RESUMEN
Objective: There is a need for low-cost, wide-reaching interventions to enhance accessibility of support for people with hazardous alcohol consumption. We assessed participant experiences of using a novel, personalised mHealth intervention offering approach bias modification (ApBM) for alcohol use in a community sample drinking at harmful levels to enable a deeper understanding of the end user and engagement. Methods: Eighteen semi-structured telephone interviews were conducted with adults in the community drinking at harmful/hazardous levels. A reflexive thematic analysis approach was used and data analysis followed iterative categorisation. Results: Engagement/Motivation and Clinical Value were overarching themes. The useable, accessible, customisable design described by participants enabled training to be readily integrated into routines, enhancing autonomy and self-efficacy beliefs, and facilitating engagement/motivation. Where autonomy or perceived self-efficacy were threatened by a rigid training schedule or lack of clarity/reminders, engagement was reduced. Training increased awareness of drinking behaviours, and encouraged participants to consider alternate goal-directed behaviours with feedback suggesting training may function as a 'circuit breaker', increasing time between alcohol craving and seeking, and enabling reflective processing, at least in the short term. Conclusions: This novel smartphone intervention for alcohol use may be a useful, accessible, 'just in time' adjunctive support tool for non-treatment seekers, meeting an important gap in the field. Findings have implications for the implementation of subsequent digital interventions, suggesting participants may stand to gain more from an intervention which enables autonomy and improves self-efficacy beliefs. Theoretically, findings speak to the role of inferential processing in behaviour change, but further research is needed to clearly elucidate ApBM training mechanisms. Practical recommendations for subsequent app iterations are suggested, along with additional opportunities worthy of consideration for future initiatives.
RESUMEN
Essentials Factor (F)VIII with an intermediate-length B-domain showed higher levels in murine gene therapy. FVIII with different B-domain lengths were analysed. FVIII variants with B-domains between 186 and 240 amino acids (aa) have extended half-life in mice. Reduced cell binding of FVIII with a 237aa B-domain may explain the extended half-life. SUMMARY: Background Factor VIII consists of the A1-domain, A2-domain, B-domain, A3-domain, C1-domain, and C2-domain. FVIII with an intermediate-length B-domain of 226 amino acids (aa) has previously been evaluated in murine gene therapy studies. Objective To characterize FVIII with intermediate-length B-domains in vitro and in vivo in F8-knockout (KO) mice. Methods and results FVIII molecules with B-domains of 186-240aa had longer half-lives in F8-KO mice than FVIII molecules with shorter or longer B-domains. FVIII with a B-domain containing the 225 N-terminal aa fused to the 12 C-terminal aa of the wild-type B-domain (FVIII-237) had a 1.6-fold extended half-life in F8-KO mice as compared with FVIII with a 21aa B-domain (FVIII-21). The in vitro and in vivo activity of FVIII-237 were comparable to those of FVIII-21, as was binding to von Willebrand factor. Cell binding to LDL receptor-related protein 1 (LRP-1)-expressing cells was markedly reduced for FVIII-237 as compared with FVIII-21, whereas the affinity for LRP-1 was not reduced in surface plasmon resonance (SPR) studies. FVIII-21 cell binding and internalization could be inhibited by a fragment consisting of the 226 N-terminal aa of the FVIII B-domain, and SPR analysis suggested that this B-domain fragment might bind with weak affinity to FVIII-21. Conclusion Reduced cell binding of FVIII-237 might explain the observed extended half-life in F8-KO mice. This may contribute to the increased FVIII levels measured in murine gene therapy studies using FVIII constructs with similar B-domain lengths.
