RESUMO
Objective: To evaluate the early diagnostic value of various indicators in the simplified JSTH score criteria for sepsis-associated disseminated intravascular coagulation (DIC). Methods: A retrospective study was conducted. Patients admitted to Intensive Care Unit (ICU) of the Second Affiliated Hospital of Kunming Medical University from January in 2017 to December in 2018 were enrolled. Totally of 365 patients were recruited, with 224 males and 132 females. The simplified JSTH score criteria was used to diagnose DIC. The patients were divided into sepsis with DIC group and sepsis without DIC group according to the diagnostic criteria of sepsis. Platelet (PLT), fibrin degradation products (FDPs), prothrombin time (PT), antithrombin (AT), acute physiology and chronic health evaluation â ¡ (APACHE â ¡) scores, sequential organ failure assessment (SOFA) scores and the simplified JSTH scores were recorded on the first ICU day. Correlation analyses were conducted.Receiver operating characteristic (ROC) curves for diagnosis of DIC with each indicator were drawn to evaluate the diagnostic efficiency and predictive ability of 28-day mortality. Results: According to the simplified JSTH score, 143 cases of sepsis complicated with DIC were diagnosed. There were significant differences in PLT, FDP, AT, PT, APACHE â ¡ score, SOFA score, 28-day mortality rate between the two groups (all P<0.01). It was shown by Pearson correlation analysis that the criteria has the best correlation with APACHEâ ¡ score and SOFA score (r=0.496 and 0.612, both P<0.01). The correlation between PLT and APACHE â ¡ score or SOFA score was the best (r=-0.440 or-0.568, both P<0.01). It was shown by ROC curve that area under ROC curve (AUC) of PLT was 0.933, and the sensitivity and specificity was 93.0% and 85.0%, respectively. The 28-day mortality was predicted by using the indicators in the criteria. The AUG of AT was 0.813, and both the sensitivity (81.6%) and specificity (73.6%) were the highest. Conclusions: The simplified JSTH score criteria can be used for early diagnosis of sepsis-associated DIC and it is positively correlated with the severity of the disease. The correlation between PLT and the severity of disease is the best, and early diagnosis efficiency of PLT is the strongest. AT has a good predictive value for 28-day mortality.
Assuntos
APACHE , Coagulação Intravascular Disseminada , Diagnóstico Precoce , Sepse , Coagulação Intravascular Disseminada/diagnóstico , Coagulação Intravascular Disseminada/etiologia , Feminino , Humanos , Unidades de Terapia Intensiva , Masculino , Prognóstico , Curva ROC , Estudos Retrospectivos , Sepse/complicações , Sepse/diagnósticoRESUMO
Lumbosacral transitional vertebrae (LSTV) is a common phenomena of developmental anomaly, which is characterized by anatomic variation and biomechanical changes. LSTV is often accompanied with low back pain, lumbar disc herniation, lumbar spinal stenosis, lumbar spondylolisthesis and other spinal diseases. The diagnosis of LSTV has a great significance for proper treatment process. Early imageological studies have limitations on distinguishing different types of LSTV from the aspect of morphological changes. This review focuses on recent studies of LSTV anatomy and variation, its influence in local biomechanics and spinal alignment, and its relationship with spinal diseases.
Assuntos
Vértebras Lombares/anatomia & histologia , Vértebras Lombares/fisiopatologia , Sacro/anatomia & histologia , Sacro/fisiopatologia , Doenças da Coluna Vertebral/fisiopatologia , Fenômenos Biomecânicos , Humanos , Dor Lombar/etiologia , Dor Lombar/fisiopatologia , Região Lombossacral/anatomia & histologia , Região Lombossacral/fisiopatologia , Doenças da Coluna Vertebral/diagnóstico por imagem , Doenças da Coluna Vertebral/etiologiaRESUMO
Postoperative delirium is a common complication after spinal surgery, and it is a complex issue involving multiple factors. However, there is currently insufficient understanding of postoperative delirium in patients undergoing spinal surgery, and there is still a lack of clear regulation in prevention and treatment. Although the literature and research on postoperative delirium have been comprehensive, there are still few studies on postoperative delirium in patients undergoing spinal surgery. This article mainly reviews the incidence, social and economic problems, risk factors, prevention and treatment of postoperative delirium in patients undergoing spinal surgery.
