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1.
Acta Chir Orthop Traumatol Cech ; 89(2): 104-107, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35621399

RESUMO

PURPOSE OF THE STUDY To evaluate a possible association between hip fracture and statin use. MATERIAL AND METHODS In this case-control study we compared the use of statins between two groups of 210 patients: the first group (case group) included patients hospitalized for hip fractures while the second group (control group) included patients who did not suffer femur bone injuries. The two groups were matched for age, sex, year of hospitalization and possible confounding factors. Inside the group of cases, we also evaluated the differences in terms of fracture type, presence of previous fragility fracture and mortality between statin users and non-users. RESULTS The use of statins was most common among patients without previous fractures (OR=0.54; 95% CI=0.33-0.89; p=0.0138), especially in older patients (OR=0.40; 95% CI=0.22-0.76). We did not find any significant difference in statin intake between men and women in the control group. In the case group, those who did not use statins were more likely to undergo a medial hip fracture (28.5% vs 16.1%). Patients from case group also presented a greater mortality (27.9% vs 19.35%) and an higher percentage of previous hip fractures (20.11% vs 9.7%). However, they didn't presented a significant higher rate of fragility fractures in other sites. DISCUSSION AND CONCLUSIONS Our study suggests a reduced hip fracture risk, especially in cases aged 80 or more, a different fracture pattern (lower percentage of medial fractures) and a reduced mortality at 9 months in patients treated with HMG-CoA reductase inhibitors, confirming the previous evidences reported in literature. Key words: statin, hip fractures, fracture risk, osteoporosis.


Assuntos
Fraturas do Quadril , Inibidores de Hidroximetilglutaril-CoA Redutases , Osteoporose , Idoso , Osso e Ossos , Estudos de Casos e Controles , Feminino , Fraturas do Quadril/epidemiologia , Fraturas do Quadril/prevenção & controle , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/efeitos adversos , Masculino
2.
J Biol Regul Homeost Agents ; 31(4 suppl 1): 121-127, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29186948

RESUMO

Bone cement implantation syndrome (BCIS) is a rare form of intraoperative pulmonary embolism (EP) that occurs during cementation. It can be explained by two main theories: the monomer mediated model and the mechanic model. Our goal is to evaluate thromboelastographic changes in patients undergoing surgery for femoral neck fractures. We recruited 32 patients with a femoral neck fracture. The average age was 81.91 years (range 62-95). The patients were divided in two different groups: cemented hip arthroplasty (CC, 13 patients) and other surgical non-cemented techniques (SC, non-cemented hip arthroplasty, osteosynthesis). The coagulation was evaluated by TEG in the early pre-operatory (time A) and post-operatory (time B), both on native blood and on blood added with Heparinase. We used the t-test to compare the differences between the two groups. The coagulation index CI was modified on hypercoagulability by surgery in both groups, but without statistical significance between the two groups (p>0.05). R parameter decreases between time A and time B in the same way in both groups (p>0.05). Parameter MA had no major variations between time A and B, without statistical significance (p>0.05). From our study it is evident that although the surgery would result in a change in the layout of the TEG toward hypercoagulability, this is similar both in cemented and non-cemented surgical interventions for femoral neck fractures in elderly patients. An altered coagulation does not appear to be the cause or a factor in determining the BCIS.

3.
J Biol Regul Homeost Agents ; 31(4 suppl 1)2017.
Artigo em Inglês | MEDLINE | ID: mdl-29186945

RESUMO

Gamma-glutamyltransferase (GGT) has been recently identified as a bone-resorbing factor. The aim of this study was to investigate the association between plasma GGT fractions levels and bone quality. Plasma GGT fractions were analysed by gel-filtration chromatography. Bone quality was established quantitatively by two micro-CT derived microarchitectural parameters: the BV/TV (mineralised bone volume/total volume), and the SMI (structure model index) that describes the rod-like (low resistant) or plate-like (high-resistant) shape of bone trabeculae. We enrolled 93 patients hospitalised for elective total hip replacement (group Arthrosis, n=46) or for proximal femoral fracture (group Fracture, n=47). Patients within the first quartile of BV/TV (Q1, osteoporotic patients, n=6) showed higher levels of b-GGT fraction [median (min-max): 3.37 (1.42­6.81)] compared to patients with normal bone density (fourth quartile Q4, n=10; 1.40 (0.83­4.36); p=0.0393]. Also, according to SMI, b-GGT value was higher in the subgroup with bone fragility [Q1, n=8: 1.36 (0.43­4.36); Q4, n=8: 5.10 (1.4 ­7.60); p=0.0117]. In conclusion, patients characterised by fragile bone structure showed specifically higher levels of plasma b-GGT activity thus suggesting fractional GGT analysis as a possible biomarker in the diagnosis of osteoporosis.

