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1.
J Ment Health ; 30(4): 526-540, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32186223

RESUMO

BACKGROUND: Electroconvulsive therapy (ECT) is a controversial treatment. Research has predominantly focused on clinician assessment of short-term efficacy and, occasionally, on participant experiences of the treatment itself. While service user accounts of the long-term impacts of ECT are reported, they are dispersed throughout the literature and typically tangential to studie's main foci. AIM: The aim of this study was to synthesise service-user accounts, within peer-reviewed literature, of long-term impacts of ECT in their daily lives. METHODS: A qualitative meta-synthesis was conducted. A systematic literature search identified qualitative articles meeting the inclusion criteria. Results sections of eligible papers were analysed thematically. RESULTS: From 16 eligible papers, the review identified 11 long-term impacts, four social influences and five strategies that people employed to navigate these long-term impacts. CONCLUSION: Limited research has examined long-term experiences of ECT from service-user perspectives. These lived experience perspectives are required to facilitate peer-to-peer learning and assist future service delivery to align with needs of people living with long-term ECT impacts.


Assuntos
Eletroconvulsoterapia , Humanos , Pesquisa Qualitativa
2.
Transgenic Res ; 29(4): 395-407, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32607872

RESUMO

Advances in genome editing tools have reduced barriers to the creation of animal models. Due to their anatomical and physiological similarities to humans, there has been a growing need for pig models to study human diseases, for xenotransplantation and translational research. The ability to determine the sex of genetically modified embryos, cells or fetuses is beneficial for every project involving the production of transgenic animals. This strategy can improve the time-efficiency and lower the production costs. Additionally, sex assessment is very useful for wildlife studies to understand population behavior and structure. Thus, we developed a simple and fast PCR-based protocol for sex determination in pigs by using a unique primer set to amplify either the DDX3X or DDX3Y gene. The sex was 100% correctly assigned when tail genomic DNA, Day-35 fetus and hair samples from pigs were used. For both blastocysts and oocytes (84.6% and 96.5% of efficacy, respectively) the unidentified samples were potentially due to a limitation in sample size. Our assay also worked for domestic sheep (Ovis aries), American bison (Bison bison) and European cattle (Bos taurus) samples and by in silico analysis we confirmed X-Y amplicon length polymorphisms for the DDX3 gene in 12 other mammalian species. This PCR protocol for determining sex in pig tissues and cells showed to be simple, specific, highly reproducible and less time consuming as well as an important tool for other livestock species and wildlife studies.


Assuntos
RNA Helicases DEAD-box/genética , Genes Ligados ao Cromossomo X , Genes Ligados ao Cromossomo Y , Variação Genética , Análise de Sequência de DNA/métodos , Análise para Determinação do Sexo/métodos , Animais , Bison , Bovinos , Feminino , Masculino , Reação em Cadeia da Polimerase , Carneiro Doméstico , Suínos
3.
Environ Sci Technol ; 50(11): 5631-40, 2016 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-27149080

RESUMO

Formic acid (HCOOH) is one of the most abundant carboxylic acids and a dominant source of atmospheric acidity. Recent work indicates a major gap in the HCOOH budget, with atmospheric concentrations much larger than expected from known sources. Here, we employ recent space-based observations from the Tropospheric Emission Spectrometer with the GEOS-Chem atmospheric model to better quantify the HCOOH source from biomass burning, and assess whether fire emissions can help close the large budget gap for this species. The space-based data reveal a severe model HCOOH underestimate most prominent over tropical burning regions, suggesting a major missing source of organic acids from fires. We develop an approach for inferring the fractional fire contribution to ambient HCOOH and find, based on measurements over Africa, that pyrogenic HCOOH:CO enhancement ratios are much higher than expected from direct emissions alone, revealing substantial secondary organic acid production in fire plumes. Current models strongly underestimate (by 10 ± 5 times) the total primary and secondary HCOOH source from African fires. If a 10-fold bias were to extend to fires in other regions, biomass burning could produce 14 Tg/a of HCOOH in the tropics or 16 Tg/a worldwide. However, even such an increase would only represent 15-20% of the total required HCOOH source, implying the existence of other larger missing sources.


