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1.
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
2.
PLoS One ; 14(12): e0224610, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31869339

RESUMO

Malaria is an infectious disease of major worldwide clinical importance that causes a variety of severe, or complicated, syndromes including cerebral malaria, which is often fatal. Leukocyte integrins are essential for host defense but also mediate physiologic responses of the innate and adaptive immune systems. We previously showed that targeted deletion of the αD subunit (αD-/-) of the αDß2 integrin, which is expressed on key leukocyte subsets in mice and humans, leads to absent expression of the integrin heterodimer on murine macrophages and reduces mortality in mice infected with Plasmodium berghei ANKA (P. berghei ANKA). To further identify mechanisms involved in the protective effect of αD deletion in this model of severe malaria we examined wild type C57BL/6 (WT) and αD-/- mice after P. berghei ANKA infection and found that vessel plugging and leukocyte infiltration were significantly decreased in the brains of αD-/- animals. Intravital microscopy demonstrated decreased rolling and adhesion of leukocytes in cerebral vessels of αD-/- mice. Flow cytometry analysis showed decreased T-lymphocyte accumulation in the brains of infected αD-/- animals. Evans blue dye exclusion assays demonstrated significantly less dye extravasation in the brains of αD-/- mice, indicating preserved blood-brain barrier integrity. WT mice that were salvaged from P. berghei ANKA infection by treatment with chloroquine had impaired aversive memory, which was not observed in αD-/- mice. We conclude that deletion of integrin αDß2 alters the natural course of experimental severe malaria, demonstrating previously unrecognized activities of a key leukocyte integrin in immune-inflammatory responses that mediate cerebral involvement.


Assuntos
Antígenos CD11/metabolismo , Cadeias alfa de Integrinas/metabolismo , Malária/fisiopatologia , Animais , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Edema Encefálico/metabolismo , Edema Encefálico/fisiopatologia , Antígenos CD11/fisiologia , Cloroquina/metabolismo , Modelos Animais de Doenças , Inflamação/metabolismo , Cadeias alfa de Integrinas/fisiologia , Integrinas/imunologia , Integrinas/metabolismo , Contagem de Leucócitos , Leucócitos/metabolismo , Leucócitos/fisiologia , Macrófagos/metabolismo , Malária/genética , Malária Cerebral/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Plasmodium berghei/metabolismo
3.
Fish Shellfish Immunol ; 94: 389-397, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31520749

RESUMO

The aim of this study was to analyze the probiotic potential, fatty acid composition and immunostimulant activities of Kluyveromyces lactis M3 isolated from a hypersaline sediment. For this purpose, K. lactis M3 resistance to different pH, salinities and bile, as well as its antioxidant capability were assayed. Furthermore, total fatty acid composition of the yeast was determined where the dominant fatty acids were palmitic, palmitoleic, oleic and linoleic acids. K. lactis M3 showed no cytotoxic effects on peripheral blood leukocytes. During an in vivo experiment in gilthead seabream (Sparus aurata), dietary K. lactis M3 supplemented at 0.55 or 1.1% of the basal diet enhanced bactericidal activity against Vibrio parahaemolyticus N16, V. harveyi Lg 16/00, and V. anguillarum CECT 43442 compared to fish fed commercial diet (control group). Finally, nitric oxide production, peroxidase activity and skin mucus lectin union levels strongly increased in fish fed K. lactis M3 with respect to the control group. The results suggested that the yeast K. lactis M3 had exhibited high antioxidant capability, and its dietary administration at 0.55 or 1% basal diet had immunostimulant activity for gilthead seabream. For all these reasons, it should be considered an appropriate probiotic candidate for the aquaculture fish industry.


Assuntos
Imunidade Inata/imunologia , Kluyveromyces/química , Muco/imunologia , Perciformes/imunologia , Probióticos/farmacologia , Pele/imunologia , Ração Animal/análise , Animais , Antibacterianos/farmacologia , Antioxidantes/metabolismo , Sobrevivência Celular , Dieta/veterinária , Ácidos Graxos/análise , Concentração de Íons de Hidrogênio , Kluyveromyces/fisiologia , Leucócitos/microbiologia , Leucócitos/fisiologia , Muco/efeitos dos fármacos , Muco/microbiologia , Distribuição Aleatória , Salinidade , Pele/efeitos dos fármacos , Pele/microbiologia
4.
Nutrients ; 11(8)2019 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-31398886