Asunto(s)
Coagulantes/farmacocinética , Factor VIII/farmacocinética , Hemofilia A/tratamiento farmacológico , Animales , Línea Celular , Coagulantes/sangre , Modelos Animales de Enfermedad , Factor VIII/genética , Técnicas de Inactivación de Genes , Semivida , Hemofilia A/sangre , Proteína 1 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Ratones Noqueados , Unión Proteica , Dominios Proteicos , Proteínas Recombinantes/farmacocinéticaRESUMEN
UNLABELLED: Essentials Von Willebrand factor (VWF) stabilizes factor VIII (FVIII) and prevents its premature clearance. Rat anatomical and hepatocellular distribution studies assessed the VWF effect on FVIII clearance. Hepatocytes and liver sinusoidal endothelial cells play a key role in FVIII clearance. Anatomical and hepatocellular distribution of FVIII is independent of high-affinity VWF binding. ABSTRACT: Background Von Willebrand factor (VWF) stabilizes factor VIII in the circulation and prevents its premature clearance. Objective To study the effects of VWF on FVIII clearance in rats with endogenous VWF. Methods Anatomical and hepatocellular distribution studies were performed in rats following intravenous administration of glycoiodinated recombinant FVIII (rFVIII) and a FVIII variant, FVIII-Y1680F, lacking high-affinity VWF binding. Radioactivity was quantified in organs, and in distinct liver cell populations. The role of VWF binding was also studied by immunohistochemical staining of rat livers perfused ex vivo with rFVIII alone or with a FVIII-binding VWF fragment. Results The liver was the predominant organ of rFVIII distribution, and a radioactivity peak was also observed in the intestines, suggesting FVIII secretion to the bile by hepatocytes. In the liver, ~60% of recovered radioactivity was associated with hepatocytes, 32% with liver sinusoidal endothelial cells (LSECs), and 9% with Kupffer cells (KCs). When calculated per cell, 1.5-fold to 3-fold more radioactivity was associated with LSECs than with hepatocytes. The importance of hepatocytes and LSECs was confirmed by immunohistochemical staining; strong staining was seen in LSECs, and less intense, punctate staining in hepatocytes. Minor staining in KCs was observed. Comparable anatomical and hepatocellular distributions were observed with rFVIII and FVIII-Y1680F, and the presence of the VWF fragment, D'D3A1, did not change the FVIII staining pattern in intact livers. Conclusions The present data support FVIII clearance via the liver, with hepatocytes and LSECs playing a key role. High-affinity VWF binding did not alter the anatomical or hepatocellular distribution of FVIII.
Asunto(s)
Células Endoteliales/metabolismo , Factor VIII/metabolismo , Hepatocitos/citología , Hígado/metabolismo , Factor de von Willebrand/metabolismo , Animales , Línea Celular Tumoral , Modelos Animales de Enfermedad , Combinación de Medicamentos , Ensayo de Inmunoadsorción Enzimática , Factor VIII/uso terapéutico , Glioblastoma/metabolismo , Hepatocitos/metabolismo , Humanos , Inmunohistoquímica , Yodo/química , Lactoperoxidasa/metabolismo , Masculino , Unión Proteica , Ratas , Ratas Sprague-Dawley , Distribución Tisular , Enfermedades de von Willebrand/sangre , Enfermedades de von Willebrand/tratamiento farmacológico , Factor de von Willebrand/uso terapéuticoAsunto(s)
Factor IX/química , Animales , ADN Complementario/metabolismo , Diseño de Fármacos , Glicosilación , Hemofilia A/tratamiento farmacológico , Hemofilia A/genética , Humanos , Ratones , Mutagénesis Sitio-Dirigida , Mutación , Polisacáridos/química , Proteínas Recombinantes/química , Factores de TiempoRESUMEN
SUMMARY: A new recombinant factor VIII (FVIII), N8, has been produced in Chinese hamster ovary (CHO) cells. The molecule consists of a heavy chain of 88 kDa including a 21 amino acid residue truncated B-domain and a light chain of 79 kDa. The two chains are held together by non-covalent interactions. The four-step purification includes capture, affinity purification using a monoclonal recombinant antibody, anion exchange chromatography and gel filtration. The specific clotting activity of N8 was 8800-9800 IU mg(-1). Sequence and mass spectrometry analysis revealed two variants of the light chain, corresponding to two alternative N-terminal sequences also known from plasma FVIII. Two variants of the heavy chain are present in the purified product, namely with and without the B-domain linker attached. This linker is removed upon thrombin activation of N8 rendering an activated FVIII (FVIIIa) molecule similar to plasma FVIIIa. All six known tyrosine sulphations of FVIII were confirmed in N8. Two N-linked glycosylations are present in the A3 and C1 domain of the light chain and two in the A1 domain of the heavy chain. The majority of the N-linked glycans are sialylated bi-antennary structures. An O-glycosylation site is present in the B-domain linker region. This site was glycosylated with a doubly sialylated GalNAc-Gal structure in approximately 65% of the product. In conclusion, the present data show that N8 is a pure and well-characterized FVIII product with biochemical properties that equal other FVIII products.
