proved by a spectrum of diverse cells (benign, borderline malignant, and malignant cells) during pathological examination. RESULTS: This is the fourth male case of PRMC in the world with a favorable outcome after hand-assisted laparoscopic excision, and this is also distinct by the longest follow-up period in this disease entity. CONCLUSIONS: Because of its low-malignant potential and recurrence rate, surgical excision is still the best choice of treatment, but the least invasion method should be adopted in front.
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PURPOSE: Although many reports advocate computed tomography (CT) as the initial surveillance tool for occult cervical spine injury (CSI) at the emergency department (ED), the role of a lateral cervical spine radiograph (LCSX) has still not been replaced. We hypothesized that the increased width of the prevertebral soft tissue on an LCSX provides helpful information for selecting the high-risk patients who need to be evaluated with more accurate diagnostic tools. METHODS: This was a retrospective and consecutive series of injured patients requiring cervical spine evaluation who were first imaged with three-view plain films at the ED. The prevertebral soft tissue thickness (PVST) and ratio of prevertebral soft tissue thickness to the cervical vertebrae diameter (PVST ratio) were calculated on the LCSX. Suspicion of CSI was confirmed by either CT or magnetic resonance imaging (MRI) scans. RESULTS: A total of 826 adult trauma patients requiring cervical spine evaluation were enrolled. The C3 PVST and PVST ratio were significantly different between patients with or without upper cervical area injury (UCAI, 8.64 vs. 5.49 mm, and 0.394 vs. 0.276, respectively), and, likewise, the C6 PVST and PVST ratio for patients with or without lower cervical area injury (LCAI, 16.89 vs. 14.66 mm, and 0.784 vs. 0.749, respectively). The specificity was greater than 90 % in predicting UCAI and LCAI when combining these two parameters. CONCLUSIONS: This method maximizes the usefulness of LCSX during the initial assessment of a conscious patient with blunt head and neck injury, especially for the identification of high-risk patients requiring prompt CT or MRI; on the other hand, it prevents the overuse of these high-cost imaging studies as initial diagnostic tools.
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Two enzymatic methods commonly used in N-terminal sequence analysis of blocked proteins are presented in this unit; one uses pyroglutamate aminopeptidase for N(alpha)-pyrrolidone carboxyl-proteins in solution or blotted onto a membrane, and the other uses acylaminoacyl-peptide hydrolase for N(alpha)-acyl-proteins blocked with other acyl groups. A Support Protocol describes a colorimetric assay for pyroglutamate aminopeptidase activity. Sequencing with acylaminoacyl-peptide hydrolase must include fragmentation of the protein before unblocking can be carried out, so procedures are provided for chemically blocking newly generated peptides with either succinic anhydride or phenylisothiocyanate/performic acid. The hydrolase is then applied to the total mixture of peptides, only one of which, the acylated N-terminal peptide, should be a substrate for hydrolase. After incubation, the mixture of peptides is subjected to sequence analysis. Protocols are also provided for unblocking N-terminally blocked proteins using acid-catalyzed hydrolysis or methanolysis, hydrazinolysis, and beta-elimination after acid-catalyzed N-O shift. Alternate protocols describe chemical removal of acetyl and longer-chain alkanoyl groups, as well as formyl groups to open the cyclic imide of pyrrolidone carboxylate.
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Bioquímica/métodos , Proteínas/química , Proteínas/metabolismo , Western Blotting , Colorimetría , Formiatos/metabolismo , Hidrazinas/metabolismo , Hidrolasas/metabolismo , Hidrólisis , Isotiocianatos/metabolismo , Piroglutamil-Peptidasa I/metabolismo , Ácido Pirrolidona Carboxílico/química , Ácido Pirrolidona Carboxílico/aislamiento & purificación , Soluciones , Anhídridos Succínicos/metabolismo , Ácido Trifluoroacético/metabolismoRESUMEN
A general method for total synthesis of natural trisubstituted gamma-lactones is developed on the basis of the chemistry of alkynyltungsten compounds. The key step in this approach involves the cycloalkenation of tungsten-eta1-(3R,4S)-pent-1-yne-3,4-diol with aldehydes to give tungsten-oxacarbenium salts, further leading to 3-alkylidene-4-hydroxy-5-methyl-gamma-lactones upon demetalation. This synthetic sequence proceeds well for alkynylaldehydes and the MOM derivative of tungsten-eta1-(3R,4S)-pent-1-yne-3,4-diol. The resulting butyrolactone products are transformed into natural trisubstituted butyrolactones including (+)-blastmycinone, (+)-blastmycinolactol, (+)-antimycinone, NFX-2, and (+)-isodihydromahubanolide A. By using the same approach based on (R)-ethyl lactate, the natural (-)-litsenolide C1 can be prepared in a few steps.