RESUMO

Cellular pathways such as inflammation or oxidative stress are the cause and triggers of disease-related malnutrition (DRM), but the influence of these markers on endoplasmic reticulum (ER) stress is unknown. The objective of this study was to analyze the relationship between mitochondrial function and ER stress parameters in a DRM population. The study population was composed of 82 outpatient subjects, of whom 45 were diagnosed with DRM and 37 were confirmed to be normonourished according to the American Society for Parenteral and Enteral Nutrition ASPEN criteria. We evaluated anthropometrical and biochemical parameters, pro-inflammatory cytokines in serum. Oxidative and ER stress markers were analyzed in leukocytes. DRM patients showed significant reductions in albumin and transferrin concerning the normonourished group, and also displayed higher levels of hsCRP, IL6, and TNFα, and the soluble adhesion molecules VCAM-1 and ICAM-1. Besides, oxygen consumption and mitochondrial membrane potential were reduced and Radical Oxygen Species ROS production was enhanced in DRM patients. In the case of ER markers, protein and mRNA expression revealed that CHOP, ATF6, and the P-eIF2α signal were enhanced in malnourished patients compared to control subjects. Correlation studies supported a relationship between weight loss and increased inflammation, mitochondrial dysfunction, and compromised function of the ER. Our results demonstrate that ER stress signaling pathways are influenced by inflammation and mitochondrial function in the leukocytes of a DRM population.


Assuntos
Estresse do Retículo Endoplasmático/fisiologia , Retículo Endoplasmático/fisiologia , Leucócitos/fisiologia , Desnutrição/fisiopatologia , Estresse Oxidativo/fisiologia , Adulto , Idoso , Antropometria , Citocinas/sangue , Feminino , Humanos , Masculino , Desnutrição/sangue , Desnutrição/etiologia , Pessoa de Meia-Idade , Estado Nutricional/fisiologia , Pacientes Ambulatoriais
5.
Arterioscler Thromb Vasc Biol ; 39(8): 1588-1601, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31294624

RESUMO

OBJECTIVE: MR (mineralocorticoid receptor) activation is associated with cardiovascular ischemia in humans. This study explores the role of the MR in atherosclerotic mice of both sexes and identifies a sex-specific role for endothelial cell (EC)-MR in vascular inflammation. Approach and Results: In the AAV-PCSK9 (adeno-associated virus-proprotein convertase subtilisin/kexin type 9) mouse atherosclerosis model, MR inhibition attenuated vascular inflammation in males but not females. Further studies comparing male and female littermates with intact MR or EC-MR deletion revealed that although EC-MR deletion did not affect plaque size in either sex, it reduced aortic arch inflammation specifically in male mice as measured by flow cytometry. Moreover, MR-intact females had larger plaques but were protected from vascular inflammation compared with males. Intravital microscopy of the mesenteric vasculature demonstrated that EC-MR deletion attenuated TNFα (tumor necrosis factor α)-induced leukocyte slow rolling and adhesion in males, while females exhibited fewer leukocyte-endothelial interactions with no additional effect of EC-MR deletion. These effects corresponded with decreased TNFα-induced expression of the endothelial adhesion molecules ICAM-1 (intercellular adhesion molecule-1) and E-selectin in males with EC-MR deletion compared with MR-intact males and females of both genotypes. These observations were also consistent with MR and estrogen regulation of ICAM-1 transcription and E-selectin expression in primary cultured mouse ECs and human umbilical vein ECs. CONCLUSIONS: In male mice, EC-MR deletion attenuates leukocyte-endothelial interactions, plaque inflammation, and expression of E-selectin and ICAM-1, providing a potential mechanism by which the MR promotes vascular inflammation. In females, plaque inflammation and leukocyte-endothelial interactions are decreased relative to males and EC-MR deletion is not protective.


Assuntos
Aterosclerose/complicações , Células Endoteliais/fisiologia , Receptores de Mineralocorticoides/fisiologia , Vasculite/etiologia , Animais , Células Cultivadas , Selectina E/genética , Feminino , Molécula 1 de Adesão Intercelular/genética , Leucócitos/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Caracteres Sexuais
6.
J Zoo Wildl Med ; 50(2): 498-502, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31260223

RESUMO

A 3.5-yr-old asymptomatic female Asian elephant (Elephas maximus) with a high load of circulating EEHV1B DNA on qPCR on a routine blood sample, showed progressive depletion of monocytes, lymphocytes, and platelets. Twice daily IV ganciclovir, plasma transfusions, and fluid therapy coincided with a decreasing viral load, which may support potential efficacy of this antiviral drug. An increase in lymphocytes followed initial treatment and preceded the onset of clinical signs. Administration of short-acting glucocorticosteroids for two consecutive days preceded a reduction of lymphocytes, recovery and maturation of monocytes, and gradually decreasing clinical signs, illustrating the potential value of glucocorticosteroids in treatment of clinical EEHV. Three subsequent subclinical episodes with high monocyte and platelet counts did not require intervention. Decision-making was led not just by quantification of viral load and clinical signs, but more specifically by interpretation of the hematological changes using easily accessible, in-house blood smear analysis.


