RESUMO
Severe corticosteroid-refractory graft-versus-host-disease (GVHD) is a major non-relapse cause of mortality and morbidity after an allogeneic hematopoietic stem cell transplantation (allo-HSCT). One of the most promising treatment options is using advanced therapy medicinal products based on mesenchymal stem cells (MSCs) immunomodulation ability. The protocols of MSC application differ in many parameters including a source of MSC, a dose, a number of doses or way of preparation of the medicinal product. The process is limited by the need for laborious and expensive manufacturing processes fraught with batch-to-batch variability. In our study, we compared the immunomodulatory effects of different MSC batches versus pooled MSC, specifically the influence on lymphocyte proliferation, the metabolic activity, and the expression of activation markers on T cells. Our goal was to determine whether the effect depends on donor-to-donor heterogeneity and if pooling of MSCs could increase their immunomodulatory ability. All tested batches showed an immunomodulatory effect, with no significant differences between the groups. Our study suggests that immunosuppressive potential is comparable in single batches and pooled products, and the use of products got from individual donors is suitable to treat corticosteroid-refractory GVHD.
Assuntos
Imunomodulação , Células-Tronco Mesenquimais/imunologia , Proliferação de Células , Separação Celular , Células Cultivadas , Técnicas de Cocultura , Doença Enxerto-Hospedeiro/etiologia , Doença Enxerto-Hospedeiro/imunologia , Doença Enxerto-Hospedeiro/terapia , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Humanos , Ativação Linfocitária , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Linfócitos T/citologia , Linfócitos T/imunologia , Doadores de TecidosRESUMO
A novel sampling scheme, using a combination of electrofishing, visual exploration by scuba divers, two types of fyke nets and longlines, was tested in four reservoirs (including their inlets and outlets) to monitor a population of burbot Lota lota. This was supplemented by fry trawling and vertical hydro-acoustics, to detect L. lota larvae in two deep reservoirs that have had a long-term stocking programme. The majority of the L. lota detected were juveniles, captured by electrofishing in the littoral zones of the reservoirs and in running waters. Older individuals were rarely captured with longlines or fyke nets in deeper zones or structured habitats within the reservoirs. A combination of multiple sampling methods provided an assessment of the whole population. Population establishment could not be demonstrated as the age structure of the sampled fish corresponded with that of the stocked fish. Low post-stocking survival, migratory behaviour, interactions with other species and warmer water temperatures are considered the potential drivers for unsuccessful establishment of L. lota populations in these reservoirs.
Assuntos
Conservação dos Recursos Naturais , Gadiformes/fisiologia , Animais , República Tcheca , Espécies em Perigo de Extinção , Pesqueiros , Larva/fisiologia , Densidade Demográfica , Dinâmica PopulacionalRESUMO
In this work, a novel force equilibrium method called distributed dielectrophoretic cytometry (2DEP cytometry) was developed. It uses a dielectrophoresis (DEP)-induced vertical translation of live cells in conjunction with particle image velocimetry (PIV) in order to measure probabilistic distribution of DEP forces acting on an entire cell population. The method is integrated in a microfluidic device. The bottom of the microfluidic channel is lined with an interdigitated electrode array. Cells passing through the micro-channel are acted on by sedimentation forces, while DEP forces either oppose sedimentation, support sedimentation, or neither, depending on the dielectric (DE) signatures of the cells. The heights at which cells stabilize correspond to their DE signature and are measured indirectly using PIV, which enables simultaneous and high-throughput collection of hundreds of single-cell responses in a single PIV frame. The system was validated using polystyrene micro-particles. Preliminary experimental data quantify the DE signatures of immortalized myelogenous leukemia cell lines K562 and KG1. We show DEP-induced cell translation along the parabolic velocity profile can be measured by PIV with sub-micron precision, enabling identification of individual cell DE signatures. DE signatures of the selected cell lines are distinguishable. Throughput of the method enables measurement of DE signatures at 10 different frequencies in almost real time.
