RESUMO
BACKGROUND: Distinguishing temporal patterns of depressive symptoms during pregnancy and after childbirth has important clinical implications for diagnosis, treatment, and maternal and child outcomes. The primary aim of the present study was to distinguish patterns of chronically elevated levels of depressive symptoms v. trajectories that are either elevated during pregnancy but then remit after childbirth, v. patterns that increase after childbirth. METHODS: The report uses latent growth mixture modeling in a large, population-based cohort (N = 12 121) to investigate temporal patterns of depressive symptoms. We examined theoretically relevant sociodemographic factors, exposure to adversity, and offspring gender as predictors. RESULTS: Four distinct trajectories emerged, including resilient (74.3%), improving (9.2%), emergent (4.0%), and chronic (11.5%). Lower maternal and paternal education distinguished chronic from resilient depressive trajectories, whereas higher maternal and partner education, and female offspring gender, distinguished the emergent trajectory from the chronic trajectory. Younger maternal age distinguished the improving group from the resilient group. Exposure to medical, interpersonal, financial, and housing adversity predicted membership in the chronic, emergent, and improving trajectories compared with the resilient trajectory. Finally, exposure to medical, interpersonal, and financial adversity was associated with the chronic v. improving group, and inversely related to the emergent class relative to the improving group. CONCLUSIONS: There are distinct temporal patterns of depressive symptoms during pregnancy, after childbirth, and beyond. Most women show stable low levels of depressive symptoms, while emergent and chronic depression patterns are separable with distinct correlates, most notably maternal age, education levels, adversity exposure, and child gender.
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Depressão Pós-Parto/diagnóstico , Depressão/diagnóstico , Adulto , Diagnóstico Diferencial , Escolaridade , Feminino , Humanos , Modelos Logísticos , Estudos Longitudinais , Masculino , Idade Materna , Gravidez , Escalas de Graduação Psiquiátrica , Resiliência Psicológica , Índice de Gravidade de Doença , Fatores Sexuais , Estresse Psicológico , Fatores de Tempo , Adulto JovemRESUMO
Stress and glucocorticoids alter glutamatergic transmission, and the outcome of stress may range from plasticity enhancing effects to noxious, maladaptive changes. We have previously demonstrated that acute stress rapidly increases glutamate release in prefrontal and frontal cortex via glucocorticoid receptor and accumulation of presynaptic SNARE complex. Here we compared the ex vivo effects of acute stress on glutamate release with those of in vitro application of corticosterone, to analyze whether acute effect of stress on glutamatergic transmission is mediated by local synaptic action of corticosterone. We found that acute stress increases both the readily releasable pool (RRP) of vesicles and depolarization-evoked glutamate release, while application in vitro of corticosterone rapidly increases the RRP, an effect dependent on synaptic receptors for the hormone, but does not induce glutamate release for up to 20 min. These findings indicate that corticosterone mediates the enhancement of glutamate release induced by acute stress, and the rapid non-genomic action of the hormone is necessary but not sufficient for this effect.
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Corticosterona/metabolismo , Lobo Frontal/patologia , Neurônios/patologia , Terminações Pré-Sinápticas/metabolismo , Estresse Psicológico/patologia , Análise de Variância , Animais , Ácido Aspártico/metabolismo , Corticosterona/farmacologia , Relação Dose-Resposta a Droga , Eletrochoque/efeitos adversos , Potenciais Pós-Sinápticos Excitadores/efeitos dos fármacos , Técnicas In Vitro , Masculino , Neurônios/metabolismo , Terminações Pré-Sinápticas/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Estresse Psicológico/etiologia , Sinapsinas/metabolismo , Vesículas Sinápticas/efeitos dos fármacos , Vesículas Sinápticas/metabolismo , Sinaptossomos/metabolismo , Trítio/farmacocinéticaRESUMO
Elevation in [Ca(2+)]i and activation of calpain-1 occur in central nervous system of SOD1(G93A) transgenic mice model of amyotrophic lateral sclerosis (ALS), but few data are available about the early stage of ALS. We here investigated the level of activation of the Ca(2+)-dependent protease calpain-1 in spinal cord of SOD1(G93A) mice to ascertain a possible role of the protease in the aetiology of ALS. Comparing the events occurring in the 120 day old mice, we found that [Ca(2+)]i and activation of calpain-1 were also increased in the spinal cord of 30 day old mice, as indicated by the digestion of some substrates of the protease such as nNOS, αII-spectrin, and the NR2B subunit of NMDA-R. However, the digestion pattern of these proteins suggests that calpain-1 may play different roles depending on the phase of ALS. In fact, in spinal cord of 30 day old mice, activation of calpain-1 produces high amounts of nNOS active species, while in 120 day old mice enhanced-prolonged activation of calpain-1 inactivates nNOS and down-regulates NR2B. Our data reveal a critical role of calpain-1 in the early phase and during progression of ALS, suggesting new therapeutic approaches to counteract its onset and fatal course.
