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1.
J Fish Dis ; 45(3): 479-483, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34843624

RESUMO

ISAV is the causative agent of the infectious salmon anaemia (ISA), a disease listed by the OIE that has caused important economic losses to the Atlantic salmon (Salmo salar) industry. ISAV variants are identified as pathogenic or non-pathogenic based on the presence or absence of a deletion in the highly polymorphic region (HPR) of segment 6 (S6). HPRΔ variants (pathogenic) are the only forms of the virus known to grow in cell culture. This is the first report of a HPR0 variant isolated in cell culture. The isolate is, however, atypical as it shows a marker of virulent variants on another segment (S5), which has never been reported for any other HPR0 variants. The significance of this finding remains unclear until more in-depth work is carried out but does challenge current knowledge.


Assuntos
Doenças dos Peixes , Isavirus , Infecções por Orthomyxoviridae , Salmo salar , Animais , Técnicas de Cultura de Células , Isavirus/genética , Infecções por Orthomyxoviridae/veterinária , Salmão
2.
Virus Evol ; 7(1): veab008, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34168895

RESUMO

[This corrects the article DOI: 10.1093/ve/veaa054.].

3.
J Med Entomol ; 58(4): 1686-1700, 2021 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-33822118

RESUMO

Approximately 80 species of mosquitoes (Diptera: Culicidae) have been documented in Canada. Exotic species such as Aedes albopictus (Skuse) (Diptera: Culicidae) are becoming established. Recently occurring endemic mosquito-borne diseases (MBD) in Canada including West-Nile virus (WNV) and Eastern Equine Encephalitis (EEE) are having significant public health impacts. Here we explore the use of DNA metabarcoding to identify mosquitoes from CDC light-trap collections from two locations in eastern Canada. Two primer pairs (BF2-BR2 and F230) were used to amplify regions of the cytochrome c oxidase subunit I (CO1) gene. High throughput sequencing was conducted using an Illumina MiSeq platform and GenBank-based species identification was applied using a QIIME 1.9 bioinformatics pipeline. From a site in southeastern Ontario, Canada, 26 CDC light trap collections of 72 to >300 individual mosquitoes were used to explore the capacity of DNA metabarcoding to identify and quantify captured mosquitoes. The DNA metabarcoding method identified 33 species overall while 24 species were identified by key. Using replicates from each trap, the dried biomass needed to identify the majority of species was determined to be 76 mg (equivalent to approximately 72 mosquitoes), and at least two replicates from the dried biomass would be needed to reliably detect the majority of species in collections of 144-215 mosquitoes and three replicates would be advised for collections with >215 mosquitoes. This study supports the use of DNA metabarcoding as a mosquito surveillance tool in Canada which can help identify the emergence of new mosquito-borne disease potential threats.


Assuntos
Biovigilância/métodos , Culicidae/classificação , Código de Barras de DNA Taxonômico , Mosquitos Vetores/classificação , Animais , Canadá , Culicidae/genética , Mosquitos Vetores/genética , Projetos Piloto
4.
Virus Evol ; 6(2): veaa054, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33381304

RESUMO

Piscine orthoreovirus (PRV-1) is a segmented RNA virus, which is commonly found in salmonids in the Atlantic and Pacific Oceans. PRV-1 causes the heart and skeletal muscle inflammation disease in Atlantic salmon and is associated with several other disease conditions. Previous phylogenetic studies of genome segment 1 (S1) identified four main genogroups of PRV-1 (S1 genogroups I-IV). The goal of the present study was to use Bayesian phylogenetic inference to expand our understanding of the spatial, temporal, and host patterns of PRV-1 from the waters of the northeast Pacific. To that end, we determined the coding genome sequences of fourteen PRV-1 samples that were selected to improve our knowledge of genetic diversity across a broader temporal, geographic, and host range, including the first reported genome sequences from the northwest Atlantic (Eastern Canada). Nucleotide and amino acid sequences of the concatenated genomes and their individual segments revealed that established sequences from the northeast Pacific were monophyletic in all analyses. Bayesian inference phylogenetic trees of S1 sequences using BEAST and MrBayes also found that sequences from the northeast Pacific grouped separately from sequences from other areas. One PRV-1 sample (WCAN_BC17_AS_2017) from an escaped Atlantic salmon, collected in British Columbia but derived from Icelandic broodstock, grouped with other S1 sequences from Iceland. Our concatenated genome and S1 analysis demonstrated that PRV-1 from the northeast Pacific is genetically distinct but descended from PRV-1 from the North Atlantic. However, the analyses were inconclusive as to the timing and exact source of introduction into the northeast Pacific, either from eastern North America or from European waters of the North Atlantic. There was no evidence that PRV-1 was evolving differently between free-ranging Pacific Salmon and farmed Atlantic Salmon. The northeast Pacific PRV-1 sequences fall within genogroup II based on the classification of Garseth, Ekrem, and Biering (Garseth, A. H., Ekrem, T., and Biering, E. (2013) 'Phylogenetic Evidence of Long Distance Dispersal and Transmission of Piscine Reovirus (PRV) between Farmed and Wild Atlantic Salmon', PLoS One, 8: e82202.), which also includes North Atlantic sequences from Eastern Canada, Iceland, and Norway. The additional full-genome sequences herein strengthen our understanding of phylogeographical patterns related to the northeast Pacific, but a more balanced representation of full PRV-1 genomes from across its range, as well additional sequencing of archived samples, is still needed to better understand global relationships including potential transmission links among regions.

