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1.
Injury ; 53(11): 3774-3780, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36045030

RESUMO

INTRODUCTION: Antegrade posterior column screw (aPCS) fixation via the anterior approach has been widely used for separated the posterior columns in acetabular fracture treatment. Although the relationship between pelvic dysmorphism and sacroiliac screws has been widely studied, no studies have reported on the clinical impact of pelvic dysmorphism on acetabular fractures. This study aimed to reveal the difference in the insertion angle and entry point of aPCS between the dysmorphic and normal pelvises. METHODS: Patients diagnosed with unilateral acetabular fractures and who underwent pelvic computed tomography scans between 2013 and 2019 in two institutes were enrolled in this study. Patients were divided into the dysmorphic and control groups according to the sacral dysmorphic score, which predicts the presence of pelvic dysmorphism, and each group enrolled 130 patients. The semitransparent 3D hemipelvis model was reconstructed using a 3D reconstruction program. The sagittal and coronal angles of a virtual cylinder that fill the safe corridor of the column screw the most were measured. The surface area of the safe corridor and distance of the optimal entry point from the anterior border of the sacroiliac joint were analyzed. The measurements were compared between the dysmorphic and control groups. RESULTS: The average sacral dysmorphic score in the normal and dysmorphic pelvis groups was 56.1 and 81.0, respectively. There were no significant differences in demographic data, including age, sex, height, weight, and body mass index, between the dysmorphic and control groups. There was a significant difference in the average sagittal insertion angle of PCs, which was 38.3° in the control group and 27.2° in the dysmorphic group (P < 0.001). The coronal insertion angles were not significantly different. The dysmorphic group presented longer straight distances (25.9 vs 24.8 mm, P = 0.026) and had a smaller aPCS surface area (685 vs 757 mm2, P < 0.001) than the control group. CONCLUSION: The present study describes a difference in the corridor of aPCS between the dysmorphic and normal pelvis. Insertion of aPCS in the dysmorphic pelvis requires a more acute angular trajectory in the sagittal plane than that in the normal pelvis.


Assuntos
Fixação Interna de Fraturas , Fraturas do Quadril , Humanos , Parafusos Ósseos , Pelve , Fraturas do Quadril/cirurgia , Articulação Sacroilíaca
2.
Mar Drugs ; 20(6)2022 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-35736148

RESUMO

Aquacultured fish are the richest natural source of protein. However, their overproduced biomass is often discarded due to production imbalance, causing considerable losses to the fishery industry. Therefore, it is necessary to utilize surplus fish and add value to overproduced fish. We performed complex enzyme-assisted hydrolysis to determine the correlation between its physical characteristics and anti-hypertensive activity in vitro and in vivo using an SHR model. Protamex-Pepsin assisted hydrolysate from Paralichthys olivaceus (POppH) produced by complex enzyme-assisted hydrolysis contained low-molecular-weight peptides and amino acids with anti-hypertensive activity. POppH regulated blood pressure and serum angiotensin II and angiotensin-I-converting enzyme levels, and histological and ultrasound image analysis revealed substantially reduced thickness and diameter of the carotid aorta in the POppH-administered SHR group. Therefore, we propose to reduce food loss due to overproduction by utilizing the anti-hypertensive activity and physical properties of POppH; the results demonstrate its application as a therapeutic agent.


Assuntos
Linguado , Hipertensão , Inibidores da Enzima Conversora de Angiotensina/química , Animais , Anti-Hipertensivos/química , Pressão Sanguínea , Peixes , Hipertensão/tratamento farmacológico , Hipertensão/patologia , Hidrolisados de Proteína/química , Hidrolisados de Proteína/farmacologia , Ratos , Ratos Endogâmicos SHR
3.
Biomed Pharmacother ; 152: 113210, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35689860

RESUMO

With the rapid increase in the elderly population worldwide, the number of people with sarcopenia has also increased significantly, and this disease is emerging as a medical and social issue. The development of pharmaceutics targeting sarcopenia is limited owing to the occurrence of side effects, and exercise therapy also has a limited scope of application. Therefore, it is necessary to develop safe and biocompatible agents to treat age-related sarcopenia. Ishige okamurae (IO), an edible brown alga, and its active substance, diphloroethohydroxycarmalol (DPHC), have been reported to have various physiological functions, including skeletal muscle regeneration ability. However, this effect has not been verified in an in vivo aging model. As an aging model, the oral IO extracts and DPHC supplemented 14-month-old female C57BL/6J mice were compared to the young group in this study; the mice model showed a substantial restoration of physical exercise ability with the imbalance of famine hormone and senescence-associated secretary phenotypes compared with those in young mice. Regarding the lean mass increase in aging mice following IO extract and DPHC administration, the muscular characteristics and molecular alterations in the gastrocnemius and soleus muscles, which are sensitive to the damage that occurs during the aging process, were significantly improved. Collectively, the current study reveals that the natural agent IO extract and its derivative DPHC can reverse sarcopenia that occurs during the process of aging by improving the imbalance of muscle regeneration in vivo.