Asunto(s)
Factor VIII/química , Secuencia de Aminoácidos , Animales , Western Blotting , Cromatografía Líquida de Alta Presión , Cromatografía por Intercambio Iónico , Cricetinae , Factor VIII/aislamiento & purificación , Glicoproteínas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Tripsina/metabolismoRESUMEN
During the last decade, various zinc salts have been used against the common cold syndrome, which is known to be initiated by respiratory viruses, particularly rhinoviruses. Using rhinovirus as the challenge virus, we investigated whether zinc salts (Zn) could potentiate the antiviral action of native human leukocyte interferon (HuIFN-alpha) and rHuIFN-gamma. We found that HuIFN-alpha was potentiated tenfold at rather low levels of IFN activity (0.6-0.8 U/ml), resulting in 100% protection. Zn alone gave only marginal protection, if any. In contrast to HuIFN-alpha, rHuIFN-gamma directly increased the cytopathic effect of rhinovirus at low levels (<2 U/ml) but protected the cells at higher IFN levels (5-20 U/ml). No potentiation was seen with Zn. HuIFN-beta protected against rhinovirus at the same doses as used with HuIFN-alpha, but in contrast to HuIFN-alpha, no potentiation was noted.
Asunto(s)
Antivirales/farmacología , Interferón-alfa/farmacología , Zinc/metabolismo , Línea Celular , Efecto Citopatogénico Viral/efectos de los fármacos , Sinergismo Farmacológico , Humanos , Rhinovirus/efectos de los fármacos , Zinc/farmacologíaRESUMEN
Vaccine strains of measles virus (MV) use CD46 as receptor and downregulate CD46 from the surface of infected cells. MVs isolated and passaged on B-lymphoid cells (wild-type MVs) seem to use another receptor and do not downregulate CD46. In the present study, we found that isolation of MV on human or marmoset B-lymphoid cells did not alter the MV haemagglutinin (H) protein relative to that in the patient. The wild-type isolates were adapted to the human epithelial HEp-2 cell line or the monkey fibroblast Vero cell line. All HEp-2 cell adapted viruses and 1 out of 4 Vero cell adapted viruses acquired the capacity to use CD46 as receptor, as measured by their ability to infect murine cells expressing human CD46. Adaptation to CD46 receptor usage was coupled to substitution of amino acid 481 of the MV H protein from asparagine to tyrosine but not to CD46 downregulation. The present study demonstrates that CD46 receptor usage can be induced by adaptation of wild-type MV to cells that do not express a wild-type receptor and suggests that a similar mechanism acted on the progenitor viruses of the present MV vaccine strains during their isolation and attenuation.
Asunto(s)
Antígenos CD/fisiología , Virus del Sarampión/fisiología , Glicoproteínas de Membrana/fisiología , Receptores Virales/fisiología , Animales , Antígenos CD/genética , Linfocitos B/inmunología , Linfocitos B/virología , Callithrix , Adhesión Celular , Línea Celular , Chlorocebus aethiops , Regulación hacia Abajo , Citometría de Flujo , Hemaglutininas Virales/fisiología , Humanos , Virus del Sarampión/genética , Virus del Sarampión/inmunología , Proteína Cofactora de Membrana , Glicoproteínas de Membrana/genética , Ratones , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales Cultivadas , Células VeroRESUMEN
The present study examines the coprecipitation of measles virus (MV) glycoproteins with host cell endoplasmic reticulum (ER) chaperone proteins. Both the haemagglutinin (H) and fusion (F) glycoproteins interacted with calnexin and GRP78, whereas interaction with calreticulin was only demonstrated for the H glycoprotein. The alpha-glucosidase inhibitor castanospermine reduced and delayed the association of F proteins with calnexin. We have previously shown that alpha-glucosidase activity is important for the functionality and antigenicity of the MV F glycoprotein and for release of MV particles from infected cells. Thus, interaction with calnexin appears vital for processing of nascent MV F protein into its functional conformation. In contrast to many other viral glycoproteins, a substantial proportion of the pulsed MV glycoproteins remained associated with ER chaperones for more than 2(1/2) h. Thus, the slow and incomplete migration of MV glycoproteins to the cell surface may result from their retention by ER chaperones, probably due to malfolding. MV infection upregulated the cellular expression of calreticulin and GRP78 and also increased their presence at the cell surface. The chaperone proteins are involved in a wide range of cellular processes, and their induction by MV may play a role for the pathogenesis of measles and its sequelae.
