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1.
Food Chem ; 308: 125576, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-31648092

RESUMO

This study investigated the effects of cold storage at different temperatures (4, -0.5, -3, and -20 °C) on protein degradation and its relationship to structural changes of black carp muscle. At -0.5 and 4 °C, major structural changes occurred, including the formation of gaps between myofibers and myofibrils, breakage of myofibrils and myofibers, and degradation of sarcoplasmic reticulum. Gel-based proteomic analysis showed that these structural changes were accompanied by degradation of a series of myofibrillar proteins, including titin, nebulin, troponin, myosin, myomesin, myosin-binding protein, and α-actinin. Loss of extractable gelatinolytic and caseinolytic protease activities was also observed. At -3 and -20 °C, formation of ice crystals was the most noticeable change. The major proteins were degraded at different locations in the black carp muscle, and gelatinolytic and caseinolytic proteases appear to contribute to the degradation of those proteins.


Assuntos
Carpas/metabolismo , Actinina/metabolismo , Animais , Proteínas de Transporte , Temperatura Baixa , Conectina/metabolismo , Cyprinidae/metabolismo , Proteínas Musculares/metabolismo , Miofibrilas/metabolismo , Miosinas/metabolismo , Proteólise , Proteômica
2.
Cell Mol Biol Lett ; 24: 38, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31182966

RESUMO

Background: Exploration of the genes with abnormal expression during the development of breast cancer is essential to provide a deeper understanding of the mechanisms involved. Transcriptome sequencing and bioinformatics analysis of invasive ductal carcinoma and paracancerous tissues from the same patient were performed to identify the key genes and signaling pathways related to breast cancer development. Methods: Samples of breast tumor tissue and paracancerous breast tissue were obtained from 6 patients. Sequencing used the Illumina HiSeq platform. All. Only perfectly matched clean reads were mapped to the reference genome database, further analyzed and annotated based on the reference genome information. Differentially expressed genes (DEGs) were identified using the DESeq R package (1.10.1) and DEGSeq R package (1.12.0). Using KOBAS software to execute the KEGG bioinformatics analyses, enriched signaling pathways of DEGs involved in the occurrence of breast cancer were determined. Subsequently, quantitative real time PCR was used to verify the accuracy of the expression profile of key DEGs from the RNA-seq result and to explore the expression patterns of novel cancer-related genes on 8 different clinical individuals. Results: The transcriptomic sequencing results showed 937 DEGs, including 487 upregulated and 450 downregulated genes in the breast cancer specimens. Further quantitative gene expression analysis was performed and captured 252 DEGs (201 downregulated and 51 upregulated) that showed the same differential expression pattern in all libraries. Finally, 6 upregulated DEGs (CST2, DRP2, CLEC5A, SCD, KIAA1211, DTL) and 6 downregulated DEGs (STAC2, BTNL9, CA4, CD300LG, GPIHBP1 and PIGR), were confirmed in a quantitative real time PCR comparison of breast cancer and paracancerous breast tissues from 8 clinical specimens. KEGG analysis revealed various pathway changes, including 20 upregulated and 21 downregulated gene enrichment pathways. The extracellular matrix-receptor (ECM-receptor) interaction pathway was the most enriched pathway: all genes in this pathway were DEGs, including the THBS family, collagen and fibronectin. These DEGs and the ECM-receptor interaction pathway may perform important roles in breast cancer. Conclusion: Several potential breast cancer-related genes and pathways were captured, including 7 novel upregulated genes and 76 novel downregulated genes that were not found in other studies. These genes are related to cell proliferation, movement and adhesion. They may be important for research into breast cancer mechanisms, particularly CST2 and CA4. A key signaling pathway, the ECM-receptor interaction signal pathway, was also identified as possibly involved in the development of breast cancer.