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Esclerose Lateral Amiotrófica/genética , Cálcio/metabolismo , Calpaína/metabolismo , Óxido Nítrico Sintase Tipo I/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Medula Espinal/metabolismo , Esclerose Lateral Amiotrófica/fisiopatologia , Animais , Modelos Animais de Doenças , Progressão da Doença , Humanos , Camundongos , Camundongos Transgênicos , Neurônios Motores/metabolismo , Óxido Nítrico Sintase Tipo I/genética , Proteólise , Receptores de N-Metil-D-Aspartato/genética , Superóxido Dismutase/genética , Superóxido Dismutase-1RESUMO
The ionization of hydrogen by a chirped XUV pulse in the presence of a few cycle infrared laser pulse has been investigated. The electron momentum distribution has been obtained by treating the interaction of the atom with the XUV radiation at the first order of the time-dependent perturbation theory and describing the emitted electron through the Coulomb-Volkov wavefunction. The results of the calculations agree with the ones found by solving numerically the time-dependent Schrödinger equation. It has been found that depending on the delay between the pulses the combined effect of the XUV chirp and of the steering action on the infrared field brings about asymmetries in the electron momentum distribution. These asymmetries may give information on both the chirp and the XUV pulse duration.
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Transporte de Elétrons , Elétrons , Transferência de Energia , Lasers , Modelos Químicos , Simulação por ComputadorRESUMO
The concentrations of trace elements were studied in roots, rhizomes, stems, and leaves of Phragmites australis stands (common reeds), and in the corresponding samples of water and sediment from the mouth of the Imera Meridionale River (Sicily, Italy), an area affected by massive urbanization and intensive agriculture. The elements considered were Ag, Al, As, B, Ba, Be, Co, Fe, Mo, Pd, Pt, Rh, Sb, Se, Sr, Tl, and V. Concentrations in belowground organs were usually higher than aboveground tissues, and the general decreasing trend of element content was root>rhizome>leaf>stem. Trace element mobility was generally higher within the organs than in sediment to plant. Regarding Al, Fe, and V, the phytotoxic levels in roots and the low plant/root mobility, may indicate that roots are inherently tolerant to these metals, and act as filters to prevent toxic distribution in the plant. The high uptake of Pd and Rh showed that emissions of catalytic converters are one of the main health hazards of the study area. P. australis showed a direct response to the environmental conditions, and its application as a biomonitor should be considered.
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Poaceae/metabolismo , Poluentes do Solo/metabolismo , Oligoelementos/metabolismo , Monitoramento Ambiental , Metais Pesados/metabolismo , Raízes de Plantas/metabolismo , Rios , Sicília , Áreas AlagadasRESUMO
Recent studies using data from the National Health and Nutrition Examination Survey (NHANES) have used inconsistent approaches to identify and categorize beverages, especially those containing low-calorie sweeteners (LCS), also referred to as low-calorie sweetened beverages (LCSBs). Herein, we investigate the approaches used to identify and categorize LCSBs in recent analyses of NHANES data. We reviewed published studies examining LCS consumption in relation to dietary and health outcomes and extracted the methods used to categorize LCS as reported by the authors of each study. We then examined the extent to which these approaches reliably identified LCSBs using the Internet Archive Wayback Machine to examine beverage ingredients lists across three NHANES cycles (2011-2016). None of the four general strategies used appeared to include all LCSBs while also excluding all beverages that did not contain LCS. In some cases, the type of sweetener in the beverage consumed could not be clearly determined; we found 9, 16, and 18 of such "mixed" beverage identifiers in the periods 2011-2012, 2013-2014, and 2015-2016, respectively. Then, to illustrate how heterogeneity in beverage categorization may impact the outcomes of published analyses, we compared results of a previously published analysis with outcomes when "mixed" beverages were grouped either all as LCSBs or all as sugary beverages. Our results suggest that caution is warranted in design and interpretation of studies using NHANES data to examine dietary and health correlates of sweetened beverage intake.