Assuntos
Poluentes Atmosféricos , Incêndios , Biomassa , Desastres , Formiatos
4.
World J Surg ; 40(8): 1859-64, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27044022

RESUMO

INTRODUCTION: In 2005, the general population of Mongolia was not aware of laparoscopic surgery and was skeptical about the safety of surgical care. A 9-year initiative to expand laparoscopic surgery was initiated by Mongolian surgeons. This study examines the current barriers to and perceptions of surgical care following laparoscopic surgical expansion countrywide. MATERIALS AND METHODS: In September 2013, interviews were conducted with 71 patients, and 39 physicians in Mongolia. Patients and physicians were interviewed using separate sets of interview questions. Questions were designed to gauge perceptions of surgical care in Mongolia evaluating for access, affordability, sustainability, barriers to care, quality, and knowledge of laparoscopy. Responses were fine coded for statistical analysis. RESULTS: 79 % of patients felt surgical care was improving in Mongolia, and 76 % would choose laparoscopy if available. Physicians (100 %) felt laparoscopic surgery had improved surgical care in Mongolia. Barriers to care for patients were time to work up and diagnosis (37 %), and funding an operation (39 %). None of the 36 % of patients who stated funding an operation would be difficult identified government sources of funding (p < 0.001). Physicians identified insufficient equipment supply (69 %), insufficient training (41 %), and cost (38 %) as barriers for laparoscopy. 74 % of physicians felt that Mongolian physicians return or stay in Mongolia after training, defying the trend of migration in low-resource settings. DISCUSSION: Improved local patient and physician perception of laparoscopy is propelling the expansion of laparoscopy in Mongolia.


Assuntos
Atitude do Pessoal de Saúde , Educação de Pós-Graduação em Medicina/organização & administração , Conhecimentos, Atitudes e Prática em Saúde , Laparoscopia/educação , Feminino , Acessibilidade aos Serviços de Saúde , Humanos , Laparoscopia/psicologia , Laparoscopia/normas , Masculino , Mongólia , Médicos/psicologia , Desenvolvimento de Programas , Avaliação de Programas e Projetos de Saúde , Melhoria de Qualidade
5.
J Mol Cell Cardiol ; 79: 295-302, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25463275

RESUMO

Rapid activation causes remodeling of atrial myocytes resembling that which occurs in experimental and human atrial fibrillation (AF). Using this cellular model, we previously observed transcriptional upregulation of proteins implicated in protein misfolding and amyloidosis. For organ-specific amyloidoses such as Alzheimer's disease, preamyloid oligomers (PAOs) are now recognized to be the primary cytotoxic species. In the setting of oxidative stress, highly-reactive lipid-derived mediators known as γ-ketoaldehydes (γ-KAs) have been identified that rapidly adduct proteins and cause PAO formation for amyloid ß1-42 implicated in Alzheimer's. We hypothesized that rapid activation of atrial cells triggers oxidative stress with lipid peroxidation and formation of γ-KAs, which then rapidly crosslink proteins to generate PAOs. To investigate this hypothesis, rapidly-paced and control, spontaneously-beating atrial HL-1 cells were probed with a conformation-specific antibody recognizing PAOs. Rapid stimulation of atrial cells caused the generation of cytosolic PAOs along with a myocyte stress response (e.g., transcriptional upregulation of Nppa and Hspa1a), both of which were absent in control, unpaced cells. Rapid activation also caused the formation of superoxide and γ-KA adducts in atriomyocytes, while direct exposure of cells to γ-KAs resulted in PAO production. Increased cytosolic atrial natriuretic peptide (ANP), and the generation of ANP oligomers with exposure to γ-KAs and rapid atrial HL-1 cell stimulation, strongly suggest a role for ANP in PAO formation. Salicylamine (SA) is a small molecule scavenger of γ-KAs that can protect proteins from modification by these reactive compounds. PAO formation and transcriptional remodeling were inhibited when cells were stimulated in the presence of SA, but not with the antioxidant curcumin, which is incapable of scavenging γ-KAs. These results demonstrate that γ-KAs promote protein misfolding and PAO formation as a component of the atrial cell stress response to rapid activation, and they provide a potential mechanistic link between oxidative stress and atrial cell injury.