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4-Butirolactona/análogos & derivados , Lactonas/síntesis química , Compuestos de Tungsteno/síntesis química , 4-Butirolactona/síntesis química , Lactonas/química , Estereoisomerismo , Compuestos de Tungsteno/químicaRESUMEN
Nurses use music therapeutically but often assume that all patients will equally appreciate the same type of music. Cultural differences in music preferences are compared across five pain studies. Music preferences for pain relief are described as the most frequently chosen type of music for each culture. Findings indicate that in four studies, musical choices were related to cultural background (p = .002 to .049). Although the majority in each group chose among the other types of music, Caucasians most frequently chose orchestra music, African Americans chose jazz, and Taiwanese chose harp music. For culturally congruent care, nurses should become aware of cultural differences in music preference and provide culturally specific selections among other music expected to have a therapeutic effect.
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Negro o Afroamericano/psicología , Características Culturales , Etnicidad/psicología , Musicoterapia , Dolor/enfermería , Satisfacción del Paciente , Población Blanca/psicología , Población Negra , Diversidad Cultural , Femenino , Humanos , Masculino , Rol de la Enfermera , Manejo del Dolor , Muestreo , Taiwán/etnología , Estados UnidosRESUMEN
Sialic acids have been implicated in a variety of complex biological regulatory and signalling events and their functional importance is reflected by their presence in a wide variety of phyla. Potentially they may inhibit intermolecular and intercellular interactions. Lectins that exhibit specificity for sialic acid or sialoglycoconjugates are ubiquitous in the body fluids of invertebrates and this has supported the assumption that these lectins are involved in defense against microbes that express sialic acids on their surfaces. This biological function has also been inferred from the absence of sialic acids in lower invertebrates. However, most invertebrate lectins are heterogeneous and may also bind other ligands. The biological significance of the different carbohydrate specificities are not yet known. We have demonstrated the presence of sialic acids in hemolymph from two marine bivalves, the Pacific oyster Crassostrea gigas (approximately 15 micrograms ml-1) and the horse mussel Modiolus modiolus (48-100 micrograms ml-1) by several different assays. The sialic acid was mostly in free form. Affinity purified lectins from the horse mussel also contained bound sialic acids (2-5 mumol g-1). Oyster hemolymph stimulated the in vitro phagocytosis of bacteria by oyster hemocytes. The stimulation by hemolymph is facilitated by a dialyzable component, that apparently is active irrespective of the binding to sialic acid (BSM). Addition of sialic acid had no significant effect on the in vitro phagocytosis of bacteria by oyster hemocytes.
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Hemolinfa/química , Lectinas/metabolismo , Moluscos/química , Ácido N-Acetilneuramínico/análisis , Aglutinación/fisiología , Animales , Eritrocitos/metabolismo , Hemocitos/fisiología , Ácido N-Acetilneuramínico/farmacología , Fagocitosis/fisiología , Vibrio/metabolismoRESUMEN
Music is a method nurses can use to help relieve pain, however little is known about its effectiveness across cultures. In this study, Western music was tested for its effectiveness in reducing postoperative pain in 38 Taiwanese patients, and its acceptability was explored. A pretest and post-test experimental design was used with visual analogue scales to measure sensation and distress of pain. Before surgery, subjects were randomly assigned to receive tape recorded music or the usual care. Those who were assigned to the music group chose among 5 types of sedative music. On postoperative Day 1 and Day 2, the effectiveness of the tape-recorded music was investigated during 15 minutes of rest in bed. Patients were interviewed on Day 3 to determine their liking for the music, its calming effects, and the helpfulness of the music. Repeated measures analysis of variance showed a significant interaction between time and group in the distress of pain on Day 1, but not on Day 2, and in pain sensation on Day 2, but not Day 1. Subjects from Taiwan were similar to subjects in a previous study in the United States in their liking for the music, and in reports of the helpfulness of the music for pain sensation and distress, but fewer Taiwanese found the music calming, and they had different choices: more chose harp music and fewer chose jazz than subjects in the U.S. study, and some would prefer Buddhist hymns or popular songs heard in Taiwan. Findings support the use of culturally acceptable music in addition to analgesic medication for the sensation and distress of postoperative pain.
