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1.
Nucleic Acids Res ; 51(D1): D942-D949, 2023 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-36420896

RESUMO

GENCODE produces high quality gene and transcript annotation for the human and mouse genomes. All GENCODE annotation is supported by experimental data and serves as a reference for genome biology and clinical genomics. The GENCODE consortium generates targeted experimental data, develops bioinformatic tools and carries out analyses that, along with externally produced data and methods, support the identification and annotation of transcript structures and the determination of their function. Here, we present an update on the annotation of human and mouse genes, including developments in the tools, data, analyses and major collaborations which underpin this progress. For example, we report the creation of a set of non-canonical ORFs identified in GENCODE transcripts, the LRGASP collaboration to assess the use of long transcriptomic data to build transcript models, the progress in collaborations with RefSeq and UniProt to increase convergence in the annotation of human and mouse protein-coding genes, the propagation of GENCODE across the human pan-genome and the development of new tools to support annotation of regulatory features by GENCODE. Our annotation is accessible via Ensembl, the UCSC Genome Browser and https://www.gencodegenes.org.


Assuntos
Biologia Computacional , Genoma Humano , Humanos , Animais , Camundongos , Anotação de Sequência Molecular , Biologia Computacional/métodos , Genoma Humano/genética , Transcriptoma/genética , Perfilação da Expressão Gênica , Bases de Dados Genéticas
2.
Nucleic Acids Res ; 49(D1): D916-D923, 2021 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-33270111

RESUMO

The GENCODE project annotates human and mouse genes and transcripts supported by experimental data with high accuracy, providing a foundational resource that supports genome biology and clinical genomics. GENCODE annotation processes make use of primary data and bioinformatic tools and analysis generated both within the consortium and externally to support the creation of transcript structures and the determination of their function. Here, we present improvements to our annotation infrastructure, bioinformatics tools, and analysis, and the advances they support in the annotation of the human and mouse genomes including: the completion of first pass manual annotation for the mouse reference genome; targeted improvements to the annotation of genes associated with SARS-CoV-2 infection; collaborative projects to achieve convergence across reference annotation databases for the annotation of human and mouse protein-coding genes; and the first GENCODE manually supervised automated annotation of lncRNAs. Our annotation is accessible via Ensembl, the UCSC Genome Browser and https://www.gencodegenes.org.


Assuntos
COVID-19/prevenção & controle , Biologia Computacional/métodos , Bases de Dados Genéticas , Genômica/métodos , Anotação de Sequência Molecular/métodos , SARS-CoV-2/genética , Animais , COVID-19/epidemiologia , COVID-19/virologia , Epidemias , Humanos , Internet , Camundongos , Pseudogenes/genética , RNA Longo não Codificante/genética , SARS-CoV-2/metabolismo , SARS-CoV-2/fisiologia , Transcrição Gênica/genética
3.
Genome Res ; 29(12): 2073-2087, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31537640

RESUMO

The most widely appreciated role of DNA is to encode protein, yet the exact portion of the human genome that is translated remains to be ascertained. We previously developed PhyloCSF, a widely used tool to identify evolutionary signatures of protein-coding regions using multispecies genome alignments. Here, we present the first whole-genome PhyloCSF prediction tracks for human, mouse, chicken, fly, worm, and mosquito. We develop a workflow that uses machine learning to predict novel conserved protein-coding regions and efficiently guide their manual curation. We analyze more than 1000 high-scoring human PhyloCSF regions and confidently add 144 conserved protein-coding genes to the GENCODE gene set, as well as additional coding regions within 236 previously annotated protein-coding genes, and 169 pseudogenes, most of them disabled after primates diverged. The majority of these represent new discoveries, including 70 previously undetected protein-coding genes. The novel coding genes are additionally supported by single-nucleotide variant evidence indicative of continued purifying selection in the human lineage, coding-exon splicing evidence from new GENCODE transcripts using next-generation transcriptomic data sets, and mass spectrometry evidence of translation for several new genes. Our discoveries required simultaneous comparative annotation of other vertebrate genomes, which we show is essential to remove spurious ORFs and to distinguish coding from pseudogene regions. Our new coding regions help elucidate disease-associated regions by revealing that 118 GWAS variants previously thought to be noncoding are in fact protein altering. Altogether, our PhyloCSF data sets and algorithms will help researchers seeking to interpret these genomes, while our new annotations present exciting loci for further experimental characterization.