Assuntos
Aldeídos/farmacologia , Amiloide/metabolismo , Átrios do Coração/metabolismo , Átrios do Coração/patologia , Dobramento de Proteína/efeitos dos fármacos , Multimerização Proteica , Aminas/farmacologia , Animais , Fator Natriurético Atrial/metabolismo , Estimulação Cardíaca Artificial , Linhagem Celular , Curcumina/farmacologia , Citosol/efeitos dos fármacos , Citosol/metabolismo , Átrios do Coração/efeitos dos fármacos , Humanos , Camundongos , Modelos Biológicos , Estresse Oxidativo/efeitos dos fármacos , Superóxidos/metabolismo
6.
J Immunol ; 190(6): 2519-26, 2013 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-23396943

RESUMO

Autoreactive B lymphocytes that are not culled by central tolerance in the bone marrow frequently enter the peripheral repertoire in a state of functional impairment, termed anergy. These cells are recognized as a liability for autoimmunity, but their contribution to disease is not well understood. Insulin-specific 125Tg B cells support T cell-mediated type 1 diabetes in NOD mice, despite being anergic to B cell mitogens and T cell-dependent immunization. Using this model, the potential of anergic, autoreactive B cells to present Ag and activate T cells was investigated. The data show that 1) insulin is captured and rapidly internalized by 125Tg BCRs, 2) these Ag-exposed B cells are competent to activate both experienced and naive CD4(+) T cells, 3) anergic 125Tg B cells are more efficient than naive B cells at activating T cells when Ag is limiting, and 4) 125Tg B cells are competent to generate low-affinity insulin B chain epitopes necessary for activation of diabetogenic anti-insulin BDC12-4.1 T cells, indicating the pathological relevance of anergic B cells in type 1 diabetes. Thus, phenotypically tolerant B cells that are retained in the repertoire may promote autoimmunity by driving activation and expansion of autoaggressive T cells via Ag presentation.


Assuntos
Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/metabolismo , Subpopulações de Linfócitos B/imunologia , Anergia Clonal/imunologia , Tolerância Imunológica , Anticorpos Anti-Insulina/uso terapêutico , Animais , Células Apresentadoras de Antígenos/patologia , Subpopulações de Linfócitos B/metabolismo , Subpopulações de Linfócitos B/patologia , Células Cultivadas , Técnicas de Cocultura , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 1/metabolismo , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Camundongos Transgênicos , Receptores de Antígenos de Linfócitos B/biossíntese , Receptores de Antígenos de Linfócitos B/fisiologia , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/patologia
7.
Int J Surg Case Rep ; 119: 109686, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38677254

RESUMO

INTRODUCTION AND IMPORTANCE: Colorectal cancer (CRC) presenting with synchronous liver metastasis is relatively common, occurring in approximately 20 % of patients1. Herein we report an atypical case of a patient who presented with a new, obstructing colon mass with synchronous liver metastasis, biopsy proven to be malignant melanoma. CASE PRESENTATION: An 81-year-old male presented to the hospital emergency department with abdominal pain, diarrhea, and 30-pound unintentional weight loss over the past 4 months. Investigations revealed an obstructing cecal mass with multiple large, hypodense hepatic masses suspicious for metastatic disease. A multidisciplinary evaluation ensued, and the decision was made to treat with palliative intent. The patient was surgically treated with a diverting stoma and an intraoperative biopsy of the hepatic masses demonstrated metastatic melanoma. The patient did report a remote history of malignant melanoma and underwent curative-intent resection a decade earlier. There was no evidence of a new primary cutaneous melanoma. A tentative plan for checkpoint inhibitor therapy was discussed, but his medical issues worsened, and the patient died before any anti-cancer therapy could be started. CLINICAL DISCUSSION: The clinical picture of obstructing colon mass with synchronous liver masses most commonly represents a colon primary with synchronous liver metastasis. The capacity for melanoma to mimic other pathologies is unusual but has been described, with case reports describing metastasis to the eye, biliary hilum, liver, pancreas, colon, small bowel, and brain. This case serves as a good reminder that melanoma may mimic a variety of oncologic presentations, even after a very long disease-free interval. CONCLUSION: Our patient suspected to have metastatic colon cancer was found instead to have metastatic melanoma, with significantly different therapeutic options and prognosis.