Assuntos
Citometria de Fluxo/instrumentação , Citometria de Fluxo/métodos , Simulação por Computador , Estimulação Elétrica , Eletroforese/instrumentação , Desenho de Equipamento , Humanos , Células K562 , Dispositivos Lab-On-A-Chip , Processos EstocásticosRESUMO
Mesenchymal stem cells (MSCs) have been reported to improve survival of cardiomyocytes (CMCs) and overall regeneration of cardiac tissue. Despite promising preclinical results, interactions of MSCs and CMCs, both direct and indirect, remain unclear. In this study, porcine bone marrow MSCs and freshly isolated porcine primary adult CMCs were used for non-contact co-culture experiments. Morphology, viability and functional parameters of CMCs were measured over time and compared between CMCs cultured alone and CMCs co-cultured with MSCs. In non-contact co-culture, MSCs improved survival of CMCs. CMCs co-cultured with MSCs maintained CMCs morphology and viability in significantly higher percentage than CMCs cultured alone. In viable CMCs, mitochondrial respiration was preserved in both CMCs cultured alone and in CMCs co-cultured with MSCs. Comparison of cellular contractility and calcium handling, measured in single CMCs, revealed no significant differences between viable CMCs from co-culture and CMCs cultured alone. In conclusion, non-contact co-culture of porcine MSCs and CMCs improved survival of CMCs with a sufficient preservation of functional and mitochondrial parameters.
Assuntos
Células-Tronco Mesenquimais/fisiologia , Mitocôndrias/fisiologia , Miócitos Cardíacos/fisiologia , Fatores Etários , Animais , Sobrevivência Celular/fisiologia , Técnicas de Cocultura/métodos , Citometria de Fluxo/métodos , SuínosRESUMO
Mesenchymal stromal cells (MSC) represent a promising treatment of graft-versus-host disease (GVHD) in patients after allogeneic haematopoietic stem cell transplantation. We performed co-cultivation experiments with non-specifically stimulated lymphocytes to characterize the immunosuppressive activity of MSC. MSC influenced expression of some activation antigens. CD25 expression was lower with MSC and reached 55.2 % vs. 84.9 % (CD4+, P = 0.0006) and 38.8 % vs. 86.6 % (CD8+, P = 0.0003) on day +4. Conversely, CD69 antigen expression remained higher with MSC (73.3 % vs. 56.8 %, P = 0.0009; 59.5 % vs. 49.7 %, ns) and its down-regulation along with the culture time was less pronounced. MSC reduced proliferation of the stimulated lymphocytes. The cell percentages detected in daughter generations were decreased (32.82 % vs. 10.68 % in generation 4, P = 0.0004 and 29.85 % vs. 10.09 % in generation 5, P = 0.0008), resulting in a lower proliferation index with MSC (1.84 vs. 3.65, P < 0.0001). The addition of MSC affected expression of some cytokines. Production of pro-inflammatory cytokines was decreased: IL-6 (19.5 vs. 16.3 MFI; P < 0.0001 in CD3+/CD4+ and 14.5 vs. 13.2 MFI; P = 0.0128 in CD3+/CD8+), IFN-γ (13.5 vs. 12.0 MFI; P = 0.0096 in CD3+/CD4+). Expression of anti-inflammatory IL-10 was only slightly increased after the addition of MSC (ns). The analysis confirmed the immunomodulatory activity of MSC. The functional tests have proved to be an important part of the quality control of the advanced therapy cellular product intended for GVHD treatment. Future research should focus on the interaction between MSC and the patient immune environment more closely.
Assuntos
Imunomodulação , Células-Tronco Mesenquimais/imunologia , Controle de Qualidade , Linfócitos T , Antígenos/genética , Proliferação de Células , Citocinas/metabolismo , Regulação da Expressão Gênica , Humanos , Ativação Linfocitária , Linfócitos T/citologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Transplante HomólogoRESUMO
OBJECTIVES: Obesity is a frequent metabolic disorder but an effective therapy is still scarce. Anorexigenic neuropeptides produced and acting in the brain have the potential to decrease food intake and ameliorate obesity but are ineffective after peripheral application. We have designed lipidized analogs of prolactin-releasing peptide (PrRP), which is involved in energy balance regulation as demonstrated by obesity phenotypes of both PrRP- and PrRP-receptor-knockout mice. RESULTS: Lipidized PrRP analogs showed binding affinity and signaling in PrRP receptor-expressing cells similar to natural PrRP. Moreover, these analogs showed high binding affinity also to anorexigenic neuropeptide FF-2 receptor. Peripheral administration of myristoylated and palmitoylated PrRP analogs to fasted mice induced strong and long-lasting anorexigenic effects and neuronal activation in the brain areas involved in food intake regulation. Two-week-long subcutaneous administration of palmitoylated PrRP31 and myristoylated PrRP20 lowered food intake, body weight and improved metabolic parameters, and attenuated lipogenesis in mice with diet-induced obesity. CONCLUSIONS: Our data suggest that the lipidization of PrRP enhances stability and mediates its effect in central nervous system. Strong anorexigenic and body-weight-reducing effects make lipidized PrRP an attractive candidate for anti-obesity treatment.
