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1.
Am J Emerg Med ; 76: 105-110, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38056055

RESUMO

BACKGROUND: The optimal management of patients taking oral anticoagulants who experience minor head injury (MHI) is unclear. The availability of validated protocols and reliable predictors of prognosis would be of great benefit. We investigated clinical factors as predictors of clinical outcomes and intracranial injury (ICI). METHODS: We conducted a single-cohort, prospective, observational study in an ED. Our structured clinical pathway included a first head CT scan, 24 h observation and a second CT scan. The primary outcome was the occurrence of MHI-related death or re-admission to ED at day +30. The secondary outcome was the rate of delayed ICI (dICI), defined as second positive CT scan after a first negative CT scan. We assessed some clinical predictors derived from guidelines and clinical prediction rules as potential risk factors for the outcomes. RESULTS: 450 patients with a negative first CT scan who underwent a second CT scan composed our 'study population'. The rate of the primary outcome was 4%. The rate of the secondary outcome was 4.7%. Upon univariate and multivariate analysis no statistically significant predictors for the outcomes were found. CONCLUSIONS: Previous retrospective studies showed a lot of negative predictive factors for anticoagulated patients suffering a minor head injury. In our prospective study no clinical factors emerged as predictors of poor clinical outcomes and dICI. So, even if we confirmed a low rate of adverse outcomes, the best management of these patients in ED remains not so clear and future trials are needed.


Assuntos
Traumatismos Craniocerebrais , Humanos , Estudos Prospectivos , Traumatismos Craniocerebrais/complicações , Traumatismos Craniocerebrais/diagnóstico por imagem , Anticoagulantes/efeitos adversos , Fatores de Risco , Estudos Retrospectivos
2.
Knee Surg Sports Traumatol Arthrosc ; 31(6): 2338-2348, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35716187

RESUMO

PURPOSE: To evaluate the mid- and long-term efficacy of autologous chondrocyte implantation (ACI) and matrix-assisted chondrocyte implantation (MACI) to treat patients with knee cartilage defects in the presence of osteoarthritis (OA). METHODS: PubMed and Cochrane databases were systematically searched for studies describing the treatment of knee OA with ACI or MACI (Kellgren-Lawrence (KL) ≥ 1, minimum follow-up 36 months). Results were reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines and included Lysholm, Western Ontario McMaster University and International Knee Documentation Committee scores. RESULTS: Of the 127 full-text articles assessed for eligibility, only five studies were selected based on inclusion/exclusion criteria (2 on ACI and 3 on MACI). In both groups, the defects were mainly located at femoral level, size 2.2-15.1 cm2 in the ACI and 2.0-7.6 cm2 in the MACI group. ACI was mostly used for patients affected by KL I, whereas MACI for patients with KL II-IV. The data obtained from 235 patients (161 ACI, 74 MACI) showed that ACI and MACI sustained stable clinical improvements up to 11 and 15 years, respectively, with a failure rate of about 10% up to 11 years. Scarce biological details regarding chondrocyte implantation were reported. CONCLUSIONS: ACI and MACI procedures for the treatment of knee cartilage lesions associated to OA showed long-term success and allowed delaying arthroplasty. Additional trials reporting homogenous data and precise patient characterization are needed to conduct an effective literature meta-analysis and identify the clinical relevance of these procedures. LEVEL OF EVIDENCE: IV.


Assuntos
Doenças das Cartilagens , Cartilagem Articular , Procedimentos Ortopédicos , Osteoartrite do Joelho , Humanos , Osteoartrite do Joelho/cirurgia , Condrócitos/transplante , Cartilagem Articular/cirurgia , Procedimentos Ortopédicos/métodos , Doenças das Cartilagens/cirurgia , Articulação do Joelho/cirurgia , Transplante Autólogo/métodos
3.
J Intern Med ; 289(5): 738-746, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33511686

RESUMO

BACKGROUND: Published reports on tocilizumab in COVID-19 pneumonitis show conflicting results due to weak designs or heterogeneity in critical methodological issues. METHODS: This open-label trial, structured according to Simon's optimal design, aims to identify factors predicting which patients could benefit from anti-IL6 strategies and to enhance the design of unequivocal and reliable future randomized trials. A total of 46 patients with COVID-19 pneumonia needing of oxygen therapy to maintain SO2 > 93% and with recent worsening of lung function received a single infusion of tocilizumab. Clinical and biological markers were measured to test their predictive values. Primary end point was early and sustained clinical response. RESULTS: Twenty-one patients fulfilled pre-defined response criteria. Lower levels of IL-6 at 24 h after tocilizumab infusion (P = 0.049) and higher baseline values of PaO2/FiO2 (P = 0.008) predicted a favourable response. CONCLUSIONS: Objective clinical response rate overcame the pre-defined threshold of 30%. Efficacy of tocilizumab to improve respiratory function in patients selected according to our inclusion criteria warrants investigations in randomized trials.