8.
Dev Biol ; 349(2): 137-46, 2011 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-20969842

RESUMO

Despite their importance to oral health, the mechanisms of minor salivary gland (SG) development are largely unexplored. Here we present in vivo and in vitro analyses of developing minor SGs in wild type and mutant mice. Eda, Shh and Fgf signalling pathway genes are expressed in these glands from an early stage of development. Developing minor SGs are absent in Eda pathway mutant embryos, and these mice exhibit a dysplastic circumvallate papilla with disrupted Shh expression. Supplementation of Eda pathway mutant minor SG explants with recombinant EDA rescues minor SG induction. Supplementation with Fgf8 or Shh, previously reported targets of Eda signalling, leads to induction of gland like structures in a few cases, but these fail to develop into minor SGs.


Assuntos
Ectodisplasinas/metabolismo , Proteínas Recombinantes/farmacologia , Glândulas Salivares Menores/embriologia , Transdução de Sinais/fisiologia , Animais , Primers do DNA/genética , Ectodisplasinas/genética , Fator 8 de Crescimento de Fibroblasto/metabolismo , Fator 8 de Crescimento de Fibroblasto/farmacologia , Genótipo , Proteínas Hedgehog/metabolismo , Proteínas Hedgehog/farmacologia , Técnicas Histológicas , Hibridização In Situ , Camundongos , Camundongos Mutantes , Reação em Cadeia da Polimerase , Proteínas Recombinantes/metabolismo , Glândulas Salivares Menores/efeitos dos fármacos
9.
Am J Physiol Heart Circ Physiol ; 302(3): H782-9, 2012 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-22101522

RESUMO

Na(+) current derived from expression of the cardiac isoform SCN5A is reduced by receptor-mediated or direct activation of protein kinase C (PKC). Previous work has suggested a possible role for loss of Na(+) channels at the plasma membrane in this effect, but the results are controversial. In this study, we tested the hypothesis that PKC activation acutely modulates the intracellular distribution of SCN5A channels and that this effect can be visualized in living cells. In human embryonic kidney cells that stably expressed SCN5A with green fluorescent protein (GFP) fused to the channel COOH-terminus (SCN5A-GFP), Na(+) currents were suppressed by an exposure to PKC activation. Using confocal microscopy, colocalization of SCN5A-GFP channels with the plasma membrane under control and stimulated conditions was quantified. A separate population of SCN5A channels containing an extracellular epitope was immunolabeled to permit temporally stable labeling of the plasma membrane. Our results demonstrated that Na(+) channels were preferentially trafficked away from the plasma membrane by PKC activation, with a major contribution by Ca(2+)-sensitive or conventional PKC isoforms, whereas stimulation of protein kinase A (PKA) had the opposite effect. Removal of the conserved PKC site Ser(1503) or exposure to the NADPH oxidase inhibitor apocynin eliminated the PKC-mediated effect to alter channel trafficking, indicating that both channel phosphorylation and ROS were required. Experiments using fluorescence recovery after photobleaching demonstrated that both PKC and PKA also modified channel mobility in a manner consistent with the dynamics of channel distribution. These results demonstrate that the activation of protein kinases can acutely regulate the intracellular distribution and molecular mobility of cardiac Na(+) channels in living cells.


Assuntos
Miocárdio/enzimologia , Proteína Quinase C/metabolismo , Transporte Proteico/fisiologia , Canais de Sódio/metabolismo , Cálcio/metabolismo , Membrana Celular/enzimologia , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Ativação Enzimática/fisiologia , Proteínas de Fluorescência Verde/genética , Células HEK293 , Humanos , Potenciais da Membrana/fisiologia , Contração Miocárdica/fisiologia , Canal de Sódio Disparado por Voltagem NAV1.5 , Canais de Sódio/genética
10.
Circ Res ; 106(8): 1342-50, 2010 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-20339120