Assuntos
Neoplasias da Mama/genética , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Receptores de Superfície Celular/metabolismo , Transdução de Sinais/genética , Regulação para Baixo/genética , Feminino , Humanos , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Reprodutibilidade dos Testes , Análise de Sequência de RNA , Transcriptoma/genética , Regulação para Cima/genética
3.
Pathol Res Pract ; 215(6): 152405, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30981459

RESUMO

To reveal novel lncRNAs and explore how could lncRNA affect the ER/PR positive type breast cancer, 16 different lncRNA transcriptomes (8 breast cancer tissues and 8 normal breast tissues) were successfully sequenced. In total, 8,954 high quality lncRNAs, including 5,516 lncRNAs reported in the previous studies and 3,438 novel lncRNAs, were annotated. The highest expressed lncRNAs were MALAT1, SCARNA10, RP11-206M11.7 and NEAT1, and the highest expressing mRNAs were RPL19, SCGB2A2, FTL and TMSB4 × . Of the 615 differentially expressed lncRNAs, 323 showed up regulated (P < 0.05) expression patterns in breast cancer, and 292 showed down regulated expression patterns. Of the 8,954 genes, 5,516 genes were upregulated in breast cancer, and 3,438 were downregulated. In total, the targets of 238 lncRNAs were confirmed by two lncRNA target prediction programs. Within these genes, Ras responsive element binding protein 1, Ras association domain family member 6, Ras association domain family member 8, Ras protein specific guanine nucleotide releasing factor 1and other 10 different Ras associated different expressed genes were predicted as targets of lncRNAs. These different expressed lncRNAs which could regulate the Ras gene families and ECM pathway may be another mechanism why the expression pattern of Ras genes changed in breast cancer. All these cancer-related genes (Ras genes) were annotated as targets of lncRNAs in the breast cancer transcriptome may provide us with a new way to understand the occurrence and development of breast cancer.


Assuntos
Neoplasias da Mama/genética , Carcinoma Ductal de Mama/genética , Genes ras/genética , RNA Longo não Codificante/genética , Adulto , Feminino , Perfilação da Expressão Gênica , Humanos , Pessoa de Meia-Idade , Receptores Estrogênicos/biossíntese , Receptores de Progesterona/biossíntese , Transcriptoma
4.
Crit Rev Food Sci Nutr ; 59(22): 3564-3578, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30040449

RESUMO

Protein oxidation readily occurs in postmortem muscle during storage and processing. Over the past decade new analytical methods have been developed and new aspects of protein oxidation in meat have been studied, such as the reaction mechanism, and impacts on eating quality and nutritional value. It is now evident that amino acid side chains in myofibrillar proteins undergoes modifications due to oxidative stress. In turn this will lead to formation of new protein-protein cross-links in structural proteins, however, also the overall level of fixed-charge groups attached to the peptide backbones is modified. Meat texture and water-holding are important quality attributes and they are affected by the oxidation of structural proteins. Different mechanisms have been suggested to explain the oxidation-induced quality changes, focusing mainly on reduced proteolysis and formation of cross-links. This review explores the current understanding of protein oxidation in fresh meat in relation to texture and water-holding. The consequences of protein oxidation at molecular level in relation to oxidation-induced cross-linking and changes in net charges of myofibrillar proteins, and the impacts on texture and water-holding are discussed.

5.
Sensors (Basel) ; 18(11)2018 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-30463259

RESUMO

This paper presents two approaches for the structural damage identification of a bridge from the dynamic response recorded from a test vehicle during its passage over the bridge. Using the acceleration response recorded by the vibration sensors mounted on a test vehicle during its passage over the bridge, along with the computed displacement response, the bending stiffness of the bridge can be determined using either: (1) the frequency-domain method based on the improved directed stiffness method with the identified frequency and corresponding mode shape, or (2) the time-domain method based on the residual vector of the least squares method with a fourth-order displacement moment. By comparing the bending stiffness values identified from the vehicle-collected data for the bridge under the undamaged and damaged states that are monitored regularly by the test vehicle, the bridge damage location and severity can be identified. Through numerical simulations and field tests, the present approaches are shown to be effective and feasible.

6.
Sensors (Basel) ; 18(1)2018 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-29346289

RESUMO

In this study, we used bands 7, 4, and 3 of the Advance Himawari Imager (AHI) data, combined with a Threshold Algorithm and a visual interpretation method to monitor the entire process of grassland fires that occurred on the China-Mongolia border regions, between 05:40 (UTC) on April 19th to 13:50 (UTC) on April 21st 2016. The results of the AHI data monitoring are evaluated by the fire point product data, the wind field data, and the environmental information data of the area in which the fire took place. The monitoring result shows that, the grassland fire burned for two days and eight hours with a total burned area of about 2708.29 km². It mainly spread from the northwest to the southeast, with a maximum burning speed of 20.9 m/s, a minimum speed of 2.52 m/s, and an average speed of about 12.07 m/s. Thus, using AHI data can not only quickly and accurately track the dynamic development of a grassland fire, but also estimate the spread speed and direction. The evaluation of fire monitoring results reveals that AHI data with high precision and timeliness can be highly consistent with the actual situation.

