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1.
Mol Cell Neurosci ; 128: 103914, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38086519

RESUMO

The exocyst protein complex is important for targeted vesicle fusion in a variety of cell types, however, its function in neurons is still not entirely known. We found that presynaptic knockdown (KD) of the exocyst component sec15 by transgenic RNAi expression caused a number of unexpected morphological and physiological defects in the synapse. These include the development of active zones (AZ) devoid of essential presynaptic proteins, an increase in the branching of the presynaptic arbor, the appearance of satellite boutons, and a decrease in the amplitude of stimulated postsynaptic currents as well as a decrease in the frequency of spontaneous synaptic vesicle release. We also found the release of extracellular vesicles from the presynaptic neuron was greatly diminished in the Sec15 KDs. These effects were mimicked by presynaptic knockdown of Rab11, a protein known to interact with the exocyst. sec15 RNAi expression caused an increase in phosphorylated Mothers against decapentaplegic (pMad) in the presynaptic terminal, an indication of enhanced bone morphogenic protein (BMP) signaling. Some morphological phenotypes caused by Sec15 knockdown were reduced by attenuation of BMP signaling through knockdown of wishful thinking (Wit), while other phenotypes were unaffected. Individual knockdown of multiple proteins of the exocyst complex also displayed a morphological phenotype similar to Sec15 KD. We conclude that Sec15, functioning as part of the exocyst complex, is critically important for proper formation and function of neuronal synapses. We propose a model in which Sec15 is involved in the trafficking of vesicles from the recycling endosome to the cell membrane as well as possibly trafficking extracellular vesicles for presynaptic release and these processes are necessary for the correct structure and function of the synapse.


Assuntos
Proteínas de Drosophila , Drosophila , Animais , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Animais Geneticamente Modificados/metabolismo , Sinapses/metabolismo , Neurônios/metabolismo , Proteínas de Transporte Vesicular/metabolismo
2.
Vet Clin North Am Food Anim Pract ; 39(2): 325-336, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37164520

RESUMO

Grazing cows undergo a similar degree of metabolic stress and immune dysregulation to those reported in high-yielding housed cows consuming total mixed rations, but the ability to manage daily nutrient intake is much less. Feed quality varies from day-to-day and weather can greatly influence amount consumed on any given day. Transition cow management, therefore, tends to revolve around pragmatism as opposed to precision. Mid- and late-gestation management of body condition score is essential to a smooth transition period.


Assuntos
Dieta , Lactação , Feminino , Gravidez , Bovinos , Animais , Lactação/fisiologia , Dieta/veterinária , Ração Animal/análise , Leite/metabolismo
3.
Reprod Fertil ; 3(1): 1-8, 2022 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-35118417

RESUMO

The efficacy of a long-acting synthetic derivative of kisspeptin (Kp) to initiate normal oestrous cycles was tested in 24 mixed-aged, Holstein-Friesian cows that were 18-25 days postpartum on the day of treatment (D0). Groups of eight cows received saline (Sal) vehicle by intramuscular injection at 8:00 and 16:00 h (Sal-Sal), Kp at 8:00 h and vehicle at 16:00 h (Kp-Sal) or Kp on both occasions (Kp-Kp). The Kp dose was 15 nmol per 60 kg body weight. The ovaries of the cows were examined daily by ultrasonography between D4 and D14. Blood samples were collected from a tail vessel at 0, 2, 4, 8, 10 and 12 h relative to the time of the first injection for luteinizing hormone (LH) and follicle-stimulating hormone assay. Additional samples were collected daily from D4 until D14 and D19, 22, 26 and 29 for progesterone assay. LH surge-like responses were observed in cows treated with Kp at 8:00 h. Ovulation was consistently induced by Kp within 48 h when a dominant ovarian follicle of at least 10 mm in diameter was observed (8/14) but in no cases (6/14) during a new wave of ovarian follicular development comprising follicles <10 mm in diameter. The subsequent ovulatory cycle was of normal length in most cases as compared with short 8- to 12-day cycles observed in spontaneously ovulating cows. We conclude that Kp treatment can induce ovulation in postpartum dairy cows, with ensuing oestrous cycles of normal length, if administered when a mature dominant follicle is present in the ovaries. LAY SUMMARY: Cow fertility is important for efficient, profitable dairy farming. Cows that take too long after calving to become fertile are problematic. We tested a synthetically made, long-acting hormone called kisspeptin (Kp) to advance the time that cows become fertile after calving. Twenty-four dairy cows that had been calved for 3-4 weeks were used. One group of eight cows received an injection of Kp at the morning milking, another eight cows received Kp at both the morning and afternoon milking, while the last group of eight cows served as untreated controls. Kp treatment caused a desirable hormone response from the cows' brain. Normal oestrous cycles resulted, but only when a mature follicle was present in the ovary. Further study is required to analyse whether the use of a long-acting Kp drug could be used as an effective treatment for stimulating dairy cows to become more fertile after calving.


