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1.
AJNR Am J Neuroradiol ; 42(11): 2009-2015, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34593379

RESUMO

BACKGROUND AND PURPOSE: The golden-angle radial sparse parallel-volumetric interpolated breath-hold (GRASP-VIBE) sequence is a recently introduced imaging technique with high resolution. This study compared the image quality between conventional fat-suppressed T1-weighted TSE and GRASP-VIBE after gadolinium enhancement in the head and neck region. MATERIALS AND METHODS: Data from 65 patients with clinical indications for head and neck MR imaging between September 2020 and January 2021 were retrospectively reviewed. Two radiologists assessed the overall image quality, overall artifacts, and image conspicuities in the oropharynx, hypopharynx, and cervical lymph nodes according to 5-point scores (best score: 5). Interobserver agreement was assessed using weighted κ statistics. The SNR and contrast-to-noise ratio were calculated and compared between the 2 sequences using a paired Wilcoxon signed rank test. RESULTS: The analysis included 52 patients (mean age, 60 [SD, 14 ] years; male, 71.2% [37/52]) who were mostly diagnosed with head and neck malignancies (94.3% [50/52]). κ statistics ranged from slight agreement in cervical lymph node conspicuity (κ = 0.18) to substantial agreement in oropharyngeal mucosal conspicuity (κ = 0.80) (κ range, 0.18-0.80). Moreover, GRASP-VIBE demonstrated significantly higher mean scores in overall image quality (4.68 [SD, 0.41] versus 3.66 [SD, 0.73]), artifacts (4.47 [SD, 0.48] versus 3.58 [SD, 0.71]), oropharyngeal mucosal conspicuity (4.85 [SD, 0.41] versus 4.11 [SD, 0.79]), hypopharyngeal mucosal conspicuity (4.84 [SD, 0.34] versus 3.58 [SD, 0.81]), and cervical lymph node conspicuity (4.79 [SD, 0.32] versus 4.08 [SD, 0.64]) than fat-suppressed T1-weighted TSE (all, P < .001). Furthermore, GRASP-VIBE demonstrated a higher SNR (22.8 [SD, 11.5] versus 11.3 [SD, 5.6], P < .001) and contrast-to-noise ratio (4.7 [SD, 5.4] versus 2.3 [SD, 2.7], P = .059) than fat-suppressed T1-weighted TSE. CONCLUSIONS: GRASP-VIBE provided better image quality with fewer artifacts than conventional fat-suppressed T1-weighted TSE for the head and neck regions.


Assuntos
Meios de Contraste , Aumento da Imagem , Artefatos , Gadolínio , Humanos , Imageamento Tridimensional , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
2.
J Food Sci ; 80(7): S1563-7, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26077386

RESUMO

UNLABELLED: Pork meat sausages were prepared using protein hydrolysates from mechanically deboned chicken meat (MDCM). In terms of the color, compared to the controls before and after storage, the redness (a*) was significantly higher in sausages containing MDCM hydrolysates, ascorbate, and sodium erythorbate. After storage, compared to the other sausage samples, the yellowness (b*) was lower in the sausages containing ascorbate and sodium erythorbate. TBARS was not significantly different among the sausage samples before storage, whereas TBARS and DPPH radical scavenging activities were significantly higher in the sausagescontainingascorbate and sodium erythorbate, compared to the other sausage samples after 4 wk of storage. In terms of sensory evaluation, the color was significantly higher in the sausages containing MDCM hydrolysates, ascorbate, and sodium erythorbate, compared to the other sausage samples after 4 wk of storage. The "off-flavor" and overall acceptability were significantly lower in the sausages containing MDCM hydrolysates than in the other sausage samples. PRACTICAL APPLICATION: In most of the developed countries, meat from spent laying hens is not consumed, leading toan urgent need for effectively utilization or disposal methods. In this study, sausages were prepared using spent laying hens and protein hydrolysates from mechanically deboned chicken meat. Sausage can be made by spent laying hens hydrolysates, although overall acceptability was lower than those of other sausage samples.


