Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 7.285
Filtrar
1.
Aging Cell ; 20(2): e13316, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33524238

RESUMO

The ageing of the global population brings about unprecedented challenges. Chronic age-related diseases in an increasing number of people represent an enormous burden for health and social care. The immune system deteriorates during ageing and contributes to many of these age-associated diseases due to its pivotal role in pathogen clearance, tissue homeostasis and maintenance. Moreover, in order to develop treatments for COVID-19, we urgently need to acquire more knowledge about the aged immune system, as older adults are disproportionally and more severely affected. Changes with age lead to impaired responses to infections, malignancies and vaccination, and are accompanied by chronic, low-degree inflammation, which together is termed immunosenescence. However, the molecular and cellular mechanisms that underlie immunosenescence, termed immune cell senescence, are mostly unknown. Cellular senescence, characterised by an irreversible cell cycle arrest, is thought to be the cause of tissue and organismal ageing. Thus, better understanding of cellular senescence in immune populations at single-cell level may provide us with insight into how immune cell senescence develops over the life time of an individual. In this review, we will briefly introduce the phenotypic characterisation of aged innate and adaptive immune cells, which also contributes to overall immunosenescence, including subsets and function. Next, we will focus on the different hallmarks of cellular senescence and cellular ageing, and the detection techniques most suitable for immune cells. Applying these techniques will deepen our understanding of immune cell senescence and to discover potential druggable pathways, which can be modulated to reverse immune ageing.


Assuntos
Senescência Celular , Imunossenescência , Leucócitos/fisiologia , Animais , Biomarcadores/metabolismo , Humanos , Estresse Oxidativo , Proteostase
2.
Arterioscler Thromb Vasc Biol ; 41(3): 1167-1178, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33441028

RESUMO

OBJECTIVE: People with diabetes are at a significantly higher risk of cardiovascular disease, in part, due to accelerated atherosclerosis. Diabetic subjects have increased number of platelets that are activated, more reactive, and respond suboptimally to antiplatelet therapies. We hypothesized that reducing platelet numbers by inducing their premature apoptotic death would decrease atherosclerosis. Approach and Results: This was achieved by targeting the antiapoptotic protein Bcl-xL (B-cell lymphoma-extra large; which is essential for platelet viability) via distinct genetic and pharmacological approaches. In the former, we transplanted bone marrow from mice carrying the Tyr15 to Cys loss of function allele of Bcl-x (known as Bcl-xPlt20) or wild-type littermate controls into atherosclerotic-prone Ldlr+/- mice made diabetic with streptozotocin and fed a Western diet. Reduced Bcl-xL function in hematopoietic cells significantly decreased platelet numbers, exclusive of other hematologic changes. This led to a significant reduction in atherosclerotic lesion formation in Bcl-xPlt20 bone marrow transplanted Ldlr+/- mice. To assess the potential therapeutic relevance of reducing platelets in atherosclerosis, we next targeted Bcl-xL with a pharmacological strategy. This was achieved by low-dose administration of the BH3 (B-cell lymphoma-2 homology domain 3) mimetic, ABT-737 triweekly, in diabetic Apoe-/- mice for the final 6 weeks of a 12-week study. ABT-737 normalized platelet numbers along with platelet and leukocyte activation to that of nondiabetic controls, significantly reducing atherosclerosis while promoting a more stable plaque phenotype. CONCLUSIONS: These studies suggest that selectively reducing circulating platelets, by targeting Bcl-xL to promote platelet apoptosis, can reduce atherosclerosis and lower cardiovascular disease risk in diabetes. Graphic Abstract: A graphic abstract is available for this article.