Assuntos
Anovulação , Kisspeptinas , Animais , Bovinos , Feminino , Humanos , Hormônio Luteinizante , Folículo Ovariano , Ovulação , Período Pós-Parto
4.
Sustainability ; 13(10): 5568, 2021 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-34164161

RESUMO

Dietary transitions, such as eliminating meat consumption, have been proposed as one way to reduce the climate impact of the global and regional food systems. However, it should be ensured that replacement diets are indeed nutritious and that climate benefits are accurately accounted for. This study uses New Zealand food consumption as a case study for exploring the cumulative climate impact of adopting the national dietary guidelines and the substitution of meat from hypothetical diets. The new GWP* metric is used as it was designed to better reflect the climate impacts of the release of methane than the de facto standard 100-year Global Warming Potential metric (GWP100). A transition at age 25 to the hypothetical dietary guideline diet reduces cumulative warming associated with diet by 7 to 9% at the 100th year compared with consuming the average New Zealand diet. The reduction in diet-related cumulative warming from the transition to a hypothetical meat-substituted diet varied between 12 and 15%. This is equivalent to reducing an average individual's lifetime warming contribution by 2 to 4%. General improvements are achieved for nutrient intakes by adopting the dietary guidelines compared with the average New Zealand diet; however, the substitution of meat items results in characteristic nutrient differences, and these differences must be considered alongside changes in emission profiles.

5.
Nat Biomed Eng ; 4(12): 1188-1196, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33122853

RESUMO

Accurate assays for the detection of antibodies to SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) are essential for the control of the COVID-19 (coronavirus disease 2019) pandemic. Here, we report antibody and antibody-avidity assays, relying on near-infrared-fluorescence amplification by nanostructured plasmonic gold substrates, for the simultaneous detection of antibodies to the S1 subunit of the spike protein and to the receptor binding domain of SARS-CoV-2 in human serum and saliva, and for quantifying immunoglobulin avidities against coronavirus antigens from SARS-CoV-2, SARS-CoV-1 and the common-cold viruses OC43, HKU1, NL63 and 229E. The antibody assay detected immunoglobulin M in 87% (52 of 60) COVID-19-positive serum samples collected 6 or more days after symptom onset (and the immunoglobulins M and G in all 33 samples collected at least 15 days after symptom onset), and correctly classified 456 out of the 457 COVID-19-negative serum samples tested (424 of them collected before the pandemic, including 73 that were positive for other viruses). We used the antibody-avidity assay to study antibody-maturation patterns, anamnestic responses, and cross-immunity to the common-cold coronaviruses.


Assuntos
Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , COVID-19/imunologia , SARS-CoV-2/imunologia , Saliva/imunologia , Idoso , Afinidade de Anticorpos/imunologia , Antígenos Virais/imunologia , COVID-19/sangue , Teste Sorológico para COVID-19/métodos , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Pandemias/prevenção & controle
6.
J Reprod Immunol ; 137: 102624, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31707221

RESUMO

Abnormalities in endometrial function contribute to poor fertility and reproductive failure. Exosomes are small lipid vesicles that contain transferable bioactive substances; they participate in intercellular signaling and may have critical roles in reproductive mechanisms, including endometrial remodeling in preparation for pregnancy. In this study, we evaluated the effects of exosomes from heifers with high and low genetic merit for fertility on inflammatory mediator expression by bovine endometrial epithelial and stromal cell lines. Co-incubation of exosomes from low, compared with high, fertility heifers upregulated the gene expression of pro-inflammatory IL1A and IL8 (CXCL8) but downregulated IL4 gene expression in epithelial cells. In contrast, stromal cells co-incubated with exosomes from low, compared with high, fertility heifers downregulated the gene expression of CXCL9, CXCL10, and CX3CL1. Our findings demonstrated that circulating exosomes from high fertility heifers did not alter endometrial inflammatory mediator gene expression. In contrast, circulating exosomes from low fertility heifers enhanced endometrial expression of inflammatory mediators, which may contribute to aberrant inflammation, leading to a reduced fertility in low fertility heifers. However, an in-depth investigation is required to elucidate the role of exosomes in regulating endometrial remodeling events required for enhanced reproductive performance and fertility in dairy cows.