7.
Meat Sci ; 135: 102-108, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28968552

RESUMO

Hypochlorous acid (HClO) is a strong oxidant that is able to mediate protein oxidation. In order to study the effect of oxidation on charges, aggregation and water-holding of myofibrillar proteins, extracted myofibrils were oxidized by incubation with different concentrations of HClO (0, 1, 5, and 10mM). Loss of free thiols, loss of histidine and formation of carbonyls were greater with increasing oxidation level and the particle size increased. Water-holding in the 5 and 10mM HClO groups were greater than in the non-oxidized control. Isoelectric focusing (IEF) showed that the isoelectric point (pI) of oxidized proteins were lower compared to non-oxidized ones. The lower pI values of oxidized proteins suggests that oxidation increased the overall net negative charge of myofibrillar proteins solubilized for IEF. Here we propose a hypothesis that oxidation-induced increase in net negative charges is the driving force for improved water-holding in myofibrils, whereas protein cross-linking and aggregation have an opposing effect by decreasing the water-holding.


Assuntos
Proteínas Musculares/fisiologia , Miofibrilas/química , Oxirredução , Aminoácidos/análise , Animais , Histidina , Ácido Hipocloroso/farmacologia , Ponto Isoelétrico , Compostos de Sulfidrila , Suínos , Água/metabolismo
8.
Artigo em Inglês | MEDLINE | ID: mdl-27834923

RESUMO

A suspended monorail transit system is a category of urban rail transit, which is effective in alleviating traffic pressure and injury prevention. Meanwhile, with the advantages of low cost and short construction time, suspended monorail transit systems show vast potential for future development. However, the suspended monorail has not been systematically studied in China, and there is a lack of relevant knowledge and analytical methods. To ensure the health and reliability of a suspended monorail transit system, the driving safety of vehicles and structure dynamic behaviors when vehicles are running on the bridge should be analyzed and evaluated. Based on the method of vehicle-bridge coupling vibration theory, the finite element method (FEM) software ANSYS and multi-body dynamics software SIMPACK are adopted respectively to establish the finite element model for bridge and the multi-body vehicle. A co-simulation method is employed to investigate the vehicle-bridge coupling vibration for the transit system. The traffic operation factors, including train formation, track irregularity and tire stiffness, are incorporated into the models separately to analyze the bridge and vehicle responses. The results show that the coupling of dynamic effects of the suspended monorail system between vehicle and bridge are significant in the case studied, and it is strongly suggested to take necessary measures for vibration suppression. The simulation of track irregularity is a critical factor for its vibration safety, and the track irregularity of A-level road roughness negatively influences the system vibration safety.


Assuntos
Ferrovias , Segurança , China , Análise de Elementos Finitos , Modelos Teóricos , Ferrovias/normas , Vibração
9.
Meat Sci ; 121: 189-195, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27341620

RESUMO

Patties were made from raw minced beef after storage for 6days in modified atmosphere (0, 20, 40, 60, and 80% oxygen) to study the combined effect of oxygen concentration and cooking temperature on hardness and color. Increased oxygen concentrations generally led to larger (P<0.01) thiobarbituric acid-reactive substances (TBARS) values, greater (P<0.01) loss of free thiols and more formation of cross-linked myosin heavy chain. Hardness of cooked patties was generally lower (P<0.01) without oxygen. Premature browning of cooked patties was observed already at a relative low oxygen concentration of 20%. The internal redness of cooked patties decreased (P<0.05) with increasing oxygen concentrations and increasing cooking temperatures. Mean particle size (D(3,2)) of homogenized cooked meat generally increased (P<0.05) with increasing cooking temperatures and increasing oxygen concentrations, and particle size was correlated (r=0.80) with hardness of cooked patties.