Assuntos
Fármacos Antiobesidade/farmacologia , Lipídeos/química , Obesidade/prevenção & controle , Hormônio Liberador de Prolactina/farmacologia , Animais , Regulação do Apetite , Ingestão de Alimentos , Metabolismo Energético , Meia-Vida , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Hormônio Liberador de Prolactina/análogos & derivados , Transdução de SinaisRESUMO
Expression of doublecortin (DCX), a 43-53 kDa microtubule binding protein, is frequently used as (i) an early neuronal marker to identify the stage of neuronal maturation of in vivo grafted neuronal precursors (NSCs), and (ii) a neuronal fate marker transiently expressed by immature neurons during development. Reliable identification of the origin of DCX-immunoreactive cells (i.e., host vs. graft) requires detailed spatial and temporal mapping of endogenous DCX expression at graft-targeted brain or spinal cord regions. Accordingly, in the present study, we analyzed (i) the time course of DCX expression in pre- and postnatal rat and porcine spinal cord, and (ii) the DCX expression in spinally grafted porcine-induced pluripotent stem cells (iPS)-derived NSCs and human embryonic stem cell (ES)-derived NSCs. In addition, complementary temporospatial GFAP expression study in porcine spinal cord was also performed. In 21-day-old rat fetuses, an intense DCX immunoreactivity distributed between the dorsal horn (DH) and ventral horn was seen and was still present in the DH neurons on postnatal day 20. In animals older than 8 weeks, no DCX immunoreactivity was seen at any spinal cord laminae. In contrast to rat, in porcine spinal cord (gestational period 113-114 days), DCX was only expressed during the pre-natal period (up to 100 days) but was no longer present in newborn piglets or in adult animals. Immunohistochemical analysis was confirmed with a comparable expression profile by western blot analysis. Contrary, the expression of porcine GFAP started within 70-80 days of the pre-natal period. Spinally grafted porcine iPS-NSCs and human ES-NSCs showed clear DCX expression at 3-4 weeks postgrafting. These data indicate that in spinal grafting studies which employ postnatal or adult porcine models, the expression of DCX can be used as a reliable marker of grafted neurons. In contrast, if grafted neurons are to be analyzed during the first 4 postnatal weeks in the rat spinal cord, additional markers or grafted cell-specific labeling techniques need to be employed to reliably identify grafted early postmitotic neurons and to differentiate the DCX expression from the neurons of the host.
Assuntos
Células-Tronco Embrionárias/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Células-Tronco Pluripotentes Induzidas/metabolismo , Proteínas Associadas aos Microtúbulos/biossíntese , Neuropeptídeos/biossíntese , Medula Espinal/metabolismo , Transplante de Células-Tronco/tendências , Animais , Células Cultivadas , Proteínas do Domínio Duplacortina , Proteína Duplacortina , Células-Tronco Embrionárias/transplante , Feminino , Humanos , Células-Tronco Pluripotentes Induzidas/transplante , Neurogênese/fisiologia , Gravidez , Ratos , Ratos Wistar , Especificidade da Espécie , Medula Espinal/crescimento & desenvolvimento , Suínos , Fatores de TempoRESUMO
Catecholamines participate in the food intake regulation, however, there are no literature data available, dealing with the activity of tyrosine hydroxylase (TH) neurons in response to stimulation or inhibition of GHS-R (growth hormone secretagogue receptor) in the hypothalamic arcuate nucleus (ARC). The present study was focused to reveal whether [Dpr(N-octanoyl) 3ghrelin], a stable GHS-R agonist, itself in doses of 5 or 10 mg/kg (s.c.) or in combination with GHS-R receptor antagonist ([DLys3]GHRP-6) in dose of 10 mg/kg (s.c.), may affect the activity of ARC TH-containing neurons in C57BL/6 male mice fed either with standard (SD) or high fat diet (HFD) that developed a diet-induced obesity (DIO). The data of the present study clearly indicate that both doses of GHS-R agonist stimulated food intake in SD mice and GHS-R antagonist significantly reduced GHS-R agonist orexinergic effect in SD mice and suppressed the voluntary food intake in HFD mice. Both doses of the GHS-R agonist stimulated Fos expression in ARC neurons in both diet groups of mice which was not abolished by GHS-R antagonist pretreatment. Moreover, both doses of the GHS-R agonist significantly influenced the activation of TH neurons in the ARC of SD mice. The GHS-R antagonist also significantly increased TH neurons activation after GHS-R agonist although this effect was less powerful in HFD mice. This is the first study demonstrating response of local ARC TH neurons to peripherally applied GHS-R agonist and antagonist. The present data point out that the response of TH neurons to GHS-R agonist and antagonist is different in normal and DIO mice and extend our knowledge about the further ARC neuronal phenotype responding to peripheral ghrelin. To bring insight into the understanding of the functional significance of the activated TH neurons in ARC, in the context of the ghrelin peripheral increase, further studies are required.
