RESUMEN
The reported incidence rate of venous and arterial thrombotic events in critically ill patients with COVID-19 infections is high, ranging from 20% to 60%. We adopted a patient-tailored thromboprophylaxis protocol based on clinical and laboratory presentations for these patients in our institution. We hypothesised that patients who received high-intensity thromboprophylaxis treatment would experience fewer thrombotic events. The aims of our study were to explore the incidence of thrombotic events in this population; to assess independent factors associated with thrombotic events and to evaluate the incidence of haemorrhagic events. A retrospective review of all adult patients with confirmed SARS-CoV-2 infection admitted to the intensive care unit (ICU) between 1 March and 29 May 2020 was performed. The primary outcome was a composite of venous and arterial thrombotic events diagnosed during the ICU stay. Multivariable logistic regression was used to identify the independent factors associated with thrombotic events. A total of 188 patients met the inclusion criteria. All received some type of thromboprophylaxis treatment except for six patients who did not receive any prophylaxis. Of the 182 patients who received thromboprophylaxis, 75 (40%) received high-intensity thromboprophylaxis and 24 (12.8%) were treated with therapeutic anticoagulation. Twenty-one patients (11.2%) experienced 23 thrombotic events (incidence rate of 12.2% (95%CI 7.9-17.8)), including 12 deep venous thromboses, 9 pulmonary emboli and 2 peripheral arterial thromboses. The multivariable logistic regression analysis showed that only D-dimer (OR 2.80, p = 0.002) and high-intensity thromboprophylaxis regimen (OR 0.20, p = 0.01) were independently associated with thrombotic events. Thirty-one patients (16.5%) experienced haemorrhagic events; among them, 13 were classified as major bleeding according to the International Society on Thrombosis and Haemostasis criteria. Therapeutic anticoagulation, but not the high-intensity thromboprophylaxis regimen, was associated with major bleeding. A proactive approach to the management of thromboembolism in critically ill COVID-19 patients utilising a high-intensity thromboprophylaxis regimen in appropriately selected patients may result in lower thrombotic events without increasing the risk of bleeding.
Asunto(s)
Anticoagulantes/uso terapéutico , COVID-19/complicaciones , Protocolos Clínicos , Cuidados Críticos/estadística & datos numéricos , Enoxaparina/uso terapéutico , Tromboembolia Venosa/complicaciones , Adulto , Anciano , Anciano de 80 o más Años , Enfermedad Crítica , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , SARS-CoV-2 , Tromboembolia Venosa/tratamiento farmacológico , Adulto JovenRESUMEN
Finite element (FE) model studies have made important contributions to our understanding of functional biomechanics of the lumbar spine. However, if a model is used to answer clinical and biomechanical questions over a certain population, their inherently large inter-subject variability has to be considered. Current FE model studies, however, generally account only for a single distinct spinal geometry with one set of material properties. This raises questions concerning their predictive power, their range of results and on their agreement with in vitro and in vivo values. Eight well-established FE models of the lumbar spine (L1-5) of different research centers around the globe were subjected to pure and combined loading modes and compared to in vitro and in vivo measurements for intervertebral rotations, disc pressures and facet joint forces. Under pure moment loading, the predicted L1-5 rotations of almost all models fell within the reported in vitro ranges, and their median values differed on average by only 2° for flexion-extension, 1° for lateral bending and 5° for axial rotation. Predicted median facet joint forces and disc pressures were also in good agreement with published median in vitro values. However, the ranges of predictions were larger and exceeded those reported in vitro, especially for the facet joint forces. For all combined loading modes, except for flexion, predicted median segmental intervertebral rotations and disc pressures were in good agreement with measured in vivo values. In light of high inter-subject variability, the generalization of results of a single model to a population remains a concern. This study demonstrated that the pooled median of individual model results, similar to a probabilistic approach, can be used as an improved predictive tool in order to estimate the response of the lumbar spine.