Assuntos
Aterosclerose/sangue , Aterosclerose/complicações , Plaquetas/patologia , Angiopatias Diabéticas/sangue , Animais , Apoptose/efeitos dos fármacos , Apoptose/genética , Aterosclerose/prevenção & controle , Compostos de Bifenilo/administração & dosagem , Plaquetas/efeitos dos fármacos , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/complicações , Feminino , Humanos , Leucócitos/patologia , Leucócitos/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Nitrofenóis/administração & dosagem , Piperazinas/administração & dosagem , Contagem de Plaquetas , Receptores de LDL/deficiência , Receptores de LDL/genética , Fatores de Risco , Sulfonamidas/administração & dosagem
3.
J Vis Exp ; (158)2020 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-32364551

RESUMO

Intravital microscopy (IVM) is widely used to monitor physiological and pathophysiological processes within the leukocyte recruitment cascade in vivo. The current protocol represents a practical and reproducible method to visualize the leukocyte endothelium interaction leading to leukocyte recruitment in skeletal muscle derived tissue within the intact organism of the mouse. The model is applicable to all fields of research that focus on granulocyte activation and their role in disease. We provide a step by step protocol to guide through the method and to highlight potential pitfalls and technical difficulties. The protocol covers the following aspects: experimental settings and required material, anesthesia of the mouse, dissection of the cremaster muscle as well as tracheal and carotid cannulation, IVM recordings and offline analysis. Data formats like adherent leukocytes, rolling flux (RF) and rolling flux fraction (RFF) are explained in detail and appropriate applications are discussed. Representative results from dystrophin deficient mdx mice are provided in the results section. IVM is a powerful tool to assess leukocyte recruitment in an in vivo setting; however, delineating for example endothelial and leukocyte function may require a combination with ex vivo setups like flow chamber experiments. Furthermore, the genetic background of animals of interest may greatly influence baseline recruitment, requiring individual fine tuning of the protocol provided. Despite its limitations, IVM may serve as a platform to readily translate in vitro findings into a living vertebrate organism.


Assuntos
Músculos Abdominais/fisiologia , Adesão Celular , Endotélio/metabolismo , Microscopia Intravital/métodos , Migração e Rolagem de Leucócitos , Leucócitos/fisiologia , Músculos Abdominais/diagnóstico por imagem , Animais , Endotélio/citologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos mdx
4.
Georgian Med News ; (299): 26-29, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32242839

RESUMO

According to the statistics of the Russian Federation, the number of patients with male infertility increased by 60.3% over the past 12 years. Objective - to evaluate the activity of peripheral blood leukocytes, the rate of their production of reactive oxygen species, and the DNA fragmentation index in men with non-obstructive azoospermia. To solve this problem, we examined 65 men with non-obstructive idiopathic azoospermia. A control group consisted of 20 healthy fertile men. All patients underwent examination of ejaculate with measurement of levels of reactive oxygen species (ROS) and DNA fragmentation. To identify additional links in the pathogenesis of infertility, we determined the functional activity of peripheral blood leukocytes by registering luminol-dependent chemiluminescence (LCH) of leukocytes. The functional activity of leukocytes in men with azoospermia was found to be 4.5 times higher than in healthy men (p<0.01). We found that with such leukocyte activity the level of ROS and DNA fragmentation was 6 times higher in patients with infertility compared to the control group.


Assuntos
Azoospermia , Fragmentação do DNA , Infertilidade Masculina/etiologia , Leucócitos/fisiologia , Espermatozoides/metabolismo , Humanos , Masculino , Espécies Reativas de Oxigênio , Federação Russa , Motilidade Espermática/fisiologia
5.
Proc Natl Acad Sci U S A ; 117(17): 9466-9476, 2020 04 28.
Artigo em Inglês | MEDLINE | ID: mdl-32295886

RESUMO

Peripheral nerves contain axons and their enwrapping glia cells named Schwann cells (SCs) that are either myelinating (mySCs) or nonmyelinating (nmSCs). Our understanding of other cells in the peripheral nervous system (PNS) remains limited. Here, we provide an unbiased single cell transcriptomic characterization of the nondiseased rodent PNS. We identified and independently confirmed markers of previously underappreciated nmSCs and nerve-associated fibroblasts. We also found and characterized two distinct populations of nerve-resident homeostatic myeloid cells that transcriptionally differed from central nervous system microglia. In a model of chronic autoimmune neuritis, homeostatic myeloid cells were outnumbered by infiltrating lymphocytes which modulated the local cell-cell interactome and induced a specific transcriptional response in glia cells. This response was partially shared between the peripheral and central nervous system glia, indicating common immunological features across different parts of the nervous system. Our study thus identifies subtypes and cell-type markers of PNS cells and a partially conserved autoimmunity module induced in glia cells.