4.
Nutr Metab Cardiovasc Dis ; 26(1): 60-6, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26643211

RESUMO

BACKGROUND AND AIMS: Childhood obesity promotes adverse changes in cardiovascular structure and function. This study evaluated whether these changes are related to intra-abdominal adiposity and associated cardiometabolic risk or to body-size induced hemodynamic overload. METHODS AND RESULTS: 55 obese children/adolescents and 35 healthy-weight controls underwent carotid, cardiac and abdominal ultrasound to assess carotid artery intima-media thickness (IMT), diameter, distension and stiffness, left ventricular (LV) dimension, mass and function and extent of intra-abdominal adiposity. As compared to controls with healthy BMI, obese children had higher systolic blood pressure (BP), stroke volume and lower total peripheral resistance (P < 0.001-0.0001), higher plasma triglycerides, glycated hemoglobin, insulin and HOMA-IR index (P = 0.01-<0.0001), higher carotid IMT, diameter and distension (P < 0.005-0.0005), higher LV diameter, wall thickness and mass (P < 0.001-0.0001), and impaired LV diastolic function assessed by myocardial longitudinal performance (P < 0.005). In entire population, independent determinants of carotid diameter, LV diameter, wall thickness and mass were fat-free mass (or stroke volume, respectively) and BP. Carotid distension was determined by carotid diameter and BP, and carotid IMT by carotid diameter, BP, HDL-cholesterol and glycated hemoglobin. LV diastolic performance was inversely related to preperitoneal fat thickness and plasma insulin levels. CONCLUSIONS: Obese youths present signs of impaired lipid and glucose metabolism, hyperdynamic circulation and cardiovascular changes. Increase in LV dimensions and mass and in carotid diameter and distension seems to reflect adaptation to body-size induced increase in hemodynamic load, changes in LV diastolic performance a negative impact of intra-abdominal adiposity and associated metabolic risk, and increase in IMT both adaptive remodeling and metabolic risk.


Assuntos
Adiposidade , Doenças Cardiovasculares/etiologia , Hemodinâmica , Gordura Intra-Abdominal/fisiopatologia , Obesidade Infantil/complicações , Adolescente , Fatores Etários , Biomarcadores/sangue , Glicemia/análise , Doenças Cardiovasculares/diagnóstico , Doenças Cardiovasculares/fisiopatologia , Doenças das Artérias Carótidas/diagnóstico , Doenças das Artérias Carótidas/etiologia , Doenças das Artérias Carótidas/fisiopatologia , Espessura Intima-Media Carotídea , Estudos de Casos e Controles , Criança , Feminino , Humanos , Hipertrofia Ventricular Esquerda/diagnóstico , Hipertrofia Ventricular Esquerda/etiologia , Hipertrofia Ventricular Esquerda/fisiopatologia , Lipídeos/sangue , Masculino , Obesidade Infantil/sangue , Obesidade Infantil/diagnóstico , Obesidade Infantil/fisiopatologia , Medição de Risco , Fatores de Risco , Disfunção Ventricular Esquerda/diagnóstico , Disfunção Ventricular Esquerda/etiologia , Disfunção Ventricular Esquerda/fisiopatologia , Função Ventricular Esquerda , Remodelação Ventricular
5.
Invest New Drugs ; 31(2): 381-9, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22555773