Asunto(s)
Retículo Endoplásmico/virología , Proteínas de Choque Térmico , Hemaglutininas Virales/fisiología , Virus del Sarampión/fisiología , Chaperonas Moleculares/fisiología , Proteínas Virales de Fusión/fisiología , Proteínas de Unión al Calcio/fisiología , Calnexina , Calreticulina , Proteínas Portadoras/fisiología , Retículo Endoplásmico/metabolismo , Chaperón BiP del Retículo Endoplásmico , Regulación de la Expresión Génica , Proteínas HSP70 de Choque Térmico/fisiología , Humanos , Proteínas de la Membrana/fisiología , Chaperonas Moleculares/genética , Ribonucleoproteínas/fisiología , Regulación hacia ArribaRESUMEN
The fusion (F) glycoprotein of respiratory syncytial virus (RSV) is synthesized as a nonfusogenic precursor protein (F(0)), which during its migration to the cell surface is activated by cleavage into the disulfide-linked F(1) and F(2) subunits. In the present study, soluble secreted human furin produced by a recombinant baculovirus cleaved RSV F(0) into proteins the size of F(1) and F(2). Furthermore, cleavage of F(0) was partially inhibited in the furin defective LoVo cell line, in calcium depleted HEp-2 cells, and in HEp-2 cells treated with the furin inhibitor decanoyl-R-V-K-R-chloromethylketon. These findings strongly suggest an important role for furin in activation of the RSV F protein. The F(0) protein could not be detected on the surface of cells, in which F protein activation was inhibited, and RSV particles did not appear to be released from these cells. It thus seems that in contrast to the F proteins of most other paramyxoviruses, the RSV F(0) protein is very inefficient in reaching the cell surface or is unable to reach the cell surface and therefore cannot be incorporated into virus particles.
Asunto(s)
Proteína HN , Infecciones por Virus Sincitial Respiratorio/virología , Virus Sincitiales Respiratorios/fisiología , Subtilisinas/metabolismo , Proteínas Virales/metabolismo , Baculoviridae/genética , Baculoviridae/metabolismo , Furina , Humanos , Pruebas de Precipitina , Virus Sincitiales Respiratorios/patogenicidad , Subtilisinas/genética , Células Tumorales Cultivadas , Proteínas del Envoltorio ViralRESUMEN
The envelope of measles virus (MV) particles contains two viral glycoproteins, the haemagglutinin (H) and the fusion (F) protein, which together induce the entry of MV into cells. In the present study, we investigated the role of oligosaccharide processing for the function and antigenicity of the MV glycoproteins by means of glycosidase inhibitors. Golgi alpha-mannosidase inhibitors (1-deoxymannojirimycin and swainsonine) prevented the oligosaccharides on the MV glycoproteins from obtaining Endo H resistance, but that did not appear to influence in vitro MV infections, indicating that conversion of oligosaccharide chains into the complex form was not required for the function of the MV glycoproteins. The alpha-glucosidase inhibitor castanospermine (CSP) quantitatively reduced the production of infectious MV particles in cells infected with both vaccine strain and wild-type MV. CSP reduced the detection of the MV F protein by certain monoclonal antibodies (MAbs) that appeared to recognize nonlinear epitopes. CSP also inhibited syncytium formation in MV infected cells, but did not affect MV induced CD46 downregulation, suggesting that CSP primarily influenced the F protein. We propose that CSP induces aberrant folding of MV glycoproteins in a manner that influences their function and antigenicity.