Assuntos
Éxons , Genoma Humano , Estudo de Associação Genômica Ampla , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta , Análise de Sequência de DNA , Animais , Humanos , Pseudogenes
4.
Proteomics ; 20(21-22): e2000009, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32937025

RESUMO

Mass spectrometry (MS)-based quantitative proteomics experiments typically assay a subset of up to 60% of the ≈20 000 human protein coding genes. Computational methods for imputing the missing values using RNA expression data usually allow only for imputations of proteins measured in at least some of the samples. In silico methods for comprehensively estimating abundances across all proteins are still missing. Here, a novel method is proposed using deep learning to extrapolate the observed protein expression values in label-free MS experiments to all proteins, leveraging gene functional annotations and RNA measurements as key predictive attributes. This method is tested on four datasets, including human cell lines and human and mouse tissues. This method predicts the protein expression values with average R2 scores between 0.46 and 0.54, which is significantly better than predictions based on correlations using the RNA expression data alone. Moreover, it is demonstrated that the derived models can be "transferred" across experiments and species. For instance, the model derived from human tissues gave a R2=0.51 when applied to mouse tissue data. It is concluded that protein abundances generated in label-free MS experiments can be computationally predicted using functional annotated attributes and can be used to highlight aberrant protein abundance values.


Assuntos
Aprendizado Profundo , Animais , Espectrometria de Massas , Camundongos , Anotação de Sequência Molecular , Proteínas , Proteômica
5.
PLoS Pathog ; 14(10): e1007406, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30365535

RESUMO

Infection with Citrobacter rodentium triggers robust tissue damage repair responses, manifested by secretion of IL-22, in the absence of which mice succumbed to the infection. Of the main hallmarks of C. rodentium infection are colonic crypt hyperplasia (CCH) and dysbiosis. In order to colonize the host and compete with the gut microbiota, C. rodentium employs a type III secretion system (T3SS) that injects effectors into colonic intestinal epithelial cells (IECs). Once injected, the effectors subvert processes involved in innate immune responses, cellular metabolism and oxygenation of the mucosa. Importantly, the identity of the effector/s triggering the tissue repair response is/are unknown. Here we report that the effector EspO ,an orthologue of OspE found in Shigella spp, affects proliferation of IECs 8 and 14 days post C. rodentium infection as well as secretion of IL-22 from colonic explants. While we observed no differences in the recruitment of group 3 innate lymphoid cells (ILC3s) and T cells, which are the main sources of IL-22 at the early and late stages of C. rodentium infection respectively, infection with ΔespO was characterized by diminished recruitment of sub-mucosal neutrophils, which coincided with lower abundance of Mmp9 and chemokines (e.g. S100a8/9) in IECs. Moreover, mice infected with ΔespO triggered significantly lesser nutritional immunity (e.g. calprotectin, Lcn2) and expression of antimicrobial peptides (Reg3ß, Reg3γ) compared to mice infected with WT C. rodentium. This overlapped with a decrease in STAT3 phosphorylation in IECs. Importantly, while the reduced CCH and abundance of antimicrobial proteins during ΔespO infection did not affect C. rodentium colonization or the composition of commensal Proteobacteria, they had a subtle consequence on Firmicutes subpopulations. EspO is the first bacterial virulence factor that affects neutrophil recruitment and secretion of IL-22, as well as expression of antimicrobial and nutritional immunity proteins in IECs.


Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Citrobacter rodentium/metabolismo , Infecções por Enterobacteriaceae/imunologia , Imunidade Inata/imunologia , Mucosa Intestinal/imunologia , Sistemas de Secreção Tipo III/metabolismo , Animais , Infecções por Enterobacteriaceae/metabolismo , Infecções por Enterobacteriaceae/microbiologia , Feminino , Mucosa Intestinal/lesões , Mucosa Intestinal/microbiologia , Camundongos , Camundongos Endogâmicos C57BL
6.
BMC Genet ; 21(1): 25, 2020 03 06.
Artigo em Inglês | MEDLINE | ID: mdl-32138667

RESUMO

BACKGROUND: POLG, located on nuclear chromosome 15, encodes the DNA polymerase γ(Pol γ). Pol γ is responsible for the replication and repair of mitochondrial DNA (mtDNA). Pol γ is the only DNA polymerase found in mitochondria for most animal cells. Mutations in POLG are the most common single-gene cause of diseases of mitochondria and have been mapped over the coding region of the POLG ORF. RESULTS: Using PhyloCSF to survey alternative reading frames, we found a conserved coding signature in an alternative frame in exons 2 and 3 of POLG, herein referred to as ORF-Y that arose de novo in placental mammals. Using the synplot2 program, synonymous site conservation was found among mammals in the region of the POLG ORF that is overlapped by ORF-Y. Ribosome profiling data revealed that ORF-Y is translated and that initiation likely occurs at a CUG codon. Inspection of an alignment of mammalian sequences containing ORF-Y revealed that the CUG codon has a strong initiation context and that a well-conserved predicted RNA stem-loop begins 14 nucleotides downstream. Such features are associated with enhanced initiation at near-cognate non-AUG codons. Reanalysis of the Kim et al. (2014) draft human proteome dataset yielded two unique peptides that map unambiguously to ORF-Y. An additional conserved uORF, herein referred to as ORF-Z, was also found in exon 2 of POLG. Lastly, we surveyed Clinvar variants that are synonymous with respect to the POLG ORF and found that most of these variants cause amino acid changes in ORF-Y or ORF-Z. CONCLUSIONS: We provide evidence for a novel coding sequence, ORF-Y, that overlaps the POLG ORF. Ribosome profiling and mass spectrometry data show that ORF-Y is expressed. PhyloCSF and synplot2 analysis show that ORF-Y is subject to strong purifying selection. An abundance of disease-correlated mutations that map to exons 2 and 3 of POLG but also affect ORF-Y provides potential clinical significance to this finding.


Assuntos
Códon de Iniciação/genética , DNA Polimerase gama/genética , Mitocôndrias/genética , Ribossomos/genética , Sequência de Aminoácidos , DNA Mitocondrial/genética , Éxons/genética , Humanos , Conformação de Ácido Nucleico , Fases de Leitura Aberta/genética
7.
J Proteome Res ; 15(12): 4686-4695, 2016 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-27786492

RESUMO

Proteogenomics leverages information derived from proteomic data to improve genome annotations. Of particular interest are "novel" peptides that provide direct evidence of protein expression for genomic regions not previously annotated as protein-coding. We present a modular, automated data analysis pipeline aimed at detecting such "novel" peptides in proteomic data sets. This pipeline implements criteria developed by proteomics and genome annotation experts for high-stringency peptide identification and filtering. Our pipeline is based on the OpenMS computational framework; it incorporates multiple database search engines for peptide identification and applies a machine-learning approach (Percolator) to post-process search results. We describe several new and improved software tools that we developed to facilitate proteogenomic analyses that enhance the wealth of tools provided by OpenMS. We demonstrate the application of our pipeline to a human testis tissue data set previously acquired for the Chromosome-Centric Human Proteome Project, which led to the addition of five new gene annotations on the human reference genome.


Assuntos
Mineração de Dados/métodos , Anotação de Sequência Molecular , Proteogenômica/métodos , Genoma Humano , Humanos , Aprendizado de Máquina , Masculino , Proteômica/métodos , Ferramenta de Busca , Software , Testículo
8.
Proteomics ; 14(9): 1011-4, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24504981

RESUMO

Protein identification by MS/MS is an important technique in proteome studies. The Open Mass Spectrometry Search Algorithm (OMSSA) is an open-source search engine that can be used to identify MS/MS spectra acquired in these experiments. Here, we present a software tool, termed OMSSAPercolator, which interfaces OMSSA with Percolator, a post-search machine learning method for rescoring database search results. We demonstrate that it outperforms the standard OMSSA scoring scheme, and provides reliable significant measurements. OMSSAPercolator is programmed using JAVA and can be readily used as a standalone tool or integrated into existing data analysis pipelines. OMSSAPercolator is freely available and can be downloaded at http://sourceforge.net/projects/omssapercolator/.