Asunto(s)
Análisis de Elementos Finitos , Vértebras Lumbares/fisiología , Modelos Teóricos , Algoritmos , Fuerza Compresiva , Humanos , Vértebras Lumbares/anatomía & histología , Postura , Presión , Probabilidad , Rango del Movimiento Articular/fisiología , Reproducibilidad de los Resultados , Rotación , Articulación Cigapofisaria/fisiologíaRESUMEN
Determination of physiological loads in human lumbar spine is critical for understanding the mechanisms of lumbar diseases and for designing surgical treatments. Computational models have been used widely to estimate the physiological loads of the spine during simulated functional activities. However, various assumptions on physiological factors such as the intra-abdominal pressure (IAP), centers of mass (COMs) of the upper body and lumbar segments, and vertebral centers of rotation (CORs) have been made in modeling techniques. Systematic knowledge of how these assumptions will affect the predicted spinal biomechanics is important for improving the simulation accuracy. In this paper, we developed a 3D subject-specific numerical model of the lumbosacral spine including T12 and 90 muscles. The effects of the IAP magnitude and COMs locations on the COR of each motion segment and on the joint/muscle forces were investigated using a global convergence optimization procedure when the subject was in a weight bearing standing position. The data indicated that the line connecting the CORs showed a smaller curvature than the lordosis of the lumbar spine in standing posture when the IAP was 0 kPa and the COMs were 10 mm anterior to the geometric center of the T12 vertebra. Increasing the IAP from 0 kPa to 10 kPa shifted the location of CORs toward the posterior direction (from 1.4 ± 8.9 mm anterior to intervertebral disc (IVD) centers to 40.5 ± 3.1 mm posterior to the IVD centers) and reduced the average joint force (from 0.78 ± 0.11 Body weight (BW) to 0.31 ± 0.07 BW) and overall muscle force (from 349.3 ± 57.7 N to 221.5 ± 84.2 N). Anterior movement of the COMs from -30 mm to 70 mm relative to the geometric center of T12 vertebra caused an anterior shift of the CORs (from 25.1 ± 8.3 mm posterior to IVD centers to 7.8 ± 6.2 mm anterior to IVD centers) and increases of average joint forces (from 0.78 ± 0.1 BW to 0.93 ± 0.1 BW) and muscle force (from 348.9 ± 47.7 N to 452.9 ± 58.6 N). Therefore, it is important to consider the IAP and correct COMs in order to accurately simulate human spine biomechanics. The method and results of this study could be useful for designing prevention strategies of spinal injuries and recurrences, and for enhancing rehabilitation efficiency.
Asunto(s)
Abdomen , Vértebras Lumbares , Fenómenos Mecánicos , Modelos Biológicos , Presión , Adulto , Fenómenos Biomecánicos , Femenino , Humanos , Vértebras Lumbares/fisiología , Masculino , MovimientoRESUMEN
In experimental and clinical research, it is difficult to directly measure responses in the human body, such as contact pressure and stress in a joint, but finite element analysis (FEA) enables the examination of in vivo responses by contact analysis. Hence, FEA is useful for pre-operative planning prior to orthopaedic surgeries, in order to gain insight into which surgical options will result in the best outcome. The present study develops a numerical simulation technique based on FEA to predict the surgical outcomes of osteotomy methods for the treatment of slipped capital femoral epiphyses. The correlation of biomechanical parameters including contact pressure and stress, for moderate and severe cases, is investigated. For severe slips, a base-of-neck osteotomy is thought to be the most reliable and effective surgical treatment, while any osteotomy may produce dramatic improvement for moderate slips. This technology of pre-operative planning using FEA can provide information regarding biomechanical parameters that might facilitate the selection of optimal osteotomy methods and corresponding surgical options.