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Bebidas Adoçadas Artificialmente/classificação , Bebidas/classificação , Ingredientes de Alimentos/análise , Inquéritos Nutricionais/estatística & dados numéricos , Edulcorantes/análise , Bebidas Adoçadas Artificialmente/análise , Bebidas/análise , Dieta/estatística & dados numéricos , Humanos , Inquéritos Nutricionais/métodos , Reprodutibilidade dos Testes , Projetos de Pesquisa , Estados UnidosRESUMO
Although many studies have unequivocally demonstrated the promise of understanding resilience to adversity and characterizing the consequences if stress is unabated, needed are dynamic theories and methods to enhance the rigor and interpretation of these assessments. From a dynamic systems perspective, the focus is not whether an individual possesses some fixed ability or unchangeable trait, but rather to understand the flexibility and responsiveness of stress regulation systems to daily hassles and adverse life events. A renewed interest in individual variability allows researchers to see trajectories of change over both short- and long-time scales to understand the developmental course. As a result, it is possible to answer questions, such as, how does the dysregulation in emotion caused by stress, to both within and between daily affect processes, relate to longitudinal trajectories (over time-scales of years) of dysfunction and disease? The overarching goal of the Notre Dame Study of Health & Well-being is to detail the types and qualities of contextual influences, in conjunction with dynamic psychobiological systems, to assess the precursors, concomitant influences and consequences of stress and resilience in the face of adversity on cognitive, health and well-being outcomes.
Assuntos
Corticosterona/metabolismo , Lobo Frontal/patologia , Ácido Glutâmico/metabolismo , Terminações Pré-Sinápticas/metabolismo , Estresse Psicológico/metabolismo , Estresse Psicológico/patologia , Animais , Corticosterona/farmacologia , Técnicas In Vitro , Terminações Pré-Sinápticas/efeitos dos fármacosRESUMO
Paramecium primaurelia expresses a significant amount of gamma-amino butyric acid (GABA). Paramecia possess both glutamate decarboxylase (GAD)-like and vesicular GABA transporter (vGAT)-like proteins, indicating the ability to synthesize GABA from glutamate and to transport GABA into vesicles. Using antibodies raised against mammalian GAD and vGAT, bands with an apparent molecular weight of about 67 kDa and 57 kDa were detected. The presence of these bands indicated a similarity between the proteins in Paramecium and in mammals. VAMP, syntaxin and SNAP, putative proteins of the release machinery that form the so-called SNARE complex, are present in Paramecium. Most VAMP, syntaxin and SNAP fluorescence is localized in spots that vary in size and density and are primarily distributed near the plasma membrane. Antibodies raised against mammal VAMP-3, sintaxin-1 or SNAP-25 revealed protein immunoblot bands having molecular weights consistent with those observed in mammals. Moreover, P. primaurelia spontaneously releases GABA into the environment, and this neurotransmitter release significantly increases after membrane depolarization. The depolarization-induced GABA release was strongly reduced not only in the absence of extracellular Ca(2+) but also by pre-incubation with bafilomycin A1 or with botulinum toxin C1 serotype. It can be concluded that GABA occurs in Paramecium, where it is probably stored in vesicles capable of fusion with the cell membrane; accordingly, GABA can be released from Paramecium by stimulus-induced, neuronal-like exocytotic mechanisms.