RESUMO

Purpose To study the influence of repeated oral administration of ketoconazole, a potent CYP3A4 inhibitor, on the plasma pharmacokinetics of eribulin mesylate administered by single-dose intravenous infusion. Eribulin mesylate is a non-taxane microtubule dynamics inhibitor that is currently under development in phase I-III trials for the treatment of solid tumors. Experimental design A randomized, open-label, two treatments, two sequences, crossover phase I study was performed in patients with advanced solid tumors. Treatments were given on day 1 and day 15 and consisted of 1.4 mg/m(2) eribulin mesylate alone or 0.7 mg/m(2) eribulin mesylate plus 200 mg ketoconazole on the day of eribulin mesylate administration and the following day. Pharmacokinetic sampling for determination of eribulin plasma concentration was performed up to 144 h following administration of eribulin mesylate. Also safety and anti-tumor activity were determined. Results Pharmacokinetic sampling and analysis was completed in ten patients. Statistical analysis of dose-normalized log-transformed AUC0-∞ and Cmax indicated that single-dose exposure of eribulin was not statistically different when co-administered with ketoconazole (ratio of geometric least square means: 0.95 (90%CI: 0.80-1.12) and 0.97 (90%CI: 0.83-1.12), respectively) in patients with solid tumors. Ketoconazole had no effect on eribulin clearance and elimination half-life. The most frequently reported treatment related adverse events were fatigue and nausea, each reported in 8/12 patients. Seven patients (58.3 %) achieved stable disease as best overall response. Conclusions The results indicate that eribulin mesylate can be safely co-administered with ketoconazole. Drug-drug interactions are not expected with other CYP3A4 inhibitors.


Assuntos
Antifúngicos/administração & dosagem , Furanos/uso terapêutico , Cetoconazol/administração & dosagem , Cetonas/uso terapêutico , Neoplasias/tratamento farmacológico , Administração Oral , Idoso , Relação Dose-Resposta a Droga , Feminino , Seguimentos , Humanos , Infusões Intravenosas , Masculino , Dose Máxima Tolerável , Pessoa de Meia-Idade , Neoplasias/metabolismo , Neoplasias/patologia , Prognóstico , Distribuição Tecidual
6.
Nanotechnology ; 24(17): 175601, 2013 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-23548801

RESUMO

A new approach to thermal decomposition of organic iron precursors is reported, which results in a simpler and more economical method to produce well crystallized γ-Fe2O3 nanoparticles (NPs) with average sizes within the 3-17 nm range. The NPs were characterized by TEM, SAED, XRD, DLS-QELS, Mössbauer spectroscopy at different temperatures, FT-IR and magnetic measurements. The obtained γ-Fe2O3 NPs are coated with oleic acid and, in a lower quantity, with oleylamine (about 1.5 nm in thickness). It was shown that changing operative variables allows us to tune the average particle diameters and obtain a very narrow or monodisperse distribution of sizes. The γ-Fe2O3 NPs behave superparamagnetically at room temperature and their magnetization saturation is reduced by about 34% in comparison with bulk maghemite. The results indicate that the distance between two neighbour NPs, generated by the coating, of about 3 nm is insufficient to inhibit interparticle magnetic interactions when the average diameter is 8.8 nm. The good quality of the NPs, obtained through the present low-cost and easy-handling process, open a new perspective for future technological applications.


Assuntos
Nanopartículas de Magnetita/química , Nanotecnologia/métodos , Nanopartículas de Magnetita/ultraestrutura , Nanotecnologia/economia , Tamanho da Partícula , Espectroscopia de Infravermelho com Transformada de Fourier , Espectroscopia de Mossbauer , Temperatura
7.
Environ Int ; 179: 108131, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37586275

RESUMO

COVID-19 pandemic had a significant impact on global public health. The spread of the disease was related to the high transmissibility of SARS-CoV-2 virus but incidence and mortality rate suggested a possible relationship with environmental factors. Air pollution has been hypothesized to play a role in the transmission of the virus and the resulting severity of the disease. Here we report a plausible in vitro toxicological mode of action by which fine particulate matter (PM2.5) could promote a higher infection rate of SARS-CoV-2 and severity of COVID-19 disease. PM2.5 promotes a 1.5 fold over-expression of the angiotensin 2 converting enzyme (ACE2) which is exploited by viral particles to enter human lung alveolar cells (1.5 fold increase in RAB5 protein) and increases their inflammatory state (IL-8 and NF-kB protein expression). Our results provide a basis for further exploring the possible synergy between biological threats and air pollutants and ask for a deeper understanding of how air quality could influence new pandemics in the future.