Assuntos
Comunicação Celular/imunologia , Citocinas/metabolismo , Endométrio/imunologia , Exossomos/metabolismo , Fertilidade/imunologia , Animais , Bovinos , Citocinas/sangue , Citocinas/imunologia , Endométrio/citologia , Exossomos/imunologia , Feminino , Fertilidade/genética , Regulação da Expressão Gênica/imunologia , Inflamação/sangue , Inflamação/genética , Inflamação/imunologia , Mediadores da Inflamação/imunologia , Mediadores da Inflamação/metabolismo , Modelos Animais , Gravidez
7.
Sci Rep ; 9(1): 13879, 2019 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-31554846

RESUMO

Disease susceptibility of dairy cows is greatest during the transition from pregnancy to lactation. Circulating exosomes may provide biomarkers to detect at-risk cows to enhance health and productivity. From 490 cows, animals at high- (n = 20) or low-risk (n = 20) of transition-related diseases were identified using plasma non-esterified fatty acid and ß-hydroxybutyrate concentrations and liver triacylglyceride concentrations during the two weeks post-calving. We isolated circulating exosomes from plasma of dairy cows at low-risk (LR-EXO) and high-risk (HR-EXO), and analyzed their proteome profiles to determine markers for metabolic dysfunction. We evaluated the effects of these exosomes on eicosanoid pathway expression by bovine endometrial stromal (bCSC) and epithelial (bEEL) cells. HR-EXO had significantly lower yield of circulating exosomes compared with LR-EXO, and unique proteins were identified in HR-EXO and LR-EXO. Exposure to LR-EXO or HR-EXO differentially regulated eicosanoid gene expression and production in bCSC and bEEL cells. In bCSC, LR-EXO exposure increased PGE2 and PGD2 production, whereas HR-EXO exposure increased PTGS2 gene expression. In bEEL, HR-EXO exposure caused a decrease in PGE2, PGF2α, PGD2, PGFM and TXB2 production. The unique presence of serpin A3-7, coiled-coil domain containing 88A and inhibin/activin ß A chain in HR-EXO, indicates potential biomarkers for cows at-risk for metabolic diseases. Our results are in line with the health status of the cow indicating a potential diagnostic role for exosomes in enhancing cows' health and fertility.


Assuntos
Biomarcadores/sangue , Biomarcadores/metabolismo , Exossomos/metabolismo , Doenças Metabólicas/sangue , Doenças Metabólicas/metabolismo , Ácido 3-Hidroxibutírico/metabolismo , Animais , Bovinos , Endométrio/metabolismo , Células Epiteliais/metabolismo , Feminino , Expressão Gênica/fisiologia , Fígado/metabolismo , Triglicerídeos/metabolismo
8.
J Anim Sci ; 97(1): 192-207, 2019 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30428048

RESUMO

During the peripartum period, dairy cows often have signs of inflammation. Various stresses, including infectious and metabolic diseases, have been discussed as causative for this inflammation. In this study, expression profiles for 17 immune markers were measured in whole blood preparations from 78 dairy cows over a time frame starting 1 wk before calving to 4 wk after calving. Additionally, the effects of far-off and close-up feeding on immune function of dairy cows during the peripartum period were investigated. Cows were assigned to 1 of 2 feeding levels in late lactation to achieve a low and high BCS at the time of dry-off (approximately 4.25 and 5.0 on a 10-point scale). Following dry-off, both herds were managed to achieve a BCS of 5.0 one month before calving; this involved controlled feeding (i.e., maintenance) and over-feeding of ME during the far-off dry period. Within each far-off feeding-level treatment, cows were offered 65, 90, or 120% of their precalving ME requirements for 3 wk precalving in a 2 × 3 factorial arrangement. Analysis of gene expression profiles from blood cells revealed effects of time indicating that the transition cow's immune system counteracts the peripartum inflammation, whereas later postcalving it becomes activated to provide protection against postpartum infections. Far-off feeding affected (P < 0.05) the expression of 2 of the investigated genes at calving. Interleukin-6 (IL-6) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in unstimulated, peripheral leukocytes were lower (P < 0.05) in animals from the Far-Off_Over-fed group compared with the Far-Off_Control-fed group. Close-up feeding had several effects on gene expression, indicating that immune function in Feed120 animals was distinct from the Feed90 and Feed65. In conclusion, feeding management precalving becomes an important intervention to ensure immunocompetence at and after calving.