Assuntos
Algoritmos , Proteoma/análise , Proteômica/métodos , Software , Mapeamento de Peptídeos , Proteínas/análise , Proteínas/química , Proteoma/química , Reprodutibilidade dos Testes , Análise de Sequência de Proteína
9.
Mol Cell Proteomics ; 11(8): 478-91, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22493177

RESUMO

Peptide identification using tandem mass spectrometry is a core technology in proteomics. Latest generations of mass spectrometry instruments enable the use of electron transfer dissociation (ETD) to complement collision induced dissociation (CID) for peptide fragmentation. However, a critical limitation to the use of ETD has been optimal database search software. Percolator is a post-search algorithm, which uses semi-supervised machine learning to improve the rate of peptide spectrum identifications (PSMs) together with providing reliable significance measures. We have previously interfaced the Mascot search engine with Percolator and demonstrated sensitivity and specificity benefits with CID data. Here, we report recent developments in the Mascot Percolator V2.0 software including an improved feature calculator and support for a wider range of ion series. The updated software is applied to the analysis of several CID and ETD fragmented peptide data sets. This version of Mascot Percolator increases the number of CID PSMs by up to 80% and ETD PSMs by up to 60% at a 0.01 q-value (1% false discovery rate) threshold over a standard Mascot search, notably recovering PSMs from high charge state precursor ions. The greatly increased number of PSMs and peptide coverage afforded by Mascot Percolator has enabled a fuller assessment of CID/ETD complementarity to be performed. Using a data set of CID and ETcaD spectral pairs, we find that at a 1% false discovery rate, the overlap in peptide identifications by CID and ETD is 83%, which is significantly higher than that obtained using either stand-alone Mascot (69%) or OMSSA (39%). We conclude that Mascot Percolator is a highly sensitive and accurate post-search algorithm for peptide identification and allows direct comparison of peptide identifications using multiple alternative fragmentation techniques.


Assuntos
Algoritmos , Peptídeos/análise , Proteômica/métodos , Software , Espectrometria de Massas em Tandem/métodos , Inteligência Artificial , Cromatografia Líquida , Bases de Dados de Proteínas , Escherichia coli/metabolismo , Proteínas de Escherichia coli/análise , Proteínas Fúngicas/análise , Humanos , Reprodutibilidade dos Testes , Leveduras/metabolismo
10.
Mol Cell Proteomics ; 11(12): 1682-9, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22949509

RESUMO

The original PRIDE Converter tool greatly simplified the process of submitting mass spectrometry (MS)-based proteomics data to the PRIDE database. However, after much user feedback, it was noted that the tool had some limitations and could not handle several user requirements that were now becoming commonplace. This prompted us to design and implement a whole new suite of tools that would build on the successes of the original PRIDE Converter and allow users to generate submission-ready, well-annotated PRIDE XML files. The PRIDE Converter 2 tool suite allows users to convert search result files into PRIDE XML (the format needed for performing submissions to the PRIDE database), generate mzTab skeleton files that can be used as a basis to submit quantitative and gel-based MS data, and post-process PRIDE XML files by filtering out contaminants and empty spectra, or by merging several PRIDE XML files together. All the tools have both a graphical user interface that provides a dialog-based, user-friendly way to convert and prepare files for submission, as well as a command-line interface that can be used to integrate the tools into existing or novel pipelines, for batch processing and power users. The PRIDE Converter 2 tool suite will thus become a cornerstone in the submission process to PRIDE and, by extension, to the ProteomeXchange consortium of MS-proteomics data repositories.


