RESUMO
SNOMED CT (Systematized NOmenclature of MEDicine Clinical Terms) is a standardized multilingual healthcare terminology. It was developed to meet the needs of our electronic world so that care can be documented and clinicians can retrieve and transmit data in electronic format. It is anticipated that SNOMED CT will provide the core general terminology for electronic health records and, as such, replace existing classification systems such as the International Statistical Classification of Diseases and Related Health Problems 10th Revision (ICD-10). At present, there is no special interest group for the hypertensive disorders of pregnancy (HDP) within the SNOMED CT initiative. We believe that members of the ISSHP, and others interested in the HDP, should take a leadership role in this regard for a number of reasons.
Assuntos
Sistemas Computadorizados de Registros Médicos , Systematized Nomenclature of Medicine , Feminino , Humanos , Hipertensão Induzida pela Gravidez , GravidezRESUMO
The use of a Rayleigh laser guide star (LGS) as a wavefront reference for adaptive optics (AO) allows complete control over the distance to, and depth of, the slice of the Rayleigh plume that is selected. By altering the LGS range-gate depth (RGD) and distance at three defined stages during closed-loop operations, the LGS performance can be optimized. For the given example of a 20 km LGS on a 4.2 m telescope the application of this technique would increase the permissible RGD from 200to930 m after optimization. This can allow either an increase in LGS altitude or an increase in wavefront sensor frame rate, thereby increasing AO system performance.
RESUMO
We have previously reported on the use of a tobacco mosaic virus (TMV) vector TMV-30B to express foreign viral antigens for use as experimental immunogens. Here we describe the development of an improved TMV-30B vector that adds a sequence of 7 histidine residues to the C-terminus of recombinant proteins expressed in the vector. We used this TMV-30B-HISc vector to express the VP8* fragment of the VP4 protein from bovine rotavirus (BRV) strain C-486 in plants. Recombinant VP8* protein was purified from N. benthamiana leaves at 7 days post-inoculation by immobilized metal affinity chromatography. The plant-produced VP8* was initially detected using anti-His tag mAb and its antigenic nature was confirmed using both monoclonal and polyclonal specific antisera directed against BRV. Adult female mice, inoculated by the intraperinoteal route with an immunogen containing 4 microg of recombinant VP8*, developed a specific and sustained response to the native VP8* from the homologous BRV. Eighty five percent of suckling mice from immunized dams that were challenged with the homologous virus at the fifth day of age were protected from virus as compared to 35% of the pups from mothers immunized with a control protein. These results demonstrate that the plant-produced VP8* was able to induce passive protection in the new born through the immunization of dams. This suggests that the technology presented here provides a simple method for using plants as an inexpensive alternative source for production of recombinant anti-rotavirus antigens.