RESUMO

RATIONALE: Voltage-gated sodium channels initiate action potentials in excitable tissues. Mice in which Scn5A (the predominant sodium channel gene in heart) has been knocked out die early in development with cardiac malformations by mechanisms which have yet to be determined. OBJECTIVE: Here we addressed this question by investigating the role of cardiac sodium channels in zebrafish heart development. METHODS AND RESULTS: Transcripts of the functionally-conserved Scn5a homologs scn5Laa and scn5Lab were detected in the gastrulating zebrafish embryo and subsequently in the embryonic myocardium. Antisense knockdown of either channel resulted in marked cardiac chamber dysmorphogenesis and perturbed looping. These abnormalities were associated with decreased expression of the myocardial precursor genes nkx2.5, gata4, and hand2 in anterior lateral mesoderm and significant deficits in the production of cardiomyocyte progenitors. These early defects did not appear to result from altered membrane electrophysiology, as prolonged pharmacological blockade of sodium current failed to phenocopy channel knockdown. Moreover, embryos grown in calcium channel blocker-containing medium had hearts that did not beat but developed normally. CONCLUSIONS: These findings identify a novel and possibly nonelectrogenic role for cardiac sodium channels in heart development.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Cardiopatias Congênitas/genética , Coração/embriologia , Miocárdio/metabolismo , Canais de Sódio/genética , Proteínas de Peixe-Zebra/genética , Peixe-Zebra/embriologia , Fatores Etários , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Células CHO , Diferenciação Celular , Cricetinae , Cricetulus , Fatores de Transcrição GATA/genética , Gastrulação/genética , Técnicas de Silenciamento de Genes , Genótipo , Cardiopatias Congênitas/embriologia , Cardiopatias Congênitas/metabolismo , Proteína Homeobox Nkx-2.5 , Potenciais da Membrana , Dados de Sequência Molecular , Morfogênese/genética , Canal de Sódio Disparado por Voltagem NAV1.5 , Oligonucleotídeos Antissenso/metabolismo , Fenótipo , RNA Mensageiro/metabolismo , Canais de Sódio/metabolismo , Fatores de Transcrição/genética , Transfecção , Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo
11.
J Clin Invest ; 118(11): 3790-7, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18846254

RESUMO

The pancreatic islets of Langerhans are highly vascularized micro-organs that play a key role in the regulation of blood glucose homeostasis. The specific arrangement of endocrine cell types in islets suggests a coupling between morphology and function within the islet. Here, we established a line-scanning confocal microscopy approach to examine the relationship between blood flow and islet cell type arrangement by real-time in vivo imaging of intra-islet blood flow in mice. These data were used to reconstruct the in vivo 3D architecture of the islet and time-resolved blood flow patterns throughout the islet vascular bed. The results revealed 2 predominant blood flow patterns in mouse islets: inner-to-outer, in which blood perfuses the core of beta cells before the islet perimeter of non-beta cells, and top-to-bottom, in which blood perfuses the islet from one side to the other regardless of cell type. Our approach included both millisecond temporal resolution and submicron spatial resolution, allowing for real-time imaging of islet blood flow within the living mouse, which has not to our knowledge been attainable by other methods.


Assuntos
Diagnóstico por Imagem/métodos , Hemodinâmica , Imageamento Tridimensional/métodos , Ilhotas Pancreáticas/irrigação sanguínea , Animais , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/metabolismo , Camundongos , Camundongos Transgênicos , Microscopia Confocal , Fatores de Tempo
12.
Psychol Med ; 41(8): 1751-61, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21134315

RESUMO

BACKGROUND: The aim was to examine barriers to initiation and continuation of treatment among individuals with common mental disorders in the US general population. METHOD: Respondents in the National Comorbidity Survey Replication with common 12-month DSM-IV mood, anxiety, substance, impulse control and childhood disorders were asked about perceived need for treatment, structural barriers and attitudinal/evaluative barriers to initiation and continuation of treatment. RESULTS: Low perceived need was reported by 44.8% of respondents with a disorder who did not seek treatment. Desire to handle the problem on one's own was the most common reason among respondents with perceived need both for not seeking treatment (72.6%) and for dropping out of treatment (42.2%). Attitudinal/evaluative factors were much more important than structural barriers both to initiating (97.4% v. 22.2%) and to continuing (81.9% v. 31.8%) of treatment. Reasons for not seeking treatment varied with illness severity. Low perceived need was a more common reason for not seeking treatment among individuals with mild (57.0%) than moderate (39.3%) or severe (25.9%) disorders, whereas structural and attitudinal/evaluative barriers were more common among respondents with more severe conditions. CONCLUSIONS: Low perceived need and attitudinal/evaluative barriers are the major barriers to treatment seeking and staying in treatment among individuals with common mental disorders. Efforts to increase treatment seeking and reduce treatment drop-out need to take these barriers into consideration as well as to recognize that barriers differ as a function of sociodemographic and clinical characteristics.