5.
J Fish Dis ; 41(3): 421-430, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28782809

RESUMO

The infectious salmon anaemia virus (ISAV) is an important viral disease of farmed Atlantic salmon that has caused considerable financial losses for salmon farmers around the world, including Atlantic Canada. It is listed as a notifiable disease by the World Organization for Animal Health, and to this day, culling of infected cages or farms remains the current practice in many countries to mitigate the spread of the virus. In Atlantic Canada, ISAV was first detected in 1996 and continues to be detected. While some outbreaks seemed to have arisen from isolated infections of unknown source, others were local clusters resulting from horizontal spread of infection. This study provides a description of the detected ISAV isolates in Atlantic Canada between 2012 and 2016, and explores the phylogenetic relatedness between these ISAV isolates. A key finding is the detection for the first time of a North American-HPR0 ISAV subtype, which was predicted to exist for many years. Through phylogenetic analysis, a scenario emerges with at least three separate incursions of ISAV in Atlantic Canada. An initial ISAV introduction follows a genotypic separation between North America and Europe which resulted in the NA and EU genotypes known today; this separation predates the salmon aquaculture industry. The second incursion of ISAV from Europe to North America led to a sublineage in Atlantic Canada consisting of EU-HPR∆ isolates detected in Nova Scotia and New Brunswick, and the predominant form of ISAV-HPR0 (EU). Finally, we observed what could be the third and most recent incursion of ISAV in Newfoundland, in the form of an isolate highly similar to ISAV EU-HPR0 isolates found in the Faroe Islands and the one isolate from Norway.


Assuntos
Doenças dos Peixes/virologia , Genótipo , Isavirus/fisiologia , Infecções por Orthomyxoviridae/veterinária , Salmão , Proteínas Virais/genética , Sequência de Aminoácidos , Animais , Canadá , Isavirus/classificação , Isavirus/genética , Infecções por Orthomyxoviridae/virologia , Filogenia , Filogeografia , Alinhamento de Sequência , Proteínas Virais/química , Proteínas Virais/metabolismo
6.
J Sports Med Phys Fitness ; 55(12): 1488-96, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25781214

RESUMO

AIM: The purpose of this investigation was to determine the effects of 3 d of creatine supplementation on thermoregulation and isokinetic muscular performance. METHODS: Fourteen males performed two exercise bouts following 3 d of creatine supplementation and placebo. Subjects exercised for 60 min at 60-65% of VO2max in the heat followed by isokinetic muscular performance at 60, 180, and 300°·s(-1). Dependent variables for pre- and postexercise included nude body weight, urine specific gravity, and serum creatinine levels. Total body water, extracellular water and intracellular water were measured pre-exercise. Core temperature was assessed every 5 min during exercise. Peak torque and Fatigue Index were used to assess isokinetic muscular performance. RESULTS: Core temperature increased during the run for both conditions. Total body water and extracellular water were significantly greater (P<0.05) following creatine supplementation. No significant difference (P>0.05) was found between conditions for intracellular water, nude body weight, urine specific gravity, and serum creatinine. Pre-exercise scores for urine specific gravity and serum creatinine were significantly less (P<0.05) versus post-exercise. No significant differences (P>0.05) were found in peak torque values or Fatigue Index between conditions for each velocity. A significant (P<0.05) overall velocity effect was found for both flexion and extension. As velocity increased, mean peak torque values decreased. CONCLUSION: Three d of creatine supplementation does not affect thermoregulation during submaximal exercise in the heat and is not enough to elicit an ergogenic effect for isokinetic muscle performance following endurance activity.