Assuntos
Antivirais/uso terapêutico , Elefantes/sangue , Famciclovir/uso terapêutico , Infecções por Herpesviridae/veterinária , Herpesviridae/classificação , Animais , Animais de Zoológico , Diferenciação Celular , DNA Viral , Elefantes/virologia , Infecções por Herpesviridae/tratamento farmacológico , Infecções por Herpesviridae/virologia , Leucócitos/fisiologia , Carga Viral , Viremia
7.
G3 (Bethesda) ; 9(9): 2977-2987, 2019 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-31352405

RESUMO

With growing interest in monitoring mutational processes in normal tissues, tumor heterogeneity, and cancer evolution under therapy, the ability to accurately and economically detect ultra-rare mutations is becoming increasingly important. However, this capability has often been compromised by significant sequencing, PCR and DNA preparation error rates. Here, we describe FERMI (Fast Extremely Rare Mutation Identification) - a novel method designed to eliminate the majority of these sequencing and library-preparation errors in order to significantly improve rare somatic mutation detection. This method leverages barcoded targeting probes to capture and sequence DNA of interest with single copy resolution. The variant calls from the barcoded sequencing data are then further filtered in a position-dependent fashion against an adaptive, context-aware null model in order to distinguish true variants. As a proof of principle, we employ FERMI to probe bone marrow biopsies from leukemia patients, and show that rare mutations and clonal evolution can be tracked throughout cancer treatment, including during historically intractable periods like minimum residual disease. Importantly, FERMI is able to accurately detect nascent clonal expansions within leukemias in a manner that may facilitate the early detection and characterization of cancer relapse.


Assuntos
Análise Mutacional de DNA/métodos , Leucemia/genética , Algoritmos , Biópsia , Medula Óssea , Reações Falso-Positivas , Biblioteca Gênica , Humanos , Janus Quinase 2/genética , Leucemia/patologia , Leucócitos/patologia , Leucócitos/fisiologia , Mutação , Neoplasia Residual , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Nucleotídeo Único , Sensibilidade e Especificidade , Imagem Individual de Molécula/métodos
8.
J Pharmacol Sci ; 140(2): 153-161, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31253430

RESUMO

A novel AMP-activated protein kinase (AMPK) activator, IMM-H007 (H007), has been reported to reduce serum lipid levels and inhibit lipid accumulation in the liver in hyperlipidemic animal models. However, how H007 ameliorates hepatic steatosis and inflammation remains unknown. In the present study, H007, at 200 mg/kg, reduced hepatic lipid levels and the levels of collagenous fiber in the liver in high-fat diet (HFD)-fed hamsters compared to those in the HFD group. Meanwhile, compared to the controls, H007 significantly inhibited sterol-regulatory element binding protein (SREBP)-1c and acetyl CoA carboxylase (ACC) expression by upregulating the AMPK activity, suppressing the saturated fatty acid accumulation and increasing polyunsaturated fatty acid synthesis in the liver. Compared to the controls, H007 treatment inhibited the expression of monocyte chemotactic protein (MCP-1) in fatty acid-treated HepG2 cells; suppressed leukocyte adherence and rolling on the liver microvasculature; and suppressed hepatic macrophage infiltration. H007 also suppressed the expression of nuclear factor-κB (NF-κB) p65 in fatty acid- and lipopolysaccharide-treated HepG2 cells compared to that in the controls by activating AMPK. These data suggested that H007 had a beneficial effect by improving the lipid composition in the liver and inhibiting inflammatory cell trafficking in the development of nonalcoholic fatty liver disease.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Adenosina/análogos & derivados , Adesão Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Ativadores de Enzimas/administração & dosagem , Ativadores de Enzimas/farmacologia , Fígado Gorduroso/tratamento farmacológico , Fígado Gorduroso/metabolismo , Leucócitos/fisiologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Adenosina/administração & dosagem , Adenosina/farmacologia , Animais , Modelos Animais de Doenças , Células Hep G2 , Humanos , Inflamação , Masculino , Mesocricetus
9.
J Anim Physiol Anim Nutr (Berl) ; 103(5): 1578-1584, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31148265