Assuntos
Poluentes Atmosféricos , Poluição do Ar , COVID-19 , Humanos , Material Particulado/toxicidade , Material Particulado/análise , SARS-CoV-2 , Pandemias , Poluentes Atmosféricos/toxicidade , Poluentes Atmosféricos/análise , Poluição do Ar/análise
8.
ESMO Open ; 7(6): 100611, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36463731

RESUMO

BACKGROUND: In ∼3%-5% of patients with metastatic disease, tumor origin remains unknown despite modern imaging techniques and extensive pathology work-up. With long diagnostic delays and limited and ineffective therapy options, the clinical outcome of patients with cancer of unknown primary (CUP) remains poor. Large-scale genome sequencing studies have revealed that tumor types can be predicted based on distinct patterns of somatic variants and other genomic characteristics. Moreover, actionable genomic events are present in almost half of CUP patients. This study investigated the clinical value of whole genome sequencing (WGS) in terms of primary tumor identification and detection of actionable events, in the routine diagnostic work-up of CUP patients. PATIENTS AND METHODS: A WGS-based tumor type 'cancer of unknown primary prediction algorithm' (CUPPA) was developed based on previously described principles and validated on a large pan-cancer WGS database of metastatic cancer patients (>4000 samples) and 254 independent patients, respectively. We assessed the clinical value of this prediction algorithm as part of routine WGS-based diagnostic work-up for 72 CUP patients. RESULTS: CUPPA correctly predicted the primary tumor type in 78% of samples in the independent validation cohort (194/254 patients). High-confidence predictions (>95% precision) were obtained for 162/254 patients (64%). When integrated in the diagnostic work-up of CUP patients, CUPPA could identify a primary tumor type for 49/72 patients (68%). Most common diagnoses included non-small-cell lung (n = 7), gastroesophageal (n = 4), pancreatic (n = 4), and colorectal cancer (n = 3). Actionable events with matched therapy options in clinical trials were identified in 47% of patients. CONCLUSIONS: Genome-based tumor type prediction can predict cancer diagnoses with high accuracy when integrated in the routine diagnostic work-up of patients with metastatic cancer. With identification of the primary tumor type in the majority of patients and detection of actionable events, WGS is a valuable diagnostic tool for patients with CUP.


Assuntos
Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Neoplasias Primárias Desconhecidas , Humanos , Neoplasias Primárias Desconhecidas/diagnóstico , Neoplasias Primárias Desconhecidas/genética , Neoplasias Primárias Desconhecidas/tratamento farmacológico , Genômica , Sequenciamento Completo do Genoma
9.
Injury ; 52(8): 2459-2462, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33731290

RESUMO

INTRODUCTION: Periprosthetic femoral fractures (PFF) actually represent a serious public health problem. They are reported to occur in 0,1-4.5% of all patients undergoing total hip replacement (THR). PFF are commonly distinguished using the Vancouver classification. This study principal aim is to evaluate results obtained using the Intrauma Iron Lady® Conical Coupling locking plate for the treatment of Vancouver type B1 periprosthetic femoral fractures. MATERIALS AND METHODS: We enrolled 32 patients affected by Vancouver B1 PFF and treated with the same device. Metal cerclages were additionally used in 12 (38%) patients. A clinical and radiographical post-operative follow-up was then planned at 1, 3 and 6 months after surgery; than the follow-up was annually fixed. RESULTS: Mean age at the moment of trauma was 76,7 years. All involved femoral stem were uncemented and the they were all radiographically and intraoperativelly judged to be stable. Mean post-operative follow-up period was 5,8 years. 29 patients (91%) presented healed fracture at 6 months follow-up. 9% patients developed a superficial surgical site infection. DISCUSSION AND CONCLUSIONS: Literature highlights that Vancouver B1 PFF should be treated with open reduction and internal fixation (ORIF) using polyaxial locking plates. However, no single technique has gained universal acceptance to be superior that the other. The current reported healing rate ranges from 40 to 100%. Using the Intrauma Iron Lady® Conical Coupling locking plate, we obtained a healing rate of 91%; this data is consistent with recent literature. Moreover, the role of cerclages in addition to femoral plating is actually controversial because they potentially damage the soft callus vascularization. Our results showed no difference in term of healing rate between patients with and without cerclages, according with some of most recent articles. A prospective study with a higher number of patients should be carried out in order to better evaluate the role of cerclages on healing rate but also the complications frequency after PFF surgical treatment.