Assuntos
Creatina/administração & dosagem , Desidratação/fisiopatologia , Exercício Físico/fisiologia , Contração Muscular/fisiologia , Músculo Esquelético/fisiologia , Esforço Físico/fisiologia , Adulto , Regulação da Temperatura Corporal , Peso Corporal , Creatina/metabolismo , Suplementos Nutricionais , Método Duplo-Cego , Teste de Esforço , Frequência Cardíaca , Humanos , Masculino , Contração Muscular/efeitos dos fármacos , Músculo Esquelético/efeitos dos fármacos , Esforço Físico/efeitos dos fármacos , Torque
7.
J Appl Physiol (1985) ; 113(10): 1604-12, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22984250

RESUMO

Cardiovascular deconditioning after a 4-h head-down bed rest (HDBR) might be a consequence of the time of day relative to pre-HDBR testing, or simply 4 h of confinement and inactivity rather than the posture change. Ten men and 11 women were studied during lower body negative pressure (LBNP) before and after 4-h HDBR and 4-h seated posture (SEAT) as a control for time of day and physical inactivity effects to test the hypotheses that cardiovascular deconditioning was a consequence of the HDBR posture, and that women would have a greater deconditioning response. Following HDBR, men and women had lower blood volume, higher heart rate with a greater increase during LBNP, a greater decrease of stroke volume during LBNP, lower central venous pressure, smaller inferior vena cava diameter, higher portal vein resistance index with a greater increase during LBNP, but lower forearm vascular resistance, lower norepinephrine, and lower renin. Women had lower vasopressin and men had higher vasopressin after HDBR, and women had lower pelvic impedance and men higher pelvic impedance. Following SEAT, brachial vascular resistance was reduced, thoracic impedance was elevated, the reduction of central venous pressure during LBNP was changed, women had higher angiotensin II whereas men had lower levels, and pelvic impedance increased in women and decreased in men. Cardiovascular deconditioning was greater after 4-h HDBR than after SEAT. Women and men had similar responses for most cardiovascular variables in the present study that tested the responses to LBNP after short-duration HDBR compared with a control condition.


Assuntos
Repouso em Cama/efeitos adversos , Descondicionamento Cardiovascular , Sistema Cardiovascular/fisiopatologia , Antebraço/irrigação sanguínea , Decúbito Inclinado com Rebaixamento da Cabeça/efeitos adversos , Hemodinâmica , Pressão Negativa da Região Corporal Inferior/efeitos adversos , Pelve/irrigação sanguínea , Adulto , Análise de Variância , Angiotensina II/sangue , Volume Sanguíneo , Artéria Braquial/fisiopatologia , Pressão Venosa Central , Tontura/fisiopatologia , Feminino , Frequência Cardíaca , Humanos , Masculino , Norepinefrina/sangue , Veia Porta/fisiopatologia , Fluxo Sanguíneo Regional , Renina/sangue , Fatores Sexuais , Volume Sistólico , Fatores de Tempo , Resistência Vascular , Vasopressinas/sangue , Veia Cava Inferior/fisiopatologia
8.
Mol Immunol ; 51(2): 197-209, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22475434

RESUMO

Following an infection with a specific pathogen, the acquired immune system of many teleostean fish, including salmonids, is known to retain a specific memory of the infectious agent, which protects the host against subsequent infections. For example, Atlantic salmon (Salmo salar) that have survived an infection with a low-virulence infectious salmon anemia virus (ISAV) isolate are less susceptible to subsequent ISAV infections. A greater understanding of the mechanisms and immunological components involved in this acquired protection against ISAV is fundamental for the development of efficacious vaccines and treatments against this pathogen. To better understand the immunity components involved in this observed resistance, we have used an Atlantic salmon DNA microarray to study the global gene expression responses of preexposed Atlantic salmon (fish having survived an infection with a low-virulence ISAV isolate) during the course of a secondary infection, 18 months later, with a high-virulence ISAV isolate. We present global gene expression patterns in both preexposed and naïve fish, following exposure by either cohabitation with infected fish or by direct intra-peritoneal injection of a high-virulence ISAV isolate. Our results show a clear reduction of ISAV viral loads in head-kidney of secondary infected fish compared to primary infected fish. Further, we note a lower-expression of many antiviral innate immunity genes in the secondary infected fish, such as the interferon induced GTP-binding protein Mx, CC-chemokine 19 and signal transducer and activator of transcription 1 (STAT 1), as well as MHC class I antigen presentation involved genes. Potential acquired immunity genes such as GILT, leukocyte antigen transcript CD37 and Ig mu chain C region membrane-bound form were observed to be over-expressed in secondary infected fish. The observed differential gene expression profile in secondary and primary infected fish head-kidney provides great insight into immunity components involved during primary and secondary ISAV infection.