Assuntos
Bases de Dados de Proteínas , Processamento Eletrônico de Dados , Espectrometria de Massas , Proteômica , Proteoma/análise , Software , Design de Software , Interface Usuário-Computador
11.
Can Vet J ; 55(1): 1255-7, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24381348

RESUMO

This study describes the effect of enteric biopsy closure orientation on circumference and volume of saline needed for leak testing. There were significant differences in circumference measurements at baseline, central circumference of longitudinally closed sites, and volume of saline for leak testing.


Effet de l'orientation de la fermeture de la biopsie entérique sur la circonférence entérique et le volume de solution saline requis pour l'essai d'étanchéité. Cette étude décrit l'effet de l'orientation de la fermeture de la biopsie entérique sur la circonférence et le volume de solution saline requis pour l'essai d'étanchéité. Il y avait des différences importantes dans les mesures de la circonférence pour les données de référence, la circonférence centrale des sites fermés longitudinalement et le volume de solution saline pour l'essai d'étanchéité.(Traduit par Isabelle Vallières).


Assuntos
Cães/cirurgia , Enteropatias/veterinária , Cloreto de Sódio/administração & dosagem , Técnicas de Fechamento de Ferimentos/veterinária , Animais , Biópsia/veterinária , Técnicas de Diagnóstico do Sistema Digestório/veterinária , Enteropatias/patologia
12.
J Vet Med Educ ; 40(3): 303-9, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23975075

RESUMO

In 2008, the US experienced a disruption in human rabies vaccine supplies, leading public health authorities to prioritize vaccine release for post-exposure prophylaxis (PEP) and limit vaccine supplies for pre-exposure prophylaxis (PreEP) in high-risk groups. In 2008, the Association of American Veterinary Medical Colleges (AAVMC) surveyed its member institutions on rabies vaccination policies and practices. Senior administrators at Colleges of Veterinary Medicine (CVMs) and departments of veterinary science and comparative medicine were asked to identify the person most knowledgeable about their institution's student rabies vaccination program. Respondents were asked to describe their policies and procedures for administering PreEP to veterinary medical students and staff and to estimate the annual demand for student and staff PreEP vaccine. Twenty-one CVMs responded. Twenty (95%) reported requiring PreEP of veterinary medical students and 16 (80%) of those 20 required vaccination upon matriculation. An estimated 7,309 doses of vaccine were required for PreEP of an estimated 2,436 first-year US veterinary medical students. Seventy-two percent of respondents administered PreEP in August, September, and October, coinciding with the highest public demand for PEP. CVMs should consider altering the timing of rabies vaccine administration to veterinary medical students and staff to other months, thereby helping to ensure that PEP rabies vaccine will be available to people with validated rabies exposures and to ensure that supplies will be available for PreEP of students and staff. AAVMC may wish to identify and support a point of coordination to facilitate the purchase and distribution of human rabies vaccine among its US member CVMs.


Assuntos
Antibioticoprofilaxia , Vacina Antirrábica/uso terapêutico , Raiva/prevenção & controle , Faculdades de Medicina Veterinária , Antibioticoprofilaxia/estatística & dados numéricos , Docentes , Política de Saúde , Humanos , Raiva/imunologia , Vacina Antirrábica/administração & dosagem , Estudantes , Inquéritos e Questionários , Estados Unidos
13.
Front Mol Biosci ; 9: 1062031, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36523653

RESUMO

Multi-omics approaches including proteomics analyses are becoming an integral component of precision medicine. As clinical proteomics studies gain momentum and their sensitivity increases, research on identifying individuals based on their proteomics data is here examined for risks and ethics-related issues. A great deal of work has already been done on this topic for DNA/RNA sequencing data, but it has yet to be widely studied in other omics fields. The current state-of-the-art for the identification of individuals based solely on proteomics data is explained. Protein sequence variation analysis approaches are covered in more detail, including the available analysis workflows and their limitations. We also outline some previous forensic and omics proteomics studies that are relevant for the identification of individuals. Following that, we discuss the risks of patient reidentification using other proteomics data types such as protein expression abundance and post-translational modification (PTM) profiles. In light of the potential identification of individuals through proteomics data, possible legal and ethical implications are becoming increasingly important in the field.