Assuntos
Vetores Genéticos , Vacinas contra Rotavirus/biossíntese , Vacinas contra Rotavirus/imunologia , Rotavirus/imunologia , Animais , Animais Lactentes/imunologia , Animais Lactentes/virologia , Antígenos Virais/análise , Bovinos , Linhagem Celular , Feminino , Imunidade Materno-Adquirida , Camundongos , Camundongos Endogâmicos BALB C , Rotavirus/genética , Infecções por Rotavirus/imunologia , Infecções por Rotavirus/prevenção & controle , Nicotiana/imunologia , Nicotiana/virologia , Vírus do Mosaico do Tabaco , Vacinas Sintéticas/biossíntese , Vacinas Sintéticas/imunologiaRESUMO
Production of structural proteins from foot-and-mouth disease virus (FMDV) and bovine herpes virus (BHV-1) in Nicotiana benthamiana through the use of a tobacco mosaic virus-based vector (TMV-30B) has been reported previously. The development of the TMV-30B-HISc vector, a new version that adds a C-terminal histidine (His) sequence to the foreign protein expressed is described. Coding sequences from the FMDV VPl protein and the core protein, p24, from a clade C HIV-1 isolate from Zambia were cloned into the new vector and infective RNAs were generated for each construct to inoculate N. benthamiana plants. His-tagged proteins were purified from inoculated leaves using immobilized metal affinity chromatography (IMAC) as detected by Coomassie blue staining and proteins were further characterized in Western blot assays using a commercial anti-6xHis mAb and specific polyclonal antisera for each protein. While yields obtained for the VPl-His protein after purification were similar to those in crude extracts obtained with the previous TMV-VPl vector, p24-His yields were 10-15 times higher than those of VPl-His. Twenty-five grams of TMV-p24-HISc inoculated leaves were processed to obtain 2.5 mg of isolated p24-His and the recombinant protein was inoculated in rabbits to test immunogenicity and antigenic integrity of the plant-produced p24-His. Animals developed a strong and specific humoral response to the p24-His after the first booster and immune sera was able to recognize the native p24 from a different clade expressed on the surface of the HIV-1 chronically infected HUT78/ARV T-cell line. Importantly, the recombinant p24-His proved its efficiency by confirming the serology of 117 samples previously tested by two rapid HIV-1 tests, thus representing an excellent alternative for production of highly specific diagnostic reagents for HIV endemic regions in the developing world.
Assuntos
Antígenos Virais/imunologia , Proteína do Núcleo p24 do HIV/genética , Proteína do Núcleo p24 do HIV/imunologia , HIV-1 , Nicotiana/metabolismo , Vírus do Mosaico do Tabaco/genética , Animais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Clonagem Molecular , Feminino , Vetores Genéticos/metabolismo , Proteína do Núcleo p24 do HIV/biossíntese , Soropositividade para HIV , Histidina , Humanos , Soros Imunes , Folhas de Planta/metabolismo , Coelhos , Proteínas Recombinantes/biossíntese , Especificidade da Espécie , Nicotiana/virologia , Vírus do Mosaico do Tabaco/metabolismoRESUMO
A tobacco mosaic virus (TMV)-based vector was utilized for expression of a cytosolic form of the bovine herpesvirus type 1 (BHV-1) protein glycoprotein D (gDc). Nicotiana benthamiana plants were harvested 7 days after inoculation with RNA transcripts derived from the TMV-gDc recombinant virus. Recombinant gDc protein of expected electrophoretic mobility accumulated in inoculated leaves to a concentration of about 20 micrograms/g of fresh leaf tissue. Oil-based vaccines were formulated with crude foliar extracts to immunize mice parentally. After a single injection, animals developed a sustained and specific response to both the isolated gD and native virus particles. Cattle vaccinated with the same gDc containing extracts developed specific humoral and cellular immune responses directed against both the viral gD and BHV-1 particles. Most importantly, animals vaccinated with the plant-produced gDc showed good levels of protection after challenge with the virulent BHV-1. Virus excretion was drastically reduced in these animals, reaching levels comparable to animals vaccinated with a commercial BHV-1 vaccine. The positive immunological characterization obtained for the gDc, indicated that an important part of the natural conformation was retained in the plant recombinant protein.