RESUMO

Asthma is a chronic inflammatory lung disease of the airway; the incidence and prevalence of asthma remain high worldwide. Astragaloside IV (AS-IV) is the main active constituent of Astragalus membranaceus. Accumulating evidence suggests that AS-IV possesses anti-inflammatory and anti-asthmatic ability, but the potential molecular mechanism is required to further clarify. In this study, the anti-asthmatic effects of AS-IV on mice with ovalbumin (OVA)-induced allergic inflammation were analysed. We analysed airway hyperresponsiveness (AHR), numbers of inflammatory cells, inflammation situation in lung tissue and cytokines level in bronchoalveolar lavage fluid (BALF) between OVA-induced mice with and without AS-IV treatment. Moreover, we explored the possible signalling pathway behind the anti-asthmatic effects. Our results revealed that AS-IV treatment ameliorates airway inflammation and AHR in an OVA-induced asthma model. Besides, AS-IV treatment inhibits the interleukin (IL)-4, -5 and -13 production, and further study indicated that AS-IV treatment downregulates the expression level of p-JAK2/p-STAT6 proteins. Taken together, the present study suggested that the inhibitory effects of AS-IV on asthma therapy are at least partially involved in inhibiting the JAK2/STAT6 signalling pathway.


Assuntos
Asma/induzido quimicamente , Regulação da Expressão Gênica/efeitos dos fármacos , Janus Quinase 2/metabolismo , Fator de Transcrição STAT6/metabolismo , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Triterpenos/farmacologia , Animais , Asma/tratamento farmacológico , Líquido da Lavagem Broncoalveolar/citologia , Feminino , Janus Quinase 2/genética , Leucócitos/fisiologia , Masculino , Cloreto de Metacolina/toxicidade , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/toxicidade , Parassimpatomiméticos/toxicidade , Fator de Transcrição STAT6/genética
10.
Infect Immun ; 87(8)2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31109947

RESUMO

Little is known about whether pathogen invasion of neural tissue is affected by immune-based mechanisms in endothelial cells. We examined the effects of endothelial cell CD40 on Toxoplasma gondii invasion of the retina and brain, organs seeded hematogenously. T. gondii circulates in the bloodstream within infected leukocytes (including monocytes and dendritic cells) and as extracellular tachyzoites. After T. gondii infection, mice that expressed CD40 restricted to endothelial cells exhibited diminished parasite loads and histopathology in the retina and brain. These mice also had lower parasite loads in the retina and brain after intravenous (i.v.) injection of infected monocytes or dendritic cells. The protective effect of endothelial cell CD40 was not explained by changes in cellular or humoral immunity, reduced transmigration of leukocytes into neural tissue, or reduced invasion by extracellular parasites. Circulating T. gondii-infected leukocytes (dendritic cells used as a model) led to infection of neural endothelial cells. The number of foci of infection in these cells were reduced if endothelial cells expressed CD40. Infected dendritic cells and macrophages expressed membrane-associated inducible Hsp70. Infected leukocytes triggered Hsp70-dependent autophagy in CD40+ endothelial cells and anti-T. gondii activity dependent on ULK1 and beclin 1. Reduced parasite load in the retina and brain not only required CD40 expression in endothelial cells but was also dependent on beclin 1 and the expression of inducible Hsp70 in dendritic cells. These studies suggest that during endothelial cell-leukocyte interaction, CD40 restricts T. gondii invasion of neural tissue through a mechanism that appears mediated by endothelial cell anti-parasitic activity stimulated by Hsp70.


Assuntos
Encéfalo/parasitologia , Antígenos CD40/fisiologia , Células Endoteliais/imunologia , Retina/parasitologia , Toxoplasma/patogenicidade , Animais , Autofagia , Movimento Celular , Proteínas de Choque Térmico HSP70/fisiologia , Leucócitos/fisiologia , Camundongos , Camundongos Endogâmicos C57BL
11.
J Clin Lab Anal ; 33(6): e22894, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31131502

RESUMO

BACKGROUND: The confirmation of clinical diagnosis, molecular remission, and sequential minimal residual disease monitoring required PML-RARα detection in acute promyelocytic leukemia (APL). The current status of PML-RARα detection in various laboratories remains unknown. METHODS: In 2018, external quality assessment (EQA) for PML-RARα detection was carried out in China. Three EQA sample panels for PML-RARα isoform L/S/V were prepared by different mock leukocyte samples. The performances of PML-RARα detection, including admission screening, and qualitative and quantitative detection by real-time quantitative reverse transcription PCR (RT-qPCR), were assessed based on APL simulated clinical case. RESULTS: The mock leukocyte samples met the requirements of a clinically qualified sample for PML-RARα EQA panel. Among the laboratories, 13/50 (26.0%) were "competent," 21/50 (42%) classified as "acceptable," and 16/50 (32.0%) classified as "improvable." One (1/50, 2.0%) laboratory reported one screening mistake. Twenty-six (26/50, 52.0%) laboratories reported 29 false-positive and 19 false-negative results. Twenty-three (23/50, 46.0%) laboratories reported 42 quantitative incorrect results. CONCLUSION: Significant differences were not found in PML-RARα detection performance among laboratories that used different extraction methods. The performances of qualitative and quantitative RT-qPCR detection were worse accurate for PML-RARα isoform V. Quantitative variation was higher for low-level samples. Further continuous external assessment and education are needed in the management of PML-RARα detection.