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Musicoterapia , Dolor Postoperatorio/terapia , Adulto , Cultura , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
OBJECTIVE: To study the efficacy of a low sodium hypertonic resuscitation fluid for resuscitation of severe hemorrhage in a pediatric animal, using the intraosseous route. DESIGN: Prospective, randomized, controlled animal study. SETTING: University physiology laboratory. SUBJECTS: Seventeen immature (6- to 9-wk-old) piglets, weighing 10.6 +/- 0.4 kg, were studied under anesthesia. INTERVENTIONS: A new 2400 mosm/L hypertonic fluid, "Isosal" was formulated with reduced (3.45%) sodium content compared with a 2400-mosm/L (7.5%) hypertonic saline solution. This formulation was accomplished by substituting glucose and mixed amino acids for sodium. Piglets were subjected to 1 hr of hemorrhage, reducing the cardiac output to 50% of baseline value. Resuscitation was carried out through the intraosseous route with an initial 6 mL/kg bolus of either hypertonic saline, Isosal, or lactated Ringer's solution. After the initial bolus, additional test fluid was given to maintain the cardiac output at baseline value for a 2-hr period. MEASUREMENTS AND MAIN RESULTS: Total resuscitation volumes, hemodynamic variables, and electrolytes were measured. Intraosseous vascular access was easily established in all animals, and fluid resuscitation was carried out effectively through this route. Resuscitation volumes were significantly lower for both of the hypertonic fluids (12.7 +/- 1.2 mL/kg for hypertonic saline, and 12.5 +/- 1.7 mL/kg for Isosal solution) compared with lactated Ringer's solution (75.3 +/- 11.6 mL/kg) (p = .01). Both hypertonic saline and Isosal solution resulted in an immediate supranormal response in cardiac output that lasted 20 mins. In contrast, when lactated Ringer's solution was used, multiple boluses were required over a 20-min period to normalize cardiac output. Serum sodium was significantly higher in the hypertonic saline group compared with the Isosal or lactated Ringer's groups (p = .001). CONCLUSIONS: Isosal solution was as effective as hypertonic saline in "small volume" resuscitation of severe hemorrhagic shock in a pediatric animal model through the intraosseous route, and produced significantly less hypernatremia when compared with hypertonic saline.
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Aminoácidos/uso terapéutico , Glucosa/uso terapéutico , Solución Salina Hipertónica/uso terapéutico , Choque Hemorrágico/terapia , Cloruro de Sodio/uso terapéutico , Aminoácidos/administración & dosificación , Análisis de Varianza , Animales , Glucemia , Modelos Animales de Enfermedad , Combinación de Medicamentos , Glucosa/administración & dosificación , Hemodinámica , Infusiones Intraóseas , Soluciones Isotónicas/administración & dosificación , Soluciones Isotónicas/uso terapéutico , Estudios Prospectivos , Distribución Aleatoria , Resucitación/métodos , Solución de Ringer , Solución Salina Hipertónica/administración & dosificación , Cloruro de Sodio/administración & dosificación , PorcinosRESUMEN