Assuntos
Doenças dos Peixes/imunologia , Expressão Gênica/imunologia , Memória Imunológica/imunologia , Infecções por Orthomyxoviridae/imunologia , Salmão/imunologia , Transcrição Gênica , Animais , Doenças dos Peixes/genética , Doenças dos Peixes/virologia , Expressão Gênica/genética , Perfilação da Expressão Gênica , Memória Imunológica/genética , Isavirus , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Salmão/genética
9.
Med Phys ; 39(6Part17): 3810, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28517460

RESUMO

PURPOSE: The biologically effective dose (BED) of temporary brachytherapy treatments is a function of both chosen radionuclide (R) and implant duration (T). This study endeavored to evaluate BED delivered to the tumor volume and surrounding ocular structures as a function of plaque position (P), prescription dose, R, and T. METHODS: Plaque-heterogeneity- corrected dose distributions were generated with MCNP5 for the range of currently-available COMS plaques using low-energy radionuclides. These physical dose distributions were imported into the Pinnacle3 TPS using the TG-43 hybrid technique and used to generate DVHs for a T=7d implant within a reference eye geometry at eight standard treatment positions. The Dale equation was employed to create biologically effective dose volume histograms (BEDVHs), allowing for BED volumetric analysis of all ROIs. Isobiologically-effective prescription doses were calculated for T=5-0.01d, with BEDVHs subsequently generated for all ROIs using correspondingly reduced prescription doses. Objective functions were created to evaluate the BEDVHs as a function of R and T. RESULTS: Reducing T from 7 to 0.01d for a 10mm plaque produced an average BED benefit of 26%, 20%, and 17% for 103 Pd, 125 I, and 131 Cs, respectively, for all P; 16mm and 22mm plaque results were more position-dependent. 103 Pd produced a 16-35% BED benefit over 125 I, whereas 131 Cs produced a 3-7% BED detriment, independent of P, T, and plaque size. Additionally, corresponding OAR physical doses were lowest using 103 Pd in all circumstances. CONCLUSIONS: Shorter implant durations may correlate with more favorable outcomes vs. 7d implants for small and medium lesions. T may be safely reduced if the prescription dose is appropriately diminished. 103 Pd offers a substantial 16- 35% radiobiological benefit over 125 I and 131 Cs irrespective of P, T, and tumor size. The objective functions used in this study can be applied to temporary or permanent brachytherapy implants for a variety of disease sites.

10.
Fish Shellfish Immunol ; 29(2): 217-32, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20371292

RESUMO

Infectious diseases among fish present an important economic burden for the aquaculture and fisheries industries around the world. For example, the infectious salmon anemia virus (ISAV) infects farmed Atlantic salmon (Salmo salar), and results in millions of dollars of lost revenue to salmon farmers. Although improved management and husbandry practices over the last few years have minimized the losses and the number of outbreaks, the risk of new virulent strains emerging is a looming threat to the viability and sustainability of this industry. A greater understanding of the host-pathogen interactions at the molecular level during the course of an infection thus remains of strategic importance for the development of molecular tools and efficient vaccines capable of minimizing losses in the eventual case of a new outbreak. Using a 32 k cDNA microarray platform (cGRASP), we have identified various signaling pathways and immune regulated genes, which are activated or repressed in Atlantic salmon head-kidney during the course of an ISAV infection. Gene expressions were measured at five different time-points: 6 h, 24 h, 3 d, 7 d and 16 d post-injection. The earliest time points showed only a few differentially expressed genes in ISAV injected fish, relative to sham injected controls, although as time progressed, many additional genes involved in key defense pathways were up-regulated including MHC type I, beta-2 microglobulin, TRIM 25 and CC chemokine 19. During the latest stage of the infection process, many genes related to oxygen transportation were under-expressed, which correlates well with the observed anemia that occurs prior to death in Atlantic salmon infected with virulent strains of ISAV.


Assuntos
Doenças dos Peixes/imunologia , Regulação da Expressão Gênica , Marcadores Genéticos/imunologia , Infecções por Orthomyxoviridae/veterinária , Salmo salar/imunologia , Animais , Perfilação da Expressão Gênica , Isavirus/imunologia , Infecções por Orthomyxoviridae/imunologia , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Salmo salar/genética
11.
J Fish Dis ; 33(3): 231-40, 2010 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-19912455

RESUMO

Finfish nodaviruses (betanodaviruses) can cause highly destructive infections in numerous species of farmed marine fish larvae and juveniles worldwide. The betanodavirus genome consists of two single-stranded positive-sense RNA molecules (RNA1 and RNA2). The virus can be classified into four genotypes based on the partial sequences of the coat protein (CP) gene (T2 and T4 regions). Currently, genomic sequence information for RNA1 regions of RNA2 outside of T2 and T4 is less well documented. This study reports on the characterization of the full RNA2 sequence of a Tunisian betanodavirus with a length of 1433 nt, containing a 339 amino acid open-reading frame encoding the CP, and typing to the redspotted grouper nervous necrosis virus Ia genotype following phylogenetic analysis. The homology of the capsid protein to other betanodaviruses or alphanodaviruses was compared. In addition, a full length RNA1 sequence of 3104 nt encoding a 982 amino acid RNA-dependent RNA polymerase was obtained.