Assuntos
Delírio/terapia , Procedimentos Neurocirúrgicos/efeitos adversos , Coluna Vertebral/cirurgia , Delírio/etiologia , Delírio/prevenção & controle , Humanos , Incidência , Fatores de Risco , Fatores SocioeconômicosRESUMO
The majority of cervical spine injuries in children occur in the upper cervical spine, of which odontoid fracture is the most common. Odontoid fracture in children is a very insidious injury. Due to the unclear language and incompatible physical examination, the disease is often missed diagnosis. Because the child axis is still in the developmental segment, including 4 synchondrosis and 6 ossification centers, there are obvious anatomical and biological differences between the child odontoid fracture and the adult. Therefore, the choice of treatment is different from that of adults. This article will introduce the development of odontoid in children, and summarize the injury characteristics, clinical classification and treatment of odontoid fracture in children.
Assuntos
Processo Odontoide/lesões , Fraturas da Coluna Vertebral/diagnóstico , Fraturas da Coluna Vertebral/terapia , Adulto , Pesquisa Biomédica , Criança , Humanos , Lesões do Pescoço/complicações , Fraturas da Coluna Vertebral/classificação , Fraturas da Coluna Vertebral/etiologiaRESUMO
Though great progress on spinal sagittal alignment has been seen recently, which focuses on the lumbar spine-pelvic region and the whole spine, while there is a few research mainly concentrated on the cervical spine. In recent years, a growing number of researchers have been exploring the changes in the compensation of cervical sagittal alignment and their effect on surgery, and the preliminary results of these researches are satisfactory. The present review focuses on the measurement of sagittal plane parameters of cervical spine, changes of sagittal alignment in cervical spine disorders, and its effect on cervical surgery.
Assuntos
Doenças da Coluna Vertebral , Humanos , Vértebras Lombares , Região Lombossacral , Pescoço , Radiografia , Doenças da Coluna Vertebral/diagnóstico por imagem , Coluna VertebralRESUMO
To establish the experts consensus on the right heart function management in critically ill patients. The panel of consensus was composed of 30 experts in critical care medicine who are all members of Critical Hemodynamic Therapy Collaboration Group (CHTC Group). Each statement was assessed based on the GRADE (Grading of Recommendations Assessment, Development, and Evaluation) principle. Then the Delphi method was adopted by 52 experts to reassess all the statements. (1) Right heart function is prone to be affected in critically illness, which will result in a auto-exaggerated vicious cycle. (2) Right heart function management is a key step of the hemodynamic therapy in critically ill patients. (3) Fluid resuscitation means the process of fluid therapy through rapid adjustment of intravascular volume aiming to improve tissue perfusion. Reversed fluid resuscitation means reducing volume. (4) The right ventricle afterload should be taken into consideration when using stroke volume variation (SVV) or pulse pressure variation (PPV) to assess fluid responsiveness.(5)Volume overload alone could lead to septal displacement and damage the diastolic function of the left ventricle. (6) The Starling curve of the right ventricle is not the same as the one applied to the left ventricle,the judgement of the different states for the right ventricle is the key of volume management. (7) The alteration of right heart function has its own characteristics, volume assessment and adjustment is an important part of the treatment of right ventricular dysfunction (8) Right ventricular enlargement is the prerequisite for increased cardiac output during reversed fluid resuscitation; Nonetheless, right heart enlargement does not mandate reversed fluid resuscitation.(9)Increased pulmonary vascular resistance induced by a variety of factors could affect right heart function by obstructing the blood flow. (10) When pulmonary hypertension was detected in clinical scenario, the differentiation of critical care-related pulmonary hypertension should be a priority. (11) Attention should be paid to the change of right heart function before and after implementation of mechanical ventilation and adjustment of ventilator parameter. (12) The pulmonary arterial pressure should be monitored timingly when dealing with critical care-related pulmonary hypertension accompanied with circulatory failure.