Assuntos
Vetores Genéticos/genética , Herpesvirus Bovino 1/imunologia , Vacinas contra Herpesvirus/biossíntese , Vacinas contra Herpesvirus/imunologia , Nicotiana/metabolismo , Vírus do Mosaico do Tabaco/genética , Proteínas Virais/biossíntese , Proteínas Virais/imunologia , Animais , Anticorpos Antivirais/análise , Anticorpos Antivirais/biossíntese , Formação de Anticorpos/imunologia , Especificidade de Anticorpos , Western Blotting , Bovinos , Ensaio de Imunoadsorção Enzimática , Feminino , Infecções por Herpesviridae/imunologia , Infecções por Herpesviridae/prevenção & controle , Imunidade Celular/imunologia , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Extratos Vegetais/imunologia , Folhas de Planta/imunologiaRESUMO
Variation within the Type and Sidney 81 strains of wheat streak mosaic virus was assessed by single-strand conformation polymorphism (SSCP) analysis and confirmed by nucleotide sequencing. Limiting-dilution subisolates (LDSIs) of each strain were evaluated for polymorphism in the P1, P3, NIa, and CP cistrons. Different SSCP patterns among LDSIs of a strain were associated with single-nucleotide substitutions. Sidney 81 LDSI-S10 was used as founding inoculum to establish three lineages each in wheat, corn, and barley. The P1, HC-Pro, P3, CI, NIa, NIb, and CP cistrons of LDSI-S10 and each lineage at passages 1, 3, 6, and 9 were evaluated for polymorphism. By passage 9, each lineage differed in consensus sequence from LDSI-S10. The majority of substitutions occurred within NIa and CP, although at least one change occurred in each cistron except HC-Pro and P3. Most consensus sequence changes among lineages were independent, with substitutions accumulating over time. However, LDSI-S10 bore a variant nucleotide (G(6016)) in NIa that was restored to A(6016) in eight of nine lineages by passage 6. This near-global reversion is most easily explained by selection. Examination of nonconsensus variation revealed a pool of unique substitutions (singletons) that remained constant in frequency during passage, regardless of the host species examined. These results suggest that mutations arising by viral polymerase error are generated at a constant rate but that most newly generated mutants are sequestered in virions and do not serve as replication templates. Thus, a substantial fraction of variation generated is static and has yet to be tested for relative fitness. In contrast, nonsingleton variation increased upon passage, suggesting that some mutants do serve as replication templates and may become established in a population. Replicated mutants may or may not rise to prominence to become the consensus sequence in a lineage, with the fate of any particular mutant subject to selection and stochastic processes such as genetic drift and population growth factors.
Assuntos
Potyviridae/genética , Triticum/virologia , Evolução Biológica , Frequência do Gene , Variação Genética , Genética Populacional , Haplótipos , Mutação , Polimorfismo Genético , Polimorfismo Conformacional de Fita Simples , Vírion/genéticaRESUMO
Type IIB fast fibres are typically demonstrated in human skeletal muscle by histochemical staining for the ATPase activity of myosin heavy-chain (MyHC) isoforms. However, the monoclonal antibody specific for the mammalian IIB isoform does not detect MyHC IIB protein in man and MyHC IIX RNA is found in histochemically identified IIB fibres, suggesting that the IIB protein isoform may not be present in man; if this is not so, jaw-closing muscles, which express a diversity of isoforms, are likely candidates for their presence. ATPase histochemistry, immunohistochemistry polyacrylamide gel electrophoresis and in situ hybridization, which included a MyHC IIB-specific mRNA riboprobe, were used to compare the composition and RNA expression of MyHC isoforms in a human jaw-closing muscle, the masseter, an upper limb muscle, the triceps, an abdominal muscle, the external oblique, and a lower limb muscle, the gastrocnemius. The external oblique contained a mixture of histochemically defined type I, IIA and IIB fibres distributed in a mosaic pattern, while the triceps and gastrocnemius contained only type I and IIA fibres. Typical of limb muscle fibres, the MyHC I-specific mRNA probes hybridized with histochemically defined type I fibres, the IIA-specific probes with type IIA fibres and the IIX-specific probes with type IIB fibres. The MyHC IIB mRNA probe hybridized only with a few histochemically defined type I fibres in the sample from the external oblique; in addition to this IIB message, these fibres also expressed RNAs for MyHC I, IIA and IIX. MyHC IIB RNA was abundantly expressed in histochemical and immunohistochemical type IIA fibres of the masseter, together with transcripts for IIA and in some cases IIX. No MyHC IIB protein was detected in fibres and extracts of either the external oblique or masseter by immunohistochemistry, immunoblotting and electrophoresis. Thus, IIB RNA, but not protein, was found in the fibres of two different human skeletal muscles. It is believed this is the first report of the substantial expression of IIB mRNA in man as demonstrated in a subset of masseter fibres, but rarely in limb muscle, and in only a few fibres of the external oblique. These findings provide further evidence for the complexity of myosin gene expression, especially in jaw-closing muscles.