Assuntos
Bovinos/fisiologia , Ingestão de Alimentos , Ingestão de Energia , Inflamação/veterinária , Transcriptoma , Animais , Bovinos/genética , Bovinos/imunologia , Feminino , Gliceraldeído-3-Fosfato Desidrogenases/genética , Imunocompetência , Interleucina-6/genética , Lactação , Período Periparto , Período Pós-Parto
9.
J Dairy Sci ; 101(7): 6462-6473, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29705424

RESUMO

The current study evaluated exosomes isolated from plasma of heifers bred to have high or low fertility through developing extreme diversity in fertility breeding values, however, key animal traits (e.g., body weight, milk production, and percentage of North American genetics) remained similar between the 2 groups. The exosomes were isolated by a combined ultracentrifugation and size exclusion chromatography approach and characterized by their size distribution (nanoparticle tracking analysis), morphology (transmission electron microscopy), and presence of exosomal markers (immunoblotting). In addition, a targeted mass spectrometry approach was used to confirm the presence of 2 exosomal markers, tumor susceptibility gene 101 and flotillin 1. The number of exosomes from plasma of high fertility heifers was greater compared with low fertility heifers. Interestingly, the exosomal proteomic profile, evaluated using mass spectrometry, identified 89 and 116 proteins in the high and low fertility heifers respectively, of which 4 and 31 were unique, respectively. These include proteins associated with specific biological processes and molecular functions of fertility. Most notably, the tetratricopeptide repeat protein 41-related, glycodelin, and kelch-like protein 8 were identified in plasma exosomes unique to the low fertility heifers. These proteins are suggested to play a role in reproduction; however, the role of these proteins in dairy cow reproduction remains to be elucidated. Their identification underscores the potential for proteins within exosomes to provide information on the fertility status and physiological condition of the cow. This may potentially lead to the development of prognostic tools and interventions to improving dairy cow fertility.


Assuntos
Bovinos/genética , Fertilidade/genética , Proteômica , Animais , Exossomos , Feminino , Plasma , Proteoma
10.
eNeuro ; 3(2)2016.
Artigo em Inglês | MEDLINE | ID: mdl-27022630

RESUMO

Synaptic signaling involves the release of neurotransmitter from presynaptic active zones (AZs). Proteins that regulate vesicle exocytosis cluster at AZs, composing the cytomatrix at the active zone (CAZ). At the Drosophila neuromuscular junction (NMJ), the small GTPase Rab3 controls the distribution of CAZ proteins across release sites, thereby regulating the efficacy of individual AZs. Here we identify Rab3-GEF as a second protein that acts in conjunction with Rab3 to control AZ protein composition. At rab3-GEF mutant NMJs, Bruchpilot (Brp) and Ca(2+) channels are enriched at a subset of AZs, leaving the remaining sites devoid of key CAZ components in a manner that is indistinguishable from rab3 mutant NMJs. As the Drosophila homologue of mammalian DENN/MADD and Caenorhabditis elegans AEX-3, Rab3-GEF is a guanine nucleotide exchange factor (GEF) for Rab3 that stimulates GDP to GTP exchange. Mechanistic studies reveal that although Rab3 and Rab3-GEF act within the same mechanism to control AZ development, Rab3-GEF is involved in multiple roles. We show that Rab3-GEF is required for transport of Rab3. However, the synaptic phenotype in the rab3-GEF mutant cannot be fully explained by defective transport and loss of GEF activity. A transgenically expressed GTP-locked variant of Rab3 accumulates at the NMJ at wild-type levels and fully rescues the rab3 mutant but is unable to rescue the rab3-GEF mutant. Our results suggest that although Rab3-GEF acts upstream of Rab3 to control Rab3 localization and likely GTP-binding, it also acts downstream to regulate CAZ development, potentially as a Rab3 effector at the synapse.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento/genética , Mutação/genética , Junção Neuromuscular/citologia , Terminações Pré-Sinápticas/fisiologia , Proteínas rab3 de Ligação ao GTP/metabolismo , Potenciais de Ação/genética , Análise de Variância , Animais , Animais Geneticamente Modificados , Drosophila , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Microscopia Confocal , Junção Neuromuscular/genética , Neurônios/fisiologia , Técnicas de Patch-Clamp , Proteínas rab3 de Ligação ao GTP/genética
11.
J Dairy Sci ; 99(3): 2329-2338, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26778312