(13) The elevation of pulmonary aterial pressure should be taken into account in critical patients with acute right heart dysfunction. (14) Prone position ventilation is an important measure to reduce pulmonary vascular resistance when treating acute respiratory distress syndrome patients accompanied with acute cor pulmonale. (15) Attention should be paid to right ventricle-pulmonary artery coupling during the management of right heart function. (16) Right ventricular diastolic function is more prone to be affected in critically ill patients, the application of critical ultrasound is more conducive to quantitative assessment of right ventricular diastolic function. (17) As one of the parameters to assess the filling pressure of right heart, central venous pressure can be used to assess right heart diastolic function. (18). The early and prominent manifestation of non-focal cardiac tamponade is right ventricular diastolic involvement, the elevated right atrial pressure should be noticed. (19) The effect of increased intrathoracic pressure on right heart diastolic function should be valued. (20) Ttricuspid annular plane systolic excursion (TAPSE) is an important parameter that reflects right ventricular systolic function, and it is recommended as a general indicator of critically ill patient. (21) Circulation management with right heart protection as the core strategy is the key point of the treatment of acute respiratory distress syndrome. (22) Right heart function involvement after cardiac surgery is very common and should be highly valued. (23) Right ventricular dysfunction should not be considered as a routine excuse for maintaining higher central venous pressure. (24) When left ventricular dilation, attention should be paid to the effect of left ventricle on right ventricular diastolic function. (25) The impact of left ventricular function should be excluded when the contractility of the right ventricle is decreased. (26) When the right heart load increases acutely, the shunt between the left and right heart should be monitored. (27) Attention should be paid to the increase of central venous pressure caused by right ventricular dysfunction and its influence on microcirculation blood flow. (28) When the vasoactive drugs was used to reduce the pressure of pulmonary circulation, different effects on pulmonary and systemic circulation should be evaluated. (29) Right atrial pressure is an important factor affecting venous return. Attention should be paid to the influence of the pressure composition of the right atrium on the venous return. (30) Attention should be paid to the role of the right ventricle in the acute pulmonary edema. (31) Monitoring the difference between the mean systemic filling pressure and the right atrial pressure is helpful to determine whether the infusion increases the venous return. (32) Venous return resistance is often considered to be a insignificant factor that affects venous return, but attention should be paid to the effect of the specific pathophysiological status, such as intrathoracic hypertension, intra-abdominal hypertension and so on. Consensus can promote right heart function management in critically ill patients, optimize hemodynamic therapy, and even affect prognosis.
Assuntos
Estado Terminal , Diástole/fisiologia , Hidratação , Insuficiência Cardíaca/diagnóstico por imagem , Hemodinâmica/fisiologia , Pressão Venosa Central , Consenso , Cuidados Críticos , Insuficiência Cardíaca/etiologia , Insuficiência Cardíaca/fisiopatologia , Humanos , Edema Pulmonar , Respiração Artificial , Síndrome do Desconforto Respiratório , Disfunção Ventricular Direita/diagnóstico por imagem , Função Ventricular EsquerdaRESUMO
The effects of goal-directed fluid therapy, with lactated Ringer's (LR) and 6% hydroxyethyl starch (HES) solution, on hemorrhagic shock dogs are unknown. We aimed to determine the optimal LR: HES ratio for the resuscitation of hemorrhagic shock dogs. Hemorrhagic shock was induced in 40 ventilated dogs by drawing an estimated 60% blood volume. The animals were randomly divided into five groups (N = 8) according to the LR: HES ratio of the resuscitation fluid (3:1, 2:1, 1:1, 1:2, and 1:3), and were then resuscitated for 24 h to reach the stroke volume variation (SVV) and hemoglobin (Hb) goals by fluid infusion and autologous blood perfusion. The extravascular lung water index (EVLWI), pH, partial pressure of oxygen (PaO2), base excess (BE), sodium, chloride, Hb and creatinine clearance (Clearcrea) were checked after 24 h (R24). The EVLWI of the 3:1 group at R24 were higher than that of the 1:3 group and the baseline value (P < 0.05), whereas the PaO2 was lower (P < 0.05). In contrast to the 3:1 group at R24 and baseline, plasma chloride and sodium in the 1:3 and 1:2 groups increased; however, pH, BE, and Clearcrea decreased (P < 0.05). No significant differences were found in the 1:1 and 2:1 groups at R24 compared with baseline (P > 0.05). Resuscitation with LR and HES at 2:1 and 1:1 ratios are superior in maintaining the acid-base, electrolyte, and lung water balances as well as renal function in hemorrhagic shock dogs than at ratios of 3:l, 1:2, and1:3.