Assuntos
Neurônios/fisiologia , Nervos Periféricos/citologia , Animais , Doenças Autoimunes/metabolismo , Biomarcadores , Comunicação Celular , Linhagem da Célula , Regulação da Expressão Gênica/fisiologia , Homeostase , Humanos , Leucócitos/fisiologia , Macrófagos/fisiologia , Camundongos , Ratos
6.
DNA Cell Biol ; 39(4): 548-554, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32155344

RESUMO

The Qing-Tibet Plateau is characterized by low oxygen pressure, which is an important biomedical and ecological stressor. However, the variation in gene expression during periods of stay on the plateau has not been well studied. We recruited eight volunteers to stay on the plateau for 3, 7, and 30 days. Human Clariom D arrays were used to measure transcriptome changes in the mRNA expression profiles in these volunteers' blood. Analysis of variance (ANOVA) indicated that 699 genes were significantly differentially expressed in response to entering the plateau during hypoxic exposure. The genes with changes in transcript abundance were involved in the terms phosphoprotein, acetylation, protein binding, and protein transport. Furthermore, numerous genes involved in hematopoietic functions, including erythropoiesis and immunoregulation, were differentially expressed in response to hypoxia. This phenomenon may be one of reasons why the majority of people entering the plateau do not have excessive erythrocyte proliferation and are susceptible to infection.


Assuntos
Aclimatação/genética , Doença da Altitude/genética , Doença da Altitude/fisiopatologia , Hipóxia/fisiopatologia , Leucócitos/fisiologia , Altitude , Eritrócitos/citologia , Eritropoese/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/genética , Humanos , Leucopoese/genética , Masculino , Oxigênio , Tibet
7.
Proc Natl Acad Sci U S A ; 117(10): 5463-5471, 2020 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-32079726

RESUMO

Chronic pain is a major clinical problem of which the mechanisms are incompletely understood. Here, we describe the concept that PI16, a protein of unknown function mainly produced by fibroblasts, controls neuropathic pain. The spared nerve injury (SNI) model of neuropathic pain increases PI16 protein levels in fibroblasts in dorsal root ganglia (DRG) meninges and in the epi/perineurium of the sciatic nerve. We did not detect PI16 expression in neurons or glia in spinal cord, DRG, and nerve. Mice deficient in PI16 are protected against neuropathic pain. In vitro, PI16 promotes transendothelial leukocyte migration. In vivo, Pi16 -/- mice show reduced endothelial barrier permeability, lower leukocyte infiltration and reduced activation of the endothelial barrier regulator MLCK, and reduced phosphorylation of its substrate MLC2 in response to SNI. In summary, our findings support a model in which PI16 promotes neuropathic pain by mediating a cross-talk between fibroblasts and the endothelial barrier leading to barrier opening, cellular influx, and increased pain. Its key role in neuropathic pain and its limited cellular and tissue distribution makes PI16 an attractive target for pain management.


Assuntos
Fibroblastos/enzimologia , Neuralgia/genética , Proteínas Secretadas Inibidoras de Proteinases/genética , Animais , Movimento Celular , Dor Crônica , Modelos Animais de Doenças , Células Endoteliais/fisiologia , Gânglios Espinais , Leucócitos/fisiologia , Meninges/citologia , Camundongos Knockout , Traumatismos dos Nervos Periféricos/fisiopatologia , Nervo Isquiático/enzimologia
8.
Gerontology ; 66(4): 315-322, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32088715