RESUMO

Precalving feeding level affects dry matter intake, postcalving energy balance, the risk of hepatic lipidosis and metabolic disease, and gene expression in liver and adipose tissue. These coincide with a higher risk of disease postpartum and, very likely, a failure to reach optimum production as well as reproductive targets. Current interpretation of the available evidence suggest that metabolic stressors affect the immune system of transition dairy cows and lead to reduced immunocompetence. The objective of the current study was to investigate the effect of precalving body condition score (BCS) and level of feeding on immunocompetence during the peripartum period. Twenty-three weeks before calving, 78 cows were allocated randomly to 1 of 6 treatment groups (n=13) in a 2 × 3 factorial arrangement: 2 precalving BCS categories (4.0 and 5.0, based on a 10-point scale) and 3 levels of energy intake during the 3 wk preceding calving (75, 100, and 125% of estimated requirements). Blood was sampled precalving and at 1, 2 and 4 wk after calving. Cells were analyzed by flow cytometry and quantitative real-time PCR. The numbers of T helper lymphocytes (CD4+), cytotoxic T lymphocytes (CD8+), natural killer cells (CD335+), and γδ T lymphocytes (WC1+) as well as their activation status [IL-2 receptor (CD25)+ cells] were highly variable between animals, but there was no evident effect of BCS, feeding level, or time. All groups presented with an increase in expression of cytokines in unstimulated blood cells in the week after calving, although this was significant only for IFNG in the BCS 4.0 group. Analysis of in vitro-stimulated cells allowed 2 general observations: (1) cows with high energy intake precalving (125%) had increased cytokine expression precalving, and (2) all cows had increased cytokine expression in the week after calving. The present study provides evidence that prepartum feed management can affect immunocompetence during the transition period. Considering the current results, optimally conditioned animals might benefit from a restricted precalving diet, whereas underconditioned cows can be fed to requirements.


Assuntos
Composição Corporal , Bovinos/fisiologia , Comportamento Alimentar , Imunocompetência , Animais , Bovinos/imunologia , Indústria de Laticínios , Feminino , Período Periparto
12.
Magn Reson Med ; 75(5): 2050-4, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26079271

RESUMO

PURPOSE: In this study, a new model of pulmonary embolism in rats was developed and tested, to examine if hyperpolarized (HP) (3) He MR images can measure impairment of the exchange of oxygen from the airspaces to the blood during pulmonary embolism. METHODS: HP (3) He MRI was used to image six treatment-group rats in which a branch of the pulmonary artery was embolized, and six control-group rats. HP (3) He MR images were used to calculate the initial partial pressure of oxygen (pO ) and the rate of oxygen depletion (R) in rat lungs. RESULTS: The pO was significantly higher in the ischemic lung than in the contralateral normal side, and pO was significantly higher in the ischemic lung than in both sides of the control lungs. Mean R in ischemic lungs was significantly lower than in the contralateral lungs, and mean R in ischemic lungs was also significantly lower than in both control lungs. CONCLUSION: These results demonstrate that pO and R, as measured by the T1 decay of HP (3) He, are sensitive to pulmonary ischemia in rats, confirming the findings in studies performed in large animal models of pulmonary ischemia.