The disulfide bonds and N-glycosylation sites in a glycoprotein from the Rathke's gland secretion of the Kemp's ridley turtle (Lepidochelys kempi) have been characterized with respect to peptide sequences and glycan structures. The glycoprotein constitutes about 70% of the total protein in the secretion, and based on partial sequence information, it shows more than 20% identity with both the catalytic (esterases) and the noncatalytic (thyroglobulin) members of the esterase/lipase family of proteins. For the determination of the disulfide locations, the glycoprotein was digested with chymotrypsin, and the three HPLC peptide peaks yielding fluorescent products after treatment with tributylphosphine (Bu3P) and 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F) were collected. The three fractions were treated with the same reagents in separate experiments, the resulting pairs of ABD-Cys-containing peptides were separated by HPLC, and the sequence of each individual peptide was determined. The peptide identity established that three disulfide bonds existed in the glycoprotein: Cys 65-Cys 91, Cys 254-Cys 265, and Cys 130-Cys 404; the first two of these are conserved in all the members of the esterase family. For the study of the glycosylation sites, the glycoprotein was reduced with Bu3P and the SH groups covalently blocked with ABD-F, and the resulting product was digested with chymotrypsin. The glycopeptides were isolated by affinity chromatography, separated by reverse-phase HPLC, and subjected to sequence analysis and fast atom bombardment mass spectrometry before and after separation of the glycans and the peptides through the action of glycoamidase. Three separate glycosylation sites were identified, each containing multiple glycans. The sugar analyses of the hydrolysates of the glycoprotein indicated that only GlcNAc and Man were present as building blocks, and the mass spectrometric data showed that Man3GlcNAc2-, GlcNAc2-4Man3GlcNAc2-, and possibly GlcNAc2Man2GlcNAc2- were the major glycan structures, distributed differently at the three sites. The three glycosylation sites match three of the nine sites glycosylated in human serum choline esterase, and one of them, Asn 106, is also found as one of two glycosylation sites in the homologous segment of thyroglobulin.
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Disulfuros/metabolismo , Glicoproteínas/metabolismo , Tortugas/metabolismo , Secuencia de Aminoácidos , Animales , Secuencia de Carbohidratos , Cromatografía Líquida de Alta Presión , Glicosilación , Datos de Secuencia Molecular , Mapeo Peptídico , Tortugas/anatomía & histologíaRESUMEN
The secretion produced by Rathke's glands of Kemp's ridley sea turtles (Lepidochelys kempi) contains the enzyme gamma-glutamyl transpeptidase. The approximately 200 kDa enzyme contains two different subunits, alpha (54 kDa) and beta (21 kDa), in an unknown stoichiometry. The enzyme transfers gamma-Glu from a number of different donors, such as glutamine, glutathione, S-Me-glutathione, N epsilon(gamma-Glu)-Lys, gamma-Glu-Ala, and other gamma-glutamyl amino acids, either to water or to a variety of acceptor substrates. It appears that a free alpha-amino group is the preferred acceptor. The enzyme is not inhibited by typical sulfhydryl reagents such as N-ethyl-maleimide, p-(chloro)mercuri-benzoate or 5,5'-dithio-bis-(2-nitrobenzoate) or by the active Ser reagent tosyl fluoride. Maleate stimulates the activity of the enzyme, and in the presence of 100 mM maleate 2 mM tosyl fluoride becomes an inactivator of the enzyme. The catalytic and molecular properties of the turtle gamma-glutamyl transpeptidase are similar to those established for mammalian gamma-glutamyl transpeptidase. Neither the physiological role of the enzyme nor the biological function of the secretion in which it occurs is understood at this time.