Asunto(s)
Toma de Decisiones Asistida por Computador , Análisis de Elementos Finitos , Osteotomía/métodos , Epífisis Desprendida de Cabeza Femoral/cirugía , Adulto , Fenómenos Biomecánicos/fisiología , Niño , Simulación por Computador , Femenino , Cabeza Femoral/cirugía , Articulación de la Cadera , Humanos , Masculino , Epífisis Desprendida de Cabeza Femoral/patologíaRESUMEN
To estimate true Brix and alcoholic strength of must and wines without distillation, a novel approach using a refractometer and a hydrometer was developed. Initial Brix (I.B.), apparent refractometer Brix (A.R.), and apparent hydrometer Brix (A.H.) of must were measured by refractometer and hydrometer, respectively. Alcohol content (A) was determined with a hydrometer after distillation and true Brix (T.B.) was measured in distilled wines using a refractometer. Strong proportional correlations among A.R., A.H., T.B., and A in sugar solutions containing varying alcohol concentrations were observed in preliminary experiments. Similar proportional relationships among the parameters were also observed in must, which is a far more complex system than the sugar solution. To estimate T.B. and A of must during alcoholic fermentation, a total of 6 planar equations were empirically derived from the relationships among the experimental parameters. The empirical equations were then tested to estimate T.B. and A in 17 wine products, and resulted in good estimations of both quality factors. This novel approach was rapid, easy, and practical for use in routine analyses or for monitoring quality of must during fermentation and final wine products in a winery and/or laboratory.
Asunto(s)
Alcoholes/análisis , Carbohidratos/análisis , Refractometría/métodos , Vino/análisis , China , Fermentación/fisiología , Manipulación de Alimentos , Corea (Geográfico) , Saccharomyces cerevisiae/metabolismo , VitisRESUMEN
The complete nucleotide sequence of pepper severe mosaic virus (PepSMV) was determined. The viral genome consisted of 9890 nucleotides, excluding a poly (A) tract at the 3' end of the genome. The PepSMV RNA genome encoded a single polyprotein of 3085 amino acid residues, resulting in ten functionally distinct potyviral proteins. The lengths of the 5' nontranslated region (NTR) and the 3' NTR were 164 and 468 nucleotides, respectively. The genome organization of the virus was typical for members of the genus Potyvirus in the family Potyviridae. The coat protein amino acid sequence identity between PepSMV and the other 45 potyviruses ranged from 53.4 to 79.7%. Sequence alignments and phylogenetic analyses of the potyviral polyprotein sequences revealed that PepSMV was the closest to potato virus Y (PVY) and closely related to members of the PVY subgroup. Our genome sequence data clearly confirmed that PepSMV belongs to a separate species in the genus Potyvirus.
Asunto(s)
Capsicum/virología , Genoma Viral , Virus del Mosaico/genética , Regiones no Traducidas 3'/genética , Regiones no Traducidas 5'/genética , Proteínas de la Cápside/química , Proteínas de la Cápside/genética , Peso Molecular , Virus del Mosaico/clasificación , Filogenia , Homología de Secuencia de Aminoácido , Especificidad de la EspecieRESUMEN
Ferritin, an iron-binding protein, was purified from the larval hemolymph of the wax moth, Galleria mellonella by KBr density ultracentrifugation and FPLC (Superose 6). The iron content of ferritin was determined by atomic emission spectroscopy and Ferene S stain. Native molecular mass of ferritin was estimated as 630 kDa. SDS-PAGE revealed that the ferritin consists of two major polypeptides of 26 and 32 kDa and one minor polypeptide of 30 kDa. An isoelectric point of ferritin was measured to be approximately 7.3 and only the 32-kDa subunit is glycosylated. The ferritin contains large amounts of lysine, glutamine, glutamic acid and leucine but tryptophan was not detected. Electron microscopic examination of negatively stained preparations showed an 11-nm particle in external diameter and 7-nm iron core. Ferritin is present in both the ovary and testis. Localization of ferritin by immunoelectron microscopy in ovary and testis revealed that the gold particles were located in vitelline membrane and yolk granules but not in follicular epithelium of ovary. In the testis, the gold particles were located in testicular fluid and lumen of vas deferens.