Assuntos
Hélio/química , Isquemia/diagnóstico por imagem , Pneumopatias/diagnóstico por imagem , Pulmão/patologia , Imageamento por Ressonância Magnética , Oxigênio/química , Artéria Pulmonar/patologia , Embolia Pulmonar/diagnóstico por imagem , Animais , Processamento de Imagem Assistida por Computador , Isquemia/patologia , Pulmão/diagnóstico por imagem , Pneumopatias/patologia , Artéria Pulmonar/diagnóstico por imagem , Embolia Pulmonar/patologia , Ratos , Ratos Sprague-Dawley
13.
PLoS One ; 10(8): e0136938, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26317909

RESUMO

At synapses, the release of neurotransmitter is regulated by molecular machinery that aggregates at specialized presynaptic release sites termed active zones. The complement of active zone proteins at each site is a determinant of release efficacy and can be remodeled to alter synapse function. The small GTPase Rab3 was previously identified as playing a novel role that controls the distribution of active zone proteins to individual release sites at the Drosophila neuromuscular junction. Rab3 has been extensively studied for its role in the synaptic vesicle cycle; however, the mechanism by which Rab3 controls active zone development remains unknown. To explore this mechanism, we conducted a mutational analysis to determine the molecular and structural requirements of Rab3 function at Drosophila synapses. We find that GTP-binding is required for Rab3 to traffick to synapses and distribute active zone components across release sites. Conversely, the hydrolytic activity of Rab3 is unnecessary for this function. Through a structure-function analysis we identify specific residues within the effector-binding switch regions that are required for Rab3 function and determine that membrane attachment is essential. Our findings suggest that Rab3 controls the distribution of active zone components via a vesicle docking mechanism that is consistent with standard Rab protein function.


Assuntos
Análise Mutacional de DNA/métodos , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Junção Neuromuscular/metabolismo , Proteínas rab3 de Ligação ao GTP/genética , Proteínas rab3 de Ligação ao GTP/metabolismo , Animais , Sítios de Ligação , Proteínas de Drosophila/química , Drosophila melanogaster/genética , Junção Neuromuscular/fisiologia , Ligação Proteica , Vesículas Sinápticas/metabolismo , Proteínas rab3 de Ligação ao GTP/química
14.
Physiol Genomics ; 47(4): 89-101, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25604124

RESUMO

Postpartum uterine inflammation (endometritis) in the dairy cow is associated with lower fertility at both the time of infection and after the inflammation has resolved. We hypothesized that aberrant DNA methylation may be involved in the subfertility associated with uterine inflammation. The objective of this study was to characterize genome-wide DNA methylation and gene expression in the endometrium of dairy cows with subclinical endometritis (SCE). Endometrial tissues were obtained at 29 days postpartum (n = 12), and microarrays were used to characterize transcription and DNA methylation. Analyses revealed 1,856 probes differentially expressed in animals with SCE (n = 6) compared with controls (CON, n = 6, P < 0.05, Storey Multiple testing correction) and 2,976 probes with significant correlation between gene expression and bacteriology score. No significant associations among DNA methylation and gene expression were detected. Analysis of transcription data using the Database for Annotation, Visualization, and Integrated Discovery and Gene Set Enrichment Analysis identified several pathways and processes enriched in SCE cows, with the majority related to the immune response. Furthermore, the top ontology terms enriched in genes that had expression data correlated to bacteriology score were: Defense response, inflammatory response, and innate immune response. Gene expression profiles in cows with subclinical endometritis in this study indicate that the immune response is activated, potentially resulting in a local proinflammatory environment in the uterus. If this period of inflammation is prolonged it could result in tissue damage or failure to complete involution of the uterus, which may create a suboptimal environment for future pregnancy.


Assuntos
Doenças dos Bovinos/imunologia , Bovinos/fisiologia , Endometrite/veterinária , Fertilidade , Reação de Fase Aguda/imunologia , Animais , Bovinos/imunologia , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/patologia , Metilação de DNA , Endometrite/imunologia , Endometrite/microbiologia , Endometrite/patologia , Feminino , Sistema Imunitário , Leite/química , Período Pós-Parto/imunologia , Útero/patologia
15.
J Dairy Sci ; 98(2): 937-46, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25483203