Assuntos
Técnicas de Laboratório Clínico/normas , Leucemia Promielocítica Aguda/genética , Proteínas de Fusão Oncogênica/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , China , Humanos , Leucócitos/fisiologia , Proteínas de Fusão Oncogênica/sangue , Isoformas de Proteínas/genética , Controle de Qualidade , RNA Ribossômico 23S
12.
Skin Pharmacol Physiol ; 32(4): 192-200, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31096247

RESUMO

BACKGROUND: Atopic diseases constitute a major health challenge for industrialized countries, and elevated levels of interleukin 4 (IL-4) frequently characterize these disorders. Previous in vitroanalyses have indicated that IL-4 strongly upregulates the expression of IL-4-sensitive genes in human monocytes. OBJECTIVE: To explore whether similar expression alterations may contribute to the pathomechanisms of atopic diseases in vivo we carried out a small-scale case-control clinical study (n = 43), in which we quantified the plasma levels of IgE and IL-4 as well as the expression of selected IL-4-sensitive genes in blood leukocytes. METHODS: 34 allergic patients suffering from allergic rhinitis (n = 11), atopic eczema (n = 11) and allergic asthma (n = 12) as well as 9 healthy control individuals were recruited. IgE and IL-4 plasma levels were determined by ELISA, and the expression of selected IL-4-sensitive gene products in blood leukocytes was quantified by qRT-PCR. In addition, the fatty acid oxygenase activity of isolated monocytes was measured by RP-HPLC analysis of the arachidonic acid oxygenation products (ex vivo activity assays). RESULTS: We found that plasma levels of IgE and IL-4 were significantly elevated in atopic patients but the degree of elevation was not sufficient to upregulate the expression of the selected IL-4-sensitive genes in circulating leukocytes. Moreover, the arachidonic acid oxygenase activity of blood monocytes was not significantly altered in atopic patients. CONCLUSION: Our data suggest that the IL-4 plasma levels of atopic patients are not high enough to impact the expression of IL-4-sensitive genes.


Assuntos
Hipersensibilidade Imediata/sangue , Hipersensibilidade Imediata/genética , Imunoglobulina E/biossíntese , Interleucina-4/biossíntese , Leucócitos/fisiologia , Adulto , Asma/sangue , Asma/genética , Estudos de Casos e Controles , Dermatite Atópica/sangue , Dermatite Atópica/genética , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Oxigenases/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Rinite Alérgica/sangue , Rinite Alérgica/genética , Regulação para Cima
13.
Biomed Res ; 40(2): 57-66, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30982801

RESUMO

The basal lamina of the villous epithelium in the small intestine has numerous fenestrations, which are produced by leukocytes for their intraepithelial migration. We previously showed that these fenestrations change due to the dynamics of migrating leukocytes in response to dietary conditions and suggested the possibility that this change is related to the regulation of the absorption of large-sized nutrients such as chylomicrons. The present study was, thus, designed to investigate structural changes in basal lamina fenestrations in response to a high-fat diet. The ultrastructure of the intestinal villi in the rat upper jejunum was investigated by electron microscopy of tissue sections in both the normal and the high-fat diet groups, and the fenestrations in the villous epithelium of rat upper jejunum were studied by scanning electron microscopy of osmium macerated/ ultrasonicated tissues. The present study showed that free cells adhering to the fenestrations increased in the upper jejunum two hours after feeding high-fat diet and the size of the fenestrations in this region also increased after feeding high-fat diet for 2 days. This enlargement of fenestrations may play an important role in increasing the efficiency of lipid absorption by facilitating the movement of chylomicrons from the intercellular space to the lamina propria.


Assuntos
Membrana Basal/ultraestrutura , Movimento Celular/fisiologia , Dieta Hiperlipídica , Mucosa Intestinal/ultraestrutura , Jejuno/ultraestrutura , Membrana Mucosa/ultraestrutura , Animais , Membrana Basal/metabolismo , Transporte Biológico/fisiologia , Quilomícrons/metabolismo , Absorção Intestinal/fisiologia , Mucosa Intestinal/metabolismo , Jejuno/metabolismo , Leucócitos/fisiologia , Leucócitos/ultraestrutura , Masculino , Microscopia Eletrônica de Varredura , Microtomia , Membrana Mucosa/metabolismo , Ratos , Ratos Wistar , Fixação de Tecidos/métodos
14.
Dev Cell ; 49(2): 206-219.e7, 2019 04 22.
Artigo em Inglês | MEDLINE | ID: mdl-30930167