Assuntos
Músculo Masseter/metabolismo , Fibras Musculares Esqueléticas/metabolismo , Cadeias Pesadas de Miosina/análise , Miosina não Muscular Tipo IIB/análise , Músculos Abdominais/metabolismo , Músculos Abdominais/ultraestrutura , Adenosina Trifosfatases , Adolescente , Adulto , Eletroforese em Gel de Poliacrilamida , Feminino , Expressão Gênica , Humanos , Immunoblotting , Hibridização In Situ , Masculino , Músculo Masseter/ultraestrutura , Pessoa de Meia-Idade , Fibras Musculares de Contração Rápida/metabolismo , Fibras Musculares de Contração Rápida/ultraestrutura , Fibras Musculares Esqueléticas/ultraestrutura , Fibras Musculares de Contração Lenta/metabolismo , Fibras Musculares de Contração Lenta/ultraestrutura , Músculo Esquelético/metabolismo , Músculo Esquelético/ultraestrutura , Cadeias Pesadas de Miosina/genética , Miosina Tipo I/análise , Miosina Tipo I/genética , Miosina Tipo II/análise , Miosina Tipo II/genética , Miosina não Muscular Tipo IIA/análise , Miosina não Muscular Tipo IIA/genética , Miosina não Muscular Tipo IIB/genética , Fenótipo , Isoformas de Proteínas/análise , Isoformas de Proteínas/genética , Sondas RNA , RNA Mensageiro/análise , RNA Mensageiro/genéticaRESUMO
Cross-protection and vector transmission bottlenecks have been proposed as mechanisms facilitating genetic isolation of sympatric viral lineages. Molecular markers were used to monitor establishment and resolution of mixed infections with genetically defined strains of wheat streak mosaic virus (WSMV). Two closely related WSMV strains from the U.S. (Type and Sidney 81) exhibited reciprocal cross-protection in wheat, confirming this classic phenomenon as a mechanism of genetic isolation. In contrast, cross-protection between either U.S. strain and the divergent El Batán 3 strain from Mexico was unilateral, erratic, and only partially effective. Distribution of WSMV strains within individual leaves of plants supporting a mixed infection of Type and Sidney 81 was spatially nonuniform. Strain distribution among individual tillers of coinfected plants also was heterogeneous, with some containing either Type or Sidney 81 alone and some containing both. Transmission by wheat curl mites, acquiring virus from source plants simultaneously infected with both Type and Sidney 81, often resulted in test plants bearing only a single WSMV strain. Spatial subdivision of virus strains within coinfected plants likely contributed to vector transmission bottlenecks during acquisition. Collectively, these three distinct mechanisms enhance genetic isolation of individual viral lineages, and together with stochastic processes, may explain generation and maintenance of genetic diversity in field populations.
Assuntos
Vírus do Mosaico/genética , Triticum/virologia , Animais , Evolução Biológica , Genótipo , México , Ácaros/virologia , Vírus do Mosaico/classificação , Vírus do Mosaico/fisiologia , Doenças das Plantas/parasitologia , Doenças das Plantas/virologia , Folhas de Planta/parasitologia , Folhas de Planta/virologia , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Triticum/parasitologia , Estados UnidosRESUMO
While human masseter muscle is known to have unusual co-expression of myosin heavy-chain proteins, cellular kinetics of individual fibers has not yet been tested. Here we examine if myosin heavy-chain protein content is closely correlated to fiber-shortening speed, as previously reported in other human muscles, or if these proteins do not correlate well to shortening speeds, as has been demonstrated previously in rat muscle. Slack-test recordings of single, skinned human masseter fibers at 15 degrees C revealed maximum shortening velocities generally slower and much more variable than those recorded in human limb muscle. The slowest fiber recorded had a maximum shortening velocity (V0) value of 0.027 muscle lengths x s(-1), several times slower than the slowest type I fibers previously measured in humans. By contrast, human limb muscle controls produced V0 measurements comparable with previously published results. Analysis by gel electrophoresis found 63% of masseter fibers to contain pure type I MyHC and the remainder to co-express mostly type I in various combinations with IIA and IIX isoforms. V0 in masseter fibers forms a continuum in which no clear relationship to MyHC isoform content is apparent.