Assuntos
Anticorpos Monoclonais Humanizados , Biomarcadores Farmacológicos/análise , COVID-19 , Monitoramento de Medicamentos/métodos , Interleucina-6 , Pneumonia Viral , Idoso , Anticorpos Monoclonais Humanizados/administração & dosagem , Anticorpos Monoclonais Humanizados/farmacocinética , COVID-19/diagnóstico , COVID-19/epidemiologia , COVID-19/fisiopatologia , COVID-19/terapia , Feminino , Humanos , Fatores Imunológicos/administração & dosagem , Fatores Imunológicos/farmacocinética , Infusões Intravenosas , Interleucina-6/antagonistas & inibidores , Interleucina-6/sangue , Itália/epidemiologia , Masculino , Oximetria/métodos , Oxigenoterapia/métodos , Pneumonia Viral/tratamento farmacológico , Pneumonia Viral/epidemiologia , Pneumonia Viral/etiologia , Valor Preditivo dos Testes , Testes de Função Respiratória/métodos , SARS-CoV-2/isolamento & purificação , Resultado do Tratamento
4.
J Appl Microbiol ; 130(1): 90-99, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32640074

RESUMO

AIMS: This work aimed to estimate the growth of Myceliophthora thermophila M.7·7 in solid-state cultivation (SSC) through quantification of N-acetyl-d-glucosamine (NAG) and enzyme activity. METHODS AND RESULTS: The fungus was cultivated in sugarcane bagasse and wheat bran. A consistent statistical analysis was done to assess the reliability of experimental data. Logistic model equation was fitted to experimental data and growth parameters were estimated. The results showed strong influence of the sample size on NAG and a minimum recommended sample size was identified. Scanning electron microscopy (SEM) was used to identify the strategy of substrate colonization. Wheat bran was attacked firstly, while sugarcane bagasse was consumed after wheat bran depletion. The biomass growth was poorly estimated by secretion kinetics of α-amylase, endoglucanase, protease and xylanase, but enzyme kinetics were important for understanding substrate colonization. CONCLUSIONS: In conclusion, the NAG concentration was strongly affected by the sample size and sampling procedure. The strategy of fungal colonization on the substrates was well characterized through SEM analysis. The colonization strategy has direct influence on the kinetic parameters of the logistic model. Myceliophthora thermophila has a well-defined dynamic of enzyme secretion to degrade the substrate, although the kinetics of enzyme secretion has shown not adequate to characterize the kinetics of fungal growth. SIGNIFICANCE AND IMPACT OF THE STUDY: The paper provides reliable growth kinetic parameters in the SSC of the cellulase producer fungus M. thermophila M.7·7, as well as a robust analysis on three indirect methods (NAG, enzymes and SEM) for estimation of fungal development.


Assuntos
Sordariales/crescimento & desenvolvimento , Acetilglucosamina/metabolismo , Biomassa , Reatores Biológicos , Celulose/metabolismo , Fibras na Dieta/metabolismo , Proteínas Fúngicas/metabolismo , Cinética , Reprodutibilidade dos Testes , Saccharum/química , Sordariales/enzimologia , Sordariales/metabolismo , Sordariales/ultraestrutura
5.
Ann Ig ; 33(4): 307-321, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33270078

RESUMO

Results: Road tunnel construction workers revealed higher frequencies of cells with genotoxic damage (i.e., MN and NBUD). MN and NBUD resulted to be Poisson distributed and counts of these genotoxicity biomarkers were then analysed by Poisson regression. The frequency ratio (FR) for MN was 1.31 (95% CI: 0.84-2.04), with an increase in the exposed subjects; this finding, though indicating a higher genotoxic risk in the exposed subjects, did not reach statistical significance. On the other hand, the FR for NBUD was 3.49 (95% CI: 1.86-6.56), with a statistically significant increased risk of chromosomal damage. Even the frequencies of binucleated cells (a marker of cell proliferation) and pyknotic cells (a cell death biomarker) were significantly higher in tunnel workers. Introduction: Tunnel construction workers are exposed to complex mixtures of toxic agents, some of which are known to be genotoxic. The aim of this study was to evaluate the genotoxic risk in this occupational setting by comparing tunnel workers with a control group for frequencies of nuclear aberrations in oral exfoliated cells. Methods: To evaluate the genotoxic effects of tunnel air pollutants, we conducted a cross-sectional, molecular epidemiological study (35 tunnel workers and 35 healthy controls) using the buccal micronucleus cytome assay. A questionnaire was administered to obtain information about demographic variables, lifestyle, dietary habits, anthropometric data, and occupational history. Buccal mucosa cells were collected by scraping the buccal mucosa with a small-headed toothbrush. Coded slides were examined blind by trained scorers for micronuclei (MN), nuclear buds (NBUD), and other nuclear abnormalities. Conclusions: Our observations provide further knowledge and understanding of the occupational hazards of tunnel workers and confirm the complexity of effects (cytotoxic and genotoxic) probably induced by fumes and dust produced in underground operations.