Assuntos
Embalagem de Alimentos , Oxigênio/análise , Carne Vermelha , Animais , Bovinos , Cor , Culinária , Concentração de Íons de Hidrogênio , Metabolismo dos Lipídeos/efeitos dos fármacos , Proteínas Musculares/análise , Tamanho da Partícula , Compostos de Sulfidrila/análise , Temperatura Ambiente , Substâncias Reativas com Ácido Tiobarbitúrico/análise
10.
Meat Sci ; 110: 174-9, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26241463

RESUMO

Pork loins were stored at 5°C for 14 days to investigate the effect of oxygen concentration in modified atmosphere packaging (MAP) on shear force and oxidation of lipids and proteins. The modified atmosphere contained 0 to 80% O2, 20% CO2, and balanced with N2. The results showed that shear force and thiobarbituric acid-reactive substances (TBARS) values increased with increasing oxygen concentration. Protein oxidation when measured as loss of free thiol groups, was greater in meat packaged under oxygen (20-80%). Myosin heavy chain (MHC) cross-linking, another marker of protein oxidation, was greater in MAP with 80% oxygen than 0% and 20% oxygen. Desmin degradation was not affected by the presence of oxygen, suggesting that the mechanism of oxygen-induced toughening of meat is through protein oxidation leading to cross-linking of structural proteins rather than through inactivation of proteolytic enzymes leading to reduced proteolysis.


Assuntos
Embalagem de Alimentos/métodos , Carne/análise , Oxigênio , Animais , Desmina , Conservação de Alimentos , Oxirredução , Resistência ao Cisalhamento , Compostos de Sulfidrila , Suínos , Substâncias Reativas com Ácido Tiobarbitúrico
11.
J Food Prot ; 77(5): 796-804, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24780335

RESUMO

The effects of different salt concentrations on the quality changes of grass carp (Ctenopharyngodon idella) fillets were evaluated in terms of biogenic amines, adenosine triphosphate and its related compounds, sensory attributes, total volatile basic nitrogen (TVB-N), and total viable counts during 4 and 20 °C storage. Grass carp fillets were brined in solutions of 2% NaCl (T1) and 10% NaCl (T2), and unsalted carp fillets were used as controls (CK). T1 and T2 showed higher sensory scores than CK. According to the TVB-N values, CK, T1, and T2 could maintain the freshness of carp for approximately 9, 12, and 27 days, respectively, when stored at 4 °C. The higher salt concentration had better inhibitory effect on the accumulation of some biogenic amines, such as tryptamine (TRM), 2-phenylethylamine (PHE), putrescine (PUT), and cadaverine (CAD). TVB-N of untreated grass carp fillets showed a significant (P < 0.05) correlation with TRM, PHE, PUT, and CAD during storage. PUT and CAD showed a significant (P < 0.05) correlation with TVB-N for T2 at 20 °C and T1 at 4 °C.


Assuntos
Aminas Biogênicas/análise , Produtos Pesqueiros/análise , Conservação de Alimentos/métodos , Animais , Carpas , Armazenamento de Alimentos , Humanos , Sais/análise , Cloreto de Sódio/análise , Paladar , Temperatura Ambiente
12.
Food Chem ; 159: 20-8, 2014 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-24767022

RESUMO

The effects of low salt and sugar dry-curing on the quality changes of black carp (Mylopharyngodon piceus) fillets stored at 4°C were evaluated by sensory, physical, chemical, and microbiological methods. Fish samples were left untreated (control), or were dry-cured with 1.5% salt (T1) or 1.5% salt+1.2% sugar (T2). Curing treatments reduced chemical changes reflected in HxR, Hx, pH, and total volatile base nitrogen (TVB-N); decreased cooking loss; and increased overall sensory quality of fish (p<0.05) compared to untreated samples. Significantly lower values of cadaverine and putrescine were observed in T1 and T2 compared to the control after the 2nd and 4th day, respectively (p<0.05). There were significant differences (p<0.05) between T1 and T2 for pH, TVB-N, total aerobic counts (TAC), and sensory characteristics. Sensory characteristics were significantly correlated with TAC, TVB-N, putrescine, and cadaverine in all samples (p<0.01).