RESUMO

BACKGROUND: Telomeres are crucial parts of chromosomes that protect the genome. They shorten every time the cell replicates, and shorter telomeres have been associated with increasing age and with many health behaviours. There is inconclusive evidence on the association between physical activity (PA) and telomere length. OBJECTIVES: To examine how leisure-time PA (LTPA) is associated with telomere length and telomere attrition during 10 years of follow-up in elderly people. DESIGN: This study is a 10-year prospective follow-up study. METHOD: For this prospective study, we examined 1,014 subjects (mean age at baseline 60.8 years) from the Helsinki Birth Cohort Study (HBCS). Relative leukocyte telomere length (LTL) was measured with a quantitative real-time PCR and LTPA with a validated questionnaire. Multiple linear regression analyses were used to assess the association between sex-specific LTPA quartiles and LTL at baseline and change in LTL over 10 years. The analyses were adjusted for age, educational attainment, smoking, body fat percentage, oestrogen exposure in women and for follow-up time when applicable. RESULTS: At baseline, volume of LTPA was not associated with LTL in men (p = 0.66) or in women (p = 0.33). Among women, however, higher volume of LTPA at baseline was associated with greater shortening of LTL (p for linearity 0.040) during the 10-year follow-up. No association was found among men (p for linearity 0.75). CONCLUSIONS: Our findings suggest that PA has a sex-specific role in regulation of telomere length in the aging process as in our study a high volume of LTPA in elderly women, but not in men, was associated with more rapid telomere attrition.


Assuntos
Exercício Físico/fisiologia , Envelhecimento Saudável/fisiologia , Encurtamento do Telômero/fisiologia , Telômero/fisiologia , Idoso , Estudos de Coortes , Feminino , Finlândia , Seguimentos , Humanos , Atividades de Lazer , Leucócitos/fisiologia , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Inquéritos e Questionários
9.
JAMA Netw Open ; 3(2): e200023, 2020 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-32101305

RESUMO

Importance: Leukocyte telomere length (LTL) is a trait associated with risk of cardiovascular disease and cancer, the 2 major disease categories that largely define longevity in the United States. However, it remains unclear whether LTL is associated with the human life span. Objective: To examine whether LTL is associated with the life span of contemporary humans. Design, Setting, and Participants: This cohort study included 3259 adults of European ancestry from the Cardiovascular Health Study (CHS), Framingham Heart Study (FHS), and Women's Health Initiative (WHI). Leukocyte telomere length was measured in 1992 and 1997 in the CHS, from 1995 to 1998 in the FHS, and from 1993 to 1998 in the WHI. Data analysis was conducted from February 2017 to December 2019. Main Outcomes and Measures: Death and LTL, measured by Southern blots of the terminal restriction fragments, were the main outcomes. Cause of death was adjudicated by end point committees. Results: The analyzed sample included 3259 participants (2342 [71.9%] women), with a median (range) age of 69.0 (50.0-98.0) years at blood collection. The median (range) follow-up until death was 10.9 (0.2-23.0) years in CHS, 19.7 (3.4-23.0) years in FHS, and 16.6 (0.5-20.0) years in WHI. During follow-up, there were 1525 deaths (482 [31.6%] of cardiovascular disease; 373 [24.5%] of cancer, and 670 [43.9%] of other or unknown causes). Short LTL, expressed in residual LTL, was associated with increased mortality risk. Overall, the hazard ratio for all-cause mortality for a 1-kilobase decrease in LTL was 1.34 (95% CI, 1.21-1.47). This association was stronger for noncancer causes of death (cardiovascular death: hazard ratio, 1.28; 95% CI, 1.08-1.52; cancer: hazard ratio, 1.13; 95% CI, 0.93-1.36; and other causes: hazard ratio, 1.53; 95% CI, 1.32-1.77). Conclusions and Relevance: The results of this study indicate that LTL is associated with a natural life span limit in contemporary humans.


Assuntos
Leucócitos/fisiologia , Expectativa de Vida , Telômero/genética , Idoso , Idoso de 80 Anos ou mais , Doenças Cardiovasculares/mortalidade , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Neoplasias/mortalidade
10.
FASEB J ; 34(1): 1745-1754, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31914612

RESUMO

Blood vessels are comprised of endothelial and smooth muscle cells. Obtaining both types of cells from vessels of living donors is not possible without invasive surgery. To address this, we have devised a strategy whereby human endothelial and smooth muscle cells derived from blood progenitors from the same donor could be cultured with autologous leukocytes to generate a same donor "vessel in a dish" bioassay. Autologous sets of blood outgrowth endothelial cells (BOECs), smooth muscle cells (BO-SMCs), and leukocytes were obtained from four donors. Cells were treated in monoculture and cumulative coculture conditions. The endothelial specific mediator endothelin-1 along with interleukin (IL)-6, IL-8, tumor necrosis factor α, and interferon gamma-induced protein 10 were measured under control culture conditions and after stimulation with cytokines. Cocultures remained viable throughout. The profile of individual mediators released from cells was consistent with what we know of endothelial and smooth muscle cells cultured from blood vessels. For the first time, we report a proof of concept study where autologous blood outgrowth "vascular" cells and leukocytes were studied alone and in coculture. This novel bioassay has usefulness in vascular biology research, patient phenotyping, drug testing, and tissue engineering.