Assuntos
Monitoramento Biológico , Exposição Ocupacional , Estudos Transversais , Análise Citogenética , Dano ao DNA , Humanos , Testes para Micronúcleos , Exposição Ocupacional/efeitos adversos
6.
Mol Psychiatry ; 23(7): 1597-1605, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29155800

RESUMO

Epidemiological studies report strong association between mood disorders and tobacco addiction. This high comorbidity requires adequate treatment but the underlying mechanisms are unknown. We demonstrate that nicotine exposure, independent of drug withdrawal effects, increases stress sensitivity, a major risk factor in mood disorders. Nicotine and stress concur to induce long-lasting cellular adaptations within the dopamine (DA) system. This interplay is underpinned by marked remodeling of nicotinic systems, causing increased ventral tegmental area (VTA) DA neurons' activity and stress-related behaviors, such as social aversion. Blocking ß2 or α7 nicotinic acetylcholine receptors (nAChRs) prevents, respectively, the development and the expression of social stress-induced neuroadaptations; conversely, facilitating α7 nAChRs activation specifically in the VTA promotes stress-induced cellular and behavioral maladaptations. Our work unravels a complex nicotine-stress bidirectional interplay and identifies α7 nAChRs as a promising therapeutic target for stress-related psychiatric disorders.


Assuntos
Neurônios Dopaminérgicos/efeitos dos fármacos , Receptores Nicotínicos/fisiologia , Animais , Dopamina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Neurônios/metabolismo , Nicotina/farmacologia , Agonistas Nicotínicos/farmacologia , Receptores Nicotínicos/efeitos dos fármacos , Receptores Nicotínicos/metabolismo , Estresse Psicológico/metabolismo , Fumar Tabaco/efeitos adversos , Fumar Tabaco/psicologia , Área Tegmentar Ventral/efeitos dos fármacos , Receptor Nicotínico de Acetilcolina alfa7/efeitos dos fármacos
7.
Pharmacogenomics J ; 18(3): 391-397, 2018 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-28696420

RESUMO

Mothers with a CYP2D6 ultrarapid metabolizer phenotype may expose their infants to risk of adverse events when taking codeine while breastfeeding, by producing more of the active metabolite, morphine. Pharmacogenetic testing may be a valuable tool to identify such mothers, but testing can be costly. The objective of the study was to determine the incremental costs of genotyping to avert neonatal adverse events during maternal pharmacotherapy. A cost-effectiveness analysis, using a decision model, was performed with a hypothetical cohort of prenatal subjects. Parameter estimates, costs and ranges for sensitivity analyses were ascertained from the literature and expert opinion. Sensitivity analyses were conducted to assess the robustness of the results. Probabilistic sensitivity analysis revealed an incremental cost-effectiveness (ICER) of $10 433 (Canadian dollars) for genotyping compared to no genotyping per adverse event averted. Results were sensitive to hospital admission costs. The ICER was lower when evaluating only subjects having caesarean deliveries or those from ethnic populations known to have a high prevalence of ultra-rapid metabolizers. Although genotyping to guide pharmacotherapy was not cost saving, the cost to avert an infant adverse event may represent good value for money in specific populations. With a growing demand for personalized medicine, these findings are relevant for decision makers, clinicians and patients.


Assuntos
Citocromo P-450 CYP2D6/genética , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/genética , Dor/tratamento farmacológico , Período Pós-Parto/genética , Aleitamento Materno/economia , Canadá , Codeína/efeitos adversos , Análise Custo-Benefício , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos/economia , Feminino , Testes Genéticos/economia , Técnicas de Genotipagem/economia , Humanos , Lactente , Morfina , Dor/economia , Dor/genética , Dor/patologia , Período Pós-Parto/efeitos dos fármacos , Gravidez
8.
J Prev Med Hyg ; 59(1): E14-E19, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29938235

RESUMO

Vaccines recommendations are available for both healthcare professionals and the general public, but although the vaccination is the most effective method to prevent infectious diseases, the coverage is still behind the recommended rate. In Italy, according to a recent study, the anti-flu vaccination rate among healthcare worker range between 9% to 30%. The aim of our study was to identify knowledge, attitude and behaviours regarding influenza vaccination among community pharmacists in order to increase the coverage rate among healthcare professional. "PErCEIVE (Pharmacist Perception on Influenza Vaccine) in Umbria" was a cross sectional survey among community pharmacists in Umbria conducted between 16th November 2015 to 29th February 2016. The questionnaire was anonymous, on-line self-administered survey. Statistical analysis were performed using STATA/SE 12 software. The response rate was 28.91% (n = 72/249). Among the studied population 76.39% (n = 55) had never performed influenza vaccine during the previous 5 years. Regarding source of information, only 15.28% of the subjects (n = 11) consulted the scientific publications, vs 52.78% (n = 38) who did not show any kind of interest upon the influenza vaccine. Our results show a low attitude to be vaccinated among pharmacists together with a low grade of awareness regarding the important role that pharmacists might play in order to reduce influenza burden, to promote health literacy among their patients and to decrease the risk of patients infection. Pharmacists might be crucial healthcare workers involved in health promotion, in vaccines' uptake and practices progression.