Assuntos
Músculo Masseter/química , Contração Muscular/fisiologia , Cadeias Pesadas de Miosina/fisiologia , Miosina Tipo I/fisiologia , Miosinas de Músculo Esquelético/fisiologia , Adolescente , Adulto , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Análise dos Mínimos Quadrados , Masculino , Músculo Masseter/metabolismo , Músculo Masseter/fisiologia , Pessoa de Meia-Idade , Fibras Musculares de Contração Lenta/química , Fibras Musculares de Contração Lenta/fisiologia , Cadeias Pesadas de Miosina/análise , Miosina Tipo I/análise , Fenótipo , Isoformas de Proteínas/biossíntese , Isoformas de Proteínas/fisiologia , Miosinas de Músculo Esquelético/análiseRESUMO
An Arabidopsis protein was found to interact specifically with the capsid protein (CP) of turnip crinkle virus (TCV) through yeast two-hybrid screening. This protein, designated TIP (for TCV-interacting protein), was found to be a member of the recently recognized NAC family of proteins. NAC proteins have been implicated in the regulation of development of plant embryos and flowers. TIP alone was able to activate expression of reporter genes in yeast if fused to a DNA binding domain, suggesting that it may be a transcriptional activator. The TIP binding region in the TCV CP has been mapped to the N-terminal 25 amino acids. Site-directed mutagenesis within this region revealed that loss of the TIP-CP interaction in the yeast two-hybrid assay correlated with loss of the ability of TCV to induce the hypersensitive response and resistance in the TCV-resistant Arabidopsis ecotype Dijon (Di-0 and its inbred line Di-17). These data suggest that TIP is an essential component in the TCV resistance response pathway.
Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Proteínas de Bactérias , Carmovirus/patogenicidade , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Arabidopsis/genética , Arabidopsis/virologia , Sequência de Bases , Capsídeo/metabolismo , Dados de Sequência Molecular , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Reação em Cadeia da Polimerase , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Homologia de Sequência de AminoácidosRESUMO
Inserts bearing the coding sequences of NPT II and beta-glucuronidase (GUS) were placed between the nuclear inclusion b (NIb) and coat protein (CP) domains of the wheat streak mosaic virus (WSMV) polyprotein ORF. The WSMV NIb-CP junction containing the nuclear inclusion a (NIa) protease cleavage site was duplicated, permitting excision of foreign protein domains from the viral polyprotein. Wheat, barley, oat and maize seedlings supported systemic infection of WSMV bearing NPT II. The NPT II insert was stable for at least 18-30 days post-inoculation and had little effect on WSMV CP accumulation. Histochemical assays indicated the presence of functional GUS protein in systemically infected wheat and barley plants inoculated with WSMV bearing GUS. The GUS constructs had greatly reduced virulence on both oat and maize. RT-PCR indicated that the GUS insert was subject to deletion, particularly when expressed as a GUS-NIb protein fusion. Both reporter genes were expressed in wheat roots at levels comparable to those observed in leaves. These results clearly demonstrate the utility of WSMV as a transient gene expression vector for grass species, including two important grain crops, wheat and maize. The results further indicate that both host species and the nature of inserted sequences affect the stability and expression of foreign genes delivered by engineered virus genomes.