Assuntos
Phaeophyceae , Sarcopenia , Idoso , Envelhecimento , Animais , Feminino , Compostos Heterocíclicos com 3 Anéis , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Sarcopenia/tratamento farmacológico
4.
Mar Drugs ; 20(5)2022 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-35621931

RESUMO

The in vitro capacity of Ishige okamurae extract (IO) to improve impaired muscle function has been previously examined. However, the mechanism underlying IO-mediated muscle protein metabolism and the role of its component, Ishophloroglucin A (IPA), in mice with dexamethasone (Dexa)-induced muscle atrophy remains unknown. In the present study, we evaluated the effect of IO and IPA supplementation on Dexa-induced muscle atrophy by assessing muscle protein metabolism in gastrocnemius and soleus muscles of mice. IO and IPA supplementation improved the Dexa-induced decrease in muscle weight and width, leading to enhanced grip strength. In addition, IO and IPA supplementation regulated impaired protein synthesis (PI3K and Akt) or degradation (muscle-specific ubiquitin ligase muscle RING finger and atrogin-1) by modulating mRNA levels in gastrocnemius and soleus muscles. Additionally, IO and IPA upregulated mRNA levels associated with muscle growth activation (transient receptor potential vanilloid type 4 and adenosine A1 receptor) or inhibition (myostatin and sirtuin 1) in gastrocnemius and soleus muscle tissues of Dexa-induced mice. Collectively, these results suggest that IO and IO-derived IPA can regulate muscle growth through muscle protein metabolism in Dexa-induced muscle atrophy.


Assuntos
Misturas Complexas , Proteínas Musculares , Atrofia Muscular , Phaeophyceae , Animais , Benzofuranos , Misturas Complexas/farmacologia , Misturas Complexas/uso terapêutico , Dexametasona/efeitos adversos , Dioxinas , Camundongos , Proteínas Musculares/metabolismo , Músculo Esquelético/efeitos dos fármacos , Músculo Esquelético/metabolismo , Atrofia Muscular/induzido quimicamente , Atrofia Muscular/tratamento farmacológico , Atrofia Muscular/metabolismo , Phaeophyceae/metabolismo , RNA Mensageiro/metabolismo
5.
Pharmaceutics ; 14(4)2022 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-35456553

RESUMO

Muscle atrophy refers to the loss of skeletal muscle mass, myofiber size, and related physical functions such as walking speed or grip strength caused by aging or a lack of physical activity due to injury or illness and can also be attributed to excessive exposure to corticosteroids. Ishige okamurae (IO) and its active component, diphlorethohydroxycarmalol (DPHC), have been known to improve glucose homeostasis by controlling the contraction of skeletal muscles. Based on this idea, we hypothesized that the effects of DPHC and IO extract on muscle metabolism are associated with their role in improving muscle physical function. This study assessed the effects of DPHC or IO extract on muscle behavioral responses with their metabolic properties in muscle atrophy induced by glucocorticoids and dexamethasone (DEX) in vivo. In addition to the improvement in muscle behavioral response by DPHC or IO extract, the loss of muscle fiber and the related metabolic properties by DEX exposure in the gastrocnemius and soleus of calf muscle was prevented. These findings suggest that IO extract and its active component DPHC can potentially prevent muscle atrophy caused by exposure to corticosteroids and could be used to treat reverse skeletal atrophy.