Asunto(s)
Ferritinas/química , Ferritinas/inmunología , Hemolinfa/química , Mariposas Nocturnas , Aminoácidos/análisis , Animales , Femenino , Ferritinas/aislamiento & purificación , Masculino , Ovario/química , Ovario/fisiología , Testículo/química , Testículo/fisiologíaRESUMEN
BACKGROUND: Controversy exists regarding the timing of thrombolytic administration and rupture rate. METHODS: Hospital records at St. Luke's-Roosevelt Hospital of the 4 study patients were reviewed and compared with those of 41 patients from a group of 537 patients concurrently admitted with a diagnosis of myocardial infarction (MI). RESULTS: Four patients experienced ventricular free wall rupture after having a MI between November 17, 1993, and July 28, 1995. All received tissue plasminogen activator. In 1 patient, pericardial effusion associated with a pseudoaneurysm was discovered in the operating room. The 3 others developed clinical pericardial tamponade before surgery. All 4 patients survived and left the hospital on postoperative days 10, 11, 11, and 82, respectively. During this same time period, 537 patients were admitted with MI, 41 of whom died; the study's 4 patients were compared with these 41. CONCLUSIONS: These data demonstrate that rupture of the ventricular free wall can occur early after thrombolytic therapy and may have a subacute course. Prompt diagnosis and surgery offer excellent chances of surviving this fatal condition.
Asunto(s)
Rotura Cardíaca Posinfarto/cirugía , Infarto del Miocardio/tratamiento farmacológico , Terapia Trombolítica , Activador de Tejido Plasminógeno/efectos adversos , Anciano , Taponamiento Cardíaco/inducido químicamente , Taponamiento Cardíaco/mortalidad , Taponamiento Cardíaco/cirugía , Femenino , Rotura Cardíaca Posinfarto/inducido químicamente , Rotura Cardíaca Posinfarto/mortalidad , Humanos , Masculino , Persona de Mediana Edad , Infarto del Miocardio/mortalidad , Estudios Retrospectivos , Tasa de Supervivencia , Activador de Tejido Plasminógeno/uso terapéuticoRESUMEN
A novel virus we call zucchini green mottle mosaic virus (ZGMMV) was isolated from zucchini squash and its properties were determined. The size and shape of its virions, and other properties suggest that the virus is a tobamovirus. The coat protein (CP) genes of ZGMMV and kyuri green mottle mosaic virus (KGMMV), which also infects zucchini squash plants, were cloned and their nucleotides sequences were determined. The CP genes of ZGMMV and KGMMV are composed of 161 amino acid residues, and they share 77.6% amino acid identity. Western blot analysis showed that the two viruses are serologically related but not identical. Comparison of the sequences with those of sixteen other tobamoviruses revealed that the two viruses had much higher identity to cucumber green mottle mosaic virus (CGMMV), another tobamovirus infectious to cucurbit plants, than other tobamoviruses. The nucleotide and amino acid sequences of ZGMMV were from 29.5 to 78.4% and from 29.3 to 77.6% identical, respectively, to those of other tobamoviruses. The predicted virion assembly origins of the two tobamoviruses were located in the CP region of the genomic RNAs, and the predicted secondary structures were more similar to that of CGMMV than those of other tobamoviruses. The seventeen tobamo-viruses could be classified into three main subgroups based on the phylogenetic tree analysis on the CP gene, and ZGMMV and KGMMV formed a third subgroup together with CGMMV and sunn-hemp mosaic virus (SHMV). These results show that ZGMMV is a previously unknown member of the Tobamovirus genus.