Assuntos
Núcleo Arqueado do Hipotálamo/metabolismo , Dieta Hiperlipídica , Receptores de Grelina/agonistas , Receptores de Grelina/antagonistas & inibidores , Tirosina 3-Mono-Oxigenase/metabolismo , Animais , Núcleo Arqueado do Hipotálamo/efeitos dos fármacos , Masculino , Camundongos Endogâmicos C57BL , Neurônios/efeitos dos fármacos , Peptídeos/farmacologia , Proteínas Proto-Oncogênicas c-fos/metabolismoRESUMO
Patients with obesity and type 2 diabetes often display high levels of the anti-diabetic factor fibroblast growth factor-21 (FGF21), suggesting that the overproduction of FGF21 may result from increased adiposity in an attempt by white adipose tissue (WAT) to counteract insulin resistance. However, the production of FGF21 diabetes in the absence of WAT has not been examined. In this study, we investigated the effects of lipodystrophy in A-ZIP F-1 mice on FGF21 production in relation to diabetes. A-ZIP F-1 mice displayed high FGF21 plasma levels resulting from enhanced FGF21 mRNA expression in the liver. Concomitant enhancement of FGF21 receptor (FGFR1) and glucose transporter 1 (GLUT-1) mRNA expression was observed in the muscles of A-ZIP F-1 mice. Furthermore, the activation of hypothalamic NPY and AgRP mRNA expression positively correlated with plasma levels of FGF21 but not active ghrelin. Our study demonstrates that an increased FGF21 plasma level in lipodystrophic A-ZIP F-1 mice results mainly from up-regulated liver production but does not suffice to overcome the lipodystrophy-induced severe type 2-diabetes and insulin resistance in the liver linked to the augmented liver fat deposition.
Assuntos
Diabetes Mellitus Experimental/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Resistência à Insulina , Lipodistrofia/metabolismo , Fígado/metabolismo , RNA Mensageiro/biossíntese , Tecido Adiposo Marrom/metabolismo , Animais , Fatores de Crescimento de Fibroblastos/biossíntese , Fatores de Crescimento de Fibroblastos/sangue , Hipotálamo/metabolismo , Camundongos , Neuropeptídeos/metabolismo , Pâncreas/metabolismo , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Baço/metabolismoRESUMO
The only peripherally released orexigenic hormone, ghrelin, plays a key role in food intake and body weight regulation. Antagonizing the ghrelin receptor, GHS-R1a, represents a promising approach for anti-obesity therapy. In our study, two novel GHS-R1a antagonists JMV4208 and JMV3002, which are trisubstituted 1,2,4-triazoles, decreased food intake in fasted lean mice in a dose-dependent manner, with ED50 values of 5.25 and 2.05 mg/kg, respectively. Both compounds were stable in mouse blood, with half-lives of 90 min (JMV4208) and 60 min (JMV3002), and disappeared from the blood 8h after administration. Fourteen days of treatment with the ghrelin antagonists (20 mg/kg twice a day) decreased food intake, body weight and adipose tissue mass in mice with diet-induced obesity (DIO). These results are likely attributable to an impact on food intake reduction and an attenuated expression of the lipogenesis-promoting enzymes (acetyl-CoA carboxylase 1 in subcutaneous fat and fatty acid synthase in subcutaneous and intraperitoneal fat). The decrease in fat mass negatively impacted circulating leptin levels. These data suggest that JMV4208 and JMV3002 could be useful therapeutic agents for the treatment of obesity.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Ácidos Picolínicos/farmacologia , Receptores de Grelina/antagonistas & inibidores , Triazóis/farmacologia , Animais , Relação Dose-Resposta a Droga , Camundongos , Camundongos Endogâmicos C57BL , Ácidos Picolínicos/química , Triazóis/químicaRESUMO