Assuntos
Atitude do Pessoal de Saúde , Vacinas contra Influenza , Influenza Humana/prevenção & controle , Farmacêuticos/psicologia , Adulto , Estudos Transversais , Feminino , Humanos , Itália , Masculino , Pessoa de Meia-Idade , Inquéritos e Questionários , Vacinação
9.
Ann Ig ; 30(6): 470-481, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30614496

RESUMO

BACKGROUND: The International Classification of Sleep disorders, the International Classification of Diseases and the Diagnostic and Statistical manual of Mental Disorders defines insomnia as an experience of insufficient or poor sleep quality, characterized by at least one of the following symptoms: difficulty in initiating or maintaining sleep, early awakenings and poor restorative sleep. In Italy, the Morfeo 1 study detects a prevalence of 20% of insomnia and a 40% of cases with day-time symptoms. The chronic sleep deprivation is responsible for cognitive disorders with effects on social life. Being common knowledge, lifestyle can also influence sleep. Some of the "sleep hygiene rules" involve a control on smoking, coffee consumption and diet. The Mediterranean Diet (MD), thanks to its high level of tryptophan, has a positive influence on sleep and can protect against stress and anxiety. STUDY DESIGN: The aim of InSOMNIA study was to determine the prevalence of sleep disorders among nursing students of the University of Perugia and, therefore, to evaluate how lifestyle, eating habits, health status and academics performance are linked to night-time and daytime symptoms of the interrupted sleep. METHODS: We adopted a cross sectional survey, collecting data from "Sleep and Daytime Habits Questionnaire" to evaluate the sleep disorders and from PREDIMED questionnaire to assess the adherence to MD. RESULTS: We found a statistical significant association between PREDIMED score and BMI (p-value=0.0127), smoking habit (p-value = 0.0125), quality of life (p-value = 0.0480) and academic progress (p-value = 0.0092). CONCLUSIONS: We found a high prevalence of sleep disturbances statistically associated with diet and poor academic progress.


Assuntos
Desempenho Acadêmico , Dieta Mediterrânea , Qualidade de Vida , Distúrbios do Início e da Manutenção do Sono/epidemiologia , Estudantes de Enfermagem/estatística & dados numéricos , Adulto , Distribuição por Idade , Consumo de Bebidas Alcoólicas , Estatura , Peso Corporal , Café , Estudos Transversais , Emprego/estatística & dados numéricos , Exercício Físico , Feminino , Nível de Saúde , Humanos , Itália/epidemiologia , Aprendizagem , Estilo de Vida , Masculino , Prevalência , Análise de Regressão , Distúrbios do Início e da Manutenção do Sono/etiologia , Distúrbios do Início e da Manutenção do Sono/prevenção & controle , Fumar/efeitos adversos , Inquéritos e Questionários , Triptofano/administração & dosagem , Adulto Jovem
10.
Clin Exp Obstet Gynecol ; 44(1): 166-168, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29714893

RESUMO

This case series demonstrates a potential new role for the use of intravenous (IV) acetaminophen. The authors reviewed two cases, whereby patients that developed intrapartum fever leading to fetal tachycardia were effectively treated with IV acetaminophen, leading to rapid reduction of maternal temperature and resolution of fetal tachycardia. Both patients had an uncomplicated vaginal delivery of healthy neonates. Intravenous acetaminophen, with its increased bioavailability and more rapid onset of action, may have benefit in the intrapartum setting by reducing adverse neonatal and maternal outcomes associated with febrile morbidity.


Assuntos
Acetaminofen/uso terapêutico , Antipiréticos/uso terapêutico , Doenças Fetais/tratamento farmacológico , Febre/tratamento farmacológico , Complicações na Gravidez/tratamento farmacológico , Taquicardia/tratamento farmacológico , Feminino , Doenças Fetais/etiologia , Humanos , Recém-Nascido , Infusões Intravenosas , Gravidez , Taquicardia/etiologia
11.
Allergy ; 71(8): 1181-91, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-26999633

RESUMO

BACKGROUND: Pollen-food syndrome (PFS) is heterogeneous with regard to triggers, severity, natural history, comorbidities, and response to treatment. Our study aimed to classify different endotypes of PFS based on IgE sensitization to panallergens. METHODS: We examined 1271 Italian children (age 4-18 years) with seasonal allergic rhinoconjunctivitis (SAR). Foods triggering PFS were acquired by questionnaire. Skin prick tests were performed with commercial pollen extracts. IgE to panallergens Phl p 12 (profilin), Bet v 1 (PR-10), and Pru p 3 (nsLTP) were tested by ImmunoCAP FEIA. An unsupervised hierarchical agglomerative clustering method was applied within PFS population. RESULTS: PFS was observed in 300/1271 children (24%). Cluster analysis identified five PFS endotypes linked to panallergen IgE sensitization: (i) cosensitization to ≥2 panallergens ('multi-panallergen PFS'); (ii-iv) sensitization to either profilin, or nsLTP, or PR-10 ('mono-panallergen PFS'); (v) no sensitization to panallergens ('no-panallergen PFS'). These endotypes showed peculiar characteristics: (i) 'multi-panallergen PFS': severe disease with frequent allergic comorbidities and multiple offending foods; (ii) 'profilin PFS': oral allergy syndrome (OAS) triggered by Cucurbitaceae; (iii) 'LTP PFS': living in Southern Italy, OAS triggered by hazelnut and peanut; (iv) 'PR-10 PFS': OAS triggered by Rosaceae; and (v) 'no-panallergen PFS': mild disease and OAS triggered by kiwifruit. CONCLUSIONS: In a Mediterranean country characterized by multiple pollen exposures, PFS is a complex and frequent complication of childhood SAR, with five distinct endotypes marked by peculiar profiles of IgE sensitization to panallergens. Prospective studies in cohorts of patients with PFS are now required to test whether this novel classification may be useful for diagnostic and therapeutic purposes in the clinical practice.