Assuntos
Exocitose/fisiologia , Paramecium/fisiologia , Ácido gama-Aminobutírico/metabolismo , Sequência de Aminoácidos , Animais , Toxinas Botulínicas/farmacologia , Exocitose/efeitos dos fármacos , Glutamato Descarboxilase/metabolismo , Dados de Sequência Molecular , Paramecium/efeitos dos fármacos , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Proteínas Qa-SNARE/metabolismo , Proteínas SNARE/genética , Proteínas SNARE/metabolismo , Alinhamento de Sequência , Proteínas de Transporte Vesicular/metabolismoRESUMO
PURPOSE: This study compared the diagnostic accuracy of magnetic resonance cholangiopancreatography (MRCP) and endoscopic ultrasonography (EUS) in evaluating the cause of extrahepatic bile duct dilatation. MATERIALS AND METHODS: Forty-five patients (26 men, mean age 57 years) with extrahepatic biliary dilatation, as shown by transabdominal ultrasound, with or without elevated biliary and pancreatic serum indices, were prospectively studied with MRCP and EUS between September 2007 and October 2008. EUS and MRCP were performed within no more than 24 h of each other to reduce the possibility of changes due to stone migration. Image analysis was carried out in a double-blind fashion. RESULTS: MRCP had 88.9% diagnostic accuracy, 91.9% sensitivity and 75% specificity, with 94.4% positive predictive value and 66.7% negative predictive value. EUS had 93.3% diagnostic accuracy, 97.3% sensitivity and 75% specificity; the positive and negative predictive values were 94.7% and 85.7%, respectively. CONCLUSIONS: MRCP and EUS do not show significant statistical differences in diagnostic accuracy. MRCP is an accurate, noninvasive modality in the study of extrahepatic biliary pathology. EUS is especially reliable in patients with extrahepatic biliary obstruction caused by endoluminal sludge.
Assuntos
Colangiopancreatografia por Ressonância Magnética/métodos , Colestase Extra-Hepática/diagnóstico , Endossonografia/métodos , Ductos Biliares Extra-Hepáticos , Colestase Extra-Hepática/diagnóstico por imagem , Meios de Contraste , Dilatação Patológica , Feminino , Humanos , Interpretação de Imagem Assistida por Computador , Masculino , Meglumina/análogos & derivados , Pessoa de Meia-Idade , Compostos Organometálicos , Valor Preditivo dos Testes , Estudos Prospectivos , Sensibilidade e EspecificidadeRESUMO
At Mount Etna volcano, the focus point of persistent tectonic extension is represented by the Summit Craters. A muographic telescope has been installed at the base of the North-East Crater from August 2017 to October 2019, with the specific aim to find time related variations in the density of volcanic edifice. The results are significant, since the elaborated images show the opening and evolution of different tectonic elements; in 2017, a cavity was detected months before the collapse of the crater floor and in 2018 a set of underground fractures was identified, at the tip of which, in June 2019, a new eruptive vent started its explosive activity, still going on (February, 2020). Although this is the pilot experiment of the project, the results confirm that muography could be a turning point in the comprehension of the plumbing system of the volcano and a fundamental step forward to do mid-term (weeks/months) predictions of eruptions. We are confident that an increment in the number of telescopes could lead to the realization of a monitoring system, which would keep under control the evolution of the internal dynamic of the uppermost section of the feeding system of an active volcano such as Mount Etna.
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Cancer stem cells have been isolated from several solid tumors including prostate, colon, liver, breast, and ovarian cancer. Stem cells isolated from nervous system and prostate express CD133 antigen, which is widely used to isolate hematopoietic stem and progenitor cells. The aims of this study were to investigate the expression of the CD133-1 and CD133-2 epitopes in primary ovarian tumors and to biologically characterize CD133(+) ovarian cancer cells, also according to clinicopathologic parameters. Tissue specimens were obtained at primary surgery from 41 ovarian carcinomas; eight normal ovaries and five benign ovarian tumors were also collected. Flow cytometry with monoclonal antibodies against CD133-1 and CD133-2 epitopes was employed. FACS (fluorescence activated cell sorting) analysis enabled the selection of CD133(+) cells, whose epithelial origin was confirmed by immunofluorescence analysis with monoclonal anti-cytokeratin 7. CD133(+) cells gave rise to a 4.7 +/- 0.9-fold larger number of colonies than that documented in CD133(-) population (P < 0.001). Moreover, CD133(+) cells showed an enhanced proliferative potential compared to CD133(-) cells. The percentages of CD133-1- and CD133-2-expressing cells were significantly lower in normal ovaries/benign tumors with respect to those in ovarian carcinoma. Both the percentages of CD133-1- and CD133-2-expressing cells were significantly lower in omental metastases than in primary ovarian cancer (P = 0.009 and 0.007 for CD133-1- and CD133-2-expressing cells, respectively). There seems not to be any difference in the distribution of the percentage of CD133-1- and CD133-2-expressing cells according to clinicopathologic parameters and response to primary chemotherapy. CD133-1 and CD133-2 may be useful in order to select and enrich the population of CD133(+) ovarian tumor cells, which are characterized by a higher clonogenic efficiency and proliferative potential.