Asunto(s)
Hemaglutininas Virales/metabolismo , Manosidasas/metabolismo , Virus del Sarampión/metabolismo , Oligosacáridos/metabolismo , Procesamiento Proteico-Postraduccional , Proteínas Virales de Fusión/metabolismo , alfa-Glucosidasas/metabolismo , 1-Desoxinojirimicina/farmacología , Inhibidores Enzimáticos/farmacología , Glicoproteínas/inmunología , Glicoproteínas/metabolismo , Inhibidores de Glicósido Hidrolasas , Hemaglutininas Virales/inmunología , Humanos , Indolizinas/farmacología , Manosidasas/antagonistas & inhibidores , Swainsonina/farmacología , Células Tumorales Cultivadas , Proteínas Virales de Fusión/inmunología , Virión/fisiología , alfa-ManosidasaRESUMEN
The fusion (F) glycoprotein gene of measles virus (MV) encodes a nonfusogenic precursor protein (F0) that is activated by cleavage into the F1 and F2 subunits during transport to the cell surface. The F protein of both the Edmonston strain and a wild-type MV was found to be cleaved in the trans-Golgi cisternae and/or the trans-Golgi network (TGN). In HEp-2 cells, B lymphoblastoid cells, and PBMC, the cleavage process required calcium, and calcium deprivation prevented syncytium formation. The calcium dependence indicated the involvement of the pro-protein convertase (PC) endoprotease family. The expression of the presently recognized members of the PC family in human cell types known to be infected during measles was examined by RT-PCR. Among the PCs residing in the TGN, only furin was expressed in all cells. Soluble secreted human furin produced by a recombinant baculovirus cleaved MV F0 into proteins the exact size of F1 and F2 and increased the titer of MV particles released from calcium-deprived or endoprotease defective infected cells. These results strongly indicate that furin is the most important and maybe the only endoprotease involved in activation of the MV F protein.
Asunto(s)
Virus del Sarampión/fisiología , Serina Endopeptidasas/metabolismo , Proteínas Virales de Fusión/metabolismo , Adenocarcinoma , Carcinoma de Células Escamosas , Furina , Aparato de Golgi/enzimología , Aparato de Golgi/virología , Humanos , Cinética , Virus del Sarampión/genética , Proteínas Recombinantes/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Serina Endopeptidasas/genética , Subtilisinas/metabolismo , Células Tumorales Cultivadas , Proteínas Virales de Fusión/genética , Ensayo de Placa ViralRESUMEN
To characterize the variability of recent field isolates of canine distemper virus (CDV) from different hosts and geographical areas, we conducted nucleotide sequence analysis of the gene encoding the haemagglutinin (H), the attachment protein of this virus. Pronounced differences between field isolates were revealed in comparison to the Convac and Onderstepoort vaccine strains. The diversity of CDV appeared to exceed that determined for measles virus. Phylogenetic analysis also separated the field isolates of CDV from the vaccine strains and provided evidence for the existence of different contemporary genotypes of CDV. Isolates from a Greenlandic sledge dog and a Siberian seal formed a distinct lineage. The remaining isolates formed a group. This group contained two European isolates from mink and ferret, a single lineage comprising three European dog isolates, and another separate lineage of North American isolates from dog, javelina, raccoon and captive leopards.
Asunto(s)
Virus del Moquillo Canino/genética , Variación Genética , Glicoproteínas/genética , Proteínas Virales/genética , Secuencia de Aminoácidos , Animales , Virus del Moquillo Canino/clasificación , Virus del Moquillo Canino/aislamiento & purificación , Perros , Datos de Secuencia Molecular , Filogenia , Phocidae , Vacunas Virales/genéticaRESUMEN
Canine angiostrongylosis was diagnosed in a whippet with typical signs of respiratory and circulatory distress. Subclinical Angiostrongylus vasorum infections were also demonstrated in two other whippets belonging to the same owner. All three dogs were given standard anthelmintic levamisole treatment combined with corticosteroids. Two days after initiation of treatment, one of the subclinically infected dogs developed severe hypovolaemic shock that required intravenous fluid therapy and corticosteroids to save its life. The shock is believed to have been caused by an anaphylactic reaction triggered by the rapid release of a large amount of worm antigen in the blood due to the rapid death of adult worms by levamisole. Thus, dog owners should be instructed to monitor dogs undergoing levamisole treatment against A vasorum. Alternatives to levamisole treatment of canine angiostrongylosis should be considered.