6.
Orthop J Sports Med ; 10(4): 23259671221087189, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-35400138

RESUMO

Background: To completely eliminate an infection, the antibiotic concentration must exceed the minimum inhibitory concentration for the causative pathogens. The antibiotic-loaded collagen sponge (ALCS) has been used to manage an infection through a high local concentration of antibiotics. Purpose: To evaluate the clinical efficacy of ALCS in the arthroscopic treatment of patients with acute septic arthritis of the native knee. Study Design: Cohort study; Level of evidence, 3. Methods: The authors retrospectively reviewed the data for 132 consecutive patients with acute septic arthritis of the knee who underwent arthroscopic debridement between 2013 and 2019. Patients with a history of surgery in the affected joint, concomitant osteomyelitis, or accompanying medical infection were excluded. The study patients were divided into 2 groups based on the use of ALCS during surgery. C-reactive protein (CRP) levels were temporally measured to normalization and the duration of intravenous antibiotic administration was recorded, and the mean length of hospital stay was compared between the groups. Independent t test and chi-square test were performed to assess differences between the groups, and univariate analysis was used to analyze factors affecting recurrence of septic arthritis. Results: Of the 132 patients, 88 were ultimately included in this study; 51 patients were managed without ALCS and 37 were treated with ALCS. There were statistically significant differences between groups in the mean period for CRP-level normalization (15.2 ± 8.2 days [ALCS] vs 26.2 ± 14.7 days [without ALCS]; P = .004), mean duration of intravenous antibiotic use (18.4 ± 7.3 days [ALCS] vs 26.6 ± 16.1 days [without ALCS]; P = .046), and mean length of hospital stay (21.1 ± 11.6 days [ALCS] vs 30.3 ± 18.3 days [without ALCS]; P = .045). The preoperative CRP level was the only significant risk factor for recurrence or treatment failure after single arthroscopic debridement. Conclusion: The results of this study indicated that ALCS was able to reduce the duration of CRP-level normalization, intravenous antibiotic use, and hospitalization in patients who underwent arthroscopic irrigation and debridement for acute septic arthritis of the native knee.

7.
Nutrients ; 11(11)2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31731817

RESUMO

It is well known that perivascular fat tissue (PVAT) dysfunction can induce endothelial cell (EC) dysfunction, an event which is related with various cardiovascular diseases. In this study, we evaluated whether Ecklonia cava extract (ECE) and pyrogallol-phloroglucinol-6,6-bieckol (PPB), one component of ECE, could attenuate EC dysfunction by modulating diet-induced PVAT dysfunction mediated by inflammation and ER stress. A high fat diet (HFD) led to an increase in the number and size of white adipocytes in PVAT; PPB and ECE attenuated those increases. Additionally, ECE and PPB attenuated: (i) an increase in the number of M1 macrophages and the expression level of monocyte chemoattractant protein-1 (MCP-1), both of which are related to increases in macrophage infiltration and induction of inflammation in PVAT, and (ii) the expression of pro-inflammatory cytokines (e.g., tumor necrosis factor-α (TNF-α) and interleukin (IL)-6, chemerin) in PVAT which led to vasoconstriction. Furthermore, ECE and PPB: (i) enhanced the expression of adiponectin and IL-10 which had anti-inflammatory and vasodilator effects, (ii) decreased HFD-induced endoplasmic reticulum (ER) stress and (iii) attenuated the ER stress mediated reduction in sirtuin type 1 (Sirt1) and peroxisome proliferator-activated receptor γ (PPARγ) expression. Protective effects against decreased Sirt1 and PPARγ expression led to the restoration of uncoupling protein -1 (UCP-1) expression and the browning process in PVAT. PPB or ECE attenuated endothelial dysfunction by enhancing the pAMPK-PI3K-peNOS pathway and reducing the expression of endothelin-1 (ET-1). In conclusion, PPB and ECE attenuated PVAT dysfunction and subsequent endothelial dysfunction by: (i) decreasing inflammation and ER stress, and (ii) modulating brown adipocyte function.


Assuntos
Adipócitos Marrons/efeitos dos fármacos , Anti-Inflamatórios/farmacologia , Células Endoteliais/efeitos dos fármacos , Doenças Vasculares Periféricas/tratamento farmacológico , Phaeophyceae , Extratos Vegetais/farmacologia , Adiponectina/metabolismo , Tecido Adiposo/metabolismo , Animais , Quimiocina CCL2/metabolismo , Dieta Hiperlipídica/efeitos adversos , Dioxinas/farmacologia , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Inflamação , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Doenças Vasculares Periféricas/etiologia , Floroglucinol/farmacologia , Pirogalol/farmacologia
8.
J Med Food ; 21(11): 1113-1119, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30457472