Assuntos
Hidratação/métodos , Derivados de Hidroxietil Amido/farmacologia , Soluções Isotônicas/farmacologia , Ressuscitação/métodos , Choque Hemorrágico/terapia , Equilíbrio Ácido-Base/efeitos dos fármacos , Animais , Transfusão de Sangue Autóloga , Cloretos/sangue , Cães , Hemoglobinas/metabolismo , Testes de Função Renal , Consumo de Oxigênio/efeitos dos fármacos , Respiração Artificial , Lactato de Ringer , Choque Hemorrágico/sangue , Choque Hemorrágico/patologia , Sódio/sangue , Volume Sistólico/efeitos dos fármacosRESUMO
BACKGROUND: Chloroplast development is coordinately regulated by plastid- and nuclear-encoding genes. Although many regulators have been reported to be involved in chloroplast development, new factors remain to be identified, given the complexity of this process. RESULTS: In this study, we characterized a rice mutant lethal albinic seedling 1(las1)form of a 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (OsHMBPP) that was targeted to the chloroplasts. The LAS1 mutation caused the albino lethal phenotype in seedlings. Transmission electron microscopy indicated that las1 were defective in early chloroplast development. LAS1 is preferentially expressed in leaves, implying its role in controlling chloroplast development. The expression levels of many chloroplast-encoded genes were altered significantly in las1. The expression levels of nuclear-encoded gene involved in Chl biosynthesis were also decreased in las1. We further investigated plastidic RNA editing in las1 and found that the edit efficiency of four chloroplast genes were markly altered. Compared with WT, las1 exhibited defective in biogenesis of chloroplast ribosomes. CONCLUSIONS: Our results show that LAS1/OsHMBPP plays an essential role in the early chloroplast development in rice.
RESUMO
Glutamic acid decarboxylase (GAD) is a pancreatic beta cell autoantigen in humans and nonobese diabetic (NOD) mice. beta Cell-specific suppression of GAD expression in two lines of antisense GAD transgenic NOD mice prevented autoimmune diabetes, whereas persistent GAD expression in the beta cells in the other four lines of antisense GAD transgenic NOD mice resulted in diabetes, similar to that seen in transgene-negative NOD mice. Complete suppression of beta cell GAD expression blocked the generation of diabetogenic T cells and protected islet grafts from autoimmune injury. Thus, beta cell-specific GAD expression is required for the development of autoimmune diabetes in NOD mice, and modulation of GAD might, therefore, have therapeutic value in type 1 diabetes.
Assuntos
Autoantígenos/imunologia , Diabetes Mellitus Tipo 1/enzimologia , Diabetes Mellitus Tipo 1/imunologia , Glutamato Descarboxilase/imunologia , Ilhotas Pancreáticas/enzimologia , Transferência Adotiva , Animais , Autoantígenos/genética , Autoantígenos/fisiologia , Autoimunidade , DNA Antissenso , Diabetes Mellitus Tipo 1/patologia , Feminino , Expressão Gênica , Glutamato Descarboxilase/genética , Glutamato Descarboxilase/fisiologia , Insulina/sangue , Insulina/metabolismo , Ilhotas Pancreáticas/imunologia , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/patologia , Transplante das Ilhotas Pancreáticas , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Camundongos Transgênicos , Linfócitos T/imunologia , TransgenesRESUMO
OBJECTIVE: To analyze the endemic situation of schistosomiasis in Wuhu City from 2009 to 2018, so as to provide evidence for adjusting and developing the control strategy. METHODS: The data regarding the endemic situation were retrospectively collected and analyzed in Wuhu City from 2009 to 2018. RESULTS: The prevalence of human Schistosoma japonicum infections reduced from 0.450% in 2009 to 0.035% in 2018 in Wuhu City, and a reduction rate of 92.22% (χ2 = 1 6128.307, P < 0.01), and the prevalence of S. japonicum infections decreased from 0.483% in 2009 to 0 in 2018 (χ2 = 27.570, P < 0.01) in livestock. The area of snail habitats increased from 1 501.07 hm2 in 2009 to 4 408.62 hm2 in 2018, with an increase of 193.70%. No infected snails were found in Wuhu City since 2012, and no egg positives were detected in humans and livestock since 2016. CONCLUSIONS: Currently, the endemic situation of schistosomiasis is at a low level in Wuhu City; however, there are still factors affecting schistosomiasis transmission. In the future, the integrated strategy with emphasis on the control of infectious source and the surveillance-response system should be intensified to consolidate the achievements of schistosomiasis control.