Assuntos
Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Serviços de Saúde Mental/estatística & dados numéricos , Adolescente , Adulto , Idoso , Feminino , Pesquisas sobre Atenção à Saúde , Humanos , Masculino , Transtornos Mentais/epidemiologia , Transtornos Mentais/psicologia , Transtornos Mentais/terapia , Pessoa de Meia-Idade , Cooperação do Paciente/estatística & dados numéricos , Fatores Socioeconômicos , Estados Unidos/epidemiologia , Adulto Jovem
13.
Transgenic Res ; 20(3): 643-53, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20872247

RESUMO

Inactivation of the endogenous pig immunoglobulin (Ig) loci, and replacement with their human counterparts, would produce animals that could alleviate both the supply and specificity issues of therapeutic human polyclonal antibodies (PAbs). Platform genetics are being developed in pigs that have all endogenous Ig loci inactivated and replaced by human counterparts, in order to address this unmet clinical need. This report describes the deletion of the porcine kappa (κ) light chain constant (Cκ) region in pig primary fetal fibroblasts (PPFFs) using gene targeting technology, and the generation of live animals from these cells via somatic cell nuclear transfer (SCNT) cloning. There are only two other targeted loci previously published in swine, and this is the first report of a targeted disruption of an Ig light chain locus in a livestock species. Pigs with one targeted Cκ allele (heterozygous knockout or ±) were bred together to generate Cκ homozygous knockout (-/-) animals. Peripheral blood mononuclear cells (PBMCs) and mesenteric lymph nodes (MLNs) from Cκ -/- pigs were devoid of κ-containing Igs. Furthermore, there was an increase in lambda (λ) light chain expression when compared to that of wild-type littermates (Cκ +/+). Targeted inactivation of the Ig heavy chain locus has also been achieved and work is underway to inactivate the pig lambda light chain locus.


Assuntos
Clonagem de Organismos , Marcação de Genes , Cadeias kappa de Imunoglobulina/genética , Técnicas de Transferência Nuclear , Deleção de Sequência , Suínos , Animais , Feminino , Fibroblastos , Genes de Imunoglobulinas/genética , Humanos , Cadeias kappa de Imunoglobulina/metabolismo , Masculino
14.
Transgenic Res ; 20(3): 625-41, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20872248

RESUMO

A poly(A)-trap gene targeting strategy was used to disrupt the single functional heavy chain (HC) joining region (J(H)) of swine in primary fibroblasts. Genetically modified piglets were then generated via somatic cell nuclear transfer (SCNT) and bred to yield litters comprising J(H) wild-type littermate (+/+), J(H) heterozygous knockout (±) and J(H) homozygous knockout (-/-) piglets in the expected Mendelian ratio of 1:2:1. There are only two other targeted loci previously published in swine, and this is the first successful poly(A)-trap strategy ever published in a livestock species. In either blood or secondary lymphoid tissues, flow cytometry, RT-PCR and ELISA detected no circulating IgM(+) B cells, and no transcription or secretion of immunoglobulin (Ig) isotypes, respectively in J(H) -/- pigs. Histochemical and immunohistochemical (IHC) studies failed to detect lymph node (LN) follicles or CD79α(+) B cells, respectively in J(H) -/- pigs. T cell receptor (TCR)(ß) transcription and T cells were detected in J(H) -/- pigs. When reared conventionally, J(H) -/- pigs succumbed to bacterial infections after weaning. These antibody (Ab)- and B cell-deficient pigs have significant value as models for both veterinary and human research to discriminate cellular and humoral protective immunity to infectious agents. Thus, these pigs may aid in vaccine development for infectious agents such as the pandemic porcine reproductive and respiratory syndrome virus (PRRSV) and H1N1 swine flu. These pigs are also a first significant step towards generating a pig that expresses fully human, antigen-specific polyclonal Ab to target numerous incurable infectious diseases with high unmet clinical need.