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Tortugas/metabolismo , gamma-Glutamiltransferasa/química , Secuencia de Aminoácidos , Animales , Activación Enzimática , Datos de Secuencia Molecular , Especificidad por Sustrato , gamma-Glutamiltransferasa/aislamiento & purificaciónAsunto(s)
Proteínas Bacterianas , Biotina/análogos & derivados , Glicopéptidos/aislamiento & purificación , Lectinas , Secuencia de Aminoácidos , Sitios de Unión , Secuencia de Carbohidratos , Cromatografía Líquida de Alta Presión/métodos , Glicopéptidos/química , Datos de Secuencia Molecular , Oligosacáridos/química , Mapeo Peptídico/métodos , TripsinaRESUMEN
The use of the reagent tributyl phosphine (Bu3P) to reduce disulfides (Ruegg, U.T., and Rudinger, J., Methods Enzymol. 47, 111-116, 1977) and of 4-(aminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (ABD-F) to block free sulphydryl groups (Toyo'oka, T., and Imai, K. Anal. Chem. 56, 2461-2464, 1984) is well established in the literature. Since the two reagents apparently do not react with each other, their combination offers a convenient and quite general method for the complete characterization of free Cys (SH) and crosslinked Cys (S-S) in proteins (Kirley, T.L., J. Biol. Chem. 264, 7185-7192, 1989). We review some of the characteristics of the reaction of these reagents with Cys in peptides and proteins and some of the properties of the ABD-Cys derivatives. The review includes reactions with model compounds (e.g., Cys and glutathione), proteins such as enolase from yeast and rabbit muscle, containing only free Cys, a protein, fetuin, containing only crosslinked Cys, and a protein, superoxide dismutase, containing both free and crosslinked Cys. In all cases the direct comparison of the tryptic or chymotryptic peptides derived from the products of parallel reactions of the protein with ABD-F alone and with ABD-F together with Bu3P permitted the determination of both free and total Cys in the protein. Sequencing the fluorescent ABD-peptides established the position of the Cys residues in the primary sequence.(ABSTRACT TRUNCATED AT 250 WORDS)
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Disulfuros/análisis , Oxadiazoles , Fragmentos de Péptidos/química , Fosfinas , Proteínas/química , Compuestos de Sulfhidrilo/análisis , Secuencia de Aminoácidos , Animales , Sitios de Unión , Cromatografía Líquida de Alta Presión/métodos , Enzimas/química , Indicadores y Reactivos , Datos de Secuencia Molecular , Fragmentos de Péptidos/aislamiento & purificación , TripsinaRESUMEN
The current study sought to establish a preliminary patient classification system by determining how much time nurses spend each shift on their various activities. In this way we hoped to determine the direct care needs of the patient. From August 11 through the 17, snap-shot observations on the Chest Medicine Ward of Kaohsiung Medical College Hospital were taken every five minutes throughout the day to evaluate nursing activities and health care needs of patients. After employing such statistical techniques as percentage comparison, One-way ANOVA on the collected data, we gained a preliminary understanding of the distribution, characteristics, extent and nature of nursing activities. The results demonstrated that the average working hours of the day shift and the evening shift are 8.13 and 8.05 hours respectively. The night shift may be as long as 8.52 hours. Our studies also revealed that the most demanding nursing activity is indirect care (50.73%), direct care (29.39%), individual time (13.73%) and related activities (6.15%) follow. In the direct care of patient, nurses devoted most of their time monitoring vital signs, administering medication, assisting in examination and treatment, caring for, and communicating with patients. On hygiene, physical movements and the clean up of body discharges were mostly devoted by patients and families.
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Enfermedades Torácicas/enfermería , Femenino , Humanos , Pacientes/clasificación , Proyectos Piloto , Factores de TiempoRESUMEN
Glycopeptide-containing fractions in HPLC peptide maps can be detected by a simple application of the microtiter plate-bound streptavidin-biotinylated glycopeptide-lectin method (M.-C. Shao, 1992, Anal. Biochem., 205, 77-82). To illustrate this application, the glycoproteins, ovalbumin and asialofetuin, reduced and S-alkylated with vinylpyridine, were digested with trypsin-L-1-p-tosylamino-2-phenylethylchloromethyl ketone and the tryptic peptides were fractionated by reverse-phase HPLC, monitoring for absorbance at 230 nm. Aliquots of the HPLC fractions (typically 0.2-0.5% of the total volume) were biotinylated and complexed with streptavidin in the wells of a microtiter plate, allowing the streptavidin-glycopeptide complex to adhere to the plate. Suitable lectins, such as concanavalin A, Datura stramonium agglutinin, and peanut agglutinin, all of which had been coupled to horse radish peroxidase, were added, and after thorough washing, only the wells containing streptavidin-bound glycopeptides retained the complementary lectin and gave a positive peroxidase reaction. Less than 1 pmol of glycopeptide can be detected. The demonstration that the glycopeptide detection could be inhibited either by addition of an excess of the appropriate sugar inhibitor to the different lectins or by digestion of the biotinylated glycopeptides with N-glycosidase F or O-glycosidase shows that the glycopeptide-lectin interaction is the basis for the reaction.