Assuntos
Proteínas do Capsídeo/genética , Doenças dos Peixes/virologia , Nodaviridae/genética , Infecções por Vírus de RNA/veterinária , RNA Polimerase Dependente de RNA/genética , Animais , Composição de Bases , Bass/virologia , Proteínas do Capsídeo/química , Dados de Sequência Molecular , Nodaviridae/classificação , Filogenia , Infecções por Vírus de RNA/virologia , RNA Polimerase Dependente de RNA/química , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Tunísia
12.
J Fish Dis ; 32(2): 157-71, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19261043

RESUMO

Infectious salmon anaemia virus (ISAV) surveillance in the Bay of Fundy has identified the existence of a large number of genetically distinct ISAV isolates which appear to be of variable virulence. Genetically distinct isolates are currently being designated based on sequencing of the hyper polymorphic region (HPR) of genomic segment 6, which encodes the haemagglutinin-esterase protein, but it has been difficult to elucidate a clear association between these molecular variations and variations in virulence. This has hampered the establishment of proactive management decisions regarding infected fish, and ISAV infections, regardless of type, must be treated as one. Field data of ISAV infections is difficult to collect and to compare between infections because of a wide range of confounding factors including time of year, fish stock, cage site location, mitigating factors and stressors. An important tool in determining the relationship between molecular differences and virulence comes from analysis of quarantine studies. The goal of this study was to compare the virulence, by co-habitation and intraperitoneal injection, of four regionally common and recent ISAV isolates in a controlled environment. We found significant differences in mortality between ISAV molecular isolates, and present data showing that survival of ISAV infection confers significant resistance to re-infection with a different ISAV isolate. These findings, if borne out in field studies, will significantly alter the way ISAV infections are managed in the Bay of Fundy and elsewhere.


Assuntos
Doenças dos Peixes/virologia , Isavirus/patogenicidade , Infecções por Orthomyxoviridae/veterinária , Salmo salar/virologia , Animais , Doenças dos Peixes/mortalidade , Isavirus/isolamento & purificação , Infecções por Orthomyxoviridae/mortalidade , Infecções por Orthomyxoviridae/virologia , Análise de Sobrevida
13.
Dis Aquat Organ ; 80(1): 27-35, 2008 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-18714681

RESUMO

As part of a disease resistance experiment, 112 apparently healthy European flat oysters Ostrea edulis L. were exported from Canada (Nova Scotia) into France to test their susceptibility to Bonamia ostreae infection. Twelve oysters died in transit and 17 others died within 2 wk of laboratory quarantine acclimation. All oysters were examined histologically, and the 17 that died during quarantine were assayed for microcells (Bonamia sp. and Mikrocytos mackini) using molecular techniques. A microcell parasite was detected in the connective tissue of 5 of the 112 oysters. Morphological appearance, tissue affinity and molecular characterization through PCR, in situ hybridization (ISH), fluorescence in situ hybridization (FISH) and sequencing revealed a protist related to M. mackini. This is the first report of a parasite of the genus Mikrocytos in a species belonging to the genus Ostrea from the Atlantic Ocean.


Assuntos
Eucariotos/isolamento & purificação , Eucariotos/fisiologia , Ostrea/parasitologia , Quarentena , Meios de Transporte , Animais , Oceano Atlântico , Sequência de Bases , Crassostrea/parasitologia , França , Coração/parasitologia , Hibridização In Situ , Hibridização in Situ Fluorescente , Dados de Sequência Molecular , Nova Escócia , Reação em Cadeia da Polimerase , Alinhamento de Sequência
14.
J Fish Dis ; 30(4): 213-23, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17394523

RESUMO

Viral haemorrhagic septicaemia virus (VHSV) was isolated from mortalities occurring in populations of mummichog, Fundulus heteroclitus, stickleback, Gasterosteus aculeatus aculeatus, brown trout, Salmo trutta, and striped bass, Morone saxatilis, in New Brunswick and Nova Scotia, Canada. The isolated viral strains produced a cytopathic effect on the epithelioma papillosum cyprini cell line. Serum neutralization indicated the virus was VHSV and sequencing identified the rhabdovirus isolates as the North American strain of VHSV. Phylogenetic analysis indicated that the isolates are closely related and form a distinguishable subgroup of North American type VHSV. To our knowledge, this is the first report of VHSV in mummichog and striped bass.