Assuntos
Antígenos CD/metabolismo , Biomarcadores Tumorais/metabolismo , Glicoproteínas/metabolismo , Invasividade Neoplásica/patologia , Neoplasias Ovarianas/mortalidade , Neoplasias Ovarianas/patologia , Peptídeos/metabolismo , Antígeno AC133 , Adulto , Idoso , Estudos de Coortes , Feminino , Citometria de Fluxo , Imunofluorescência , Regulação Neoplásica da Expressão Gênica , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neoplasias Ovarianas/genética , Probabilidade , Prognóstico , Valores de Referência , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Medição de Risco , Sensibilidade e Especificidade , Análise de SobrevidaRESUMO
Aim of the study is the appraisal of stress in a group of the 500 anaestheticians and in a control group. Two anonymous self-administered questionnaires were utilized, one for the subjective appraisal of the perceived various risk related to job, and one for the subjective appraisal of stress: VRS questionnaire of Tarsitani and Biondi. The results of the study showed that all the anaestheticians identified the perception of the emotional aspects related to work as a risk factor. They put it mainly in relation with the forensic implications. The total score of scale VRS and the score for cluster depression, anxiety, aggressiveness are higher in the anesthetists than the control group.
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Anestesiologia , Transtornos Mentais/etiologia , Doenças Profissionais/etiologia , Estresse Psicológico/etiologia , Adulto , Agressão , Ansiedade/etiologia , Estudos Transversais , Depressão/etiologia , Emoções , Feminino , Medicina Legal , Humanos , Itália , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Inquéritos e Questionários , Recursos HumanosRESUMO
BACKGROUND: Although the stomach is the most frequent site of intestinal lymphomas, few data are available on both clinical endoscopic presentation of gastric lymphoma and possible differences between low-grade and high-grade lymphomas. METHODS: Clinical, histological and endoscopic records of consecutive patients with primary low-grade or high-grade lymphoma diagnosed were retrieved. Symptoms were categorized as 'alarm' or 'not alarm'. The endoscopic findings were classified as 'normal' or 'abnormal'. RESULTS: Overall, 144 patients with primary gastric lymphoma were detected, including 74 low-grade and 70 high-grade lymphoma. Alarm symptoms, particularly persistent vomiting and weight loss, were more frequently present in patients with high-grade lymphoma than in those with low-grade lymphoma (54% vs. 28%; P = 0.002). Low-grade lymphomas presented as 'normal' appearing mucosa (20% vs. 0%; P = 0.0004) or petechial haemorrhage in the fundus (9% vs. 0%; P = 0.02) more frequently than high-grade lymphomas, being also more often confined to the antrum (47% vs. 27%, P = 0.03) and associated with Helicobacter pylori infection (88% vs. 52%, P < 0.0001). On the contrary, high-grade lymphomas presented more commonly as ulcerative type (70% vs. 52%; P = 0.03), being also more frequently diagnosed in stage >I when compared with low-grade lymphomas (70% vs. 21%, P < 0.0001). CONCLUSIONS: The overall prevalence of alarm symptoms is quite low and may be absent in more than 70% of patients with low-grade lymphoma.