Ghrelin and agonists of its receptor GHS-R1a are potential substances for the treatment of cachexia. In the present study, we investigated the acute and long term effects of the GHS R1a agonist JMV 1843 (H Aib-DTrp-D-gTrp-CHO) on food intake, body weight and metabolic parameters in lean C57BL/6 male mice. Additionally, we examined stability of JMV 1843 in mouse blood serum. A single subcutaneous injection of JMV 1843 (0.01-10 mg/kg) increased food intake in fed mice in a dose-dependent manner, up to 5-times relative to the saline-treated group (ED(50)=1.94 mg/kg at 250 min). JMV 1843 was stable in mouse serum in vitro for 24 h, but was mostly eliminated from mouse blood after 2 h in vivo. Ten days of treatment with JMV 1843 (subcutaneous administration, 10 or 20 mg/kg/day) significantly increased food intake, body weight and mRNA expression of the orexigenic neuropeptide Y and agouti-related peptide in the medial basal hypothalamus and decreased the expression of uncoupling protein 1 in brown adipose tissue. Our data suggest that JMV 1843 could have possible future uses in the treatment of cachexia.
Assuntos
Proteína Relacionada com Agouti/metabolismo , Estimulantes do Apetite/farmacologia , Ingestão de Alimentos/efeitos dos fármacos , Grelina/agonistas , Hipotálamo/efeitos dos fármacos , Neuropeptídeo Y/metabolismo , Oligopeptídeos/farmacologia , Aumento de Peso/efeitos dos fármacos , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Marrom/metabolismo , Proteína Relacionada com Agouti/genética , Animais , Estimulantes do Apetite/administração & dosagem , Estimulantes do Apetite/farmacocinética , Relação Dose-Resposta a Droga , Grelina/metabolismo , Hipotálamo/metabolismo , Indóis , Injeções Subcutâneas , Canais Iônicos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Mitocondriais/metabolismo , Neuropeptídeo Y/genética , Oligopeptídeos/administração & dosagem , Oligopeptídeos/farmacocinética , RNA Mensageiro/metabolismo , Receptores de Grelina/agonistas , Receptores de Grelina/metabolismo , Transdução de Sinais/efeitos dos fármacos , Triptofano/análogos & derivados , Proteína Desacopladora 1 , Regulação para CimaRESUMO
Many peripheral substances, including ghrelin, induce neuronal activation in the brain. In the present study, we compared the effect of subcutaneously administered ghrelin and its three stable agonists: Dpr(3)ghr ([Dpr(N-octanoyl)(3)] ghrelin) (Dpr - diaminopropionic acid), YA GHRP-6 (H-Tyr-Ala-His-DTrp-Ala-Trp-DPhe-Lys-NH(2)), and JMV1843 (H-Aib-DTrp-D-gTrp-CHO) on the Fos expression in food intake-responsive brain areas such as the hypothalamic paraventricular (PVN) and arcuate (ARC) nuclei, the nucleus of the solitary tract (NTS), and area postrema (AP) in male C57BL/6 mice. Immunohistochemical analysis showed that acute subcutaneous dose of each substance (5mg/kg b.w.), which induced a significant food intake increase, elevated Fos protein expression in all brain areas studied. Likewise ghrelin, each agonist tested induced distinct Fos expression overall the PVN. In the ARC, ghrelin and its agonists specifically activated similarly distributed neurons. Fos occurrence extended from the anterior (aARC) to middle (mARC) ARC region. In the latter part of the ARC, the Fos profiles were localized bilaterally, especially in the ventromedial portions of the nucleus. In the NTS, all substances tested also significantly increased the number of Fos profiles in neurons, which also revealed specific location, i.e., in the NTS dorsomedial subnucleus (dmNTS) and the area subpostrema (AsP). In addition, cells located nearby the NTS, in the AP, also revealed a significant increase in number of Fos-activated cells. These results demonstrate for the first time that ghrelin agonists, regardless of their different chemical nature, have a significant and similar activating impact on specific groups of neurons that can be a part of the circuits involved in the food intake regulation. Therefore there is a real potency for ghrelin agonists to treat cachexia and food intake disorders. Thus, likewise JMV1843, the other ghrelin agonists represent substances that might be involved in trials for clinical purposes.