Assuntos
Artroplastia de Quadril , Placas Ósseas , Fraturas do Fêmur , Fraturas Periprotéticas , Fraturas do Fêmur/diagnóstico por imagem , Fraturas do Fêmur/cirurgia , Fixação Interna de Fraturas , Consolidação da Fratura , Humanos , Fraturas Periprotéticas/diagnóstico por imagem , Fraturas Periprotéticas/cirurgia , Estudos Prospectivos , Reoperação , Estudos Retrospectivos
10.
Anal Chim Acta ; 1142: 201-210, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33280698

RESUMO

There is an increasing interest in determining the concentration of furanic compounds naturally formed in food aqueous matrices, by in situ, fast and low-cost methods. A sensor presenting such characteristics is here proposed, and characterized. It is based on a molecularly imprinted polymer (MIP) as a receptor with electrochemical transduction on a screen printed cell (SPC). The molecularly imprinted polymer has been developed for a particular furanic derivative, 2-furaldehyde (2-FAL). The detection bases on the reduction of 2-FAL selectively adsorbed on the polymer layer in contact with the working electrode. The polymer layer is simply formed by in situ polymerization, directly over the SPC and it was characterized by IR, SEM and electrochemical methods. Even if based on an easy and fast preparation procedure, the layer sufficiently adheres to the cell surface giving a reusable sensor. Square wave voltammetry (SWV) was applied as the signal acquisition method. The sensor performance in aqueous solution (NaCl 0.1 M) was tested, obtaining that the dose-response curve is fitted by the Langmuir adsorption isotherm. The sensitivity, and so the limit of detection, were noticeably improved by a chemometric approach based on the Design of experiment method. (optimized conditions: Estep = 0.03 V, Epulse = 0.066 V, f = 31 s-1). In water solution at pH around neutrality the dynamic range was from about 50 µM to 20 mM. Similar results were obtained for a white wine containing 12% ethanol, which has been considered as a typical example of beverage possibly containing furhaldehydes. The higher limit of quantification can be modulated by the amount of MIP deposited, while the lower detection limit by the conditions of the electrochemical measurement.


Assuntos
Impressão Molecular , Bebidas , Técnicas Eletroquímicas , Eletrodos , Furaldeído , Limite de Detecção , Polímeros Molecularmente Impressos
12.
Clin Pharmacol Drug Dev ; 9(8): 929-937, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-31970932

RESUMO

Microdose studies are exploratory trials to determine early drug pharmacokinetics in humans. In this trial we examined whether the pharmacokinetics of gemcitabine at a therapeutic dose could be predicted from the pharmacokinetics of a microdose. In this prospective, open-label microdosing study, a gemcitabine microdose (100 µg) was given intravenously to participants on day 1, followed by a therapeutic dose (1250 mg/m2 ) on day 2. Gemcitabine and its metabolite 2',2'-difluorodeoxyuracil (dFdU) were quantified in plasma and intracellularly by using liquid chromatography-mass spectrometry). Noncompartmental pharmacokinetic analysis was performed. Ten patients participated in this study. The mean area under the plasma concentration-time curve (AUC0-8 ) of gemcitabine after microdosing was 0.00074 h·mg/L and after therapeutic dosing was 16 h·mg/L. The mean AUC0-8 of dFdU following the microdose and therapeutic dose were 0.022 h·mg/L and 169 h·mg/L, respectively. Exposure to gemcitabine after the therapeutic dose was within 2-fold of the exposure following a microdose, when linearly extrapolated to 1250 mg/m2 . However, the shape of the concentration-time curve was different, as reflected by poor scalability in volume of distribution (939 L versus 222 L). Furthermore, intracellularly phosphorylated gemcitabine and phosphorylated dFdU levels could not be predicted from the microdose. The AUC0-8 of gemcitabine at therapeutic dose was accurately predicted by the pharmacokinetics of a microdose, when linearly extrapolated to 1250 mg/m2 . Volume of distribution, elimination rate constant, and intracellular pharmacokinetics of the therapeutic dose could not be predicted from the microdose, which demonstrates limitations of the microdose approach in this case.


Assuntos
Antimetabólitos Antineoplásicos/farmacocinética , Cromatografia Líquida/métodos , Desoxicitidina/análogos & derivados , Relação Dose-Resposta a Droga , Espectrometria de Massas/instrumentação , Administração Intravenosa , Idoso , Antimetabólitos Antineoplásicos/administração & dosagem , Antimetabólitos Antineoplásicos/sangue , Antimetabólitos Antineoplásicos/uso terapêutico , Área Sob a Curva , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Desoxicitidina/administração & dosagem , Desoxicitidina/sangue , Desoxicitidina/farmacocinética , Desoxicitidina/uso terapêutico , Feminino , Humanos , Masculino , Mesotelioma/tratamento farmacológico , Pessoa de Meia-Idade , Projetos Piloto , Valor Preditivo dos Testes , Estudos Prospectivos , Timoma/tratamento farmacológico , Gencitabina
13.
Audiol Neurootol ; 14(5): 290-5, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19372646