Assuntos
Grão Comestível/genética , Técnicas de Transferência de Genes , Vírus do Mosaico/genética , Simportadores , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Grão Comestível/metabolismo , Grão Comestível/virologia , Genes Reporter , Vetores Genéticos , Glucuronidase/genética , Glucuronidase/metabolismo , Raízes de Plantas/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas Cotransportadoras de Sódio-FosfatoRESUMO
Infectious transcripts were successfully derived from full-length cDNA clones of foxtail mosaic potexvirus (FoMV). Full-length clones were constructed by RT-PCR whereby 5' and 3' genomic segments of 2.7 and 3.4 kb, respectively, were ligated into Bluescript II KS. The in vitro RNA transcripts were infectious to moncotyledonous (barley) and dicotyledonous (Chenopodium amaranticolor) plant species. Individual mutation studies on clones of each of the five major ORFs confirmed predicted gene function for the polymerase, TGB (triple gene block), and coat protein (CP) genes. Protoplast studies on expression of a unique open reading frame, ORF 5A, which initiates 143 nts upstream of the CP before it "reads through" the CP, revealed that the 5A protein was produced in vivo. Mutation analysis of the 5A ORF indicated, however, that it was not required for either replication or for productive infection of plants. However, the nucleic acid sequences encoding the extended CP segment were shown to be important for CP expression. Additional mutations in 5A had no effect on FoMV replication in protoplasts but rendered the virus noninfectious to plants. A correlation with diminished CP production from both mutant clones implies that synthesis of subgenomic CP mRNA was compromised, and this limited systemic infection.
Assuntos
Plantas/virologia , Potexvirus/genética , RNA Viral/genética , Regiões 3' não Traduzidas , Regiões 5' não Traduzidas , Western Blotting , Capsídeo/análise , Capsídeo/genética , Clonagem Molecular , Primers do DNA , RNA Polimerases Dirigidas por DNA/genética , Eletroporação , Genes Virais , Mutação , Fases de Leitura Aberta , Potexvirus/metabolismo , Protoplastos/virologia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , TransfecçãoRESUMO
Turnip crinkle virus (TCV) is a single-stranded positive-sense RNA virus of the Carmovirus genus. Two of its five open reading frames (ORFs), encoding proteins of 8 and 9 kDa, are required for cell-to-cell movement of the virus. These movement proteins (MPs) were fused to green fluorescent protein (GFP) to determine their cellular localization. In protoplasts, p9-GFP, like GFP itself, is found throughout the cytoplasm, as well as in cell nuclei. In contrast, p8-GFP was confined to the cell nucleus. Similar localization patterns were observed when specific small peptide epitopes were fused to p8 and p9 proteins instead of GFP. The cytoplasmic localization of p9-GFP and nuclear localization of p8-GFP were also detected in leaves after particle bombardment of DNA encoding these fusion proteins or after overexpression of p8-GFP in transgenic Arabidopsis seedlings. The expression of the GFP fusion proteins by recombinant TCV viruses in infected protoplasts or on inoculated Arabidopsis leaves produced similar patterns. Unlike TMV-MP and other MPs studied to date, no obvious punctuate expression in the cell wall or association with the cytoskeleton was detected. The sequence analysis of p8 revealed two unique nuclear localization signals (NLSs), which were not conserved within p8 homologues of other viruses in the genus Carmovirus. Mutation in either of these NLSs did not disrupt the nuclear localization of p8-GFP. However, when both NLSs were mutated, p8-GFP expression was no longer restricted to cell nuclei. The NLSs are not required for cell-to-cell movement; TCV recombinant viruses mutated in one or both NLSs could still facilitate cell-to-cell movement of the virus. The nuclear localization of p8 suggests a novel function for this protein in the cell nucleus.