14.
Am J Vet Res ; 71(9): 997-1002, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20807137

RESUMO

OBJECTIVE: To establish a protocol to collect temporal-spatial gait analysis variables by use of a portable walkway system in Labrador Retrievers at a walk and to determine reference values. ANIMALS: 56 healthy Labrador Retrievers. PROCEDURES: 6 passes across the walkway (3 passes in each direction) were recorded. Inclusion criteria for a pass were that the dog was at a walk (velocity, 60.0 to 90.0 cm/s) and had minimal head turning. The first 3 passes that met the inclusion criteria were analyzed for each dog. RESULTS: Mean stride length was 88.4 cm. Mean stance time (ST) of forelimbs and hind limbs was 0.62 and 0.56 seconds, respectively. Mean stance time percentage (ST%; proportion of stance time to total gait cycle time) for forelimbs and hind limbs was 55.6% and 50.2%, respectively. Mean total pressure index (TPI) of forelimbs and hind limbs was 27.1 and 17.4, respectively. Mean number of sensors (NS) activated by each paw strike of forelimbs and hind limbs was 17 and 13, respectively. Mean forelimb-to-hind limb symmetry ratios were 1.11 (ST), 1.10 (ST%), 1.62 (TPI), and 1.37 (NS). Symmetry ratios for left limbs to right limbs, left forelimb to right forelimb, and left hind limb to right hind limb were 1.00. CONCLUSIONS AND CLINICAL RELEVANCE: A protocol for collection of temporal-spatial gait analysis variables with a portable walkway system in Labrador Retrievers at a walk was developed, and reference values for variables and symmetry ratios were reported. Further research will determine the extent to which symmetry ratios differ in dogs with orthopedic disorders.


Assuntos
Cães/fisiologia , Marcha/fisiologia , Caminhada/fisiologia , Animais , Fenômenos Biomecânicos , Feminino , Membro Anterior/fisiologia , Membro Posterior/fisiologia , Locomoção/fisiologia , Masculino , Orquiectomia , Ovariectomia , Postura , Valores de Referência
15.
J Environ Health ; 72(7): 14-8; quiz 32, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20235404

RESUMO

Animal hoarding is an under-recognized problem that exists in most communities and adversely impacts the health, welfare, and safety of humans, animals, and the environment. These guidelines address public health and worker safety concerns in handling situations where animal hoarding or other dense concentrations of animals have caused unhealthy and unsafe conditions. Because animal hoarding situations are often complex, a full response is likely to be prolonged and require a cross-jurisdictional multiagency effort. Each animal hoarding case has unique circumstances related to the types and numbers of animals involved, the physical structure(s) where they are being kept, and the health status of the animals, among other factors that must be taken into account in planning a response. Some general public health considerations and associated recommendations for personal protective equipment use are presented that apply to all cases, however.


Assuntos
Poluição do Ar em Ambientes Fechados , Comportamento Compulsivo/epidemiologia , Exposição Ambiental/legislação & jurisprudência , Transtornos Mentais/epidemiologia , Saúde Pública/legislação & jurisprudência , Zoonoses/epidemiologia , Bem-Estar do Animal , Animais , Exposição Ambiental/efeitos adversos , Vínculo Humano-Animal , Humanos , Estados Unidos/epidemiologia , Zoonoses/transmissão
16.
Front Vet Sci ; 7: 579967, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33195592

RESUMO

There are several non-steroidal intra-articular therapeutics (NSIATs) available for use by equine practitioners for the treatment of performance-limiting joint-related pathology. Information is limited on perceived clinical efficacy, recommended treatment protocols, and associated complications. Our objective with this cross-sectional survey was to investigate the current clinical usage of NSIATs by equine practitioners. An electronic cross-sectional convenience survey inquiring about the use of steroidal and NSIATS (platelet-rich plasma, autologous conditioned serum, autologous protein solution, cellular therapies, and polyacrylamide hydrogel) was distributed internationally to equine practitioners. A total of 353 surveys were completed. NSIATs were used by 87.5% of the participants. Corticosteroids and hyaluronic acid remain the intra-articular therapeutic of choice among practitioners, followed by autologous conditioned serum, platelet-rich plasma and autologous conditioned protein. Polyacrylamide hydrogel was the least used. Practitioners were more likely to use NSIATs if their caseload was > 50% equine (P < 0.001), they treated more than 10 horses intra-articularly per month (P < 0.001), and horses treated were considered English sport horses (P = 0.02). Years in practice and practice location did not influence the use of NSIATs. One of the most common reasons why NSIATs were chosen was to treat acute articular pathologies. As survey limitations, answers to questions regarding clinical response and complication rates were based on subjective estimation and practitioners recall, not clinical records. In conclusion, corticosteroids remain the most widely used intra-articular therapeutic. Among the NSIATs, blood-based products are more commonly used by practitioners, followed by cellular and synthetic products. Equine practitioners frequently use NSIATs, choosing to treat acute joint pathology more than previously reported.