Assuntos
Células Endoteliais/fisiologia , Leucócitos/fisiologia , Músculo Liso Vascular/fisiologia , Miócitos de Músculo Liso/fisiologia , Bioensaio/métodos , Células Cultivadas , Técnicas de Cocultura/métodos , Citocinas/metabolismo , Descoberta de Drogas/métodos , Células Endoteliais/metabolismo , Humanos , Interleucina-6/metabolismo , Leucócitos/metabolismo , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Fenótipo , Engenharia Tecidual/métodos , Fator de Necrose Tumoral alfa/metabolismo
11.
Ann Neurol ; 87(3): 466-479, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31899551

RESUMO

OBJECTIVE: Bridging the gap between experimental stroke and patients by ischemic blood probing during the hyperacute stage of vascular occlusion is crucial to assess the role of inflammation in human stroke and for the development of adjunct treatments beyond recanalization. METHODS: We prospectively observed 151 consecutive ischemic stroke patients with embolic large vessel occlusion of the anterior circulation who underwent mechanical thrombectomy. In all these patients, we attempted microcatheter aspiration of 3 different arterial blood samples: (1) within the core of the occluded vascular compartment and controlled by (2) carotid and (3) femoral samples obtained under physiological flow conditions. Subsequent laboratory analyses comprised leukocyte counting and differentiation, platelet counting, and the quantification of 13 proinflammatory human chemokines/cytokines. RESULTS: Forty patients meeting all clinical, imaging, interventional, and laboratory inclusion criteria could be analyzed, showing that the total number of leukocytes significantly increased under the occlusion condition. This increase was predominantly driven by neutrophils. Significant increases were also apparent for lymphocytes and monocytes, accompanied by locally elevated plasma levels of the T-cell chemoattractant CXCL-11. Finally, we found evidence that short-term clinical outcome (National Institute of Health Stroke Scale at 72 hours) was negatively associated with neutrophil accumulation. INTERPRETATION: We provide the first direct human evidence that neutrophils, lymphocytes, and monocytes, accompanied by specific chemokine upregulation, accumulate in the ischemic vasculature during hyperacute stroke and may affect outcome. These findings strongly support experimental evidence that immune cells contribute to acute ischemic brain damage and indicate that ischemic inflammation initiates already during vascular occlusion. Ann Neurol 2020;87:466-479.


Assuntos
Leucócitos/fisiologia , Acidente Vascular Cerebral/imunologia , Idoso , Idoso de 80 Anos ou mais , Plaquetas/fisiologia , Contagem de Células/estatística & dados numéricos , Diferenciação Celular/fisiologia , Transtornos Cerebrovasculares/sangue , Transtornos Cerebrovasculares/complicações , Quimiocinas/sangue , Feminino , Humanos , Mediadores da Inflamação/sangue , Masculino , Trombólise Mecânica , Estudos Prospectivos , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral/complicações , Resultado do Tratamento
12.
J Dairy Sci ; 103(2): 1908-1913, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31837777

RESUMO

The aim of this study was to investigate changes in the abundance of genes involved in leukocyte function between cows highly specialized for milk production (Holstein, n = 12) and cows selected for meat and milk (Simmental, n = 13). Blood was collected on d 3 after calving in PAXgene tubes (Preanalytix, Hombrechtikon, Switzerland) to measure mRNA abundance of 33 genes. Normalized gene abundance data were subjected to MIXED model ANOVA using SAS (SAS Institute Inc. Cary, NC). Simmental cows had greater transcript abundance of proinflammatory cytokines and receptor genes (IL1B, TNF, IL1R, TNFRSF1A), cell migration- and adhesion-related genes (CX3CR1, ITGB2, CD44, LGALS8), and the antimicrobial IDO1 gene. In contrast, compared with Holstein cows, Simmental cows had lower abundance of the toll-like receptor (TLR) recognition-related gene TLR2, the antimicrobial-related gene LTF, and S100A8, which is involved in cell maturation, regulation of inflammatory processes, and immune response. These results revealed that breed plays an important role in the modulation of genes involved in immune adaptation and inflammatory response, and the immune system of Simmental cows could potentially have a more acute response in early lactation. In turn, this might be beneficial for mounting a more efficient response after calving and allow for a smoother homeorhetic adaptation to lactation.