Assuntos
Aminas Biogênicas/análise , Carpas , Conservação de Alimentos/métodos , Alimentos Marinhos/análise , Animais , Carboidratos/farmacologia , Temperatura Baixa , Concentração de Íons de Hidrogênio , Nucleotídeos/análise , Alimentos Marinhos/microbiologia , Cloreto de Sódio/farmacologia
13.
Sensors (Basel) ; 14(2): 3207-26, 2014 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-24553086

RESUMO

In this paper, an Urban Light Index (ULI) is constructed to facilitate analysis and quantitative evaluation of the process of urbanization and expansion rate by using DMSP/OLS Nighttime Light Data during the years from 1992 to 2010. A unit circle urbanization evaluation model is established to perform a comprehensive analysis of the urbanization process of 34 prefecture-level cities in Northeast China. Furthermore, the concept of urban light space is put forward. In this study, urban light space is divided into four types: the core urban area, the transition zone between urban and suburban areas, suburban area and fluorescent space. Proceeding from the temporal and spatial variation of the four types of light space, the pattern of morphologic change and space-time evolution of the four principal cities in Northeast China (Harbin, Changchun, Shenyang, Dalian) is analyzed and given particular attention. Through a correlation analysis between ULI and the traditional urbanization indexes (urban population, proportion of the secondary and tertiary industries in the regional GDP and the built-up area), the advantages and disadvantages as well as the feasibility of using the ULI in the study of urbanization are evaluated. The research results show that ULI has a strong correlation with urban built-up area (R2 = 0.8277). The morphologic change and history of the evolving urban light space can truly reflect the characteristics of urban sprawl. The results also indicate that DMSP/OLS Nighttime Light Data is applicable for extracting urban space information and has strong potential to urbanization research.

14.
Guang Pu Xue Yu Guang Pu Fen Xi ; 33(4): 1057-60, 2013 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-23841428

RESUMO

The corn in the grain filling stage fell over in the central region of Jilin province by the Typhoon Bolaven influence. In order to determine the impact of falling over corn canopy on the reflected information, the hyperspectral reflectance was detected at different viewing zenith angles, at the same time, the polarized reflection was also measured. The results from the analysis by combining the reflection and polarization from corn canopy showed that the reflection of falling over corn is low in visible, while increases in the near infrared wavelength. The reflection from falling over corn canopy was more anisotropic than stand-up corn canopy. The reflected light was highly polarized, the polarization of corn canopy provided the probability for distinguishing between falling over corn and stand-up corn. This research provides a basis for estimating the disaster area and lost units.


Assuntos
Biomassa , Produtos Agrícolas/crescimento & desenvolvimento , Refratometria/métodos , Análise Espectral/métodos , Vento , China , Produtos Agrícolas/química , Desastres , Previsões , Tecnologia de Sensoriamento Remoto/métodos
15.
Food Chem ; 138(2-3): 1476-82, 2013 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-23411270

RESUMO

The effects of different freezing treatments on the quality changes of bighead carp heads were evaluated in terms of pH value, TBARS, TVB-N, K-value, biogenic amine, total aerobic counts (TACs), drip loss, cooking loss and electrical conductivity (EC) during ice storage. Fish heads were stored at -40°C (T1), -40°C for 12h and then -18°C (T2), -18°C (T3) for 3months prior to ice storage. No significant differences were observed among T1, T2 and T3 for drip loss, cooking loss and EC (p>0.05). T2 showed lower TACs, pH value, TBARS and TVB-N than T3 did. Significant lower value of spermine and spermidine were observed in T1, T2 and T3 than those of control group (fresh) from 9th to 18th day (p<0.05). Drip loss was significantly correlated with TBARS, pH value, TVB-N, and TACs in groups T1, T2 and T3 (p<0.05).