Assuntos
Alérgenos/imunologia , Conjuntivite Alérgica/diagnóstico , Hipersensibilidade Alimentar/diagnóstico , Alimentos/efeitos adversos , Pólen/imunologia , Rinite Alérgica Sazonal/diagnóstico , Adolescente , Idade de Início , Criança , Pré-Escolar , Análise por Conglomerados , Comorbidade , Conjuntivite Alérgica/epidemiologia , Conjuntivite Alérgica/imunologia , Feminino , Hipersensibilidade Alimentar/epidemiologia , Hipersensibilidade Alimentar/imunologia , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Itália/epidemiologia , Masculino , Vigilância da População , Rinite Alérgica Sazonal/epidemiologia , Rinite Alérgica Sazonal/imunologia , Fatores de Risco , Estações do Ano , Testes Cutâneos , Síndrome
12.
Calcif Tissue Int ; 99(2): 209-23, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27075029

RESUMO

Large bone defects still represent a major burden in orthopedics, requiring bone-graft implantation to promote the bone repair. Along with autografts that currently represent the gold standard for complicated fracture repair, the bone tissue engineering offers a promising alternative strategy combining bone-graft substitutes with osteoprogenitor cells able to support the bone tissue ingrowth within the implant. Hence, the optimization of cell loading and distribution within osteoconductive scaffolds is mandatory to support a successful bone formation within the scaffold pores. With this purpose, we engineered constructs by seeding and culturing autologous, osteodifferentiated bone marrow mesenchymal stem cells within hydroxyapatite (HA)-based grafts by means of a perfusion bioreactor to enhance the in vivo implant-bone osseointegration in an ovine model. Specifically, we compared the engineered constructs in two different anatomical bone sites, tibia, and femur, compared with cell-free or static cell-loaded scaffolds. After 2 and 4 months, the bone formation and the scaffold osseointegration were assessed by micro-CT and histological analyses. The results demonstrated the capability of the acellular HA-based grafts to determine an implant-bone osseointegration similar to that of statically or dynamically cultured grafts. Our study demonstrated that the tibia is characterized by a lower bone repair capability compared to femur, in which the contribution of transplanted cells is not crucial to enhance the bone-implant osseointegration. Indeed, only in tibia, the dynamic cell-loaded implants performed slightly better than the cell-free or static cell-loaded grafts, indicating that this is a valid approach to sustain the bone deposition and osseointegration in disadvantaged anatomical sites.


Assuntos
Osso e Ossos/efeitos dos fármacos , Durapatita/farmacologia , Osseointegração/fisiologia , Engenharia Tecidual , Animais , Substitutos Ósseos/metabolismo , Transplante Ósseo/métodos , Osso e Ossos/metabolismo , Células Cultivadas , Células-Tronco Mesenquimais/citologia , Osteogênese/fisiologia , Ovinos , Engenharia Tecidual/métodos , Alicerces Teciduais
13.
Eur Cell Mater ; 31: 395-406, 2016 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-27232666

RESUMO

Quantitative gene expression analysis is widely used to evaluate the expression of specific tissue markers. To obtain reliable data it is essential to select stable housekeeping genes whose expression is not influenced by the anatomical origin of cells or by the culture conditions. No studies have evaluated housekeeping gene stability in intervertebral disc (IVD) cells and only few studies using cartilaginous endplate (CEP) and articular cartilage (AC) cells are present in the literature. We analysed the stability of four candidate housekeeping genes (GAPDH, TBP, YWHAZ and RPL13A) in human cells isolated from nucleus pulposus (NP) and annulus fibrosus (AF), CEP and AC. Cell isolation, expansion, cryoconservation, and differentiation in 3D pellets were tested. GeNorm, NormFinder, BestKeeper tools and the comparative ΔCt method were used to evaluate housekeeping gene stability. In each cell population, TBP alone or combined with YWHAZ was identified as the best normaliser in both monolayer and 3D pellets. GAPDH was the best performer only for AC cells in monolayer. In most culture conditions considering groups of two or more cell types, TBP was the most stable and YWHAZ was the second choice. GAPDH was the best performer only in 3D pellets with factors for AC and AF combined with CEP cells. RPL13A was the most stable only for AF with CEP cells at isolation. Our findings will be useful to properly design the experimental set-up of studies involving IVD, CEP or AC cells in different culture conditions, in order to obtain accurate and high quality data from quantitative gene expression analysis.