Assuntos
Regulação do Apetite/efeitos dos fármacos , Regulação do Apetite/fisiologia , Química Encefálica/efeitos dos fármacos , Grelina/agonistas , Grelina/fisiologia , Neurônios/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-fos/biossíntese , Regulação para Cima/efeitos dos fármacos , Animais , Biomarcadores/metabolismo , Química Encefálica/fisiologia , Transtornos da Alimentação e da Ingestão de Alimentos/tratamento farmacológico , Transtornos da Alimentação e da Ingestão de Alimentos/fisiopatologia , Grelina/análogos & derivados , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neurônios/fisiologia , Regulação para Cima/fisiologiaRESUMO
Mesenchymal stem cells (MSCs) have been repeatedly shown to be able to repair bone defects. The aim of this study was to characterize the osteogenic differentiation of miniature pig MSCs and markers of this differentiation in vitro. Flow-cytometrically characterized MSCs were seeded on cultivation plastic (collagen I and vitronectin coated/uncoated) or plasma clot (PC)/plasma-alginate clot (PAC) scaffolds and differentiated in osteogenic medium. During three weeks of differentiation, the formation of nodules and deposition of calcium were visualized by Alizarin Red Staining. In addition, the production of alkaline phosphatase (ALP) activity was quantitatively detected by fluorescence. The expression of osteopontin, osteonectin and osteocalcin were assayed by immunohistochemistry and Western Blot analysis. We revealed a decrease of osteopontin expression in 2D and 3D environment during differentiation. The weak initial osteonectin signal, culminating on 7(th) or 14(th) day of differentiation, depends on collagen I and vitronectin coating in 2D system. The highest activity of ALP was detected on 21(th) day of osteogenic differentiation. The PC scaffolds provided better conditions for osteogenic differentiation of MSCs than PAC scaffolds in vitro. We also observed expected effects of collagen I and vitronectin on the acceleration of osteogenic differentiation of miniature pig MSC. Our results indicate similar ability of miniature pig MSCs osteogenic differentiation in 2D and 3D environment, but the expression of osteogenic markers in scaffolds and ECM coated monolayers started earlier than in the monolayers without ECM.
Assuntos
Diferenciação Celular , Células-Tronco Mesenquimais/citologia , Osteogênese/fisiologia , Animais , Antraquinonas , Corantes , Matriz Extracelular/metabolismo , Imuno-Histoquímica , Células-Tronco Mesenquimais/metabolismo , Osteocalcina/metabolismo , Osteonectina/metabolismo , Osteopontina/metabolismo , Sus scrofa , Alicerces TeciduaisRESUMO
BACKGROUND: The capacity to focus and concentrate or to direct attention supports many aspects of cognitive functioning including short-term memory and higher-level cognitive functions. The purpose was to assess attentional networks in euthymic patients with unipolar depression using the Attentional Network Test (ANT). MATERIALS AND METHODS: We investigated performance of attention by virtue of ANT during remission from unipolar depressive disorder and tested a hypothesis that there are no differences between outpatient group (euthymic patients, N=32) and matched controls in attentional variables, the relationship of attentional networks and everyday cognitive failures. RESULTS: No differences between the groups in attentional networks were found and no relationship between attentional networks and cognitive failures was found. LIMITATIONS: One assessment during remission could be insufficient to recognize long-term pattern of cognitive functions. CONCLUSIONS: These data show non-impaired attentional networks possibly explained by sufficient level of remission and ameliorated influence of high education on cognition.
Assuntos
Atenção , Cognição , Transtorno Depressivo/psicologia , Desempenho Psicomotor , Adulto , Idoso , Estudos de Casos e Controles , Transtorno Depressivo/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Testes Neuropsicológicos , Pacientes Ambulatoriais , Reconhecimento Visual de Modelos , Escalas de Graduação Psiquiátrica , Recidiva , Fatores de TempoRESUMO
The aim of our study was to investigate the parthenogenetic activation of in vitro matured pig oocytes after their combined treatment with calcium ionophore A 23187 and the inhibitor of protein kinases, 6-dimethylaminopurine (6-DMAP) and to study the further embryonic development of oocytes activated using this treatment. The oocytes were exposed to ionophore (10, 25 or 50 microM) for 0.5, 1, 3, 5 or 7 min and then cultured with 6-DMAP (0 or 2 microM) The highest activation rate (up to 88% of the activated eggs reached the pronuclear stage) was observed after combined treatment of the oocytes with 50 microM ionophore and 6-DMAP. The highest rate of embryonic development was observed after treatment with 25 microM ionophore without 6-DMAP, when up to 51% of the eggs developed beyond two-cell stage, 2% of the eggs developed up to the stage of morula and up to 3% of the eggs reached the stage of blastocyst. When 50 microM ionophore was used, the embryonic development of the activated eggs was arrested before the morula and blastocyst stage. After treatment of the activated eggs with 6-DMAP, we did not observe any development beyond the stage of 16 blastomeres. We can conclude that combined treatment with calcium ionophore A 23187 and 6-DMAP increases the activation rate in pig oocytes matured in vitro, but this combined treatment exerts a detrimental effect on further embryonic development of the activated eggs.