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Glicopéptidos/análisis , Mapeo Peptídico/métodos , Secuencia de Aminoácidos , Asialoglicoproteínas/análisis , Biotina , Secuencia de Carbohidratos , Cromatografía Líquida de Alta Presión/métodos , Fetuínas , Glicosilación , Lectinas , Microquímica , Datos de Secuencia Molecular , Ovalbúmina/análisis , Fragmentos de Péptidos/análisis , Peroxidasas , Polisacáridos/análisis , Sensibilidad y Especificidad , alfa-Fetoproteínas/análisisRESUMEN
The enzyme acylaminoacyl-peptide hydrolase represents an attractive reagent for the removal of acetylamino acids from the N-terminus of proteins prior to sequencing. However, the enzyme will not accept intact proteins as substrates, and a blocked protein must consequently be fragmented to generate a relative short blocked peptide, and all the newly generated amino termini must be blocked with an hydrolase-resistant reagent before the enzyme can be used to specifically unblock the N-terminus. When a number of N-acetylated proteins (enolase, alpha-crystallin, ovalbumin, cytochrome c, parvalbumin, superoxide dismutase, and myelin basic protein) were subjected to fragmentation with proteases or cyanogen bromide, treatment with succinic anhydride and exhaustive extraction with ether, and the resulting salt-free, succinylated peptides were incubated with the hydrolase, the N-terminal sequence was specifically unblocked. An aliquot of the entire peptide mixture was applied to the protein sequencer, and a single sequence, corresponding to the known N-terminal sequence starting at residue 2, was obtained. When another aliquot of the same hydrolase-treated peptide mixture was treated with the enzyme acylase I, the liberated acetylamino acid was cleaved, and the N-terminal amino acid (residue 1) could be identified by amino acid analysis. The amount of sequence information obtained from different proteins with different fragmentation methods varied considerably; in the case of parvalbumin a sequence of 12 residues was obtained, while for myelin basic protein, only 3 residues could be identified; the other proteins yielded from 5- to 9-residue sequences.(ABSTRACT TRUNCATED AT 250 WORDS)
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Amidohidrolasas , Proteínas/química , Acetilación , Secuencia de Aminoácidos , Animales , Endopeptidasas , Datos de Secuencia Molecular , Músculos/enzimología , Fragmentos de Péptidos/química , Conejos , RatasRESUMEN
The primary amino acid sequence of rabbit muscle enolase has been determined by standard spinning-cup sequencing techniques applied to peptides produced by chemical (cyanogen bromide and mild acid hydrolysis) and enzymatic fragmentation of the enzyme. The 433 amino acid sequence has been compared to other available enolase sequences from eukaryotic and prokaryotic sources, confirming a high degree of conserved sequence identity; the three mammalian muscle sequences (mouse and rat deduced from c-DNA sequences and rabbit) show 94% identity.
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Músculos/enzimología , Fosfopiruvato Hidratasa/química , Secuencia de Aminoácidos , Animales , Sitios de Unión , Bromuro de Cianógeno , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Fragmentos de Péptidos/química , Conejos , SolubilidadRESUMEN
This study has two objectives: (1) to establish the norm of three graphical projective tests (Children's Social-Self Test, Children's Attitude Inventory and Children's Self-Concept Inventory), and (2) to explore the influence of sex, order of birth, social economic status of family, and leadership of class that effect self-concept of the school-age children. These subjects are selected from elementary schools in Kaohsiung, twenty-two classes are drawn out from each grade by systemic-random sampling. Then two classes are drawn out from 22 classes at random to arrange retest for reliability. The total subjects are 6207 students. From October 1987 to June 1988, six researchers are divided into three group to conduct the test. Besides establishing the norm, we find results from analysis of data as follow: 1. The reliability and validity of three graphical projective tests are good, they can be used for assessing the self-concept of school-age children. 2. The grade, sex, order of birth, social economic status of family, leadership of class all influence the self-concept of school-age children. 3. The self-concept and the grade has negative correlation. 4. The self-concept and the social economic status of family has positive correlation. 5. The girls have higher self-concept than the boys. 6. The order of first-born or second-born has higher self-concept than the others. 7. The ones who are leaders of class have higher self-concept than the others.