RESUMO

Peripartum, and especially during the transition period, dairy cows undergo dramatic physiological changes. These coincide with an increased risk of disease during the first 2 wk after calving and have been linked to dairy cows failing to achieve production as well as reproductive targets. Previous evidence suggests that these physiological changes affect the immune system and that transition dairy cows experience some form of reduced immunocompetence. However, almost all of these studies were undertaken in high-production, housed dairy cows. Grazing cows have much lower levels of production and this study aimed to provide clarity whether or not the dysfunctional attributes of the peripartum immune system reported in high production housed cows are evident in these animals. Therefore, cell culture techniques, flow cytometry, and quantitative PCR were applied to analyze the cellular composition of peripheral blood mononuclear cells from transition dairy cows as well as the performance of these cells in an in vitro assay. First, a combination of in vitro stimulation and quantitative PCR for cytokines was validated as a quantifiable immunocompetence assay in 29 cattle and a correlation of quantitative PCR and ELISA demonstrated. Second, the relative number of T helper cells, cytotoxic T cells, B cells, γδ T cells, natural killer cells, and monocytes in peripheral blood was measured, of which B cells and natural killer cells increased in number postcalving (n=29) compared with precalving. Third, following in vitro stimulation cytokine profiles indicated decreased expression of IFNγ, tumor necrosis factor, and IL-17 and increased expression of IL-10 wk 1 after calving, which later all returned to precalving values (n=39). Additionally, treatment of transition cows with a nonsteroidal anti-inflammatory drug (i.e., carprofen) administered on d 1, 3, and 5 postcalving (n=19; untreated control n=20) did not affect the cytokine expression at any time point. In conclusion, an immunocompetence assay has been developed that highlights a characteristic expression pattern for IFNγ, tumor necrosis factor, IL-17, and IL-10 that reflects a state of reduced immunocompetence in moderate-yielding pasture-based transition cows after calving, which is similar to that described for higher-yielding housed cows.


Assuntos
Bovinos/fisiologia , Interferon gama/metabolismo , Interleucina-10/metabolismo , Interleucina-17/metabolismo , Período Pós-Parto/fisiologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Feminino , Regulação da Expressão Gênica/fisiologia , Interferon gama/genética , Interleucina-10/genética , Interleucina-17/genética , Leucócitos Mononucleares , Monócitos/metabolismo , Fator de Necrose Tumoral alfa/genética
16.
Acad Radiol ; 21(12): 1524-9, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25172411

RESUMO

RATIONALE AND OBJECTIVES: Hyperpolarized (HP) gas magnetic resonance imaging (MRI) is an advanced imaging technique that provides high-resolution regional information on lung function without using ionizing radiation. Before this modality can be considered for assessing clinical or investigational interventions, baseline repeatability needs to be established. We assessed repeatability of lung function measurement using HP helium-3 MRI (HP (3)He MRI) in a small cohort of patients with cystic fibrosis (CF). MATERIALS AND METHODS: We examined repeatability of HP (3)He MR images of five patients with CF in four scanning sessions over a 4-week period. We acquired images on a Philips 3.0 Tesla Achieva MRI scanner using a quadrature, flexible, wrap-around, (3)He radiofrequency coil with a fast gradient-echo pulse sequence. We determined ventilation volume and ventilation defect volume using an advanced semiautomatic segmentation algorithm and also quantified ventilation heterogeneity. RESULTS: There were no significant differences in total ventilation volume, ventilation defect volume, ventilation defect percentage, or mean ventilation heterogeneity (repeated-measures analysis of variance, P = .2116, P = .2825, P = .2871, and P = .7265, respectively) in the patients across the four scanning sessions. CONCLUSIONS: Our results indicate that total ventilation volume, ventilation defect volume, ventilation defect percentage, and mean ventilation heterogeneity as assessed by HP gas MRI in CF patients with stable health are reproducible over time. This repeatability and the technique's capability to provide noninvasive high-resolution data on regional lung function without ionizing radiation make (3)He MRI a potentially useful outcome measure for CF-related clinical trials.


Assuntos
Fibrose Cística/fisiopatologia , Imageamento por Ressonância Magnética/métodos , Algoritmos , Hélio , Humanos , Isótopos , Ventilação Pulmonar , Reprodutibilidade dos Testes , Testes de Função Respiratória
17.
Bioinform Biol Insights ; 8: 45-63, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24578603