Assuntos
Carmovirus/metabolismo , Núcleo Celular/química , Proteínas Virais/análise , Sequência de Aminoácidos , Arabidopsis , Análise Mutacional de DNA , Dados de Sequência Molecular , Peso Molecular , Folhas de Planta/química , Proteínas do Movimento Viral em Plantas , Plantas Geneticamente Modificadas , Plantas Tóxicas , Plasmídeos , Proteínas Recombinantes de Fusão/análise , Nicotiana/citologia , Nicotiana/virologiaRESUMO
U.S. military forces are frequently deployed with little warning to regions of the world where chloroquine-resistant malaria is endemic. Doxycycline is often used for malaria chemoprophylaxis in these environments. The use of doxycycline can be complicated by esophageal injury. Two cases of esophageal ulceration will be discussed, followed by a review of the literature. Doxycycline causes esophageal injury through a combination of drug-specific factors, the circumstances of drug administration, and individual patient conditions. Patients with dysphagia attributable to esophageal ulceration are managed by intravenous fluid support and control of gastric acid reflux until their symptoms resolve over 5 to 7 days. The risk of esophageal injury can be minimized by use of fresh capsules, drug administration in the upright position well before lying down to sleep, and drinking at least 100 ml of water after swallowing the medication.
Assuntos
Antibacterianos/efeitos adversos , Transtornos de Deglutição/induzido quimicamente , Doxiciclina/efeitos adversos , Esofagite/induzido quimicamente , Militares , Úlcera/induzido quimicamente , Adulto , Transtornos de Deglutição/terapia , Esofagite/terapia , Feminino , Humanos , Malária/tratamento farmacológico , Masculino , Medicina Militar/métodos , Úlcera/terapia , Estados UnidosRESUMO
Mammalian skeletal muscle cells are composed of repeated sarcomeric units containing thick and thin filaments of myosin and actin, respectively. Excitation of the myosin ATPase enzyme is possible only with presence of Mg-ATP and Ca(2+). Skeletal muscle fibres may be classified into several types according to the isoform of myosin they contain. Nine isoforms of myosin heavy chain are known to exist in mammalian skeletal muscle including type I, IIA, IIB, IIX, IIM, alpha, neonatal, embryonic, and extra-ocular. Healthy adult human limb skeletal muscle contains type I, IIA, IIB, and IIX myosin heavy chains. The jaw-closing muscles of most carnivores and primates have tissue-specific expression of the type IIM or 'type II masticatory' myosin heavy chain. Adult human jaw-closing muscles, however, do not contain IIM myosin. Rather, they express type I, IIA, IIX (as in human limb muscle), and myosins typically expressed in developing or cardiac muscle. The morphology of human jaw-closing muscle fibres is also unusual in that the type II fibres are of smaller diameter that type I fibres, except in cases of increased function and hypertrophy. This paper describes the relationship of fibre types and motor unit function to changes in human occlusion and masticatory activity. Refereed Scientific Paper
Assuntos
Músculos da Mastigação/química , Fibras Musculares Esqueléticas/química , Fibras Musculares Esqueléticas/fisiologia , Músculo Esquelético/fisiologia , Adulto , Oclusão Dentária , Humanos , Mastigação , Músculos da Mastigação/fisiologia , Neurônios Motores/fisiologia , Contração Muscular , Músculo Esquelético/química , Cadeias Pesadas de Miosina/química , Fenótipo , Isoformas de ProteínasRESUMO
The presence of translational control elements and cap structures has not been carefully investigated for members of the Carmovirus genus, a group of small icosahedral plant viruses with positive-sense RNA genomes. In this study, we examined both the 5' and 3' untranslated regions (UTRs) of the turnip crinkle carmovirus (TCV) genomic RNA (4 kb) as well as the 5' UTR of the coat protein subgenomic RNA (1.45 kb) for their roles in translational regulation. All three UTRs enhanced translation of the firefly luciferase reporter gene to different extents. Optimal translational efficiency was achieved when mRNAs contained both 5' and 3' UTRs. The synergistic effect due to the 5'-3' cooperation was at least fourfold greater than the sum of the contributions of the individual UTRs. The observed translational enhancement of TCV mRNAs occurred in a cap-independent manner, a result consistent with the demonstration, using a cap-specific antibody, that the 5' end of the TCV genomic RNA was uncapped. Finally, the translational enhancement activity within the 5' UTR of 1.45-kb subgenomic RNA was shown to be important for the translation of coat protein in protoplasts and for virulent infection in Arabidopsis plants.