Assuntos
Aminas Biogênicas/análise , Conservação de Alimentos/métodos , Alimentos Marinhos/análise , Animais , Aminas Biogênicas/metabolismo , Carpas/anatomia & histologia , Carpas/metabolismo , Congelamento , Cabeça/anatomia & histologia
16.
J Food Prot ; 75(12): 2228-33, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23212022

RESUMO

The formation of eight biogenic amines (BAs), total volatile base nitrogen (TVB-N), aerobic plate counts (APC), and sensory evaluation in crucian carp (Carassius auratus) during storage in ice and at 4°C were investigated. The sum of contents of all eight biogenic amines (SBA) in crucian carp increased from 49.82 to 197.09 mg/kg on day 36 when stored in ice and from 49.82 to 219.35 mg/kg on day 24 when stored at 4°C. TVB-N content increased with storage time; good relationships between TVB-N content and SBA were observed during storage in ice and at 4°C (R(2) = 0.95 and 0.93, respectively). APC of all samples significantly (P < 0.05) increased with storage time in the first 12 days and toward the end of the storage period, there were only minor changes; good relationships between APC and SBA were observed during storage in ice and at 4°C (R(2) = 0.96 and 0.58, respectively). According to sensory evaluation results, the crucian carp became unacceptable on days 24 and 16 during storage in ice and at 4°C, respectively. Putrescine and tyramine contents could be good quality evaluation indices for crucian carp during storage.


Assuntos
Aminas Biogênicas/biossíntese , Carpas/metabolismo , Contaminação de Alimentos/análise , Conservação de Alimentos/métodos , Alimentos Marinhos/análise , Animais , Aminas Biogênicas/análise , Carpas/microbiologia , Temperatura Baixa , Contagem de Colônia Microbiana , Qualidade de Produtos para o Consumidor , Microbiologia de Alimentos , Humanos , Gelo , Alimentos Marinhos/microbiologia , Alimentos Marinhos/normas , Fatores de Tempo
17.
Gene ; 510(1): 78-86, 2012 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-22951807

RESUMO

CapZ (ß-actinin) and tropomodulin (Tmod) are capping proteins involved in the maintenance of thin filaments in vertebrate skeletal muscles. In this study, we focused on amphioxus, the most primitive chordate. We searched for CapZ and Tmod genes in the amphioxus genome and determined their primary structures. Amphioxus possess one CapZα gene (CAPZA) and one CapZß gene (CAPZB), and the transcripts of these genes were found to be 67%-85% identical to those of human CapZ genes. On the other hand, amphioxus contain one Tmod gene (TMOD), and the product of this gene has an identity of approximately 50% with human Tmod genes 1-4. However, helix 2 of amphioxus Tmod, which is involved in protein-binding to tropomyosin, was highly conserved with approximately 74% identity to human Tmod genes. Western blotting indicated the presence of CapZ and Tmod in the striated muscle of amphioxus. These results suggest that unlike most of vertebrates, such as fish, amphibian, bird, and mammal, CapZ from amphioxus striated muscle is derived from two genes CAPZA and CAPZB, and Tmod is derived from one TMOD gene.


Assuntos
Proteína de Capeamento de Actina CapZ/genética , Cordados/genética , Músculo Esquelético/metabolismo , Tropomodulina/genética , Actinina/classificação , Actinina/genética , Actinina/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Proteína de Capeamento de Actina CapZ/classificação , Proteína de Capeamento de Actina CapZ/metabolismo , Cordados/metabolismo , DNA Complementar/química , DNA Complementar/genética , Variação Genética , Humanos , Dados de Sequência Molecular , Filogenia , Isoformas de Proteínas/classificação , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Tropomodulina/classificação , Tropomodulina/metabolismo
18.
Int J Environ Res Public Health ; 9(6): 2057-74, 2012 06.
Artigo em Inglês | MEDLINE | ID: mdl-22829790

RESUMO

In this study, an experiment was performed to assess the trip difficulty for urban residents of different age groups walking in various depths of water, and the data were corroborated with the real urban rainstorm waterlogging scenarios in downtown (Daoli district) Ha-Erbin (China). Mathematical models of urban rainstorm waterlogging were constructed using scenario simulation methods, aided by the GIS spatial analysis technology and hydrodynamic analysis of the waterway systems in the study area. Then these models were used to evaluate the impact of waterlogging on the safety of residents walking in the affected area. Results are summarized as: (1) for an urban rainstorm waterlogging scenario reoccurring once every 10 years, three grid regions would have waterlogging above 0.5 m moving at a velocity of 1.5 m/s. Under this scenario, waterlogging would accumulate on traffic roads only in small areas, affecting the safety and mobility of residents walking in the neighborhood; (2) for an urban rainstorm waterlogging scenario reoccurring once every 20 years, 13 grids experienced the same waterlogging situation affecting a larger area of the city; (3) for an urban rainstorm waterlogging scenario reoccurring once every 50 years, 86 grid regions were affected (waterlogging above 0.5 m moving at 1.5 m/s), and those areas would become impassable for residents.