Assuntos
Anticorpos/metabolismo , Linfócitos B/metabolismo , Modelos Animais de Doenças , Marcação de Genes , Cadeias Pesadas de Imunoglobulinas/genética , Isotipos de Imunoglobulinas/genética , Poli A/genética , Animais , Animais Recém-Nascidos , Anticorpos/genética , Anticorpos/imunologia , Linfócitos B/imunologia , Infecções Bacterianas/imunologia , Células Cultivadas , Fibroblastos , Engenharia Genética/métodos , Humanos , Cadeias Pesadas de Imunoglobulinas/metabolismo , Isotipos de Imunoglobulinas/metabolismo , Imuno-Histoquímica , Suínos , Linfócitos T/imunologia , Linfócitos T/metabolismo , Transfecção
15.
Med Vet Entomol ; 25(3): 311-9, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21219372

RESUMO

The diversity of ectoparasites in Southeast Asia and flea-host associations remain largely understudied. We explore specialization and interaction patterns of fleas infesting non-volant small mammals in Bornean rainforests, using material from a field survey carried out in two montane localities in northwestern Borneo (Sabah, Malaysia) and from a literature database of all available interactions in both lowland and montane forests. A total of 234 flea individuals collected during our field survey resulted in an interaction network of eight flea species on seven live-captured small mammal species. The interaction network from all compiled studies currently includes 15 flea species and 16 small mammal species. Host specificity and niche partitioning of fleas infesting diurnal treeshrews and squirrels were low, with little difference in specialization among taxa, but host specificity in lowland forests was found to be higher than in montane forests. By contrast, Sigmactenus alticola (Siphonaptera: Leptopsyllidae) exhibited low host specificity by infesting various montane and lowland nocturnal rats. However, this species exhibited low niche partitioning as it was the only commonly recorded flea from rats on Borneo. Overall complementary specialization was of intermediate intensity for both networks and differed significantly from random association; this has important implications for specific interactions that are also relevant to the potential spread of vector-borne diseases.


Assuntos
Ecossistema , Ectoparasitoses/veterinária , Mamíferos/parasitologia , Sifonápteros/fisiologia , Árvores , Animais , Bornéu , Ectoparasitoses/parasitologia , Interações Hospedeiro-Parasita , Especificidade da Espécie
16.
Dev Dyn ; 239(10): 2674-84, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20803597

RESUMO

Hypohidrotic ectodermal dysplasia (HED) is characterized by defective ectodermal organ development. This includes the salivary glands (SGs), which have an important role in lubricating the oral cavity. In humans and mice, HED is caused by mutations in Ectodysplasin A (Eda) pathway genes. Various phenotypes of the mutant mouse Eda(Ta/Ta), which lacks the ligand Eda, can be rescued by maternal injection or in vitro culture supplementation with recombinant EDA. However, the response of the SGs to this treatment has not been investigated. Here, we show that the submandibular glands (SMGs) of Eda(Ta/Ta) mice exhibit impaired branching morphogenesis, and that supplementation of Eda(Ta/Ta) SMG explants with recombinant EDA rescues the defect. Supplementation of Edar(dlJ/dlJ) SMGs with recombinant Sonic hedgehog (Shh) also rescues the defect, whereas treatment with recombinant Fgf8 does not. This work is the first to test the ability of putative Eda target molecules to rescue Eda pathway mutant SMGs.


Assuntos
Ectodisplasinas/metabolismo , Proteínas Hedgehog/metabolismo , Glândulas Salivares/metabolismo , Animais , Ectodisplasinas/genética , Receptor Edar/genética , Receptor Edar/metabolismo , Proteína de Domínio de Morte Associada a Edar/genética , Proteína de Domínio de Morte Associada a Edar/metabolismo , Genótipo , Proteínas Hedgehog/genética , Hibridização In Situ , Camundongos , Camundongos Mutantes , Morfogênese/genética , Morfogênese/fisiologia , Técnicas de Cultura de Órgãos , Glândulas Salivares/embriologia , Transdução de Sinais/genética , Transdução de Sinais/fisiologia
18.
Vet Comp Orthop Traumatol ; 24(6): 450-6, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21975493

RESUMO

OBJECTIVE: To evaluate a cohort of dogs undergoing tibial plateau levelling osteotomy (TPLO) implant removal to determine key clinical features, prevalence, and indications for implant removal. METHODS: Medical records of dogs undergoing TPLO implant removal at a private referral practice (Dallas Veterinary Surgical Center) between 2004-2008 were reviewed. Patient signalment, implant type, presence of concurrent medical disease, surgeon, antibiotic use, aerobic bacterial culture result, and operative findings were recorded. Data were analyzed using paired t-test, Fisher's exact test, and Wilcoxon-rank sum test. Statistical significance was set at p <0.05. RESULTS: The TPLO implants were removed from 126 dogs (n = 129, 4.8% of TPLO procedures) during the study period. Average time interval from TPLO to implant removal was 16.0 ± 17.8 months. The most common clinical signs were the presence of an open wound (n = 80), draining tract (n = 64), and lameness (n = 59). Culture of tissue or fluid from the implant bed or implants was positive for bacterial growth in 95/115 cases. A significantly greater proportion of the implants removed were Slocum TPLO plates (n = 109; 6.1%) when compared to other TPLO plate types (n = 20; 2.3%) (p <0.0001). No association was identified between a positive bacterial culture and measured variables. CLINICAL SIGNIFICANCE: Local bacterial infection and clinical signs of inflammation were the most common reasons for TPLO implant removal. There may be an increased implant-associated complication rate for Slocum TPLO plates in the study population.