RESUMO

Black-fruited chokeberries (Aronia melanocarpa), growing mainly in the Central and Eastern European countries, have health benefits due to the high concentrations of polyphenolic compounds. However, a strong bitter taste of chokeberries limits its usage as functional food. We hypothesized that the fermented A. melanocarpa with a reduced bitter taste would improve insulin sensitivity and/or ameliorate weight gain induced by high-fat diet (HFD) in male C57BL/6J mice. The mice were administered with HFD together with the 100 mg/kg of natural A. melanocarpa (T1) or the fermented A. melanocarpa (T2) for 8 weeks. The treatment with T2 (100 mg/kg body weight, p.o.) markedly attenuated the weight gain and the increase in serum triglyceride level induced by HFD. The T2-treated group had better glucose tolerance and higher insulin sensitivity as measured by oral glucose tolerance test and intraperitoneal insulin tolerance test in comparison to the T1-treated group. Phytochemical analysis revealed that the main constituents of T2 were cyanidin-3-xyloside and 1-(3',4'-dihydroxycinnamoyl)cyclopenta-2,3-diol, and the content of cyanidin glycosides (3-glucoside, 3-xyloside) was significantly reduced during the fermentation process. From the above results, we postulated that antiobesity effect of black chokeberry was not closely correlated with the cyanidin content. Fermented chokeberry might be a viable dietary supplement rich in bioactive compounds useful in preventing obesity.


Assuntos
Acetobacter/metabolismo , Fármacos Antiobesidade/metabolismo , Alimentos Fermentados/análise , Obesidade/dietoterapia , Photinia/microbiologia , Extratos Vegetais/metabolismo , Saccharomyces/metabolismo , Animais , Antocianinas/administração & dosagem , Antocianinas/química , Antocianinas/metabolismo , Fármacos Antiobesidade/química , Dieta Hiperlipídica/efeitos adversos , Fermentação , Alimentos Fermentados/microbiologia , Frutas/química , Frutas/metabolismo , Frutas/microbiologia , Humanos , Insulina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Obesidade/etiologia , Obesidade/metabolismo , Photinia/química , Photinia/metabolismo , Extratos Vegetais/química
9.
Physiol Plant ; 148(2): 189-99, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23039825

RESUMO

R2R3-type MYB transcription factors (TFs) play important roles in transcriptional regulation of anthocyanins. The R2R3-type IbMYB1 is known to be a key regulator of anthocyanin biosynthesis in the storage roots of sweetpotato. We previously showed that transient expression of IbMYB1a led to anthocyanin pigmentation in tobacco leaves. In this article, we generated transgenic Arabidopsis plants expressing the IbMYB1a gene under the control of CaMV 35S promoter, and the sweetpotato SPO and SWPA2 promoters. Overexpression of IbMYBa in transgenic Arabidopsis produced strong anthocyanin pigmentation in seedlings and generated a deep purple color in leaves, stems and seeds. Reverse transcription-polymerase chain reaction analysis showed that IbMYB1a expression induced upregulation of several structural genes in the anthocyanin biosynthetic pathway, including 4CL, CHI, F3'H, DFR, AGT, AAT and GST. Furthermore, overexpression of IbMYB1a led to enhanced expression of the AtTT8 (bHLH) and PAP1/AtMYB75 genes. high-performance liquid chromatography analysis revealed that IbMYB1a expression led to the production of cyanidin as a major core molecule of anthocyanidins in Arabidopsis, as occurs in the purple leaves of sweetpotato (cv. Sinzami). This result shows that the IbMYB1a TF is sufficient to induce anthocyanin accumulation in seedlings, leaves, stems and seeds of Arabidopsis plants.


Assuntos
Antocianinas/metabolismo , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Ipomoea batatas/genética , Fatores de Transcrição/genética , Antocianinas/análise , Arabidopsis/metabolismo , Expressão Gênica , Especificidade de Órgãos , Proteínas Associadas a Pancreatite , Fenótipo , Pigmentação , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Caules de Planta/genética , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas/genética , Plântula/genética , Plântula/metabolismo , Sementes/genética , Sementes/metabolismo , Fatores de Transcrição/metabolismo , Transcrição Gênica
10.
Biotechnol Lett ; 33(8): 1663-6, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21487781

RESUMO

A new zymogram method, silver-stained fibrin zymography, for separation of protease bands and activity detection using a single substrate gel, was developed. The method takes advantage of the nanoscale sensitivity of both zymography and silver staining. After SDS-PAGE in a gel containing fibrin, the gel was incubated in enzyme reaction buffer and the zymogram was silver-stained. Bands with protease activity were stained with silver in clear areas where the protein substrate had been degraded. The molecular sizes of proteases were accurately determined. Furthermore, proteases of high molecular weight were clearly and sharply resolved.