Asunto(s)
Cápside/genética , Cucurbitaceae/virología , Tobamovirus/clasificación , Tobamovirus/genética , Secuencia de Bases , Cápside/química , Clonación Molecular , ADN Complementario , Genoma Viral , Datos de Secuencia Molecular , Filogenia , ARN Viral/análisis , ARN Viral/genética , Análisis de Secuencia de ADN , Tobamovirus/ultraestructura , Virión/ultraestructuraRESUMEN
In order to determine the detection limit for chemically treated virions by gel electrophoresis, reverse transcription-polymerase chain reaction (RT-PCR) and infectivity assay, tobacco mosaic virus (TMV) exposed to various concentrations of chemicals was studied. When virions were exposed to 0.2 N HCl for 30 mins, partially degraded TMV particles were observed by gel electrophoresis. Under the same exposure, a major RT-PCR amplified DNA product corresponding to the target size of 806 bp, which decreased as a function of time, could be detected for up to 60 mins of exposure. When virions were treated with NaOH (0.02 N or higher normality) for 5 mins, partially degraded virions were detected by gel electrophoresis, exhibiting multiple band patterns. Exposure of the virions to 0.1 N NaOH for 5 mins revealed severely degraded viral RNA, but disappearance of the amplified RT-PCR products was apparent during 30-60 mins of exposure. Therefore, these data showed clearly the difference in the detection limit of gel electrophoresis and that of RT-PCR for the degraded viral RNA. In addition, the infectivity assay showed that the number of local lesions in Nicotiana rustica were significantly reduced by more than 95% when the virus was exposed to 0.2 N HCl for 15 mins or 0.1 N NaOH for 10 mins. From these results we conclude that loss of infectivity was not related to that of PCR product. Other chemical disinfectants such as phenol or formalin were also found to be effective to reduce the virus infectivity, but a corresponding degradation of viral RNA was detected by neither gel electrophoresis nor RT-PCR.
Asunto(s)
Virus del Mosaico del Tabaco/efectos de los fármacos , Electroforesis en Gel de Agar , Formaldehído/farmacología , Ácido Clorhídrico/farmacología , Fenol/farmacología , Enfermedades de las Plantas/virología , Plantas Tóxicas , ARN Viral/análisis , ARN Viral/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Hidróxido de Sodio/farmacología , Factores de Tiempo , Nicotiana/virología , Virus del Mosaico del Tabaco/patogenicidad , Virus del Mosaico del Tabaco/efectos de la radiación , Virión/aislamiento & purificación , Virión/metabolismoRESUMEN
Reverse transcription and polymerase chain reaction (RT-PCR) was used for detection and identification of three cucumoviruses (cucumber mosaic virus, CMV; peanut stunt virus, PSV; tomato aspermy virus, TAV) in various plants sources with a single pair of primers, designed as CPTALL-3 and CPTALL-5. The pair of cucumovirus genus-specific primers that flank the coat protein gene were designed and used to amplify a DNA fragment of approximately ranging from 938 to 966 bp. The RT-PCR with the set of primers specifically amplified the target size of DNA fragment in all the tested cucumoviruses (CMV S-IA, S-IB and S-II, PSV and TAV). No DNA product of any length was produced when brome mosaic virus or tobacco mosaic virus RNA was used as templates. The cucumoviruses examined were differentiated by PCR-restriction fragment length polymorphism with different enzymes. This indicates that the designed primers are only specific for the cucumoviruses and useful for reliable information of identification of members of the Cucumovirus genus.