Assuntos
Esquistossomose Japônica , Animais , China/epidemiologia , Humanos , Gado , Estudos Retrospectivos , Schistosoma/fisiologia , Esquistossomose Japônica/epidemiologia , Esquistossomose Japônica/prevenção & controle , Esquistossomose Japônica/transmissãoRESUMO
One benefit of clonal integration is that resource translocation between connected ramets enhances the growth of the ramets grown under stressful conditions, but whether such resource translocation reduces the performance of the ramets grown under favourable conditions has not produced consistent results. In this study, we tested the hypothesis that resource translocation to recipient ramets may reduce the performance of donor ramets when resources are limiting but not when resources are abundant. We grew Mikania micrantha stolon fragments (each consisting of two ramets, either connected or not connected) under spatially heterogeneous competition conditions such that the developmentally younger, distal ramets were grown in competition with a plant community and the developmentally older, proximal ramets were grown without competition. For half of the stolon fragments, slow-release fertiliser pellets were applied to both the distal and proximal ramets. Under both the low and increased soil nutrient conditions, the biomass, leaf number and stolon length of the distal ramets were higher, and those of the proximal ramets were lower when the stolon internode was intact than when it was severed. For the whole clone, the biomass, leaf number and stolon length did not differ between the two connection treatments. Connection did not change the biomass of the plant communities competing with distal ramets of M. micrantha. Although clonal integration may promote the invasion of M. micrantha into plant communities, resource translocation to recipient ramets of M. micrantha will induce a cost to the donor ramets, even when resources are relatively abundant.
Assuntos
Mikania/metabolismo , Biomassa , Ecossistema , Mikania/fisiologia , Fotossíntese/fisiologia , Folhas de Planta/metabolismo , Folhas de Planta/fisiologiaRESUMO
Optical branch waveguides are one of the most important optical elements and have been widely exploited for optical communication systems. However, prevailing devices are typically solid and have limit in tunability. Liquid optical devices have attracted more interest for the advantage of tunability of liquid media, but their signals suffer serious leakage if the refractive index (RI) of liquid is smaller than that of solid channels. This paper demonstrates the tunable three-dimensional (3D) optofluidic Y-branch waveguides in plannar microchannels by simply introducing Dean flow. This device can reconfigure 3D Y-branch profiles and separate the intensity of light as tunable ratio from 0 to 1 by adjusting the flow rates with low loss. Different from the prevailing 2D liquid counterparts, the 3D configuration offer much more freedom in the selection of liquid media as liquid's RI can be totally independent to the solid channel structure. The transmission loss through the device is estimated to 0.97 db when the splitting angle is 10°, which shows the light is confined better in the 3D liquid structures than traditional 2D liquid counterparts. The Y-branch waveguides show potential in applications of integrated optofluidic devices.