Assuntos
Produtos da Carne/análise , Carne/análise , Hidrolisados de Proteína/química , Animais , Ácido Ascórbico/análise , Galinhas , Cor , Análise de Alimentos , Manipulação de Alimentos , Armazenamento de Alimentos , Humanos , Concentração de Íons de Hidrogênio , Suínos , Paladar , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Compostos Orgânicos Voláteis/análise
3.
Free Radic Res ; 47(12): 1009-15, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24074442

RESUMO

This study investigated the effects of onion (Allium cepa, L.) extract on the antioxidant activity of lipids in low-and high-fat-fed mouse brain lipids and its structural change during in vitro human digestion. The onion extracts were passed through an in vitro human digestion model that simulated the composition of the mouth, stomach, and small intestine juice. The brain lipids were collected from low- and high-fat-fed mouse brain and then incubated with the in vitro-digested onion extracts to determine the lipid oxidation. The results confirmed that the main phenolics of onion extract were kaempferol, myricetin, quercetin, and quercitrin. The quercetin content increased with digestion of the onion extract. Antioxidant activity was strongly influenced by in vitro human digestion of both onion extract and quercetin standard. After digestion by the small intestine, the antioxidant activity values were dramatically increased, whereas the antioxidant activity was less influenced by digestion in the stomach for both onion extract and quercetin standard. The inhibitory effect of lipid oxidation of onion extract in mouse brain lipids increased after digestion in the stomach. The inhibitory effect of lipid oxidation of onion extract was higher in the high-fat-fed mouse brain lipids than that in the low-fat-fed mouse brain lipids. The major study finding is that the antioxidative effect of onion extract may be higher in high-fat-fed mouse brain lipids than that in low-fat-fed mouse brain lipids. Thus, dietary onion may have important applications as a natural antioxidant agent in a high-fat diet.


Assuntos
Antioxidantes/farmacologia , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Cebolas/química , Extratos Vegetais/farmacologia , Animais , Dieta , Gorduras na Dieta/administração & dosagem , Feminino , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Quercetina/análogos & derivados , Quercetina/farmacologia
4.
Asian-Australas J Anim Sci ; 26(12): 1781-9, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25049769

RESUMO

Many studies have been carried out with respect to packaging methods and temperature conditions of beef. However, the effects of packaging methods and temperature conditions on the quality characteristics have not been extensively studied in low-grade beef. Low-grade beef samples were divided into 3 groups (C: ziplock bag packaging, T1: vacuum packaging, and T2: modified atmosphere packaging (MAP), CO2/N2 = 3:7) and samples were stored at 4°C for 21 days. The water-holding capacity (WHC) was significantly lower in T1 than in the other samples up to 14 days of storage. The thiobarbituric acid reactive substances and volatile basic nitrogen values were significantly lower in T1 and T2 than in C after 7 to 14 days of storage. The total bacterial counts were significantly lower in T1 and T2 than in C after 14 days of storage. In a sensory evaluation, tenderness and overall acceptability were significantly higher in T1 and T2 than in C at the end of the storage period (21 days). We propose that the MAP method can improve beef quality characteristics of low-grade beef during cold storage. However, the beneficial effects did not outweigh the cost increase to implement MAP.

5.
Asian-Australas J Anim Sci ; 26(7): 1047-53, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25049884

RESUMO

Five hundred and forty crossbred (Korean native black pig×Landrace) F2 were selected at a commercial pig farm and then divided into six different coat color groups: (A: Black, B: White, C: Red, D: White spot in black, E: Black spot in white, F: Black spot in red). Birth weight, 21st d weight, 140th d weight and carcass weight varied among the different coat color groups. D group (white spot in black coat) showed a significantly higher body weight at each weigh (birth weight, 140th d weight and carcass weight) than did the other groups, whereas the C group (red coat color) showed a significantly lower body weight at finishing stage (140th d weight and carcass weight) compared to other groups. Meat quality characteristics, shear force, cooking loss and meat color were not significantly different among the different coat color groups, whereas drip loss was significantly higher in F than in other groups. Most blood characteristics were not significantly different among the different groups, except for the red blood cells.