Assuntos
Cartilagem Articular/citologia , Cartilagem Articular/metabolismo , Perfilação da Expressão Gênica , Genes Essenciais , Disco Intervertebral/citologia , Disco Intervertebral/metabolismo , Idoso , Células Cultivadas , Regulação da Expressão Gênica , Estudos de Associação Genética , Humanos , Reprodutibilidade dos Testes , Análise de Célula Única
14.
Eur Cell Mater ; 32: 58-73, 2016 07 08.
Artigo em Inglês | MEDLINE | ID: mdl-27386840

RESUMO

Tendon ruptures and/or large losses remain to be a great clinical challenge and often require full replacement of the damaged tissue. The use of auto- and allografts or engineered scaffolds is an established approach to restore severe tendon injuries. However, these grafts are commonly related to scarce biocompatibility, site morbidity, chronic inflammation and poor biomechanical properties. Recently, the decellularisation techniques of allo- or xenografts using specific detergents have been studied and have been found to generate biocompatible substitutes that resemble the native tissue. This study aims to identify a novel decellularisation protocol for large equine tendons that would produce an extracellular matrix scaffold suitable for the regeneration of injured tendons in humans. Specifically, equine tendons were treated either with tri (n-butyl) phosphate alone, or associated to multiple concentrations of peracetic acid (1, 3 and 5 %), which has never before been tested in vitro.Samples were then analysed by histology and with biochemical, biomechanical, and cytotoxicity tests. The best decellularisation protocol, resulting from these examinations, was selected and the chosen scaffold was re-seeded with murine fibroblasts. Resulting grafts were tested for cell viability, histologic analysis, DNA and collagen content. The results identified 1 % tri (n-butyl) phosphate combined with 3 % peracetic acid as the most suitable decellularised matrix in terms of biochemical and biomechanical properties. Moreover, the non-cytotoxic nature of the decellularised matrix allowed for good fibroblast reseeding, thus demonstrating a biocompatible matrix that will be suitable for tendon tissue engineering and hopefully as substitutes in severe tendon damages.


Assuntos
Materiais Biocompatíveis/farmacologia , Tendões/citologia , Engenharia Tecidual/métodos , Animais , Fenômenos Biomecânicos , Morte Celular/efeitos dos fármacos , Colágeno/metabolismo , DNA/metabolismo , Módulo de Elasticidade , Matriz Extracelular/metabolismo , Glicosaminoglicanos/metabolismo , Cavalos , Estresse Mecânico , Sulfatos/metabolismo
15.
Eur J Paediatr Dent ; 17(2): 93-9, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27377105

RESUMO

AIM: To evaluate the causes of Early Childhood Caries. STUDY DESIGN: A statistical comparison of two groups of patients, aged between 3 and 6, and their parents, in the 3-year period 2013-2015 was performed. Two groups of patients were selected: 40 children with early childhood caries (ECC group) and 40 children with no decay (controls). Parents were asked to fill out a questionnaire on food habits, oral hygiene habits, fluoride prophylaxis and family history of caries. A dental visit of children and parents was performed (collection of dmft and DMFT index). The saliva features of the two groups of children (concentration of Streptococcus mutans, pH of the oral cavity and buffer capability of stimulated saliva) were examined by means of salivary tests. The distribution of the population was analysed and parametric and non-parametric tests for continuous and non-continuous variables, were used when appropriate. The statistical significance was set at p > 0.005. RESULTS AND STATISTICS: The average age, gender and BMI index of children of the two groups was similar. The difference between the two groups is statistically significant for pH (p=0.000) and buffer capability (p=0.001). The DMFT index in the group of mothers of ECC children is higher compared to the mothers of controls (p=0.004). The comparison among the means, for the parameters connected to the consumption of fruit juice, tea, soft drinks, candies, use of pacifier dipped in sugar or honey and the length (in months) of consumption of candies show statistically significant differences (p>0.05). The pH is higher in those children (24) who never consumed fruit juice, tea, soft drinks before the information received (p<0.05). CONCLUSIONS: Children developing Early Childhood Caries have a diet characterised by high free sugars intake. Diet and, in particular, the consumption of drinks containing free sugars is the most important factor in determining the onset of ECC, being able to affect the oral environment and, in particular, saliva. There is a relationship between maternal and child's oral health, with a DMFT higher in mothers of ECC subjects than in controls' mothers.


Assuntos
Cárie Dentária/epidemiologia , Dieta , Criança , Pré-Escolar , Cárie Dentária/etiologia , Cárie Dentária/genética , Humanos , Fatores de Risco , Estados Unidos
16.
Osteoarthritis Cartilage ; 23(11): 1853-7, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26521731

RESUMO

OBJECTIVE: Synovitis with an increased presence of macrophages is observed in osteoarthritis (OA) and rheumatoid arthritis (RA). Given the important role of macrophages in arthritis, we investigated the influence of OA and RA synovial fluid (SF) on primary human monocytes (Mo), their lineage precursors. METHOD: Adherent monocytes without any stimulation (Mo(-)) or stimulated with IFN-γ and TNF-α (Mo(IFN-γ/TNF-α)) or IL-4 (Mo(IL-4)) were exposed to SF from 6 donors without any known joint disease (SF-Ctrl), 10 OA donors (SF-OA), and 10 RA donors (SF-RA). The transcriptional expression of IL6, IL1B, TNFA, IL10, CCL18, CD206, and IL1RA was analyzed. RESULTS: Mo(-) exposed to SF-RA had a lower expression of IL10 and a higher expression of IL1RA than when exposed to SF-Ctrl. Mo(IL-4) exposed to SF-RA had a lower expression of IL10 and CCL18 than when exposed to SF-Ctrl and Mo(IFN-γ/TNF-α) were not affected by SF-RA. Mo exposed to SF-OA also expressed less IL10, but only upon stimulation with IL-4, and expressed more IL1RA than when exposed to SF-Ctrl in any condition. CONCLUSION: A lower expression of IL10 may be regarded as a response to less inflammatory conditions since IL10 expression is higher in response to IFN-γ/TNF-α stimulation, probably as a feedback mechanism. Therefore, the lower expression of IL10 and the higher expression of IL1RA in Mo exposed to arthritic than to non-arthritic SF suggest that arthritic SF is mainly reducing the inflammatory responses in Mo. This may mimic the response of monocytes/macrophages recruited to the joint, where feedback mechanisms counteract pro-inflammatory processes.