Asunto(s)
Antihelmínticos/uso terapéutico , Enfermedades de los Perros/etiología , Levamisol/uso terapéutico , Choque/veterinaria , Infecciones por Strongylida/veterinaria , Corticoesteroides/uso terapéutico , Anafilaxia/complicaciones , Anafilaxia/etiología , Anafilaxia/veterinaria , Angiostrongylus/inmunología , Angiostrongylus/aislamiento & purificación , Animales , Anticuerpos Antihelmínticos/sangre , Contraindicaciones , Enfermedades de los Perros/sangre , Enfermedades de los Perros/inmunología , Perros , Quimioterapia Combinada , Masculino , Choque/sangre , Choque/etiología , Infecciones por Strongylida/tratamiento farmacológico , Infecciones por Strongylida/inmunologíaRESUMEN
DMV, dolphin morbillivirus, a paramyxovirus of uncertain origin recently emerged in Mediterranean dolphins. This study presents the complete nucleotide sequence of the hemagglutinin (H) gene including the gene boundaries. The single open reading frame of the DMV H gene encodes a protein of 604 residues which exhibits overall sequence characteristics similar to the H genes of other morbilliviruses. When compared to its closest homologues, measles virus (MV) and rinderpest virus (RPV), DMV has, respectively, 44 and 46% of amino acid residues in identical positions. The primary sequence of the DMV H protein is markedly less conserved than that of the fusion protein. The comparative data at the genomic level correspond with cross-neutralization studies with different morbilliviruses. Retrospective serogical studies dating back to 1983 indicate DMV-like infections in whales of the eastern Atlantic. The presented data support and extend previous studies suggesting that this novel morbillivirus is one of the phylogenetically oldest morbilliviruses known to circulate today. The relationship of DMV and established morbilliviruses to the newly emerged candidate morbillivirus infecting horse and man is discussed.
Asunto(s)
Delfines/virología , Hemaglutininas Virales/genética , Morbillivirus/genética , Secuencia de Aminoácidos , Animales , Anticuerpos Antivirales/inmunología , Secuencia de Bases , ADN Viral , Hemaglutininas Virales/inmunología , Datos de Secuencia Molecular , Morbillivirus/clasificación , Morbillivirus/inmunología , Filogenia , ARN Mensajero , ARN Viral , Homología de Secuencia de AminoácidoRESUMEN
The nucleotide sequence of the phosphoprotein (P) gene of a dolphin morbillivirus (DMV) isolate was determined. Like those of other morbilliviruses the DMV P gene encoded P and C proteins in overlapping open reading frames and V protein by editing the P gene transcript. Among P mRNA based clones the editing site variants GGGC, GGGG, GAGC and GGGGGGC predicting a P protein, and the variants GGGGC and GGGGGG predicting a V protein, were found. Surprisingly, the three variants GGGC, GGGG and GAGC were also found among clones generated from genomic RNA of the DMV isolate. Thus, more than one viral genome type appeared to be present in cells infected with the DMV isolate. By a similar analysis of the virus genomes in the tissue from which the DMV isolate was obtained, only the GGGC type was found, indicating that the GGGG and GAGC types arose during adaptation of the virus to growth in cell cultures. No editing site variants likely to have arisen by editing the GAGC type were encountered, and it remains ot be determined whether mRNA encoding V protein can be transcribed from genomes with this editing site. Using antisera raised against the common N terminus and unique C termini of the predicted P and V proteins, the in vivo expression of these proteins was demonstrated.