Assuntos
Eletroforese em Gel de Poliacrilamida/métodos , Peptídeo Hidrolases/isolamento & purificação , Proteínas de Bactérias/química , Proteínas de Bactérias/isolamento & purificação , Proteínas de Bactérias/metabolismo , Precursores Enzimáticos , Fibrina/química , Fibrina/metabolismo , Peso Molecular , Peptídeo Hidrolases/química , Peptídeo Hidrolases/metabolismo , Coloração pela Prata
11.
Phytother Res ; 25(9): 1415-7, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21433153

RESUMO

Gout is a clinical syndrome in which tissue damage is induced by a chronic metabolic disorder associated with increased concentrations of uric acid in the blood. The study investigated the hypouricemic effects of anthocyanin extracts from purple sweet potato (APSP), and allopurinol, on serum uric acid levels in hyperuricemic mice. It was found that administration of a single oral dose of 100 mg/kg APSP to such animals reduced the serum uric acid concentration to 4.10 ± 0.04 mg/dL, compared with a concentration of 10.25 ± 0.63 mg/dL in the hyperuricemic control group.


Assuntos
Antocianinas/farmacologia , Hiperuricemia/tratamento farmacológico , Ipomoea batatas/química , Extratos Vegetais/farmacologia , Alopurinol/farmacologia , Animais , Modelos Animais de Doenças , Hiperuricemia/induzido quimicamente , Masculino , Camundongos , Camundongos Endogâmicos ICR , Ácido Oxônico , Ácido Úrico/sangue
12.
Biotechnol Lett ; 33(4): 783-6, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21188617

RESUMO

Bioconversion of quercetin glucosides using four generally recognized as safe (GRAS) organisms (Aspergillus oryzae, Bacillus subtilis, Lactobacillus plantarum, and Saccharomyces cerevisiae) was evaluated by measuring changes in the levels of quercetin compounds of onion. Of the four organisms, S. cerevisiae increased the content of quercetin-3-O-ß-D-glucoside (III; isoquercitrin) and quercetin (IV), whereas decreasing quercetin-3,4'-O-ß-D-glucoside (I) and quercetin-4'-O-ß-D-glucoside (II). Also, S. cerevisiae converted authentic compound I to III, and II to IV, respectively. These results suggest that S. cerevisiae can be used to increase the levels of isoquercitrin (III), the most bioavailable quercetin compound in onion.


Assuntos
Glucosídeos/metabolismo , Cebolas/química , Quercetina/metabolismo , Saccharomyces cerevisiae/metabolismo , Glucosídeos/análise , Glucosídeos/química , Estrutura Molecular , Quercetina/análogos & derivados , Quercetina/análise , Quercetina/química , Estereoisomerismo , Regulação para Cima
13.
J Med Food ; 13(6): 1532-6, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20954802

RESUMO

Chives have been used both as food and as medicine. Previously, two fibrinolytic enzymes, ATFE-I (90 kDa) and ATFE-II (55 kDa), were identified in chives (Allium tuberosum), a perennial herb. In the present work, ATFE-II was purified by ion-exchange chromatography followed by gel filtration. In addition, the enzyme properties of ATFE-I and ATFE-II were compared. The molecular mass and isoelectric point (pI value) of ATFE-II were 55 kDa and pI 4.0, respectively, as revealed using one- or two-dimensional fibrin zymography. ATFE-II was optimally active at pH 7.0 and 45°C. ATFE-II degraded the Aα-chain of human fibrinogen but did not hydrolyze the Bß-chain or the γ-chain, indicating that the enzyme is an α-fibrinogenase. The proteolytic activity of ATFE-II was completely inhibited by 1 mM leupeptin, indicating that the enzyme belongs to the cysteine protease class. ATFE-II was also inhibited by 1 mM Fe²(+). ATFE-II exhibited high specificity for MeO-Suc-Arg-Pro-Tyr-p-nitroaniline (S-2586), a synthetic chromogenic substrate of chymotrypsin. Thus proteolytic enzymes from A. tuberosum may be useful as thrombolytic agents.