6.
Poult Sci ; 90(8): 1799-808, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21753218

RESUMO

Imitation crab stick (ICS) samples were divided into 5 treatments, a control composed of commercial ICS containing no breast meat from spent laying hens, and treatments 1, 2, 3, and 4, in which 5, 10, 15, and 20% batter from breast meat of whole spent laying hens was substituted for Alaska pollack surimi, respectively. Imitation crab stick samples containing spent laying hen breast meat batter showed significantly (P < 0.05) higher moisture levels than the control sample. However, the myoglobin and metmyoglobin levels did not differ significantly (P > 0.05) among ICS samples. During storage, whiteness was greater in the control sample than in the ICS samples containing spent laying hen breast meat batter. The saturated and monounsaturated fatty acids increased, whereas the polyunsaturated fatty acids decreased in response to substituting surimi with spent laying hen breast meat batter. The moisture content and pH were increased as the amount of spent laying hen breast meat batter increased. The lipid oxidation value (TBA-reactive substances) and protein degradation value (volatile basic nitrogen) tended to increase during storage as the amount of spent laying hen breast meat batter increased. None of the sensory evaluation items differed among ICS samples during storage, although the color of the final products, mechanical color (by colorimeter), and textural properties did differ among samples. These results indicate that substituting laying hen breast meat batter for Alaska pollack surimi is a very useful method for the production of ICS because it enables the use of a simple production process that does not require steps, such as washing or pH adjustment, for myofibrillar protein recovery.


Assuntos
Manipulação de Alimentos , Produtos da Carne/normas , Animais , Galinhas , Cor , Ácidos Graxos/química , Gadiformes , Miofibrilas , Mioglobina
7.
J Food Sci ; 74(1): C1-5, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19200079

RESUMO

Discoloration characteristics of 3 major muscles (LD, Longissimus dorsi; PM, Psoas major; SM, Semimemebranosus) from Korean native cattle (Hanwoo) were monitored during 7 d of cold storage at 4 degrees C. The muscles were obtained from 12 Hanwoo carcasses at 24 h postmortem. Meat color (CIE L*, a*, b*), myoglobin (Mb) concentration, chemical form, metmyoglobin (MetMb) reducing ability (MRA), mitochondria concentration, and thiobarbituric acid reactive substance (TBARS) were measured at 1, 3, 5, and 7 d of storage. Although there were no significant differences in CIE a* and b*-values between the 3 muscles at day 1, the values of PM muscle were significantly (P < 0.05) lower than those of LD and SM muscles at day 5 and 7. PM muscle showed a rapid decrease in the oxymyoglobin (OxyMb) and an increase in MetMb, which resulted in a significantly (P < 0.05) higher percentage of MetMb in PM muscle compared to LD and SM muscles. Also, the Mb and mitochondria concentration of PM muscle was significantly (P < 0.05) higher than those of LD and SM muscles. However, there were no significant differences in MRA, pH, or TBARS between the 3 muscles during 7 d of cold storage. It was concluded that rapid discoloration (that is, MetMb accumulation) in PM muscle of Hanwoo could be due to its higher contents of Mb and mitochondria.


Assuntos
Temperatura Baixa , Conservação de Alimentos/métodos , Carne/normas , Músculo Esquelético/fisiologia , Mioglobina/análise , Mioglobina/metabolismo , Animais , Bovinos , Concentração de Íons de Hidrogênio , Metamioglobina/análise , Metamioglobina/metabolismo , Mitocôndrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Oxirredução , Pigmentação , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Fatores de Tempo
8.
J Food Sci ; 74(9): C653-7, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-20492097

RESUMO

The purpose of this study was to examine the effect of various fiber additions on lipid digestion during the in vitro digestion of beef patties. The control patties were prepared with 90.5% lean meat and 9.5% tallow. Treatments consisted of 90% lean meat with 9.5% tallow and either 0.5% cellulose, 0.5% chitosan, or 0.5% pectin. The beef patties were then passed through an in vitro digestion model that simulated the composition of the mouth, stomach, and small intestine juices. The change in structure and properties of the lipid droplets was monitored by laser scanning confocal fluorescence microscopy. In general, there was a decrease in lipid droplet diameter as the droplets moved from mouth to stomach to small intestine. The amount of free fatty acid dramatically increased after in vitro digestion in all beef patties. The amount of free fatty acid was, however, lower in beef patties containing chitosan and pectin than other beef patties after in vitro digestion. Beef patties containing various fibers had lower thiobarbituric acid-reactive substances (TBARS) values than samples with no fibers. Among the samples to which fibers were added, chitosan and pectin had lower TBARS than beef patties with cellulose. The cholesterol content decreased after in vitro digestion in all beef patties but was not different among the beef patties before and after in vitro digestion. These results enhance our understanding of the physicochemical and structural changes that occur to ground beef within the gastrointestinal tract.