Assuntos
Linfoma/patologia , Neoplasias Gástricas/patologia , Endoscopia Gastrointestinal/métodos , Feminino , Mucosa Gástrica/patologia , Infecções por Helicobacter/complicações , Infecções por Helicobacter/patologia , Helicobacter pylori/isolamento & purificação , Humanos , Linfoma/complicações , Linfoma/microbiologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Estudos Retrospectivos , Estômago/patologia , Neoplasias Gástricas/complicações , Neoplasias Gástricas/microbiologiaRESUMO
We describe the case of a nurse who showed eczematous lesions wide-spread over the neck and face. She worked as a attendant to a surgical chamber and wore vinyl gloves because of previous episodes of contact dermatitis caused by rubber gloves. The allergologic examination demonstrated that total IgE was 69.8 kU/1, both the Skin prick tests and the patch test for latex were positive. Persisting her dermatological symptomatology, in the suspicion of a airborne contact dermatitis, the nurse was removed and addressed to administrative functions for detecting the clinical follow-up. Fortunately, the eczematous manifestations spontaneously disappeared immediately after her transfer. In this case the presence of eczema mainly on air-exposed areas was interpreted as a sign of airborne contact dermatitis to latex. The case is reported in that the clinical picture is not very common.
Assuntos
Poluição do Ar em Ambientes Fechados/efeitos adversos , Dermatite de Contato/etiologia , Dermatite Ocupacional/etiologia , Hipersensibilidade ao Látex/complicações , Adulto , Feminino , HumanosRESUMO
A comprehensive bibliographical retrieval of case reports on "airborne contact dermatitis" (ACD) was performed. The present review deals with the first cases published in 1986, 1991, 2001 by Huygens as well as by Dooms-Goossens, and continues with the other pertaining clinical presentations until to day. Solid particles, rather than gases or droplets, are the most frequently reported causes of ACD.
Assuntos
Poluentes Ocupacionais do Ar/efeitos adversos , Poluentes Atmosféricos/efeitos adversos , Dermatite de Contato/etiologia , Dermatite Ocupacional/etiologia , Dermatite Alérgica de Contato/etiologia , Humanos , Exposição Ocupacional/efeitos adversosRESUMO
PURPOSE: The aim of our study was to assess the smoking habits in health care workers. MATERIALS AND METHODS: In concomitance with a medical examination for Health Care Surveillance requested by the Italian Laws (D. Lgs. 626/94), our operative units interviewed 2,000 persons (47.9% males, 52.1% females, mean age = 45 yrs (SD+/-9.41 yrs) working in two General Hospitals of Eastern Sicily. RESULTS: The prevalence of smokers was found to be higher in men (34.5%) than in women (33.6%), in Health Care Operators (36.4%) and professional nurses (36.2%) than in medical doctors (27.7%), in night-time workers (34,5%) then in day-time-workers (28,7%). The compliance with smoking restriction was found to be poor; in fact, 60% out of smokers declared to smoke during the working hours. CONCLUSIONS: This investigation has revealed the significant prevalence of tobacco smoking among the health care workers in general. We think that the inclusion of an anonymous questionnaire on smoking habits within the Services of Sanitary Surveillance may help in the national campaign against cigarette smoking.
Assuntos
Pessoal de Saúde , Recursos Humanos em Hospital , Fumar/epidemiologia , Adulto , Fatores Etários , Idoso , Distribuição de Qui-Quadrado , Feminino , Hospitais Gerais , Humanos , Entrevistas como Assunto , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores Sexuais , Inquéritos e QuestionáriosRESUMO
BACKGROUND: Mesenchymal stromal cells (MSCs) have been largely investigated, in the past decade, as potential therapeutic strategies for various acute and chronic pathological conditions. MSCs isolated from different sources, such as bone marrow (BM), umbilical cord tissue (UCT) and adipose tissue (AT), share many biological features, although they may show some differences on cumulative yield, proliferative ability and differentiation potential. The standardization of MSCs growth and their functional amplification is a mandatory objective of cell therapies. The aim of this study was to evaluate the cumulative yield and the ex vivo amplification potential of MSCs obtained from various sources and different subjects, using defined culture conditions with a standardized platelet lysate (PL) as growth stimulus. METHODS: MSCs isolated from BM, UCT and AT and expanded in human PL were compared in terms of cumulative yield and growth potential per gram of starting tissue. MSCs morphology, phenotype, differentiation potential, and immunomodulatory properties were also investigated to evaluate their biological characteristics. RESULTS: The use of standardized PL-based culture conditions resulted in a very low variability of MSC growth. Our data showed that AT has the greater capacity to generate MSC per gram of initial tissue, compared to BM and UCT. However, UCT-MSCs replicated faster than AT-MSCs and BM-MSCs, revealing a greater proliferation capacity of this source irrespective of its lower MSC yield. All MSCs exhibited the typical MSC phenotype and the ability to differentiate into all mesodermal lineages, while BM-MSCs showed the most prominent immunosuppressive effect in vitro. CONCLUSIONS: The adoption of standardized culture conditions may help researchers and clinicians to reveal particular characteristics and inter-individual variability of MSCs sourced from different tissues. These data will be beneficial to set the standards for tissue collection and MSCs clinical-scale expansion both for cell banking and for cell-based therapy settings.