RESUMO
We have previously demonstrated that rat cardiac troponin T (TnT) is expressed as two different isoforms during development, the larger, more acidic embryonic isoform and the smaller, more basic adult isoform, which appear to be generated from a common transcript of the cardiac TnT gene by alternative RNA splicing. In this study, Southern blot analysis confirmed the existence of a single copy of cardiac TnT gene in the rat genome. For investigation of the molecular mechanism of isoform switch and the control of this gene expression in myocardial development, several overlapping genomic clones were isolated from a rat genomic library. Complete nucleotide sequences were determined from these genomic clones and revealed a 19,186 base-pair DNA fragment containing 16 exons of rat cardiac TnT gene. Its DNA sequence and exon organization appeared to differ from that of the rat fast skeletal muscle TnT gene or chicken cardiac TnT gene. Comparison of genomic and cDNA clones also confirmed that the cardiac TnT isoform switching was due to the inclusion or exclusion of exon 4 during RNA processing. Sequence analysis allowed us to further identify the other alternatively spliced exon containing only nine nucleotides in size (exon 12). The inclusion and complete or partial exclusion of this exon may be responsible for generating three classes of mRNAs detected by our cDNA clones. The functional significance of this variation in TnT isoforms remained unknown, but its splicing pattern did not appear to link to the developmental changes. The 5' upstream structure was very similar to that in chicken cardiac TnT gene but differed from that in the rat fast skeletal muscle TnT gene, suggesting a similar regulatory mechanism for mammalian and avian cardiac TnT expression.
Assuntos
Miocárdio/metabolismo , Troponina/genética , Processamento Alternativo , Animais , Sequência de Bases , Southern Blotting , DNA , Éxons , Regulação da Expressão Gênica , Coração/embriologia , Dados de Sequência Molecular , Ratos , Mapeamento por Restrição , Troponina/química , Troponina TRESUMO
We have identified and purified from bovine brain a novel protein kinase which catalyzes in vitro phosphorylation of neurofilament proteins NF-H and NF-M and tau proteins at sites implicating the enzyme in the regulation of neurocytoskeleton dynamics and in Alzheimer pathology. The protein kinase displays a phosphorylation site specificity similar or identical to the cell cycle regulatory kinase, cdc2 kinase. The purified kinase is a heterodimer of a cdc2-like catalytic subunit, called cdk5, and a 25 kDa regulatory subunit. The regulatory subunit is essential for kinase activity, and it is derived from a 35 kDa protein, p35 by proteolysis. Northern blot analysis of tissue distribution indicates that cdk5 is widely distributed but especially rich in brain, whereas p35 expression is only found in brain. The protein kinase is therefore termed neuronal cdc2-like kinase. The neuron-specificity of the enzyme appears to be conferred by the regulatory subunit. During cell division, cdc2 kinase is regulated by complex phosphorylation mechanisms involving a network of specific protein kinases. Some of these kinases or their homologs have been found in mammalian brains and they may be involved in the regulation of neuronal cdc2-like kinase.
Assuntos
Proteína Quinase CDC2/metabolismo , Quinases Ciclina-Dependentes , Neurônios/enzimologia , Proteínas Serina-Treonina Quinases/metabolismo , Sequência de Aminoácidos , Animais , Bovinos , Quinase 5 Dependente de Ciclina , Humanos , Dados de Sequência Molecular , Fosforilação , Proteínas Serina-Treonina Quinases/químicaRESUMO
Three muscle type-specific troponin T (TnT) genes are present in vertebrate to encode a number of protein isoforms via alternative mRNA splicing. While the genomic structures of cardiac and fast skeletal muscle TnT genes have been documented, this study cloned and characterized the slow skeletal muscle TnT (sTnT) gene. Complete nucleotide sequence and genomic organization revealed that the mouse sTnT gene spans 11.1kb and contains 14 exons, which is smaller and simpler than the fast skeletal muscle and cardiac TnT genes. Potentially representing a prototype of the TnT gene family, the 5'-region of the sTnT gene contains fewer unsplit large exons, among which two alternatively spliced exons are responsible for the NH2-terminal variation of three sTnT isoforms. The sTnT gene structure shows that the alternatively spliced central segment found in human sTnT cDNAs may be a result from splicing using an alternative acceptor site at the intron 11-exon 12 boundary. Together with the well-conserved protein structure, the highly specific expression of sTnT in slow skeletal muscles indicates a differentiated function of this member of the TnT gene family. The determination of genomic structure and alternative splicing pathways of sTnT gene lays a foundation to further understand the TnT structure-function evolution as well as contractile characteristics of different types of muscle fiber.