Assuntos
Bovinos/fisiologia , Comunicação Celular , Redes Reguladoras de Genes , Inflamação/veterinária , Leite/metabolismo , Animais , Cruzamento , Bovinos/genética , Bovinos/imunologia , Citocinas/metabolismo , Feminino , Lactação , Leucócitos/fisiologia
13.
Fish Physiol Biochem ; 46(2): 629-640, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31840217

RESUMO

We evaluated the immune response of pacu fed with a ß-glucan diet (0.5%) for 10 days. After the feeding period, fish were subjected to handling and 3 h after, inoculated with Aeromonas hydrophila. Fish were sampled before handling (baseline condition), 3, 6, and 24 h and 1 week after inoculation. A higher level of blood glucose was found in fish treated with ß-glucan in baseline conditions. Handling and bacterial inoculation increased the circulating levels of cortisol and glucose and promoted the acute inflammatory response (lymphopenia and neutrophilia). ß-Glucan prevented the decrease in the respiratory activity of leukocytes observed in the control group at 3 h sampling. ß-Glucan did not affect the complement and lysozyme, which were activated 24 h after the bacterial challenge in control fish. A reduction in the number of leukocytes was found in fish treated with ß-glucan 1 week after the challenge. We suggest two plausible hypotheses for this event: (1) it could be attributed to a depletion of the immune responses or (2) it could be due to a mobilization of the leukocytes to the spleen for antigen presenting/processing. In general, ß-glucan avoided the reduction of the activity of leukocytes after stress and the bacterial challenge and increased the baseline glucose levels. Our findings confirm the immunomodulatory action of glucan and add evidence showing that glucan can have a role in stress response.


Assuntos
Caraciformes/fisiologia , Leucócitos/fisiologia , beta-Glucanas/metabolismo , Aeromonas hydrophila , Animais , Glicemia , Doenças dos Peixes/microbiologia , Infecções por Bactérias Gram-Negativas/veterinária
14.
J Nutr Health Aging ; 24(1): 48-54, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31886808

RESUMO

BACKGROUND AND OBJECTIVES: Sex hormone concentrations and telomere length are age related responses of human body, while whether there is a direct relation between sex hormone and telomere length is uncertain. Therefore, we used the data of National Health and Nutrition Examination Survey (NHANES) to quantify their direct association. RESEARCH DESIGN AND METHODS: A total of 710 women aged 35-60 years and 539 men aged 20-85 years were included from two cycles of the NHANES (1999-2002). Telomere length relative to standard reference DNA (T/S ratio) was measured using quantitative polymerase chain reaction method. Seven hormones in serum (5 in men and 2 in women) were assayed. Logistic regressions were used to calculate the odds ratios to evaluate the telomere length-sex hormones association. RESULTS: Men with vigorous physical activity (71.1%) and without history of cardiovascular diseases, diabetes, and lipid-lowering drugs using tended to have a longer telomere length (all P-values < 0.05); while women with longer sedentary time, smaller pregnant or live birth, and with older ages of firth/last birth were likely with longer telomere length (all P-values < 0.05). After adjusted for potential confounders, only anti-Mullerian hormone was positively and stably associated with short leukocytes telomere length in men (OR: 1.098; 95% CI: 1.034, 1.165). We did not detect any significant association of short telomere length with sex hormones in men and women. Discussion and Implications: Serum anti-Mullerian hormone in men was positively and stably associated with telomere length. More large-scaled and well-designed prospective studies are warranted to reconfirm our conclusions.