Asunto(s)
Cucumovirus/genética , Cucumovirus/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Virología/métodos , Secuencia de Bases , Cápside/genética , Cucumovirus/clasificación , Cartilla de ADN/genética , ADN Viral/genética , ADN Viral/aislamiento & purificación , Estudios de Evaluación como Asunto , Genes Virales , Polimorfismo de Longitud del Fragmento de Restricción , Especificidad de la EspecieRESUMEN
Unlike other pathogens, plant viruses are hardly controlled by chemical agents. Potato virus Y (PVY) is distributed around the world, and causes a great loss economically. In an attempt to minimize the damage by viruses, the PVY coat protein (CP) gene was introduced into tobacco by Agrobacterium-mediated transformation. A significant proportion of the transgenic plants displayed resistance to PVY and showed substantially decreased CP transgene expression at both protein and steady-state mRNA levels compared to susceptible transgenic or nontransgenic plants. A resistant plant was selected and self-fertilized for several generations until T4 progenitor lines were obtained. Most of these T4 plants accumulated extremely low levels of CP protein and steady-state mRNA, and exhibited almost complete resistance to PVY. DNA gel blot analysis revealed that the transgenic plants typically had two or three copies of the transgene. These results are characteristic of pathogen-derived resistance, in which the resistance against virus is the consequence of post-transcriptional gene silencing directed by homologous transgenes. To uncover factors that may play roles in gene silencing, sequences in the 3' part of the transcribed region of the CP gene were transcribed in vitro and the RNA fragments were incubated with cell extracts from transgenic plants. A ribonuclease activity was detected that appeared to be specific for this transcript in the PVY-resistant transgenic plants.
Asunto(s)
Proteínas de la Cápside , Cápside/genética , Nicotiana/genética , Plantas Modificadas Genéticamente/genética , Plantas Tóxicas , Ensayo de Inmunoadsorción Enzimática , Regulación de la Expresión Génica de las Plantas , Regulación Viral de la Expresión Génica , Modelos Genéticos , Procesamiento Postranscripcional del ARN , ARN Mensajero/análisis , Ribonucleasas/metabolismo , Supresión Genética , Transformación GenéticaRESUMEN
Man-made mineral fibers (MMMF), most of which are referred to as man-made vitreous fibers (MMVF), are mostly amorphous silicates manufactured from glass, rock, or other minerals. Analysis for MMMF have been restricted largely to the measurement of total airborne mass concentrations, but more recently to the determination of airborne fiber levels by phase contrast optical microscopy. In Korea, many small factories are related with manufacturing and using MMMF without any special evaluation of environmental measurements. Though MMMF are known as the substitute of asbestos and their toxicity are regarded as very low, MMMF do not totally excluded from the respiratory and/or skin diseases now. Therefore, we evaluated the environments of many workplaces with total dust and fiber concentration. Most dust and fiber concentrations were below threshold limit value (TLV) at various industries and working processes. However, these data showed a slight relationship between total dust and fiber concentration.
Asunto(s)
Contaminantes Ocupacionales del Aire/análisis , Polvo/análisis , Fibras Minerales/análisis , Análisis de Varianza , Monitoreo del Ambiente/métodos , Humanos , Industrias , Exposición Profesional , Tamaño de la PartículaRESUMEN
The 3'-terminal regions of the genomic RNAs of two Korean isolates of the lily symptomless Carlavirus (LSV), LSV-Ko and LSV-KII, were cloned and their nucleotide sequences were determined. The nucleotide sequence analysis and protein analysis by the Western blot revealed that E. coli expressed a 32-kDa protein that is the viral coat protein (CP) for the LSV. The two Korean strains share 98.4% and 98.3% sequence identities at the nucleotide and amino acid levels, respectively. The CP gene of LSV-Ko showed 99.1% and 87.0% nucleotide sequence identities, and 99.0% and 96.6% amino acid sequence identities with those of the Netherlands and the Japanese LSV strains, respectively. A pairwise amino acid sequence comparison revealed a sequence similarity of 29.6% to 69.8% between LSV-Ko and other species of the carlavirus. The 16 kDa protein of LSV-Ko shares 17.6% to 42.7% amino acid similarity with those of 8 other the carlaviruses, and they are variable in the N-terminal region. The Cys repeated zinc finger nucleic acid binding domain was found in the 16 kDa protein for all of the LSV strains. Sequence comparisons of the 7 kDa protein of LSV in the strain level showed significant identities from 100.0% to 98.4%. LSV-Ko shares 21.9% to 42.2% amino acid similarity with those of 8 other carlaviruses, 4 members of the potexviruses, and a closterovirus. LSV is closely related to blueberry scorch virus (BISV) based upon the phylogenetic tree analyses of the three proteins, indicating LSV to be a quite distinct member of the genus Carlavirus.