Assuntos
Fibras na Dieta , Digestão , Metabolismo dos Lipídeos , Produtos da Carne/análise , Animais , Bovinos , Celulose , Quitosana , Colesterol/análise , Ácidos Graxos não Esterificados/análise , Microscopia Confocal , Pectinas , Substâncias Reativas com Ácido Tiobarbitúrico/análise
9.
Poult Sci ; 86(12): 2676-84, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18029816

RESUMO

The sausage samples were made from pork with spent laying hen breast surimi. The samples were divided into 4 groups [sausage made from pork (control) and sausage made from pork with 20% (T1), 40% (T2), and 60% (T3) of spent laying hen breast surimi]. In proximate compositions, the moisture and ash contents of the control were higher than sausage containing spent laying hen surimi samples in all storage periods. The pH and cooking loss were higher in T3 compared with other sausage samples. However, there was no significant difference in water-holding capacity among the sausage samples, whereas shear force was significantly higher in T2. In meat color, sausage containing spent laying hen surimi samples (T1, T2, and T3) have shown to have higher lightness (L) compared with control, and redness (a) was significantly higher in control and T1. Total amino acid content and essential amino acids were increased in sausage containing spent laying hen surimi samples at 0 d of storage. In fatty acid composition, saturated fatty acid was higher in control than sausage containing spent laying hen surimi samples. Thiobarbituric acid reactive substances value was lower in sausage containing spent laying hen surimi samples than control at 2 and 4 wk of storage. Cholesterol content was lower in control compared with sausage containing spent laying hen surimi samples. In sensory evaluation, most test items were not significantly different among the sausage samples although tenderness was higher in T2 and T3 at 0 d of storage.


Assuntos
Manipulação de Alimentos , Produtos da Carne/normas , Aminoácidos , Animais , Galinhas , Colesterol , Cor , Ácidos Graxos , Feminino , Oviposição , Sensação , Suínos , Substâncias Reativas com Ácido Tiobarbitúrico
10.
J. venom. anim. toxins incl. trop. dis ; 12(4): 560-577, 2006. tab, ilus
Artigo em Inglês | LILACS, VETINDEX | ID: lil-453688

RESUMO

In Korea, antivenoms for the treatment of patients bitten by venomous snakes have been imported from Japan or China. Although there is cross-reactivity between these antibodies and venoms from snakes indigenous to Korea (e.g. Agkistrodon genus), protection is not optimal. Antivenoms specifically prepared to neutralize Korean snake venoms could be more effective, with fewer side effects. To this end, we established an infrastructure to develop national standards and created a standardized method to evaluate the efficacy of two horse-derived antivenoms using mouse lethal toxin test. Additionally, we determined the antivenoms neutralizing activity against lethal doses (LD50) of Agkistrodon halys (from Japan) and Jiangzhe Agkistrodon halys (from China) venoms. We also performed cross-neutralization tests using probit analysis on each pairing of venom and antivenom in order to check the possibility of using Jiangzhe A. halys venom as a substitute for A. halys venom, the current standard. Slope of A. halys venom with A. halys antivenom was 10.2 and that of A. halys venom with Jiangzhe A. halys antivenom was 9.6. However, Slope of Jiangzhe A. halys venom with A. halys antivenom was 4.7 while that of Jiangzhe A. halys venom with Jiangzhe A. halys antivenom was 11.5. Therefore, the significant difference in slope patterns suggests that Jiangzhe A. halys venom cannot be used as a substitute for the standard venom to test the anti-lethal toxin activity of antivenoms (p<0.05).(AU)


Assuntos
Animais , Venenos de Serpentes , Testes de Neutralização , Agkistrodon , Anticorpos , Padrões de Referência
11.
Meat Sci ; 66(4): 771-5, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22061007

RESUMO

The effects of conjugated linoleic acid (CLA) on color and lipid oxidation of beef patties were investigated. Ground beef was divided into three batches. The control patties were prepared with 90% lean meat and 10% tallow. The second treatment consisted of 90% lean meat with 9.5% tallow+0.5% CLA sources. The third treatment consisted of 90% lean meat with 8% tallow+2% CLA sources. The patties were wrap-packaged and then stored at 4° for 14 days. The CLA concentration significantly increased (P<0.05) by substituting CLA sources for fat. Storage of the patties did not alter the CLA concentration in beef patties. The treatment substituted with CLA sources had significantly lower TBARS (2-thiobarbituric acid-reactive substances) values (P<0.05) than the control. For oxymyoglobin contents and a* value, substituted CLA sources treatments had significantly higher values than the control. However, L* value significantly increased by substituting CLA sources for fat.