RESUMO

Transcriptome alterations in liver and adipose tissue of cows with subclinical endometritis (SCE) at 29 d postpartum were evaluated. Bioinformatics analysis was performed using the Dynamic Impact Approach by means of KEGG and DAVID databases. Milk production, blood metabolites (non-esterified fatty acids, magnesium), and disease biomarkers (albumin, aspartate aminotransferase) did not differ greatly between healthy and SCE cows. In liver tissue of cows with SCE, alterations in gene expression revealed an activation of complement and coagulation cascade, steroid hormone biosynthesis, apoptosis, inflammation, oxidative stress, MAPK signaling, and the formation of fibrinogen complex. Bioinformatics analysis also revealed an inhibition of vitamin B3 and B6 metabolism with SCE. In adipose, the most activated pathways by SCE were nicotinate and nicotinamide metabolism, long-chain fatty acid transport, oxidative phosphorylation, inflammation, T cell and B cell receptor signaling, and mTOR signaling. Results indicate that SCE in dairy cattle during early lactation induces molecular alterations in liver and adipose tissue indicative of immune activation and cellular stress.

18.
Med J Aust ; 199(3): 214, 2013 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-24066363
19.
Vet Clin North Am Food Anim Pract ; 29(2): 323-36, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23809894

RESUMO

Body condition score (BCS) is an assessment of a cow's body fat (and muscle) reserves, with low values reflecting emaciation and high values equating to obesity. The intercalving profile of BCS is a mirror image of the milk lactation profile. The BCS at which a cow calves, her nadir BCS, and the amount of BCS lost after calving are associated with milk production, reproduction, and health. Genetics, peripartum nutrition, and management are factors that likely interact with BCS to determine the risk of health disorders.


Assuntos
Composição Corporal/fisiologia , Doenças dos Bovinos/prevenção & controle , Bovinos/fisiologia , Lactação/fisiologia , Doenças Metabólicas/veterinária , Fenômenos Fisiológicos da Nutrição Animal , Animais , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/metabolismo , Feminino , Doenças Metabólicas/diagnóstico , Doenças Metabólicas/metabolismo , Doenças Metabólicas/prevenção & controle , Leite/metabolismo , Necessidades Nutricionais , Período Pós-Parto/fisiologia , Gravidez , Reprodução/fisiologia
20.
J Dairy Res ; 80(2): 197-204, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23445567

RESUMO

The objective of this study was to determine the effect of cow genetic strain on fatty acid (FA) profiles in adipose tissue and milk. Adipose samples from two subcutaneous (shoulder and tail-head) and three visceral (kidney channel, mesenteric and omental) depots were obtained post mortem from New Zealand (NZ; n = 8) and North American (NA; n = 8) Holstein-Friesian cows. At the time of slaughter cows were in similar body condition (NZ: 4.0 ± 0.03, NA: 4.0 ± 0.02; ± SD) and stage of lactation (NZ: 90 ± 11.2 d; NA: 83 ± 4.3 d; ± SD). Milk was collected during the a.m. milking prior to slaughter and milk fat was extracted. Adipose and milk fat FA were quantified using gas chromatography. NZ cows had a lower proportion of saturated FA in shoulder, tail-head and omental adipose tissue and a greater proportion of mono-unsaturated FA and an elevated Δ9-desaturase index in shoulder and tail-head adipose tissue. The proportions of individual FA differed between adipose depots, with proportions of de-novo FA greater in subcutaneous compared with visceral adipose depots. Milk from NZ cows contained greater concentrations of short chain FA (C8 : 0-12 : 0) and CLA, and less cis-9 18 : 1 than milk from NA cows. Regression analysis identified moderate associations between milk FA and shoulder adipose tissue FA for 18 : 2 (R(2) = 0.24), 18 : 3 n - 3 (R(2) = 0.39), and polyunsaturated fatty acids (R(2) = 0.38). Results from this study support the hypothesis that genetic strain dictates FA profiles in adipose tissue and milk and may alter the metabolic status of the various adipose depots differently. The data further support the premise that genetic strain affects the metabolic status of the various adipose depots differently. Elucidating the mechanisms that regulate the different adipose depots in the NZ and NA strains will increase our understanding of tissue mobilization and replenishment.


Assuntos
Tecido Adiposo/química , Bovinos/genética , Gorduras/química , Ácidos Graxos/análise , Leite/química , Animais , Bovinos/metabolismo , Ácidos Graxos Monoinsaturados/análise , Feminino , Gordura Intra-Abdominal/química , Lactação , Nova Zelândia , Especificidade da Espécie , Estearoil-CoA Dessaturase/análise , Gordura Subcutânea/química
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