Assuntos
Músculo Esquelético/metabolismo , Troponina T/genética , Processamento Alternativo/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Éxons/genética , Íntrons/genética , Camundongos , Dados de Sequência Molecular , Contração Muscular/genética , Fibras Musculares de Contração Lenta/metabolismo , Isoformas de Proteínas/genética , RNA Mensageiro/genéticaRESUMO
We have cloned and sequenced full-length cDNAs encoding mouse slow skeletal muscle troponin T (sTnT). Alternative mRNA splicing-generated two high Mr isoforms and one low Mr sTnT isoform differing in the NH2-terminal primary structure have been identified by Western blotting, reverse transcription-polymerase chain reaction and cDNA cloning/expression analyses. Together with a 5'-alternative exon that was also found in human sTnT encoding an 11-amino-acid acidic segment, the results revealed a novel alternative splicing pathway to include or exclude a three-base segment to generate additional sTnT isoforms with NH2-terminal charge variations. Overriding the phylogenetic divergence, primary structure of sTnT is better conserved between mammalian and avian species than that of cardiac, fast and skeletal muscle TnTs from one species. Western blots demonstrate four expression patterns of sTnT during postnatal skeletal muscle development: (1) a decrease to a non-detectable level in mouse masseter, (2) an increase to become the sole TnT in sheep masseter, (3) an increase of the total level as well as the proportion of the low Mr isoform in sheep diaphragm and, (4) no significant change in total level or high/low Mr isoform ratio in sheep gastrocnemius. The highly conserved primary structure and fiber type-specific and developmentally regulated expression of sTnT indicate a physiological importance of this under-studied member of the TnT gene family.
Assuntos
Troponina/genética , Processamento Alternativo , Sequência de Aminoácidos , Animais , Galinhas , Clonagem Molecular , Sequência Conservada , DNA Complementar/genética , Escherichia coli/genética , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Camundongos , Dados de Sequência Molecular , Desenvolvimento Muscular , Fibras Musculares de Contração Lenta/metabolismo , Músculo Esquelético/crescimento & desenvolvimento , Músculo Esquelético/metabolismo , Filogenia , Reação em Cadeia da Polimerase , Troponina TRESUMO
The present study describes a significant amino acid sequence homology between neuronal Cdk5 activator (nck5a) and an open reading frame of an unknown gene on the yeast S. cerevisiae chromosome III. A cDNA encoding a 25 kDa fragment of this yeast protein, the region containing homologous sequence to nck5a was cloned and expressed in E. coli as a glutathine-S-transferase fusion protein (GST-p25Y). GST-p25Y was found to block the in vitro activation of Cdk5 by nck5a and to affinity precipitate Cdk5 from bovine brain extract. The observations indicate that the yeast protein is capable of specific and high affinity association with Cdk5.
Assuntos
Quinases Ciclina-Dependentes , Proteínas Fúngicas/química , Proteínas do Tecido Nervoso/química , Proteínas Serina-Treonina Quinases/metabolismo , Saccharomyces cerevisiae/genética , Sequência de Aminoácidos , Animais , Química Encefálica , Bovinos , Quinase 5 Dependente de Ciclina , Ativação Enzimática , Escherichia coli/genética , Proteínas Fúngicas/genética , Genes Fúngicos , Glutationa Transferase/genética , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Saccharomyces cerevisiae/química , Homologia de SequênciaRESUMO
Cultured airway smooth muscle cells subjected to cyclic deformational strain have increased cell content of myosin light chain kinase (MLCK) and myosin and increased formation of actin filaments. To determine how these changes may increase cell contractility, we measured isometric force production with changes in cytosolic calcium in individual permeabilized cells. The pCa for 50% maximal force production was 6.6+/-0.4 in the strain cells compared with 5.9+/-0.3 in control cells, signifying increased calcium sensitivity in strain cells. Maximal force production was also greater in strain cells (8.6+/-2.9 vs. 5.7+/-3.1 microN). The increased maximal force production in strain cells persisted after irreversible thiophosphorylation of myosin light chain, signifying that increased force could not be explained by differences in myosin light chain phosphorylation. Cells strained for brief periods sufficient to increase cytoskeletal organization but insufficient to increase contractile protein content also produced more force, suggesting that strain-induced cytoskeletal reorganization also increases force production.