12.
Meat Sci ; 63(2): 249-55, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22062185

RESUMO

Dietary conjugated linoleic acid (CLA) treatment reduced color a*- and b*-values of cooked chicken breast rolls. Sensory panels rated the color of cooked chicken rolls with CLA treatments darker than the control. The production of carbon monoxide (CO) in cooked chicken rolls increased dramatically after irradiation and was correlated with the increased redness of cooked chicken rolls after irradiation. Consumer test indicated that the color of cooked chicken rolls after irradiation was preferred to the nonirradiated, but no preference for the color among the three CLA treatments was found. Irradiation greatly increased volatile production and induced a metallic off-flavor in chicken rolls. Sensory evaluation indicated that the hardness of chicken rolls increased and juiciness decreased as the dietary level of CLA increased.

13.
Meat Sci ; 60(1): 9-15, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22063100

RESUMO

Skinless breast fillets were harvested from broilers fed with 0, 0.25, 0.5, or 1.0% conjugated linoleic acid (CLA) for 3 weeks. Fillets were either vacuum or aerobically packaged, then irradiated at 0 or 3.0 kGy using a Linear Accelerator. Breast fillets were analyzed for thiobarbituric acid reactive substances (TBARS), volatile profiles, and color at 0 and 7 days of storage at 4°C. Dietary CLA reduced TBARS, but had no effect on volatile profiles and color of breast fillets. Color a* value of breast fillets increased after irradiation. Irradiation also induced production of many volatiles, mainly alkanes and alkenes, which could be the breakdown products of unsaturated fatty acids and amino acids. High amount of dimethyl disulfide was detected in the volatiles of irradiated fillets. Low level of hexanal and pentanal in volatiles, together with low TBARS values, indicated that lipid oxidation of breast fillets after irradiation is not a big concern.

14.
Meat Sci ; 61(1): 49-54, 2002 May.
Artigo em Inglês | MEDLINE | ID: mdl-22063912

RESUMO

Raw breast fillets were divided into two groups and either vacuum or aerobically packaged. The fillets in each group were subdivided equally into two groups and then irradiated at 0 or 3 kGy using a Linear Accelerator. After 0, 3 and 7 days of storage at 4 °C, fillets were cooked in an 85 °C water bath (cook-in-bag) to an internal temperature of 74 °C. Oxidation-reduction potential (ORP) of raw fillets was measured before cooking, and color and sensory characteristics were analyzed after cooking. Irradiation decreased the ORP of meat, but the potential in aerobically packaged fillets increased during storage. After cooking, color a*-value of irradiated fillets was higher than that of the non-irradiated. Irradiation of raw meat also changed color L* and b* values after cooking. Aerobic storage reduced the redness of cooked meat induced by irradiation. Irradiated raw broiler fillets stored for 0 day and 3 day under aerobic conditions before cooking produced a oxidized chicken-like odor. The odor, however, disappeared after 7 days of storage under aerobic conditions before cooking. For raw broiler samples stored under vacuum conditions, significant differences in color and odor between irradiated and non-irradiated fillets remained throughout the 7-day storage period after cooking. Irradiation had only a minor influence on lipid oxidation of raw breast fillets as indicated by low TBARS values. This study indicates that the effect of irradiation on color and odor of broiler breast fillets after cooking can be reduced significantly through shelf-display of raw fillets under aerobic conditions. Storage under vacuum conditions before cooking is not effective in reducing irradiation-induced changes in the color and odor of breast fillet after cooking.