Assuntos
Brassica/virologia , Carmovirus/genética , Biossíntese de Proteínas , Capuzes de RNA , RNA Mensageiro/genética , RNA Viral/genética , Regiões 3' não Traduzidas/genética , Regiões 3' não Traduzidas/metabolismo , Regiões 5' não Traduzidas/genética , Regiões 5' não Traduzidas/metabolismo , Sequência de Bases , Capsídeo/genética , Capsídeo/metabolismo , Regulação Viral da Expressão Gênica , Genoma Viral , Luciferases/metabolismo , Dados de Sequência Molecular , Estabilidade de RNA , RNA Mensageiro/metabolismo , RNA Viral/metabolismoRESUMO
A tobacco mosaic virus (TMV)-based vector has been used to express in plants the complete open reading frame coding for VP1, the major immunogenic protein of foot and mouth disease virus (FMDV). In vitro RNA transcripts were inoculated into Nicotiana benthamiana plants and detectable amounts of recombinant VP1 were identified by Western blot as soon as 4 days postinfection. Foliar extracts prepared from infected leaves were injected intraperitoneally into mice and all of the immunized animals developed a specific antibody response to both the complete virus particle and the major immunogenic region as determined by ELISA and Western blot analysis. Most importantly, all immunized mice developed a protective immune response against experimental challenge with virulent FMDV. To our knowledge, this is the first report showing the expression of a complete open reading frame of an antigenic foreign protein in plants, using a recombinant plant virus, in sufficient quantity to permit use of the crude plant extract as an experimental immunogen to protect animals against virus challenge.
Assuntos
Aphthovirus/imunologia , Capsídeo/imunologia , Febre Aftosa/imunologia , Plantas/virologia , Vacinas Sintéticas , Vacinas Virais , Animais , Anticorpos Antivirais/sangue , Formação de Anticorpos , Aphthovirus/genética , Capsídeo/genética , Proteínas do Capsídeo , Ensaio de Imunoadsorção Enzimática , Febre Aftosa/prevenção & controle , Camundongos , Vírus do Mosaico do Tabaco/genéticaRESUMO
We have designed a DNA cassette to transcribe defective interfering (DI) RNAs of tomato bushy stunt virus (TBSV) and have investigated their potential to protect transgenic Nicotiana benthamiana plants from tombusvirus infections. To produce RNAs with authentic 5' and 3' termini identical to those of the native B10 DI RNA, the DI RNA sequences were flanked by ribozymes (RzDI). When RzDI RNAs transcribed in vitro were mixed with parental TBSV transcripts and inoculated into protoplasts or plants, they became amplified, reduced the accumulation of the parental RNA, and mediated attenuation of the lethal syndrome characteristic of TBSV infections. Analysis of F1 and F2 RzDI transformants indicated that uninfected plants expressed the DI RNAs in low abundance, but these RNAs were amplified to very high levels during TBSV infection. By two weeks postinoculation with TBSV, all untransformed N. benthamiana plants and transformed negative controls died. Although infection of transgenic RzDI plants initially induced moderate to severe symptoms, these plants subsequently recovered, flowered, and set seed. Plants from the same transgenic lines also exhibited broad-spectrum protection against related tombusviruses but remained susceptible to a distantly related tombus-like virus and to unrelated viruses.