Assuntos
Cidades/estatística & dados numéricos , Inundações/estatística & dados numéricos , Modelos Teóricos , População Urbana/estatística & dados numéricos , Caminhada/estatística & dados numéricos , Adolescente , Criança , China , Sistemas de Informação Geográfica , Humanos , Transportes , Adulto Jovem
19.
Methods ; 56(1): 18-24, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22245513

RESUMO

Determining the complete primary structure of large proteins is difficult because of the large sequence size and low sequence homology among animals, as is the case with connectin (titin)-like proteins in invertebrate muscles. Conventionally, large proteins have been investigated using immuno-screenings and plaque hybridization screenings that require significant time and labor. Recently, however, the genomic sequences of various invertebrates have been determined, leading to changes in the strategies used to elucidate the complete primary structures of large proteins. In this paper, we describe our methods for determining the sequences of large proteins by elucidating the primary structure of connectin from the ascidian Ciona intestinalis as an example. We searched for genes that encode connectin-like proteins in the C. intestinalis genome using the BLAST search program. Subsequently, we identified some domains present in connectin and connectin-like proteins, such as immunoglobulin (Ig), fibronectin type 3 (Fn) and kinase domains in C. intestinalis using the SMART program and manual estimation. The existence of these domains and the unique sequences between each domain were confirmed using RT-PCR. We also examined the localization of mRNA using whole-mount in situ hybridization (WISH) and protein expression using SDS-PAGE. These analyses indicate that the domain structure and molecular weight of ascidian connectin are similar to those of vertebrate connectin and that ascidian connectin is also expressed in heart muscle, similarly to vertebrate connectin. The methods described in this study can be used to determine the primary structures of large proteins, such as novel connectin-like proteins in invertebrates.


Assuntos
Ciona intestinalis/química , Genômica , Proteínas Musculares/química , Proteínas Quinases/química , Animais , Ciona intestinalis/genética , Conectina , Eletroforese em Gel de Poliacrilamida , Peso Molecular , Proteínas Musculares/genética , Filogenia , Proteínas Quinases/genética , Estrutura Terciária de Proteína , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Software
20.
J Mol Biol ; 409(3): 415-26, 2011 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-21510959

RESUMO

Connectin is an elastic protein found in vertebrate striated muscle and in some invertebrates as connectin-like proteins. In this study, we determined the structure of the amphioxus connectin gene and analyzed its sequence based on its genomic information. Amphioxus is not a vertebrate but, phylogenetically, the lowest chordate. Analysis of gene structure revealed that the amphioxus gene is approximately 430 kb in length and consists of regions with exons of repeatedly aligned immunoglobulin (Ig) domains and regions with exons of fibronectin type 3 and Ig domain repeats. With regard to this sequence, although the region corresponding to the I-band is homologous to that of invertebrate connectin-like proteins and has an Ig-PEVK region similar to that of the Neanthes sp. 4000K protein, the region corresponding to the A-band has a super-repeat structure of Ig and fibronectin type 3 domains and a kinase domain near the C-terminus, which is similar to the structure of vertebrate connectin. These findings revealed that amphioxus connectin has the domain structure of invertebrate connectin-like proteins at its N-terminus and that of vertebrate connectin at its C-terminus. Thus, amphioxus connectin has a novel structure among known connectin-like proteins. This finding suggests that the formation and maintenance of the sarcomeric structure of amphioxus striated muscle are similar to those of vertebrates; however, its elasticity is different from that of vertebrates, being more similar to that of invertebrates.


Assuntos
Cordados não Vertebrados/metabolismo , Proteínas Musculares/química , Proteínas Quinases/química , Animais , Sequência de Bases , Cordados não Vertebrados/genética , Conectina , Éxons , Fibronectinas/genética , Humanos , Dados de Sequência Molecular , Proteínas Musculares/genética , Proteínas Quinases/genética , Homologia de Sequência
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