Assuntos
Artrite Reumatoide/genética , Regulação da Expressão Gênica , Proteína Antagonista do Receptor de Interleucina 1/genética , Interleucina-10/genética , Monócitos/metabolismo , Osteoartrite/genética , Líquido Sinovial/metabolismo , Idoso , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Feminino , Humanos , Proteína Antagonista do Receptor de Interleucina 1/biossíntese , Interleucina-10/biossíntese , Masculino , Pessoa de Meia-Idade , Osteoartrite/metabolismo , Osteoartrite/patologia , Reação em Cadeia da Polimerase , RNA/genética , Líquido Sinovial/citologia
17.
Ecology ; 95(4): 1010-21, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24933819

RESUMO

The installation of green roofs, defined here as rooftops with a shallow soil cover and extensive vegetation, has been proposed as a possible measure to mitigate the loss of green space caused by the steady growth of cities. However, the effectiveness of green roofs in supporting arthropod communities, and the extent to which they facilitate connectivity of these communities within the urban environment is currently largely unknown. We investigated the variation of species community composition (beta diversity) of four arthropod groups with contrasting mobility (Carabidae, Araneae, Curculionidae, and Apidae) on 40 green roofs and 40 extensively managed green sites on the ground in the city of Zurich, Switzerland. With redundancy analysis and variation partitioning, we (1) disentangled the relative importance of local environmental conditions, the surrounding land cover composition, and habitat connectivity on species community composition, (2) searched for specific spatial scales of habitat connectivity for the different arthropod groups, and (3) discussed the ecological and functional value of green roofs in cities. Our study revealed that on green roofs community composition of high-mobility arthropod groups (bees and weevils) were mainly shaped by habitat connectivity, while low-mobility arthropod groups (carabids and spiders) were more influenced by local environmental conditions. A similar but less pronounced pattern was found for ground communities. The high importance of habitat connectivity in shaping high-mobility species community composition indicates that these green roof communities are substantially connected by the frequent exchange of individuals among surrounding green roofs. On the other hand, low-mobility species communities on green roofs are more likely connected to ground sites than to other green roofs. The integration of green roofs in urban spatial planning strategies has great potential to enable higher connectivity among green spaces, so that eventually even communities of low-mobility species become connected. Furthermore, improving the design of green roofs (composition and configuration of vegetation and soil types) could enhance the ecological value, particularly for low-mobility species.


Assuntos
Artrópodes/classificação , Artrópodes/fisiologia , Ecossistema , Animais , Cidades , Materiais de Construção , Demografia , Monitoramento Ambiental , Habitação , Suíça , Fatores de Tempo
18.
Eur Cell Mater ; 27: 298-311, 2014 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-24760577

RESUMO

Cell-based therapies have recently been proposed for the treatment of degenerative articular pathologies, such as early osteoarthritis, with an emphasis on autologous mesenchymal stem cells (MSCs), as an alternative to terminally differentiated cells. In this study, we performed a donor-matched comparison between infrapatellar fat pad MSCs (IFP-MSCs) and knee subcutaneous adipose tissue stem cells (ASCs), as appealing candidates for cell-based therapies that are easily accessible during surgery. IFP-MSCs and ASCs were obtained from 25 osteoarthritic patients undergoing total knee replacement and compared for their immunophenotype and differentiative potential. Undifferentiated IFP-MSCs and ASCs displayed the same immunophenotype, typical of MSCs (CD13+/CD29+/CD44+/CD73+/CD90+/CD105+/CD166+/CD31-/CD45-). IFP-MSCs and ASCs showed similar adipogenic potential, though undifferentiated ASCs had higher LEP expression compared to IFP-MSCs (p<0.01). Higher levels of calcified matrix (p<0.05) and alkaline phosphatase (p<0.05) in ASCs highlighted their superior osteogenic commitment compared to IFP-MSCs. Conversely, IFP-MSCs pellets showed greater amounts of glycosaminoglycans (p<0.01) and superior expression of ACAN (p<0.001), SOX9, COMP (p<0.001) and COL2A1 (p<0.05) compared to ASCs pellets, revealing a superior chondrogenic potential. This was also supported by lower COL10A1 (p<0.05) and COL1A1 (p<0.01) expression and lower alkaline phosphatase release (p<0.05) by IFP-MSCs compared to ASCs. The observed dissimilarities between IFP-MSCs and ASCs show that, despite expressing similar surface markers, MSCs deriving from different fat depots in the same surgical site possess specific features. Furthermore, the in vitro peculiar commitment of IFP-MSCs and ASCs from osteoarthritic donors towards the chondrogenic or osteogenic lineage may suggest a preferential use for cartilage and bone cell-based treatments, respectively.