Assuntos
Cebolinha-Francesa/enzimologia , Cisteína Endopeptidases , Descoberta de Drogas , Fibrinólise , Fibrinolíticos , Componentes Aéreos da Planta/enzimologia , Proteínas de Plantas , Cisteína Endopeptidases/química , Cisteína Endopeptidases/isolamento & purificação , Cisteína Endopeptidases/metabolismo , Inibidores de Cisteína Proteinase/farmacologia , Compostos Ferrosos/farmacologia , Fibrinogênio/metabolismo , Fibrinolíticos/química , Fibrinolíticos/isolamento & purificação , Fibrinolíticos/metabolismo , Concentração de Íons de Hidrogênio , Hidrólise , Ponto Isoelétrico , Isoenzimas/antagonistas & inibidores , Isoenzimas/química , Isoenzimas/isolamento & purificação , Isoenzimas/metabolismo , Leupeptinas/farmacologia , Peso Molecular , Oligopeptídeos/metabolismo , Proteínas de Plantas/antagonistas & inibidores , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Especificidade por Substrato , Temperatura , Terapia Trombolítica , Trombose/tratamento farmacológico
14.
J Ind Microbiol Biotechnol ; 36(3): 417-26, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19104859

RESUMO

A novel fibrinolytic enzyme (AJ) was purified from Staphylococcus sp. strain AJ screened from Korean salt-fermented Anchovy-jeot. Relative molecular weight of AJ was determined as 26 kDa by using SDS-PAGE and fibrin zymography. Based on a 2D gel, AJ was found to consist of three active isoforms (pI 5.5-6.0) with the same N-terminal amino acid sequence. AJ exhibited optimum pH and temperature at 2.5-3.0 and 85 degrees C, respectively. AJ kept 85% of the initial activity after heating at 100 degrees C for 20 min on the zymogram gel. The Michaelis constant (K (m)) and K (cat) values of AJ towards alpha-casein were 0.38 mM and 19.73 s(-1), respectively. AJ cleaved the A alpha-chain of fibrinogen but did not affect the B beta- and gamma-chains, indicating that it is an alpha-fibrinogenase. The fibrinolytic activity was inhibited by diisopropyl fluorophosphate, indicating AJ is a serine protease. Interestingly, AJ was very stable at acidic condition, SDS, and heat (100 degrees C), whereas it was easily degraded at neutral and alkaline conditions. In particular, AJ formed an active homo-dimer in the pH range from 7.0 to 8.0. To our knowledge, a similar combination of acid and heat stability has not yet been reported for other fibrinolytic enzymes.


Assuntos
Proteínas de Bactérias , Estabilidade Enzimática , Fibrina/metabolismo , Peixes/microbiologia , Temperatura Alta , Cloreto de Sódio , Staphylococcus/enzimologia , Sequência de Aminoácidos , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Fermentação , Fibrinólise , Microbiologia de Alimentos , Humanos , Concentração de Íons de Hidrogênio , Coreia (Geográfico) , Dados de Sequência Molecular , Análise de Sequência de DNA , Staphylococcus/genética , Staphylococcus/isolamento & purificação
15.
J Biochem Mol Biol ; 38(6): 763-5, 2005 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-16336793

RESUMO

A new simplified procedure for identifying human plasmin was developed using a DTT copolymerized agarose stacking gel (ASG) system. Agarose (1 %) was used for the stacking gel because DTT inhibits the polymerization of acrylamide. Human plasmin showed the lowest activity at pH 9.0. There was a similar catalytically active pattern observed under acidic conditions (pH 3.0) to that observed under alkaline conditions (pH 10.0 or 11.0). Using the ASG system, the primary structure of the heavy chain could be established at pH 3.0. This protein was found to consist of three fragments, 45 kDa, 23 kDa, and 13 kDa. These results showed that the heavy chain has a similar structure to the autolysed plasmin (Wu et al., 1987b) but there is a different start amino acid sequence of the N-termini.


Assuntos
Bioquímica/métodos , Ditiotreitol/farmacologia , Fibrinolisina/química , Sequência de Aminoácidos , Dissulfetos/química , Ditiotreitol/química , Eletroforese em Gel de Ágar/métodos , Eletroforese em Gel Bidimensional , Eletroforese em Gel de Poliacrilamida , Fibrinogênio/química , Humanos , Concentração de Íons de Hidrogênio , Plasminogênio/química , Estrutura Terciária de Proteína , Sefarose/química , Trombina/química
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