Asunto(s)
Delfines/virología , Genes Virales/genética , Morbillivirus/genética , Fosfoproteínas/genética , Edición de ARN/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Chlorocebus aethiops , Clonación Molecular , Variación Genética/genética , Datos de Secuencia Molecular , Peso Molecular , Fosfoproteínas/biosíntesis , Fosfoproteínas/química , ARN Mensajero/genética , ARN Viral/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Células Vero , Proteínas no Estructurales Virales/biosíntesis , Proteínas no Estructurales Virales/química , Proteínas no Estructurales Virales/genética , Proteínas Estructurales Virales/biosíntesis , Proteínas Estructurales Virales/química , Proteínas Estructurales Virales/genéticaRESUMEN
Morbilliviruses have been isolated from stranded dolphins and porpoises. The present paper describes the cloning and sequencing of the porpoise morbillivirus (PMV) F gene and of the dolphin morbillivirus (DMV) M and F genes and their flanking regions. The gene order of the DMV genome appeared to be identical to that of other morbilliviruses. A genomic untranslated region of 837 nucleotides was found between the translated DMV M and F gene regions. The predicted DMV M protein were highly conserved with those of other morbilliviruses. Both the deduced PMV and DMV F0 proteins exhibited three major hydrophobic regions as well as a cysteine rich region, a leucine zipper motif and a cleavage motif allowing cleavage of the F0 protein into F1 and F2 subunits. Apparently the DMV F0 cleavage motif was not modified by adaptation of DMV to Vero cells. The predicted PMV and DMV F proteins were 94% identical. Comparisons with the corresponding sequences of other morbilliviruses demonstrated that the cetacean morbillivirus does not derive from any known morbillivirus but represents an independent morbillivirus lineage.
Asunto(s)
Morbillivirus/genética , Proteínas Virales de Fusión/genética , Proteínas de la Matriz Viral/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cetáceos/virología , Chlorocebus aethiops , ADN Viral , Delfines/virología , Genes Virales , Datos de Secuencia Molecular , Morbillivirus/clasificación , Filogenia , ARN , ARN Viral , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Células VeroRESUMEN
Angiostrongylus vasorum has been recognised as a cause of respiratory and circulatory distress among dogs in southwestern France for more than a decade, and the nematode now appears to be of increasing importance in the British Isles and Denmark. The aim of this review is to give a concise account of present knowledge of this intriguing parasitosis.
Asunto(s)
Angiostrongylus/anatomía & histología , Angiostrongylus/fisiología , Enfermedades de los Perros/parasitología , Infecciones por Strongylida/veterinaria , Animales , Enfermedades de los Perros/epidemiología , Perros , Europa (Continente)/epidemiología , Infecciones por Strongylida/diagnóstico , Infecciones por Strongylida/epidemiología , Infecciones por Strongylida/prevención & control , Infecciones por Strongylida/terapiaRESUMEN
A morbillivirus of uncertain origin recently killed hundreds of Mediterranean dolphins. This is the first report of the nucleotide and deduced amino acid sequence of a dolphin morbillivirus (DMV) gene. The sequence of the nucleocapsid (N) gene including boundaries was determined. When the DMV N gene coding region was compared with the corresponding sequences of other morbilliviruses a distant evolutionary relationship between these viruses and DMV was apparent. Phylogenetic analysis of the sequence data provided further evidence that DMV is not closely related to any known morbillivirus, whereas phocine distemper virus exhibits a relatively close relationship to canine distemper virus.
Asunto(s)
Evolución Biológica , Cápside/genética , Genes Virales , Sarampión/genética , Morbillivirus/genética , Rumiantes/virología , Proteínas del Núcleo Viral/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cartilla de ADN , Delfines/virología , Genoma Viral , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Homología de Secuencia de AminoácidoRESUMEN
In 1990 an epidemic caused by a morbillivirus was noticed among Mediterranean dolphins. RNA was extracted from the tissues of dolphins and from cell cultures infected with a corresponding dolphin morbillivirus isolate. By nucleic acid hybridization this RNA was compared to RNA extracted from animal tissue or cell cultures infected with canine distemper virus (CDV), phocine distemper virus (PDV) or measles virus (MV). The presence of morbillivirus RNA in the dolphin tissue was demonstrated. Morbillivirus N, P, M and F gene mRNAs were detected in the RNA from dolphin morbillivirus infected cells. These mRNA species seemed to be of approximately the same size as the corresponding mRNA species of CDV, PDV and MV. The results of the comparison demonstrated that the dolphin morbillivirus is genetically different from CDV, PDV and MV. No indication of a close relationship between the dolphin isolate and either CDV, PDV or MV was found.