17.
J Feline Med Surg ; 22(8): 685-695, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-31566070

RESUMO

OBJECTIVES: Phage-gonadotropin-releasing hormone (GnRH) constructs with potential contraceptive properties were generated in our previous study via selection from a phage display library using neutralizing GnRH antibodies as selection targets. In mice, these constructs invoked the production of antibodies against GnRH and suppressed serum testosterone. The goal of this study was to evaluate this vaccine against GnRH for its potential to suppress reproductive characteristics in cats. METHODS: Sexually mature male cats were injected with a phage-GnRH vaccine using the following treatment groups: (1) single phage-GnRH vaccine with adjuvant; (2) phage-GnRH vaccine without adjuvant and half-dose booster 1 month later; or (3) phage-GnRH vaccine with adjuvant and two half-dose boosters with adjuvant 3 and 6 months later. Anti-GnRH antibodies and serum testosterone, testicular volume and sperm characteristics were evaluated monthly for 7-9 months. RESULTS: All cats developed anti-GnRH antibodies following immunization. Serum antibody titers increased significantly after booster immunizations. In group 3, serum testosterone was suppressed 8 months after primary immunization. Total testicular volume decreased in group 1 by 24-42% and in group 3 by 15-36% at 7 months after immunization, indicating potential gonadal atrophy. Vacuolation of epididymides was observed histologically. Although all cats produced sperm at the conclusion of the study, normal morphology was decreased as much as 38%. Phage alone produced no local or systemic reactions. Immunization of phage with AdjuVac produced unacceptable injection site reactions. CONCLUSIONS AND RELEVANCE: Our phage-based vaccine against GnRH demonstrated a potential for fertility impairment in cats. Future research is required to optimize vaccine regimens and identify animal age groups most responsive to the vaccine. If permanent contraception (highly desirable in feral and shelter cats) cannot be achieved, the vaccine has a potential use in zoo animals or pets where multiple administrations are more practical and/or reversible infertility is desirable.


Assuntos
Bacteriófagos , Gatos , Anticoncepção/veterinária , Hormônio Liberador de Gonadotropina/administração & dosagem , Vacinação/veterinária , Vacinas Anticoncepcionais/administração & dosagem , Animais , Bacteriófagos/imunologia , Anticoncepção/métodos , Fertilidade , Masculino
18.
BMC Genomics ; 10: 61, 2009 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-19193216

RESUMO

BACKGROUND: Proteomic data is a potentially rich, but arguably unexploited, data source for genome annotation. Peptide identifications from tandem mass spectrometry provide prima facie evidence for gene predictions and can discriminate over a set of candidate gene models. Here we apply this to the recently sequenced Aspergillus niger fungal genome from the Joint Genome Institutes (JGI) and another predicted protein set from another A.niger sequence. Tandem mass spectra (MS/MS) were acquired from 1d gel electrophoresis bands and searched against all available gene models using Average Peptide Scoring (APS) and reverse database searching to produce confident identifications at an acceptable false discovery rate (FDR). RESULTS: 405 identified peptide sequences were mapped to 214 different A.niger genomic loci to which 4093 predicted gene models clustered, 2872 of which contained the mapped peptides. Interestingly, 13 (6%) of these loci either had no preferred predicted gene model or the genome annotators' chosen "best" model for that genomic locus was not found to be the most parsimonious match to the identified peptides. The peptides identified also boosted confidence in predicted gene structures spanning 54 introns from different gene models. CONCLUSION: This work highlights the potential of integrating experimental proteomics data into genomic annotation pipelines much as expressed sequence tag (EST) data has been. A comparison of the published genome from another strain of A.niger sequenced by DSM showed that a number of the gene models or proteins with proteomics evidence did not occur in both genomes, further highlighting the utility of the method.