Asunto(s)
Cápside/genética , Carlavirus/genética , Liliaceae/virología , Secuencia de Aminoácidos , Secuencia de Bases , Cápside/química , Carlavirus/clasificación , Carlavirus/aislamiento & purificación , Clonación Molecular , Cartilla de ADN/genética , ADN Complementario/genética , ADN Viral/genética , Escherichia coli/genética , Expresión Génica , Genes Virales , Corea (Geográfico) , Datos de Secuencia Molecular , Peso Molecular , Filogenia , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Homología de Secuencia de AminoácidoRESUMEN
We have reported the sequences of four novel proteins derived from extracts of human aortic tissue and a cDNA library from human aortic adventitia. These proteins are immunoreactive with serum immunoglobulins from patients with abdominal aortic aneurysms (AAA), and they have homologies of amino acid sequence with microfibrillar proteins of connective tissue. We are reporting separately that two of these proteins are artery-specific antigenic proteins (ASAPs) in man. The present work investigates the regional distribution of these two proteins (AAAP-40 and MatCAM-1) in mouse (E-beta-b). Antibodies were raised in rabbit against polypeptides encoding novel amino acid sequences, unique to these proteins (e.g., not reported in GenBank). Immunohistochemical studies with these two specific antibodies show conspicuous immunoreactivity of collagen-associated microfibrils in the aortic adventitia of the murine abdominal and thoracic aorta. Immunoreactive peptides were not present in brain, muscle, or kidney. These findings support the hypothesis that proteins occur in the mouse that are homologous to a unique family of aortic microfibrillar proteins in man.
Asunto(s)
Aorta/metabolismo , Moléculas de Adhesión Celular/metabolismo , Proteínas Contráctiles/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Citoesqueleto de Actina/metabolismo , Animales , Anticuerpos/metabolismo , Especificidad de Anticuerpos , Unión Competitiva , Moléculas de Adhesión Celular/inmunología , Tejido Elástico/metabolismo , Proteínas de la Matriz Extracelular/inmunología , Humanos , Inmunoglobulinas/inmunología , Inmunohistoquímica , Hígado/metabolismo , Ratones , Factores de Empalme de ARN , Conejos , Bazo/metabolismo , Distribución TisularRESUMEN
The coat protein (CP) genes of the genomic RNA of two Korean isolates of sweet potato feathery mottle potyvirus (SPFMV), SPFMV-K1 and SPFMV-K2, were cloned and their complete nucleotide sequences were determined. Sequence comparisons of the two Korean isolates showed 97.8% amino acid identity in the CP cistron, and 79.9% to 99.0% identity with those of 6 other known SPFMV strains. Of 74 amino acid changes totally among the SPFMV strains, 39 changes were located at the N-terminal region. Pairwise amino acid sequence comparison revealed sequence similarities of 48.6 to 70.2% between SPFMV and 20 other potyviruses, indicating SPFMV to be a quite distinct species. Multiple alignment of the CP cistrons from other potyviruses showed that most of the conserved amino acid residues of the genus Potyvirus are well preserved in the corresponding locations.