Assuntos
Plaquetas/citologia , Proliferação de Células/fisiologia , Células-Tronco Mesenquimais/citologia , Tecido Adiposo/citologia , Adulto , Medula Óssea/fisiologia , Células da Medula Óssea/citologia , Técnicas de Cultura de Células/métodos , Diferenciação Celular/fisiologia , Linhagem da Célula/fisiologia , Células Cultivadas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Cordão Umbilical/citologiaRESUMO
Alternatives to the use of fetal bovine serum (FBS) have been investigated to ensure xeno-free growth condition. In this study we evaluated the efficacy of human platelet lysate (PL) as a substitute of FBS for the in vitro culture of some human cell lines. PL was obtained by pools of pathogen inactivated human donor platelet (PLT) concentrates. Human leukemia cell lines (KG-1, K562, JURKAT, HL-60) and epithelial tumor cell lines (HeLa and MCF-7) were cultured with either FBS or PL. Changes in cell proliferation, viability, morphology, surface markers and cell cycle were evaluated for each cell line. Functional characteristics were analysed by drug sensitivity test and cytotoxicity assay. Our results demonstrated that PL can support growth and expansion of all cell lines, although the cells cultured in presence of PL experienced a less massive proliferation compared to those grown with FBS. We found a comparable percentage of viable specific marker-expressing cells in both conditions, confirming lineage fidelity in all cultures. Functionality assays showed that cells in both FBS- and PL-supported cultures maintained their normal responsiveness to adriamycin and NK cell-mediated lysis. Our findings indicate that PL is a feasible serum substitute for supporting growth and propagation of haematopoietic and epithelial cell lines with many advantages from a perspective of process standardization, ethicality and product safety.
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BACKGROUND: Tissue homeostasis is guaranteed by stem proliferating reserve, depending on dynamic changes in gene expression. A high plasticity is shown by the haematopoietic stem cells, potential source for liver regeneration. AIM: We aimed to evaluate the gene expression modifications induced by human haematopoietic stem cell therapy after liver injury in rats. SUBJECTS: Rats were sorted as follows: (A) human-haematopoietic stem cell injection after allyl alcohol liver damage; (B) only haematopoietic stem cell injection; (C) only allyl alcohol injection; and (D) sacrifice without any treatment. METHODS: Livers, spleens and bone marrows were analysed with flow-cytometry. Livers were also studied by reverse-transcription PCR, histology, immunohistochemistry and microarray analysis; selected genes were confirmed by real-time PCR. RESULTS: In subset A, haematopoietic stem cells were selectively recruited by liver, with respect to the group B, and they improved the liver regeneration process compared to group C. As regards microarrays, haematopoietic stem cell infusion upregulates 265 genes and downregulates 149 genes. Differentially regulated genes belong to a broad range of functional pathways, including proliferation, differentiation, adhesion/migration and transcripts related to oval-cell activation. Real-time PCR validated array results. CONCLUSIONS: Our study confirmed the capacity of haematopoietic stem cells to contribute to liver regeneration. Moreover, microarray analysis led to the identification of genes whose regulation strongly correlates with a more efficient process of liver repair after haematopoietic stem cell injection.