Assuntos
Aspergillus niger/genética , Genoma Fúngico , Modelos Genéticos , Proteômica/métodos , Sequência de Aminoácidos , Análise por Conglomerados , Bases de Dados de Proteínas , Dados de Sequência Molecular , Alinhamento de Sequência , Espectrometria de Massas em Tandem
19.
Am J Vet Res ; 69(10): 1262-7, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18828680

RESUMO

OBJECTIVE: To determine whether Labrador Retrievers participating in field trials develop respiratory alkalosis and hypocapnia primarily in conditions of high ambient temperatures. ANIMALS: 16 Labrador Retrievers. PROCEDURES: At each of 5 field trials, 5 to 10 dogs were monitored during a test (retrieval of birds over a variable distance on land [1,076 to 2,200 m]; 36 assessments); ambient temperatures ranged from 2.2 degrees to 29.4 degrees C. For each dog, rectal temperature was measured and a venous blood sample was collected in a heparinized syringe within 5 minutes of test completion. Blood samples were analyzed on site for Hct; pH; sodium, potassium, ionized calcium, glucose, lactate, bicarbonate, and total CO2 concentrations; and values of PvO2 and PvCO2. Scatterplots of each variable versus ambient temperature were reviewed. Regression analysis was used to evaluate the effect of ambient temperature (< or = 21 degrees C and > 21 degrees C) on each variable. RESULTS: Compared with findings at ambient temperatures < or = 21 degrees C, venous blood pH was increased (mean, 7.521 vs 7.349) and PvCO2 was decreased (mean, 17.8 vs 29.3 mm Hg) at temperatures > 21 degrees C; rectal temperature did not differ. Two dogs developed signs of heat stress in 1 test at an ambient temperature of 29 degrees C; their rectal temperatures were higher and PvCO2 values were lower than findings in other dogs. CONCLUSIONS AND CLINICAL RELEVANCE: When running distances frequently encountered at field trials, healthy Labrador Retrievers developed hyperthermia regardless of ambient temperature. Dogs developed respiratory alkalosis and hypocapnia at ambient temperatures > 21 degrees C.


Assuntos
Alcalose Respiratória/veterinária , Doenças do Cão/fisiopatologia , Transtornos de Estresse por Calor/veterinária , Hipocapnia/veterinária , Esforço Físico/fisiologia , Alcalose Respiratória/fisiopatologia , Animais , Análise Química do Sangue , Glicemia/metabolismo , Temperatura Corporal , Cães , Transtornos de Estresse por Calor/fisiopatologia , Hematócrito , Temperatura Alta , Hipocapnia/fisiopatologia
20.
J Vet Med Educ ; 35(2): 231-4, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18723809

RESUMO

To meet long-term needs, many veterinary colleges and schools are participating in dual-degree DVM/MPH programs. Auburn University's College of Veterinary Medicine and the School of Public Health at the University of Alabama at Birmingham have developed a coordinated-degree curriculum in which the DVM and the MPH are not necessarily awarded simultaneously. Other opportunities at Auburn include Public Health Careers Day, trips to the Centers for Disease Control and Prevention, several elective courses related to veterinary epidemiology, and online access to the Emerging and Exotic Diseases of Animals course available from the Veterinary Information Network. We have been able to increase our students' exposure to the role of the veterinarian in public health and to develop a program to augment their training in public practice.


Assuntos
Currículo , Educação Profissional em Saúde Pública , Educação em Veterinária , Alabama , Doenças Transmissíveis Emergentes , Epidemiologia , Humanos , Internet , Faculdades de Saúde Pública , Faculdades de Medicina Veterinária , Universidades
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