Asunto(s)
Cápside/genética , Potyvirus/genética , Solanaceae/virología , Secuencia de Bases , ADN Viral , Corea (Geográfico) , Datos de Secuencia Molecular , Filogenia , Potyvirus/clasificación , Análisis de Secuencia de ADNRESUMEN
The 3'-terminal regions of the genomic RNAs of two Korean isolates of sweet potato feathery mottle potyvirus (SPFMV) were cloned and their nucleotide sequences of full-length coat protein (CP) gene and 3' noncoding region (NCR) were determined. The CP of the two Korean isolates contained 315 amino acid residues. The CP cistron sequences of the Korean isolates exhibit 72.7% to 98.7% nucleotide sequence identity and 79.9% to 99.0% amino acid identity when compared with those of 8 other known SPFMV strains. Pairwise comparison revealed sequence similarities of 47.4% to 62.1% at the nucleotide level, and 48.6% to 70.2% at the amino acid level between SPFMV and 21 other potyviruses. SPFMV CP has extensive amino acid sequence similarity to the other members of the genus Potyvirus throughout its central and C-terminal regions. The 3' NCR of the SPFMV showed 42.5% to 99.1% nucleotide sequence identities among the strains. The 3' NCR of SPFMV revealed 19.9% to 63.6% sequence similarities to those of 21 other potyviruses. These results support the assignment of SPFMV as a distinct member of the genus Potyvirus of the family Potyviridae.
Asunto(s)
Regiones no Traducidas 3'/genética , Cápside/genética , Potyvirus/genética , Secuencia de Aminoácidos , Secuencia de Bases , ADN Complementario/química , ADN Complementario/genética , Corea (Geográfico) , Datos de Secuencia Molecular , Potyvirus/química , Potyvirus/ultraestructura , Alineación de Secuencia , Análisis de Secuencia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Solanum tuberosum/virologíaRESUMEN
Transglutaminase (TGase) is a calcium-dependent enzyme which catalyzes the iso-peptide cross-link between peptide-bound glutamine and lysine in vivo. Though the cross-link is developed as a barrier function in the skin system, overexpression of this could invoke skin hyperkeratosis in psoriasis and roughness in aged skin. In former research, many strong irreversible TGase inhibitors failed application because of high cytotoxicity. We selected one peptide after primary screening of six synthetic peptides designed from domains of known TGase substrates. Then we attempted to reduce the size and finally obtained two tetrameric peptides. When we treated keratinocyte with these TGase inhibitors under calcium-induced differentiation, the formation of a cornified cell envelope (CE) was decreased to the same level of CE under proliferating conditions without cytotoxic effect. Therefore, we propose that these TGase inhibitors may be useful for solving the physiological hypercross-linking problems for pharmaceutical or cosmetic purposes.
Asunto(s)
Inhibidores Enzimáticos/farmacología , Queratinocitos/efectos de los fármacos , Transglutaminasas/antagonistas & inhibidores , Secuencia de Aminoácidos , Células Cultivadas , Humanos , Queratinocitos/enzimología , Cinética , Proteínas de la Membrana/biosíntesis , Datos de Secuencia Molecular , Transglutaminasas/metabolismoRESUMEN
The odontoglossum ringspot Tobamovirus (ORSV) movement and coat proteins genes were selected for the design of oligonucleotide primers for amplification of a 1,085 bp fragment. A combined assay of reverse transcription and the polymerase chain reaction (RT-PCR) was performed with 20-mer ORSV-specific primers and crude nucleic acid extracts from virus-infected orchids for rapid detection of the virus. The lowest concentration of template viral RNA required for detection was 10 fg. The RT-PCR is a 10(3) times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. No PCR product was observed when cymbidium mosaic potexvirus or a crude extract of healthy Cymbidium sp. were used as a template in RT-PCR with the same primers. The specificity of the primers was verified using other tobamoviruses RNAs.
Asunto(s)
Tobamovirus/aislamiento & purificación , Secuencia de Bases , Cartilla de ADN , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad , Tobamovirus/genéticaRESUMEN
The 3'-terminal nucleotide (nt) sequence of the coat protein (CP)-encoding gene of the Korean isolate of cymbidium mosaic potexvirus (CyMV-K) has been determined. The sequence contained a full-length ORF coding for the viral CP. It comprises 663 nt and a polypeptide chain of 220 amino acids with a M(r) of 23,718. The stop codon (TGA) is followed by a 3'-untranslated region (3'-UTR) of 79 nt and a poly(A) tract. The putative polyadenylation signal, AATAAA motif, is found in the terminus of the 3'-UTR.