Assuntos
Peixes/virologia , Septicemia Hemorrágica Viral/virologia , Novirhabdovirus/genética , Novirhabdovirus/isolamento & purificação , Sequência de Aminoácidos/genética , Animais , Bass/virologia , Canadá/epidemiologia , Ciprinodontiformes/virologia , Primers do DNA/química , Genes Virais/genética , Dados de Sequência Molecular , Testes de Neutralização/veterinária , Nucleoproteínas/genética , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Alinhamento de Sequência/veterinária , Smegmamorpha/virologia , Truta/virologia
15.
J Gravit Physiol ; 14(1): P137-8, 2007 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18372737

RESUMO

We have determined the gain of the cardiopulmonary baroreflex (CPBR) by measuring the central venous pressure (CVP) by venous catheter and calculating the total peripheral resistance (TPR) from mean arterial pressure (MAP) by Finometer and cardiac output (QD) by Doppler ultrasound. We tested the hypothesis that the BeatScope software of the Finometer, which calculates cardiac stroke volume (SVF) and cardiac output (QF) from the arterial pulse wave, could provide data with which to estimate CVP and TPR. The estimate of QF was linearly related to QD with a correction factor. Further, we found linear relationships between CVP and SVF that allowed us to establish a prediction of CVP from the SVF as the new input to the CPBR. To test the ability of this method to monitor changes in CPBR we are testing the subjects of the WISE-2005 study before and after 50-days of bed rest. We conclude that the TPR can be assessed with the Finometer and without any invasive method to record the CVP.


Assuntos
Barorreflexo , Repouso em Cama/efeitos adversos , Monitorização Ambulatorial da Pressão Arterial/métodos , Descondicionamento Cardiovascular , Tontura/fisiopatologia , Ultrassonografia Doppler , Pressão Sanguínea , Monitorização Ambulatorial da Pressão Arterial/instrumentação , Débito Cardíaco , Pressão Venosa Central , Tontura/diagnóstico por imagem , Tontura/etiologia , Desenho de Equipamento , Feminino , Decúbito Inclinado com Rebaixamento da Cabeça/efeitos adversos , Humanos , Pressão Negativa da Região Corporal Inferior , Masculino , Modelos Cardiovasculares , Reprodutibilidade dos Testes , Processamento de Sinais Assistido por Computador , Software , Voo Espacial , Fatores de Tempo , Resistência Vascular , Simulação de Ausência de Peso
16.
J Fish Dis ; 28(2): 101-10, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15705155

RESUMO

Reverse transcriptase polymerase chain reaction (RT-PCR), virus isolation (VI) and indirect fluorescent antibody tests (IFAT) are three assays currently used by the salmon industry to identify fish infected with infectious salmon anaemia virus (ISAV). However, no data are available on the repeatability (within-laboratory consistency) and reproducibility (between-laboratory consistency) of these assays and very limited information is available on the effect of freezing samples on test results. In order to evaluate these assays, five laboratories participated in a blinded study of 400 kidney samples representing four populations of farmed Atlantic salmon with different prevalence of ISAV. Each laboratory used its own testing protocols. Repeatability and reproducibility were evaluated using kappa as the measure of agreement. The effect of freezing was evaluated using the McNemar test. Freezing did not affect VI but improved the sensitivity of RT-PCR. The repeatability and reproducibility of VI was almost perfect. There was a substantial difference in repeatability of RT-PCR among the three laboratories with kappa ranging from 0.5 to 0.96. The repeatability for RT-PCR was generally low. The repeatability of IFAT was moderate when the results were analysed using 1+ and above as a positive result. The results of the study show the need to standardize the protocol and interpretation of RT-PCR.