Assuntos
Hormônios Esteroides Gonadais/análise , Homeostase do Telômero/fisiologia , Encurtamento do Telômero/fisiologia , Telômero/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/sangue , Feminino , Humanos , Leucócitos/fisiologia , Masculino , Pessoa de Meia-Idade , Inquéritos Nutricionais , Gravidez , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Comportamento Sedentário , Estados Unidos , Adulto Jovem
15.
Fertil Steril ; 113(1): 217-223, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31594634

RESUMO

OBJECTIVE: To investigate whether telomere length (TL) in granulosa cells (GC) or cumulus cells (CC) correlates with TL in leukocytes (L). DESIGN: Prospective noninterventional study. SETTING: Private assisted reproductive technology center. PATIENT(S): Thirty-five egg donors were included in the study. INTERVENTIONS(S): None. MAIN OUTCOME MEASURE(S): Average relative leukocyte telomere length (LTL), cumulus cell telomere length (CCTL), and granulosa cell telomere length (GCTL) measurements from each study subject. RESULT(S): Participants had a mean age of 25.43 ± 4.57 years, antimüllerian hormone level of 1.90 ± 0.92 ng/mL, antral follicle count of 23.29 ± 5.11, and the mean number of mature oocytes retrieved was 23.29 ± 9.13. No significant association between these variables and GCTL, CCTL, or LTL was found. In addition, no correlation was observed between TL measurements of L vs. CC, L vs. GC, or CC vs. GC. Interestingly, CCTL was significantly higher than LTL (1.54-fold), although no significant differences were found between GCTL vs. CCTL or GCTL vs. LTL. CONCLUSION(S): CC from mature follicles have significantly longer telomeres than L, suggesting that the follicular environment could possess different mechanisms to cope against telomere shortening compared with other somatic tissues. Furthermore, these data do not support the utility of telomere DNA measurement in L as an estimate of TL in follicular cells.


Assuntos
Células do Cúmulo/fisiologia , Leucócitos/fisiologia , Estudo de Prova de Conceito , Reprodução/fisiologia , Homeostase do Telômero/fisiologia , Adolescente , Adulto , Feminino , Humanos , Doação de Oócitos/métodos , Estudos Prospectivos , Telômero/fisiologia , Adulto Jovem
16.
Methods Mol Biol ; 2041: 345-349, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31646502

RESUMO

Extracellular nucleotides are potent damage-associated molecular patterns that shape the immune response to cell stress and tissue damage. These nucleotides are sensed by purinergic receptors and mediate a wide range of cellular effects. Among the best characterized of these effects is cellular migration. While the motility responses of leukocytes to nucleotides can be achieved by microscopic live-cell imaging approaches, such systems are time-consuming and require costly equipment and analysis tools not readily available to all researchers. Transwell migration chambers are a widely used alternative to microscopy due to their relatively low cost and moderate through-put capacity. However, extracellular nucleotides are labile and rapidly degraded in serum-containing cell cultures due to the presence of phosphohydrolases. Thus, evaluating leukocyte migration to nucleotides presents a number of challenges not seen with more stable classes of chemoattractants like proteins and lipids. Here we describe a method to measure leukocyte migration to nucleotides that is cost-effective, rapid and produces robust and reproducible migration of leukocytes using transwell migration chambers.


Assuntos
Movimento Celular , Fatores Quimiotáticos/metabolismo , Leucócitos/fisiologia , Monócitos/fisiologia , Nucleotídeos/metabolismo , Receptores Purinérgicos/metabolismo , Células Cultivadas , Humanos , Leucócitos/citologia , Monócitos/citologia
17.
J Vasc Res ; 57(1): 8-15, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31505501