RESUMO

Cell polarization is important for various biological processes. However, its regulation, particularly initiation, is incompletely understood. Here, we investigated mechanisms by which neutrophils break their symmetry and initiate their cytoskeleton polarization from an apolar state in circulation for their extravasation during inflammation. We show here that a local increase in plasma membrane (PM) curvature resulting from cell contact to a surface triggers the initial breakage of the symmetry of an apolar neutrophil and is required for subsequent polarization events induced by chemical stimulation. This local increase in PM curvature recruits SRGAP2 via its F-BAR domain, which in turn activates PI4KA and results in PM PtdIns4P polarization. Polarized PM PtdIns4P is targeted by RPH3A, which directs PIP5K1C90 and subsequent phosphorylated myosin light chain polarization, and this polarization signaling axis regulates neutrophil firm attachment to endothelium. Thus, this study reveals a mechanism for the initiation of cell cytoskeleton polarization.


Assuntos
Polaridade Celular/fisiologia , Neutrófilos/fisiologia , Actinas/metabolismo , Animais , Adesão Celular , Membrana Celular/metabolismo , Membrana Celular/fisiologia , Movimento Celular/fisiologia , Junções Célula-Matriz , Citoesqueleto/metabolismo , Endotélio/metabolismo , Feminino , Proteínas Ativadoras de GTPase/metabolismo , Proteínas Ativadoras de GTPase/fisiologia , Células HEK293 , Humanos , Leucócitos/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Antígenos de Histocompatibilidade Menor/metabolismo , Cadeias Leves de Miosina/metabolismo , Neutrófilos/metabolismo , Fosfatos de Fosfatidilinositol/metabolismo , Fosforilação , Fosfotransferases (Aceptor do Grupo Álcool)/metabolismo , Transdução de Sinais
15.
Curr Med Sci ; 39(2): 343-348, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31016508

RESUMO

Since X-linked chronic granulomatosis disease (X-CGD) exhibits no specific clinical symptoms at an early stage, early diagnosis is difficult and depends predominantly on neonatal screening. Therefore, the aim of this study was to explore routine biomarkers for X-CGD in children and provide clues for early diagnosis. The cases of 10 children with X-CGD diagnosed at Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from 2013 to 2016 and 122 Chinese children with X-CGD reported in the literature were summarized. Serum biomarkers and clinical symptoms at acute infection were organized. A total of 132 children with X-CGD were enrolled in this study. For 55.8% of the patients, the diagnosis was delayed more than one year after the onset of the first symptoms because no typical clinical symptoms manifested. Children with X-CGD at an acute infection stage showed three recurrent signs in terms of serum biomarkers: (1) the total number of white blood cells (especially N%) was increased significantly, accompanied by anemia in some cases; (2) C-reactive protein (CRP) levels were increased significantly; and (3) most of the patients exhibited very high serum IgG levels (>12 g/L). Diagnosis of X-CGD at an early age is difficult because of its nonspecific clinical features. Our study suggested children with X-CGD suffering acute infection show increases in three typical serum biomarkers, which can provide clues for early diagnosis.


Assuntos
Biomarcadores/sangue , Doença Granulomatosa Crônica/sangue , Doença Granulomatosa Crônica/diagnóstico , Proteína C-Reativa/metabolismo , Pré-Escolar , Doença Crônica , Diagnóstico Precoce , Feminino , Humanos , Imunoglobulina G/sangue , Lactente , Leucócitos/fisiologia , Masculino
16.
Fish Shellfish Immunol ; 89: 378-383, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30978448

RESUMO

Viral replicon particles are single-cycle viruses defective for function(s) needed for viral replication, which allow them to be recognized as a safer form for the vaccination of animals compared to attenuated live viruses. However, deletion of genes that are critical for the induction of protective immunity can diminish the vaccine potential of viral replicon particles. Therefore, the manipulation of viral replicon particles to produce a molecular adjuvant can be a way to increase immunogenicity of vaccines based on viral replicon particles. Chemokines are a class of chemotactic cytokines that control the migration of diverse cells of vertebrates. CXC chemokine ligand 12 (CXCL12) binds to a receptor CXCR4, and CXCL12-CXCR4 signaling plays an important role in the migration of hematopoietic cells during embryogenesis and the attraction of leukocytes. In the present study, to evaluate the possible use of CXCL12 as a molecular adjuvant for an rVHSV-ΔG vaccine and to know differences between CXCL12a and CXCL12b in the adjuvant ability, we rescued VHSV replicon particles that are expressing olive flounder CXCL12a, CXCL12b, or eGFP (rVHSV-ΔG-CXCL12a, rVHSV-ΔG-CXCL12b, or rVHSV-ΔG-eGFP), and compared the ability to attract olive flounder leucocytes and to induce protection against a VHSV challenge. In the leukocytes migration assay, supernatants collected from cells infected with rVHSV-ΔG-CXCL12a and rVHSV-ΔG-CXCL12b showed significantly higher ability to attract olive flounder leukocytes than the supernatant of cells infected with rVHSV-ΔG-eGFP. Moreover, the significantly higher number of leukocytes were attracted to rVHSV-CXCL12a supernatant compared to rVHSV-CXCL12b supernatant, suggesting that CXCL12a would be more appropriate for the induction of immunity than CXCL12b in olive flounder. In the immunization experiment, olive flounder immunized with rVHSV-ΔG-CXCL12a showed significantly higher survival rate than fish immunized with rVHSV-ΔG-CXCL12b or rVHSV-ΔG-eGFP. In addition, fish immunized with rVHSV-ΔG-CXCL12a showed the highest serum neutralization activity. These results suggest the availability of CXCL12a for a molecular adjuvant of vaccines based on VHSV replicon particles.