15.
Poult Sci ; 80(12): 1748-53, 2001 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11771891

RESUMO

Chicken breast fillets were equally divided into three groups. One group was vacuum packaged, cooked in a water bath (cooked-in-bag) at 82 C for 25 min, and then irradiated at 0 or 3 kGy with a linear accelerator (V-C-I). The other two groups were irradiated at 0 or 3.0 kGy in vacuum packaging (V-I-C) or aerobic packaging (A-I-C). After 3 d of storage at 4 C, the irradiated meats were cooked in a water bath (cooked-in-bag) at 82 C for 25 min. After being cooked, meats were repackaged under vacuum and stored at 4 C. Breast fillets were analyzed at 0 and 21 d after cooking and analyzed for lipid oxidation, color, and volatiles. Irradiation accelerated lipid oxidation of breast fillets. Three days of storage of raw meat in aerobic conditions after irradiation had only minor influences on lipid oxidation after cooking. However, irradiation had a significant effect on the volatile production in meat. Dimethyl disulfide, related to irradiation odor, was significantly higher in irradiated fillets than in nonirradiated fillets for V-C-I and V-I-C, whereas it was only slightly higher for A-I-C. Other volatiles, such as 3-methyl butanal and 2-methyl butanal, were also produced in significant amounts after irradiation, especially in V-C-I and V-I-C. These results showed that irradiating cooked meat induced slightly more changes in volatiles than irradiating raw meat and then cooking. The amount of dimethyl disulfide between irradiated and nonirradiated samples for A-I-C was not different, because the dimethyl disulfide produced by irradiation disappeared during the 3 d in aerobic storage before cooking. Color a* value of irradiated fillets was higher than that of nonirradiated fillets. Irradiation also induced color L* and b* value changes. After 3 d of aerobic storage after irradiation of raw meat, the influence of irradiation on color after cooking was reduced. No significant lipid oxidation occurred during storage as shown by the low values for TBA-reactive substances.


Assuntos
Manipulação de Alimentos/métodos , Irradiação de Alimentos , Conservação de Alimentos/métodos , Carne/análise , Aldeídos/análise , Animais , Galinhas , Culinária , Dissulfetos/análise , Irradiação de Alimentos/métodos , Peroxidação de Lipídeos/efeitos da radiação , Carne/efeitos da radiação , Indústria de Embalagem de Carne , Odorantes , Pigmentação/efeitos da radiação , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Fatores de Tempo
16.
Methods Mol Med ; 17: 231-6, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-21380674

RESUMO

Lambda gt11 cDNA libraries are routinely used for gene isolation with antibody and oligomer probes (1-3). Briefly, a recombinant DNA library from different types of cells is constructed in a λgt 11 expression vector. The foreign peptides (antigens) are expressed as a ß-galactosidase fusion protein in E. coli. Because the lacZ gene has a strong promoter, the fusion proteins comprise from 0.1-4% of the total E. coli proteins (1-3). The expressed fusion antigens are transferred onto nitrocellulose filters and then probed with antibodies to identify the targets containing the specific, cognate epitope.

17.
Biochem Biophys Res Commun ; 166(2): 852-9, 1990 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-2154223

RESUMO

An extracellular myeloperoxidase precursor of HL-60 cells was purified from the culture supernatant by ammonium sulfate precipitation, DEAE-Sepharose chromatography, and monoclonal antibody affinity chromatography. The purified protein was a glycoprotein of approximately 89 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The amino-terminal amino acid sequence of the protein began at amino acid residue 49 of the 745-amino acid sequence deduced from a myeloperoxidase cDNA, suggesting that the protein consisted of 697 amino acid residues. The implications of the precursor in the processing of myeloperoxidase are discussed.


Assuntos
Precursores Enzimáticos/isolamento & purificação , Leucemia Mieloide/enzimologia , Peroxidase/metabolismo , Sequência de Aminoácidos , Espaço Extracelular/enzimologia , Humanos , Dados de Sequência Molecular , Peso Molecular , Testes de Precipitina , Análise Espectral , Células Tumorais Cultivadas
18.
J Biol Chem ; 264(15): 8542-8, 1989 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-2542279

RESUMO

Extracellular myeloperoxidase of human myeloid leukemia HL-60 cells was purified to homogeneity from its culture supernatant by ammonium sulfate fractionation, CM-Sepharose column chromatography, and monoclonal antibody-Sepharose affinity column chromatography. The yield of enzyme activity was 38% that of the ammonium sulfate fraction. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the purified preparation gave a single band of approximately 84 kDa. Analysis of protein blot with antibodies specific for the light and heavy chains of myeloperoxidase indicated that the enzyme contained a light and a heavy chain in a single polypeptide. The amino-terminal amino acid sequence of the enzyme began at amino acid residue 155 of the 745-amino acid sequence predicted from myeloperoxidase cDNA, indicating that the enzyme consisted of 591 amino acids. Sucrose density gradient centrifugation of the enzyme showed that the enzyme was a monomeric form. In pulse-chase experiments on HL-60 cells with [35S]methionine, pulse-labeled myeloperoxidase precursors were shown to be processed to a light chain and a heavy chain of cellular enzyme. During a 3-day chase period, newly formed processed monomeric enzyme was converted to a dimeric form.