Assuntos
Cálcio/farmacologia , Fibras Musculares Esqueléticas/efeitos dos fármacos , Músculo Liso/fisiologia , Traqueia/fisiologia , Actinas/análise , Actinas/metabolismo , Processamento Alternativo/fisiologia , Animais , Células Cultivadas , Citoesqueleto/metabolismo , Expressão Gênica/fisiologia , Contração Isométrica/fisiologia , Contração Muscular/efeitos dos fármacos , Contração Muscular/fisiologia , Fibras Musculares Esqueléticas/citologia , Fibras Musculares Esqueléticas/enzimologia , Músculo Liso/citologia , Cadeias Pesadas de Miosina/genética , Cadeias Pesadas de Miosina/metabolismo , Cadeias Leves de Miosina/metabolismo , Quinase de Cadeia Leve de Miosina/metabolismo , Fosforilação , RNA Mensageiro/análise , Estresse Mecânico , Enxofre/metabolismo , Traqueia/citologiaRESUMO
From the snake venom of Agkistrodon acutus, two proteases, acuthrombin-A and acuthrombin-C, were isolated and purified to homogeneity. They can cleave the human fibrinogen to release the fibrinopeptide A and fibrinopeptide B with specific activity of 120 and 370 NIH units/mg, respectively; the fibrinogen-clotting activity can be inhibited distinctly by PMSF or DFP or EDTA, but not by heparin. The two proteases show also arginine-esterase activity hydrolyzing some synthetic substrates such as TAME and BAEE. Additionally, they are glycoproteins with an average content of 2.4% (acuthrombin-A) and 2.1% (acuthrombin-C) neutral carbohydrates, respectively. Acuthrombin-A has a MW of 13,900 as estimated by SDS-PAGE under reduced or nonreduced conditions and 28,000 as determined by gel filtration. For acuthrombin-C, there were two protein bands corresponding to MW of 13,900 and 14,800 on SDS-PAGE with different darkness under reduced or nonreduced conditions, while its MW was estimated to be 69,000 by gel filtration. The isoelectric points were 7.5 for acuthrombin-A and 5.0 for acuthrombin-C by isoelectric focusing. Neither acuthrombin-A nor acuthrombin-C has haemorrhagic or lethal activity. Acuthrombin-A has also a small amount of activity to activate the Factor XIII.
Assuntos
Agkistrodon/metabolismo , Coagulação Sanguínea/efeitos dos fármacos , Endopeptidases/farmacologia , Fibrinogênio/efeitos dos fármacos , Venenos de Serpentes/enzimologia , Animais , Cromatografia em Gel , Endopeptidases/isolamento & purificação , Esterases/metabolismo , Fator XIII/metabolismo , Fibrinogênio/metabolismo , Fibrinopeptídeo A/metabolismo , Fibrinopeptídeo B/metabolismo , Hemorragia/induzido quimicamente , Humanos , Hidrólise , Camundongos , Oxirredução , Peptídeo Hidrolases/farmacologia , Coelhos , Venenos de Serpentes/química , Trombina/farmacologia , Trombina/toxicidadeRESUMO
In contrast to skeletal muscles that simultaneously express multiple troponin T (TnT) isoforms, normal adult human cardiac muscle contains a single isoform of cardiac TnT. To understand the significance of myocardial TnT homogeneity, we examined the effect of TnT heterogeneity on heart function. Transgenic mouse hearts overexpressing a fast skeletal muscle TnT together with the endogenous cardiac TnT was investigated in vivo and ex vivo as an experimental system of concurrent presence of two classes of TnT in the adult cardiac muscle. This model of myocardial TnT heterogeneity produced pathogenic phenotypes: echocardiograph imaging detected age-progressive reductions of cardiac function; in vivo left ventricular pressure analysis showed decreased myocardial contractility; ex vivo analysis of isolated working heart preparations confirmed an intrinsic decrease of cardiac function in the absence of neurohumoral influence. The transgenic mice also showed chronic myocardial hypertrophy and degeneration. The dominantly negative effects of introducing a fast TnT into the cardiac thin filaments to produce two classes of Ca(2+) regulatory units in the adult myocardium suggest that TnT heterogeneity decreases contractile function by disrupting the synchronized action during ventricular contraction that is normally activated as an electrophysiological syncytium.