Assuntos
Quimiocina CXCL12/imunologia , Doenças dos Peixes/prevenção & controle , Proteínas de Peixes/imunologia , Linguados/imunologia , Septicemia Hemorrágica Viral/prevenção & controle , Novirhabdovirus/imunologia , Vacinas Virais/administração & dosagem , Animais , Movimento Celular , Doenças dos Peixes/imunologia , Doenças dos Peixes/virologia , Septicemia Hemorrágica Viral/imunologia , Septicemia Hemorrágica Viral/virologia , Leucócitos/imunologia , Leucócitos/fisiologia , Distribuição Aleatória , Replicon/imunologia , Vacinas Virais/imunologia
17.
Res Vet Sci ; 124: 328-333, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31035221

RESUMO

Telomeres are short and repetitive sequences at the ends of linear chromosomes which shorten with every cell-division in vitro. Telomere length (TL) is reported to decrease with age and stress. The domesticated water buffalo (Bubalus bubalis) is the second most important milk producing animal worldwide. The productive lifespan of water buffalo cows is reported to be longer than that of dairy cows (Bos taurus). With this background, we aimed to compare TL in leukocytes obtained from blood samples from water buffaloes across different ages. In addition, we tested the suitability of assessing TL in DNA derived from nasal and vaginal epithelial cells via swabs as potential non-invasive alternatives to blood sampling Samples were collected from 20 calves (3 months of age), 20 heifers (2 years old), 20 cows (1st lactation, 3 years old), and 13 cows (3rd lactation, about 5 years old). We found that TL in leukocytes from water buffalo calves, heifers, and from cows in their first lactation was not different, but shorter telomeres were observed in cows in their third lactation. The results thus support an age-dependent decrease of TL in water buffaloes. Leukocyte TL was weakly correlated with TL measured in DNA from nasal epithelial cells (r = 0.327; P = .025), but not with TL from vaginal epithelial cells. Due to the poor correlation between epithelial cell and leukocyte TL and to the difficulties with collecting nasal swabs, we conclude that they are no suitable alternatives to blood samples for telomere studies in water buffaloes.


Assuntos
Búfalos/fisiologia , Células Epiteliais/fisiologia , Leucócitos/fisiologia , Mucosa Nasal/fisiologia , Encurtamento do Telômero , Vagina/fisiologia , Animais , Feminino , Homeostase do Telômero
18.
Toxicon ; 164: 1-9, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30902683

RESUMO

Scorpion envenomation has been considered a public health issue around the world. Tityus serrulatus represents a specie of major medical importance in Brazil due to mortality rates of approximately 1% among children and elderly populations. The aim of this work was to evaluate the in vivo anti-inflammatory potential of aqueous extract from Hancornia speciosa fruits, its fractions and its phenolic compounds against T. serrulatus envenomation. After receiving the T. serrulatus venom (TsV, 0.8 mg/kg) intraperitoneally, the animals were treated intravenously with the aqueous extract (20, 30 and 40 mg/kg), the arachnid antivenom (50 µL/animal), the dichloromethane, ethyl acetate and n-butanol fractions (20 mg/kg) as well as rutin and chlorogenic acid (2, 2.5 and 5 mg/kg). The treatment with the aqueous extract, fractions and phenolic compounds decreased the migration of leukocytes to the peritoneal cavity and reduced the levels of IL-1ß, IL-6 and IL-12. Moreover, the pulmonary histopathologic analysis showed a reduction in both interstitial and alveolar edema, as well as in the leukocytes infiltration and vascular ectasia in the mice's lungs, which evidences a protective effect attributed to H. speciosa. This is the first study that demonstrates the inhibitory potential of the aqueous extract from H. speciosa fruits against inflammation induced by TsV. These findings suggest that the bioactive compounds from the aqueous extract, especially chlorogenic acid and rutin, are responsible for the reported anti-inflammatory activity of H. speciosa.