Assuntos
Peroxidase/isolamento & purificação , Sequência de Aminoácidos , Western Blotting , Linhagem Celular , Cromatografia de Afinidade , Cromatografia por Troca Iônica , Meios de Cultura , Eletroforese em Gel de Poliacrilamida , Humanos , Leucemia Promielocítica Aguda , Dados de Sequência Molecular , Peso Molecular , Peroxidase/metabolismo
19.
Biochemistry ; 27(16): 5906-14, 1988 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-2903767

RESUMO

Three clones of full-length cDNA encoding human myeloperoxidase were isolated from a human leukemia HL-60 cell cDNA library in lambda gt10 and characterized. Analysis of the nucleotide sequence of one of the cDNA clones, lambda MP-H17, indicated that the cDNA contained 3207 bp with an open reading frame of 2238 bp, a 5' noncoding region of 159 bp, a 3' noncoding region of 800 bp, and a poly(A) tail of 10 bp. cDNA of the two other clones, lambda MP-H7 and lambda MP-H14, each contained insertions with shorter sequences of 96 and 82 bp, respectively, on the open reading frame of lambda MP-H17 cDNA. A myeloperoxidase genomic clone was isolated, and the structure of its 5' region was determined and compared with the structures of these cDNAs. The comparison revealed that the three cDNAs were derived from myeloperoxidase mRNAs produced by alternative splicing from a transcript of the single gene. Nucleotide sequence analysis of the 3' region of the cDNAs of several clones indicated that the mRNAs were polyadenylated at five different sites. Amino acid sequence determination of the amino-terminal and carboxy-terminal portions of the myeloperoxidase light and heavy chains revealed that, during processing of a precursor polypeptide into the mature protein, the amino-terminal polypeptide, the small peptide between the light and heavy chains, and the carboxy-terminal amino acid were excised.


Assuntos
Peroxidase/genética , RNA Mensageiro/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA/genética , Humanos , Dados de Sequência Molecular , Poli A/genética , Splicing de RNA , Células Tumorais Cultivadas/enzimologia
20.
Arch Biochem Biophys ; 255(1): 147-55, 1987 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-2884926

RESUMO

A cDNA encoding the carboxyl-terminal fragment of the human myeloperoxidase heavy chain was isolated and characterized. It was then used to determine the locations of the myeloperoxidase light and heavy chains in the polypeptide precursor. A cDNA library from poly(A)+ RNA from human leukemia HL-60 cells was constructed in pBR322 and screened by differential hybridization with enriched and depleted cDNA probes and then by hybridization with an oligonucleotide probe. A cDNA clone containing 1278 bp with an open reading frame of 474 bp and a 3' noncoding region of 804 bp was isolated. The amino acid sequence deduced from the nucleotide sequence consisted of 158 residues including a sequence of 14 amino acids known to be present in the heavy chain of the molecule. The cDNA also included a stop codon of TAG followed by a noncoding sequence that included a potential recognition site for polyadenylylation and a poly(A) tail. RNA transfer blot analysis with the cDNA probe indicated that myeloperoxidase mRNA was approximately 3.3 kb in length. In vitro translation of the mRNA selected by cDNA hybridization revealed preferential synthesis of a 74,000-Da polypeptide precursor that could be precipitated with anti-myeloperoxidase IgG. Antibodies specific for the heavy and light chains of myeloperoxidase were isolated from antiserum by affinity chromatography employing Sepharose columns covalently bound to the heavy or light chains. Antibodies specific for the light chain or the heavy chain readily precipitated the 74,000-Da precursor polypeptide. These results indicated that myeloperoxidase is synthesized as a single chain which undergoes processing into a light and heavy chain. Furthermore, the heavy chain of myeloperoxidase originates from the carboxyl terminus of the precursor polypeptide.


Assuntos
Clonagem Molecular , DNA/isolamento & purificação , Peroxidase/genética , Sequência de Aminoácidos , Sequência de Bases , Precursores Enzimáticos/isolamento & purificação , Humanos , Hibridização de Ácido Nucleico , Fragmentos de Peptídeos/isolamento & purificação , Poli A/metabolismo , Biossíntese de Proteínas , RNA Mensageiro/metabolismo
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