Assuntos
Placas Ósseas/veterinária , Parafusos Ósseos/veterinária , Doenças do Cão/cirurgia , Osteotomia/veterinária , Tíbia/cirurgia , Animais , Antibacterianos/uso terapêutico , Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/etiologia , Placas Ósseas/efeitos adversos , Parafusos Ósseos/efeitos adversos , Cães , Estudos Retrospectivos
19.
Reprod Toxicol ; 105: 44-52, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34407461

RESUMO

Pigs have become an important model for agricultural and biomedical purposes. The advent of genomic engineering tools, such as the CRISPR/Cas9 system, has facilitated the production of livestock models with desired modifications. However, precise site-specific modifications in pigs through the homology-directed repair (HDR) pathway remains a challenge. In mammalian embryos, the use of small molecules to inhibit non-homologous end joining (NHEJ) or to improve HDR have been tested, but little is known about their toxicity. The compound RS-1 stimulates the activity of the RAD51 protein, which plays a key role in the HDR mechanism, demonstrating enhancement of HDR events in rabbit and bovine zygotes. Thus, in this study, we evaluated the dosage and temporal effects of RS-1 on porcine embryo development and viability. Additionally, we assessed the effects of its vehicle, DMSO, during embryo in vitro culture. Transient exposure to 7.5 µM of RS-1 did not adversely affect early embryo development and was compatible with subsequent development to term. Additionally, low concentrations of its vehicle, DMSO, did not show any toxicity to in vitro produced embryos. The transient use of RS-1 at 7.5 µM during in vitro culture seems to be the best protocol of choice to reduce the potentially toxic effects of RS-1 while attempting to improve HDR in the pig. Direct injection of the CRISPR/Cas9 system, combined with strategies to increase the frequency of targeted modifications via HDR, have become an important tool to simplify and accelerate the production of genetically modified livestock models.


Assuntos
Benzamidas/farmacologia , Dimetil Sulfóxido/farmacologia , Desenvolvimento Embrionário/efeitos dos fármacos , Rad51 Recombinase , Sulfonamidas/farmacologia , Animais , Transferência Embrionária , Embrião de Mamíferos/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Suínos , Técnicas de Cultura de Tecidos
20.
Hisp Health Care Int ; 19(1): 47-54, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-32466687

RESUMO

INTRODUCTION: Latinos are at higher risk of colorectal cancer (CRC) mortality than non-Hispanic Whites due, in part, to disparities in cancer screening. There is a need to evaluate community-based CRC interventions as they may reach underinsured communities and those at highest risk for CRC. This article describes the development of a group-based CRC intervention (Juntos contra el Cancer). METHOD: Purposive sampling was used to recruit Latino men and women aged 50 to 75 years not-up-to-date with CRC screening. The development of the intervention was guided by the socioecologic framework, a community needs assessment, literature reviews, five focus groups (n = 39) from the target community and feedback from a Community Advisory Board. RESULTS: Findings from focus groups suggested that a group-based, promotor or community health worker (CHW) led, cancer prevention education with linkages to care would address barriers to CRC screening. CONCLUSION: Development of community-based CRC screening interventions should be informed by early and sustained community engagement. Interventions led by CHWs with linkages to care are feasible and can reach populations not connected to health care settings.


Assuntos
Neoplasias Colorretais , Detecção Precoce de Câncer , Idoso , Neoplasias Colorretais/diagnóstico , Neoplasias Colorretais/prevenção & controle , Agentes Comunitários de Saúde , Feminino , Educação em Saúde , Hispânico ou Latino , Humanos , Masculino , Programas de Rastreamento , Pessoa de Meia-Idade
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