RESUMO

BACKGROUND: The cause of about 30% of bilateral sensorineural hearing loss (SNHL) is still unknown. A viral etiology is among the most frequently proposed ones and the supposed diagnosis is only based upon few clinical and laboratory data. The detection of viral presence within a damaged compartment may represent a way to supply interesting data for confirmation of viral etiology and to explain pathogenic mechanisms. OBJECTIVES: The aim of our study was to identify the possible presence of pathogenic viruses in the inner ear extracellular compartment in patients with bilateral severe sensorineural deafness of unknown etiology who underwent cochlear implant surgery. METHODS: 4 patients, aged from 2 to 7 years and affected by SNHL underwent cochlear implantation surgery and, at the same time, endolabyrinthine fluid sampling. The samples were subsequently sent for viral nucleic acid extraction and polymerase chain reaction (PCR) treatment: multiplex PCR and realtime-PCR were used. In each endolabyrinthine fluid sample, cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella-zoster virus (VZV), herpes simplex virus type 1 and 2 (HSV-1, HSV-2) and enterovirus genomes were searched for. RESULTS: One patient was positive for intracochlear CMV, as confirmed by another base-pair segment PCR. EBV, VZV, HSV and enterovirus were detected in none of the 4 patients. CONCLUSIONS: Our finding of CMV genome within the cochlea of a deaf patient without any evidence of acute and prenatal CMV infection suggests its possible role in postnatal inner ear injury through reactivation of latent virus within the cochlea. This hypothesis could also be considered valid for some patients with anti-CMV-IgG-positive serology and absence of endolabyrinthine viral genome since viruses can be in an inactive state at the time of fluid collection. PCR has proved to be a very useful tool in order to investigate infectious causes of deafness even for more than one virus type at a time and in a limited quantity of sample, such as the small volume of endolabyrinthine liquid collected from children during cochlear implant surgery.


Assuntos
Infecções por Citomegalovirus/complicações , Citomegalovirus/genética , Surdez/virologia , Endolinfa/virologia , Perda Auditiva Neurossensorial/virologia , Criança , Pré-Escolar , Cóclea/virologia , Implante Coclear , Citomegalovirus/crescimento & desenvolvimento , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/virologia , Surdez/cirurgia , Genoma Viral , Perda Auditiva Neurossensorial/cirurgia , Humanos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Latência Viral
14.
Oncogene ; 26(17): 2445-58, 2007 Apr 12.
Artigo em Inglês | MEDLINE | ID: mdl-17043649

RESUMO

Imatinib targets the Bcr-Abl oncogene that causes chronic myelogenous leukemia (CML) in humans. Recently, we demonstrated that besides triggering apoptosis in K562 cells, imatinib also mediated their erythroid differentiation. Although both events appear to proceed concomitantly, it is not known at present whether or not imatinib-induced apoptosis and differentiation are interdependent processes. Hence, we investigated the requirements for Bcr-Abl inhibitor-mediated apoptosis and erythroid differentiation in several established and engineered CML cell lines. Imatinib triggered apoptosis and erythroid differentiation of different CML cell lines, but only apoptosis exhibited sensitivity to ZVAD-fmk inhibition. Conversely, the p38 mitogen-activated protein (MAP) kinase inhibitor, SB202190, significantly slowed down erythroid differentiation without affecting caspase activation. Furthermore, imatinib and PD166326, another Bcr-Abl inhibitory molecule, triggered erythroid differentiation of K562 cell clones, nevertheless resistant to Bcr-Abl inhibitor-induced apoptosis. Finally, short hairpin RNA inhibitor (shRNAi) silencing of caspase 3 efficiently inhibited caspase activity but had no effect on erythroid differentiation, whereas silencing of Bcr-Abl mimicked imatinib or PD166326 treatment, leading to increased apoptosis and erythroid differentiation of K562 cells. Taken together, our findings not only demonstrate that Bcr-Abl inhibitor-mediated apoptosis and differentiation are fully distinguishable events, but also that caspases are dispensable for erythroid differentiation of established CML cell lines.