Assuntos
Anti-Inflamatórios/farmacologia , Apocynaceae/química , Fenóis/farmacologia , Extratos Vegetais/farmacologia , Pneumonia/tratamento farmacológico , Venenos de Escorpião/toxicidade , Animais , Anti-Inflamatórios/uso terapêutico , Antivenenos/farmacologia , Movimento Celular , Ácido Clorogênico/farmacologia , Feminino , Frutas/química , Leucócitos/efeitos dos fármacos , Leucócitos/fisiologia , Pulmão/efeitos dos fármacos , Pulmão/patologia , Masculino , Camundongos Endogâmicos BALB C , Fenóis/uso terapêutico , Extratos Vegetais/uso terapêutico , Pneumonia/induzido quimicamente , Pneumonia/patologia , Edema Pulmonar/induzido quimicamente , Edema Pulmonar/tratamento farmacológico , Edema Pulmonar/patologia , Rutina/farmacologia
19.
PLoS One ; 14(3): e0212273, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30840638

RESUMO

BACKGROUND: HIV-mediated inflammation and immune activation can accelerate telomere attrition. In addition, antiretrovirals can inhibit telomerase, possibly shortening telomeres. We examined the longitudinal dynamics of leukocyte telomere length (LTL) during pregnancy in a unique cohort of women living with HIV (WLWH) treated with combination antiretroviral therapy (cART), and HIV-negative control women. METHODS: Blood was collected at three visits during pregnancy, at 13-23, >23-30, and >30-40 weeks of gestation, and for WLWH only, at 6 weeks post-partum. LTL was measured by qPCR and both cross-sectional and longitudinal (MANOVA) models were used to examine possible predictors of LTL among participants who attended all three visits during pregnancy. RESULTS: Among WLWH (n = 64) and HIV-negative women (n = 41), within participant LTL were correlated throughout pregnancy (p<0.001). LTL was shorter among WLWH at first visit, but this difference waned by the second visit. WLWH who discontinued cART post-partum experienced a decrease in LTL. Longitudinally, LTL was similar in both groups and increased as gestation progressed, a change that was more pronounced among women under 35 years. Among WLWH, both smoking throughout pregnancy (p = 0.04) and receiving a ritonavir-boosted protease inhibitor-based regimen (p = 0.03) were independently associated with shorter LTL. CONCLUSIONS: LTL increases as pregnancy progresses; the reasons for this are unknown but may relate to changes in blood volume, hormones, and/or cell subset distribution. While our observations need confirmation in an independent cohort, our data suggest that although some cART regimens may influence LTL, being on cART appears overall protective and that stopping cART post-partum may negatively impact LTL. The effect of smoking on LTL is clearly negative, stressing the importance of smoking cessation.


Assuntos
Infecções por HIV/genética , Encurtamento do Telômero/genética , Telômero/genética , Adolescente , Adulto , Estudos Transversais , Feminino , Humanos , Leucócitos/fisiologia , Estudos Longitudinais , Análise Multivariada , Gravidez , Estudos Prospectivos , Fumar/efeitos adversos , Telomerase/genética , Adulto Jovem
20.
Fish Shellfish Immunol ; 89: 12-17, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30890431

RESUMO

Diazinon (DZN) is an organophosphate pesticide characterized by inhibiting the enzyme acetylcholinesterase (AChE) (E.C. 3.1.1.7), affecting the nervous system. There is currently enough evidence proving this pesticide also affects the immune response; however, the immunotoxicity mechanisms through which these substances exerts toxic effects remain unclear. For that reason, this work evaluated the effect of diazinon on the intracellular calcium flux, ERK1/2 phosphorylation (pERK1/2), apoptosis, senescence, and mitochondrial membrane potential (ΔΨm) in spleen mononuclear cells (SMNC) of Nile tilapia, a teleost fish of commercial and ecological relevance. The results obtained indicate that diazinon causes significant damage in all evaluated parameters, which play an essential role in intracytoplasmic signaling of immune cells, suggesting these signal pathways could be related with the immunotoxicity mechanism of these type of pesticides.


Assuntos
Apoptose/efeitos dos fármacos , Ciclídeos/fisiologia , Diazinon/efeitos adversos , Inseticidas/efeitos adversos , Leucócitos/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Poluentes Químicos da Água/efeitos adversos , Animais , Cálcio/metabolismo , Senescência Celular/efeitos dos fármacos , Leucócitos/fisiologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Baço/efeitos dos fármacos , Baço/fisiologia
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