Assuntos
Tecido Adiposo/patologia , Condrogênese , Células-Tronco Mesenquimais/citologia , Osteoartrite/patologia , Osteogênese , Patela/patologia , Doadores de Tecidos , Adipogenia , Tecido Adiposo/citologia , Idoso , Idoso de 80 Anos ou mais , Agrecanas/genética , Agrecanas/metabolismo , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Antígenos CD/genética , Antígenos CD/metabolismo , Cálcio/metabolismo , Proteína de Matriz Oligomérica de Cartilagem/genética , Proteína de Matriz Oligomérica de Cartilagem/metabolismo , Células Cultivadas , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Cadeia alfa 1 do Colágeno Tipo I , Colágeno Tipo X/genética , Colágeno Tipo X/metabolismo , Feminino , Glicosaminoglicanos/metabolismo , Humanos , Leptina/genética , Leptina/metabolismo , Masculino , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Patela/citologia , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo
19.
Ecol Appl ; 24(7): 1583-95, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-29210224

RESUMO

To ensure viable species populations in fragmented landscapes, individuals must be able to move between suitable habitat patches. Despite the increased interest in biodiversity assessment in urban environments, the ecological relevance of habitat connectivity in highly fragmented landscapes remains largely unknown. The first step to understanding the role of habitat connectivity in urban ecology is the challenging task of assessing connectivity in the complex patchwork of contrasting habitats that is found in cities. We developed a data-based framework, minimizing the use of subjective assumptions, to assess habitat connectivity that consists of the following sequential steps: (1) identification of habitat preference based on empirical habitat-use data; (2) derivation of habitat resistance surfaces evaluating various transformation functions; (3) modeling of different connectivity maps with electrical circuit theory (Circuitscape), a method considering all possible pathways across the landscape simultaneously; and (4) identification of the best connectivity map with information-theoretic model selection. We applied this analytical framework to assess habitat connectivity for the European hedgehog Erinaceus europaeus, a model species for ground-dwelling animals, in the city of Zurich, Switzerland, using GPS track points from 40 individuals. The best model revealed spatially explicit connectivity "pinch points," as well as multiple habitat connections. Cross-validation indicated the general validity of the selected connectivity model. The results show that both habitat connectivity and habitat quality affect the movement of urban hedgehogs (relative importance of the two variables was 19.2% and 80.8%, respectively), and are thus both relevant for predicting urban animal movements. Our study demonstrates that even in the complex habitat patchwork of cities, habitat connectivity plays a major role for ground-dwelling animal movement. Data-based habitat connectivity maps can thus serve as an important tool for city planners to identify habitat corridors and plan appropriate management and conservation measures for urban animals. The analytical framework we describe to model such connectivity maps is generally applicable to different types of habitat-use data and can be adapted to the movement scale of the focal species. It also allows evaluation of the impact of future landscape changes or management scenarios on habitat connectivity in urban landscapes.


Assuntos
Conservação dos Recursos Naturais/métodos , Ecossistema , Ouriços/fisiologia , Animais , Cidades , Monitoramento Ambiental , Mapeamento Geográfico , Suíça
20.
Conserv Biol ; 28(2): 414-26, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24372620

RESUMO

Surrogates, such as umbrella species, are commonly used to reduce the complexity of quantifying biodiversity for conservation purposes. The presence of umbrella species is often indicative of high taxonomic diversity; however, functional diversity is now recognized as an important metric for biodiversity and thus should be considered when choosing umbrella species. We identified umbrella species associated with high taxonomic and functional biodiversity in urban areas in Switzerland. We analyzed 39,752 individuals of 574 animal species from 96 study plots and 1397 presences of 262 plant species from 58 plots. Thirty-one biodiversity measures of 7 taxonomic groups (plants, spiders, bees, ground beetles, lady bugs, weevils and birds) were included in within- and across-taxa analyses. Sixteen measures were taxonomical (species richness and species diversity), whereas 15 were functional (species traits including mobility, resource use, and reproduction). We used indicator value analysis to identify umbrella species associated with single or multiple biodiversity measures. Many umbrella species were indicators of high biodiversity within their own taxonomic group (from 33.3% in weevils to 93.8% in birds), to a lesser extent they were indicators across taxa. Principal component analysis revealed that umbrella species for multiple measures of biodiversity represented different aspects of biodiversity, especially with respect to measures of taxonomic and functional diversity. Thus, even umbrella species for multiple measures of biodiversity were complementary in the biodiversity aspects they represented. Thus, the choice of umbrella species based solely on taxonomic diversity is questionable and may not represent biodiversity comprehensively. Our results suggest that, depending on conservation priorities, managers should choose multiple and complementary umbrella species to assess the state of biodiversity.


Assuntos
Artrópodes , Biodiversidade , Aves , Cidades , Plantas , Animais , Conservação dos Recursos Naturais , Suíça
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