Assuntos
Apoptose/efeitos dos fármacos , Inibidores de Caspase , Diferenciação Celular/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Leucemia Mielogênica Crônica BCR-ABL Positiva/enzimologia , Proteínas Tirosina Quinases/antagonistas & inibidores , Benzamidas , Caspases/metabolismo , Linhagem Celular Tumoral , Ativação Enzimática/efeitos dos fármacos , Células Eritroides/citologia , Células Eritroides/enzimologia , Células Eritroides/patologia , Proteínas de Fusão bcr-abl , Humanos , Mesilato de Imatinib , Células K562 , Leucemia Mielogênica Crônica BCR-ABL Positiva/sangue , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Piperazinas/farmacologia , Proteínas Tirosina Quinases/sangue , Piridinas/farmacologia , Pirimidinas/farmacologia
15.
Phys Rev E Stat Nonlin Soft Matter Phys ; 77(2 Pt 1): 021910, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18352054

RESUMO

In order to investigate the protein folding-unfolding process, dynamic light scattering (DLS) and atomic force microscopy (AFM) imaging were used to study two fragments of the muscle cardiac protein beta-connectin, also known as titin. Both fragments belong to the I band of the sarcomer, and they are composed of four domains from I(27) to I(30) (tetramer) and eight domains from I(27) to I(34) (octamer). DLS measurements provide the size of both fragments as a function of temperature from 20 up to 86 degrees C, and show a thermal denaturation due to temperature increase. AFM imaging of both fragments in the native state reveals a homogeneous and uniform distribution of comparable structures. The DLS and AFM techniques turn out to be complementary for size measurements of the fragments and fragment aggregates. An unexpected result is that the octamer folds into a smaller structure than the tetramer and the unfolded octamer is also smaller than the unfolded tetramer. This feature seems related to the significance of the hydrophobic interactions between domains of the fragment. The longer the fragment, the more easily the hydrophobic parts of the domains interact with each other. The fragment aggregation behavior, in particular conditions, is also revealed by both DLS and AFM as a process that is parallel to the folding-unfolding transition.


Assuntos
Modelos Químicos , Modelos Moleculares , Proteínas Musculares/química , Proteínas Musculares/ultraestrutura , Miocárdio/metabolismo , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/ultraestrutura , Proteínas Quinases/química , Proteínas Quinases/ultraestrutura , Simulação por Computador , Conectina , Humanos , Microscopia de Força Atômica/métodos , Conformação Proteica , Refratometria/métodos
16.
Oncogene ; 25(5): 781-94, 2006 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-16186797

RESUMO

The K562 cell line serves as a model to study the molecular mechanisms associated with leukemia differentiation. We show here that cotreatment of K562 cells with PMA and low doses of SB202190 (SB), an inhibitor of the p38 MAPK pathway, induced a majority of cells to differentiate towards the megakaryocytic lineage. Electronic microscopy analysis showed that K562 cells treated with PMA+SB exhibited characteristic features of physiological megakaryocytic differentiation including the presence of vacuoles and demarcation membranes. Differentiation was also accompanied by a net increase in megakaryocytic markers and a reduction of erythroid markers, especially when both effectors were present. PMA effect was selectively mediated by new PKC isoforms. Differentiation of K562 cells by the combination of PMA and SB required Erk1/2 activation, a threshold of JNK activation and p38 MAPK inhibition. Interestingly, higher concentrations of SB, which drastically activated JNK, blocked megakaryocytic differentiation, and considerably increased cell death in the presence of PMA. c-DNA microarray membranes and PCR analysis allow us to identify a set of genes modulated during PMA-induced K562 cell differentiation. Several gene families identified in our screening, including ephrins receptors and some angiogenic factors, had never been reported so far to be regulated during megakaryocytic differentiation.