Assuntos
Técnica Indireta de Fluorescência para Anticorpo/métodos , Isavirus/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Salmo salar/virologia , Animais , Aquicultura/métodos , Criopreservação/métodos , Isavirus/genética , Isavirus/imunologia , Rim/virologia , Reprodutibilidade dos Testes
17.
Dis Aquat Organ ; 62(3): 181-9, 2004 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-15672873

RESUMO

Nodaviruses (NNV) are responsible for causing disease outbreaks mainly in hatchery-reared larvae and juveniles of a wide variety of fishes throughout the world. This disease has seriously limited the culture of marine fishes over the last decade. In the Atlantic provinces of Canada, disease caused by a nodavirus was first reported in juvenile Atlantic cod being reared in Nova Scotia, in 1999. More recently, disease outbreaks caused by nodavirus have been identified in hatchery-reared Atlantic cod and haddock in Newfoundland and New Brunswick, respectively, and along the east coast of the USA. The presence of NNV in wild Atlantic cod adults and wild winter flounder has also been reported. Nodaviruses were isolated from cultured Atlantic cod and haddock, as well as from wild winter flounder from a variety of geographical localities, and their virus coat (capsid) protein genes were partially sequenced. An analysis of the data indicates that all of the nodaviruses isolated from eastern North America were closely related to one another, but that they were distinct from the European isolates already sequenced. Regardless of host species, isolates from close geographical localities were more similar than those from distant geographical areas. At the protein level, differences in coat protein sequences were seen only for strains isolated from Atlantic cod originating from Newfoundland. Our results suggest that NNV may have been present in the Atlantic off Canada and on the east coast of the USA for some time, and has evolved to form a monophyletic group, distinct from other isolates found in cold-water species. Non-lethal methods for detection of NNV are necessary to develop management strategies for this disease, and would be an asset to diagnosticians and producers. Based on the results of this study, new primers were designed and developed for an improved RT-PCR assay able to detect North Atlantic nodaviruses in ovarian fluids, eggs and other tissues. The application of this test to field samples is discussed.


Assuntos
Surtos de Doenças , Doenças dos Peixes/epidemiologia , Doenças dos Peixes/virologia , Peixes , Nodaviridae/genética , Filogenia , Infecções por Vírus de RNA/veterinária , Sequência de Aminoácidos , Animais , Aquicultura , Oceano Atlântico , Sequência de Bases , Encéfalo/virologia , Canadá/epidemiologia , Proteínas do Capsídeo/genética , Primers do DNA , Doenças dos Peixes/genética , Funções Verossimilhança , Modelos Genéticos , Dados de Sequência Molecular , Infecções por Vírus de RNA/epidemiologia , Infecções por Vírus de RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Análise de Sequência de DNA
18.
Sex Transm Dis ; 28(8): 484-91, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11473223

RESUMO

BACKGROUND: Sexually transmitted diseases (STDs) caused by HIV, herpes simplex virus (HSV), and other pathogens are spreading dramatically. The need to develop active products and vehicles to reduce this epidemic is urgent. GOAL: The efficacy of a thermoreversible gel formulation as a possible barrier to prevent the transmission of pathogens causing STDs was evaluated. STUDY DESIGN: This evaluation investigated the ability of the gel formulation to prevent infection of susceptible cells by HIV-1 and HSV-2 in vitro, the diffusion of radiolabeled herpes virus and micelles of polymer through an insertion membrane, and the electron microscopic appearance of herpes virus and gel alone or mixed together. RESULTS: The gel formulation prevents infection of susceptible cells by HIV-1 and HSV-2. It acts as an effective artificial physical barrier against the herpes virus within the first 4 hours of incubation. Herpes virus is coated by the gel or entrapped within micelles of the gel, which could hinder its attachment to target cells and inhibit its infectivity. CONCLUSION: This thermoreversible gel formulation represents an attractive matrix for the incorporation of microbicides to prevent the spread of STDs.


Assuntos
Géis/farmacologia , HIV-1/efeitos dos fármacos , Herpesvirus Humano 2/efeitos dos fármacos , Infecções Sexualmente Transmissíveis/prevenção & controle , Radioisótopos de Carbono , Linhagem Celular/efeitos dos fármacos , Infecções por HIV/prevenção & controle , Herpes Simples/prevenção & controle , Humanos , Polímeros/farmacologia
19.
J Clin Endocrinol Metab ; 85(5): 2048-56, 2000 May.
Artigo em Inglês | MEDLINE | ID: mdl-10843195