Assuntos
Adenina/análogos & derivados , Adenina/farmacologia , Calcimicina/farmacologia , Inibidores Enzimáticos/farmacologia , Ionóforos/farmacologia , Oócitos/efeitos dos fármacos , Suínos/fisiologia , Animais , Células Cultivadas , Fase de Clivagem do Zigoto/efeitos dos fármacos , Fase de Clivagem do Zigoto/fisiologia , Relação Dose-Resposta a Droga , Feminino , Oócitos/crescimento & desenvolvimento , Oócitos/fisiologia , Partenogênese , Inibidores de Proteínas Quinases , Fatores de TempoRESUMO
In vitro matured pig oocytes were activated using a combined treatment of calcium ionophore A 23187 with cycloheximide. The oocytes were exposed to ionophore (10, 25 or 50 microM) for 0.5, 1, 3, 5 or 7 min and then cultured with cycloheximide (0 or 10 microg/ml) for 6 h. Cycloheximide treatment significantly increased the activation rate of oocytes and the percentage of oocytes that were able to develop after activation. The highest activation rate was observed after treatment with 50 microM ionophore. The highest percentage of developing eggs was observed after combined treatment of ionophore (25 microM) with cycloheximide. The percentage of oocytes developing up to the morula and blastocyst stage was not significantly increased after cycloheximide treatment.
Assuntos
Calcimicina/farmacologia , Cicloeximida/farmacologia , Ionóforos/farmacologia , Oócitos/efeitos dos fármacos , Inibidores da Síntese de Proteínas/farmacologia , Suínos , Animais , Células Cultivadas , Fase de Clivagem do Zigoto/efeitos dos fármacos , Feminino , Oócitos/fisiologia , PartenogêneseRESUMO
The authors investigated the collagen architecture of the sclerocorneal trabeculum in relation to age in eight eyes within the range from a premature infant to a 90-year-old man. All specimens were investigated using a scanning electron microscope of Jeol Co. The appearance of fibrils does not change substantially in relation to age, however, their pattern and distribution change. In a 10-year-old boy Schlemm's canal is wide, oval with thin trabecular plates and wide open spaces surrounding them. In the 90-year-old man the canal is open but the plates on its luminous side are thickened, wrinkled and compressed. On the albuminous side there is a compact wall of compressed collagen layers.
Assuntos
Envelhecimento/patologia , Colágeno/ultraestrutura , Malha Trabecular/ultraestrutura , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Córnea/ultraestrutura , Humanos , Lactente , Masculino , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Esclera/ultraestruturaRESUMO
The authors induced 105 deliveries by extraamniotic administration of PGE2 (prostin Upjohn). The initial dose was 1-2 tablets, depending on the maturity of the portio uteri. If the contractions did not start within two hours, the dose was repeated. The sac was disrupted when the contractions were regular and the os uteri was larger than 2 cm. If necessary uterine activity was stimulated by small doses of Oxytocin (in 29%). Indication for induction was a programmed delivery (44.7%), protraced pregnancy (31.5%), diabetes mellitus (10.5%), a period of more than 24 hours after drainage of amniotic fluid without contractions (5.7%), hypertension or renal disease during gestation (4.8%) and hypotrophy of the foetus (2.8%). Inductions were successful in 96.2% of the patients. The parity of the patients influenced the interval between the onset of induction and the onset of uterine contractions, the duration of the first and second stage of labour and the consumption of Prostin tablets. The age of the patient, occupation, obesity and operation on the uterus did not affect the success of induction. There were no serious pathological findings during the third stage of labour, nor serious side-effects. The condition of the neonates was satisfactory.