RESUMO

OBJECTIVE: The aim of this study was to apply an innovative methodology to incident dark-field (IDF) imaging in coronary artery bypass grafting (CABG) patients for the identification and quantification of rolling leukocytes along the sublingual microcirculatory endothelium. METHODS: This study was a post hoc analysis of a prospective study that evaluated the perioperative course of the sublingual microcirculation in CABG patients. Video images were captured using IDF imaging following the induction of anesthesia (T0) and cardiopulmonary bypass (CPB) (T1) in 10 patients. Rolling leukocytes were identified and quantified using frame averaging, which is a technique that was developed for correctly identifying leukocytes. RESULTS: The number of rolling leukocytes increased significantly from T0 (7.5 [6.4-9.1] leukocytes/capillary-postcapillary venule/4 s) to T1 (14.8 [13.2-15.5] leukocytes/capillary-postcapillary venule/4 s) (p < 0.0001). A significant increase in systemic leukocyte count was also detected from 7.4 ± 0.9 × 109/L (preoperative) to 12.4 ± 4.4 × 109/L (postoperative) (p < 0.01). CONCLUSION: The ability to directly visualize leukocyte-endothelium interaction using IDF imaging facilitates the diagnosis of a systemic inflammatory response after CPB via the identification of rolling leukocytes. Integration of the frame averaging algorithm into the software of handheld vital microscopes may enable the use of microcirculatory leukocyte count as a real-time parameter at the bedside.


Assuntos
Ponte de Artéria Coronária , Endotélio/fisiologia , Leucócitos/fisiologia , Soalho Bucal/irrigação sanguínea , Idoso , Ponte Cardiopulmonar , Feminino , Humanos , Masculino , Microcirculação , Pessoa de Meia-Idade , Estudos Prospectivos
18.
Exp Cell Res ; 386(1): 111710, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31693873

RESUMO

Physiological cyclic stretch (CS), caused by artery deformation following blood pressure, plays important roles in the homeostasis of endothelial cells (ECs). Here, we detected the effect of physiological CS on endothelial microvesicles (EMVs) and their roles in leukocyte recruitment to ECs, which is a crucial event in EC inflammation. The results showed compared with the static treatment, pretreatment of 5%-CS-derived EMVs with ECs significantly decreased the adherence level of leukocytes. Comparative proteomic analysis revealed 373 proteins differentially expressed between static-derived and 5%-CS-derived EMVs, in which 314 proteins were uniquely identified in static-derived EMVs, 34 proteins uniquely in 5%-CS-derived EMVs, and 25 proteins showed obvious differences. Based on the proteomic data, Ingenuity Pathways Analysis predicted intercellular adhesion molecule 1 (ICAM1) in EMVs might be the potential molecule involved in EC-leukocyte adhesion. Western blot and flow cytometry analyses confirmed the significant decrease of ICAM1 in 5%-CS-derived EMVs, which subsequently inhibited the phosphorylation of VE-cadherin at Tyr731 in target ECs. Moreover, leukocyte adhesion was obviously decreased after pretreatment with ICAM1 neutralizing antibody. Our present research suggested that physiological stretch changes the components of EMVs, which in turn inhibits leukocyte adhesion. ICAM1 expressed on CS-induced EMVs may play an important role in maintaining EC homeostasis.


Assuntos
Adesão Celular , Micropartículas Derivadas de Células/metabolismo , Células Endoteliais/metabolismo , Molécula 1 de Adesão Intercelular/metabolismo , Leucócitos/fisiologia , Animais , Caderinas/metabolismo , Células Cultivadas , Células Endoteliais/citologia , Células Endoteliais/fisiologia , Endotélio Vascular/citologia , Leucócitos/metabolismo , Masculino , Ratos , Ratos Sprague-Dawley , Estresse Mecânico
20.
Hum Genet ; 139(3): 401-407, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31134332

RESUMO

The extent of aneuploidy of the sex chromosomes increases with age in human leukocytes. Here, we re-explore the dynamics of normal loss of the Y chromosome (LOY) with age based on microarray data using two exponential models and two different ways to estimate the fraction of LOY. This analysis shows the existence of a significant correlation between the fraction of LOY estimated from molecular cytogenetics and genotyping microarray data. Although the specific estimates of the parameters for the two exponential models are different from those derived from cytogenetics data, the present analysis in an independent dataset of normal individuals confirms that X0 cells have a selective advantage over XY cells. Moreover, patients with age-related macular degeneration display higher fraction of LOY values and seem to have a predisposition to lose their Y chromosome even at young ages compared to control individuals. As there are no data available for the same individuals at different time points, the parameters reported here are average values drawn from population analyses.


Assuntos
Envelhecimento/genética , Cromossomos Humanos Y/genética , Degeneração Macular/genética , Aneuploidia , Deleção Cromossômica , Genótipo , Humanos , Leucócitos/fisiologia , Masculino
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...