RESUMO

It has been previously shown that adrenocortical tumors (ACT) in adults exhibit structural abnormalities in tumor DNA in approximately 30% of cases. These abnormalities involve chromosome 11p15 and include loss of heterozygosity, paternal isodisomy, and overexpression of the gene for insulin-like growth factor II (IGF2), correlating with DNA demethylation at this locus. It has been hypothesized that these events occur late in the tumorigenic process in adults and seem to correlate with a worse prognosis. We present 4 pediatric cases of ACT diagnosed at 2.5 yr, 10 months, 12 yr, and 2.2 yr. All 4 patients presented with virilization, and 1 patient also showed signs and symptoms of glucocorticoid excess. The youngest patient's maternal aunt had surgical excision of a more than 15-cm ACT 18 yr previously, but the aunt is doing well at age 23 yr. They all had surgical removal of their tumors. The 2.5-yr-old child also received chemotherapy and radiotherapy because of capsular rupture and, after 3 local recurrences, died 3.3 yr after initial presentation. We investigated all 4 tumors for chromosome 11 structural abnormalities (11p15.5 to 11q23), IGF2 and H19 expression by competitive RT-PCR analysis, and IGF2 methylation patterns by Southern analysis. All 4 tumors (100%) showed a combination of structural abnormalities at the 11p15 locus with mosaic loss of heterozygosity involving 11p. All tumors also had significantly increased IGF2 messenger ribonucleic acid levels relative to normal adrenal (up to 36-fold) and significant IGF2 demethylation (mean, 87%). H19 messenger ribonucleic acid levels were undetectable in 3 of 4 tumors, explained in part by mosaic loss of the actively expressed maternal allele for this imprinted gene. By immunohistochemistry we were able to confirm increased IGF-II peptide levels within the tumor tissue in 10 pediatric patients, including the 4 patients described above. Concomitantly, we also observed nuclear accumulation of p53, suggesting somatic mutations. For the 10-month-old patient, sequencing revealed a p53 germline mutation. We therefore conclude that in pediatric ACT, structural abnormalities of tumor DNA and IGF2 overexpression as well as p53 mutations are very common and are therefore less useful for prognosis than in adults. Our findings support the theory that pediatric ACT, whose IGF2 expression and steroidogenesis evoke the phenotype of the fetal adrenal cortex, may arise because of defective apoptosis.


Assuntos
Neoplasias do Córtex Suprarrenal/genética , Cromossomos Humanos Par 11 , Regulação Neoplásica da Expressão Gênica , Fator de Crescimento Insulin-Like II/genética , Perda de Heterozigosidade , RNA não Traduzido , Neoplasias do Córtex Suprarrenal/patologia , Neoplasias do Córtex Suprarrenal/terapia , Adulto , Criança , Pré-Escolar , Mapeamento Cromossômico , Feminino , Genes Supressores de Tumor , Impressão Genômica , Haplótipos , Humanos , Lactente , Masculino , Proteínas Musculares/genética , Núcleo Familiar , RNA Longo não Codificante , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Virilismo/etiologia , Virilismo/genética
20.
J Clin Endocrinol Metab ; 84(7): 2502-6, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10404827

RESUMO

Permanent primary congenital hypothyroidism (CH) can be caused by abnormal thyroid differentiation (athyreosis), migration (ectopy), or function (leading to goiter). Goiters follow an autosomal recessive pattern of inheritance, whereas ectopy and athyreosis are considered as a single sporadic entity with a female preponderance. On the other hand, a high prevalence of extrathyroidal malformations has been reported in CH, but without linking specific defects to specific types of CH. On the basis of TSH screening, 273 newborns were referred to an academic pediatric endocrinology clinic in the province of Quebec between 1988 and 1997. Of 230 patients with permanent primary CH who had scintigraphy at diagnosis, 141 had ectopy (104 girls), 36 had athyreosis (21 girls), 42 had goiter (18 girls), 10 (3 girls) had a normal scan, and 1 girl had hemiagenesis. Only in the ectopies was the proportion of girls significantly higher than 0.5 (P<0.001). Isolated cardiac malformations were observed in 7 patients (3.0%), a prevalence 5-fold higher than that in the general population; this was largely due to atrial and ventricular septal defects, which were only observed in ectopy and athyreosis. Our data suggest that the molecular mechanisms that lead to complete absence of thyroid differentiation or defective thyroid migration 1) may be similar, but are modulated by the genetic makeup of the embryo and/or the hormonal milieu of the fetus; and 2) may also be involved in septation of the embryonic heart.


Assuntos
Hipotireoidismo Congênito , Hipotireoidismo/genética , Proteínas Nucleares , Caracteres Sexuais , Glândula Tireoide/anormalidades , Anormalidades Múltiplas , Fatores de Transcrição Hélice-Alça-Hélice Básicos , Proteínas de Ligação a DNA/genética , Feminino , Fatores de Transcrição Forkhead , Proteínas de Homeodomínio/genética , Humanos , Hipotireoidismo/etiologia , Recém-Nascido , Masculino , Mutação , Fator de Transcrição PAX8 , Fatores de Transcrição Box Pareados , Cintilografia , Proteínas Repressoras/genética , Glândula Tireoide/diagnóstico por imagem , Hormônios Tireóideos/sangue , Transativadores/genética , Fatores de Transcrição HES-1
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