Assuntos
Diferenciação Celular , DNA Complementar/genética , Leucemia/genética , Leucemia/patologia , Megacariócitos/citologia , Análise de Sequência com Séries de Oligonucleotídeos , Transdução de Sinais , Sequência de Bases , Diferenciação Celular/efeitos dos fármacos , Primers do DNA , Inibidores Enzimáticos/farmacologia , Humanos , Imidazóis/farmacologia , Células K562 , Leucemia/enzimologia , Leucemia/metabolismo , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Reação em Cadeia da Polimerase , Piridinas/farmacologia , Acetato de Tetradecanoilforbol/farmacologia
17.
G Ital Med Lav Ergon ; 29(3 Suppl): 272-4, 2007.
Artigo em Italiano | MEDLINE | ID: mdl-18409682

RESUMO

Our Institute has been following for 20 years a group of workers of a ferroalloy industry in order to evaluate neurobehavioral effects due to manganese exposure. Five years after the last study we have planned another one, to evaluate differences in neuromotor e cognitive functions between exposed and controls and to perform a longitudinal evaluation of the results. Environmental and biological sampling were collected, liver and kidney functionality, haemochrome, iron metabolism and sieric prolactine were evaluated. Several tests were administered: postural evaluation, tremor, four tests of the SPES battery, Pursuing Aiming, five tests of the Luria Nebraska Motor Battery, Raven Progressive Matrices, Trail Making Test, Mood Scale, Brief Symptoms Inventory, neuropsychological symptoms questionnaire. Personal habits and working, living and clinical histories were collected. We evaluated 43 exposed workers and 40 controls. Exposure indicators resulted all significantly higher in exposed workers. Neuropsychological examination showed differences in Raven Progressive Matrices and Pursuit Aiming, higher tremor values and differences in postural evaluation between exposed and controls.


Assuntos
Comportamento/efeitos dos fármacos , Cognição/efeitos dos fármacos , Manganês/efeitos adversos , Metalurgia , Sistema Nervoso/efeitos dos fármacos , Exposição Ocupacional/efeitos adversos , Adulto , Humanos , Estudos Longitudinais , Pessoa de Meia-Idade , Vigilância da População
18.
G Ital Med Lav Ergon ; 29(3 Suppl): 698-700, 2007.
Artigo em Italiano | MEDLINE | ID: mdl-18409911

RESUMO

To compare data obtained in a pilot study (2002) which evaluated the risk from exposure to stress and burnout in health care workers, the same three subjective questionnaires were administered: the Job Content Questionnaire (JCQ), the State-Trait Anxiety Inventory (STAI), the Maslach Burnout Inventory (MBI). The evaluation considered 294 workers employed in eight units: two of the six previous units were reorganized in four subunits. Preliminary data showed that questionnaires' scores have a concordant trend in the different units, as observed in 2002, confirming the validity of the instruments adopted. Results indicated a decreased level of the perceived stress in all the units, especially in the Third Division. We can hypothesize that the new organization, workers' turnover during the 5 years and the effect due to individual variables could have contribute to the observed variation. No significant associations, compared to 2002, between questionnaires' scores and task were found. Further evaluations, including measurement of objective parameters, will be carried out to complete the follow-up study and to determine which variables could have a role in the variation of the levels of stress and burnout's subjective perception.


Assuntos
Esgotamento Profissional/epidemiologia , Pessoal de Saúde , Doenças Profissionais/epidemiologia , Adulto , Feminino , Seguimentos , Humanos , Masculino , Estresse Psicológico/epidemiologia , Inquéritos e Questionários
19.
J Phys Chem B ; 110(48): 24761-5, 2006 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-17134241

RESUMO

By means of combined calorimetric and dynamic light-scattering measurements, we have investigated the conformational behavior of DNA chains after thermal melting in the presence of a cationic surfactant at different concentrations, up to a surfactant-to-phosphate group molar ratio close to unity. Both the specific heat capacity, C(ex)(p) and the hydrodynamic radius R of the DNA chains provide support for the existence of two structural arrangements with different thermal stabilities, coexisting in the bulk solution. Although a component remains an elongated unfolded DNA chain originated in the thermal denaturation, the second component, consisting of DNA-surfactant complexes, assumes a compact structure with an average size of about 80 nm, whose thermal denaturation occurs at temperatures higher than 100 degrees C.


Assuntos
DNA/química , Tensoativos/química , Animais , Varredura Diferencial de Calorimetria , Cátions/química , Bovinos , Cetrimônio , Compostos de Cetrimônio/química , Dicroísmo Circular , Luz , Temperatura de Transição
20.
Biophys Chem ; 121(1): 7-13, 2006 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-16380204

RESUMO

The radiowave dielectric dispersions of DNA in different water-organic co-solvent mixtures have been measured in the frequency range from 100 kHz to 100 MHz, where the polarization mechanism is generally attributed to the confinement of counterions within some specific lengths, either along tangential or perpendicular to the polyion chain. The dielectric dispersions have been analyzed on the basis of two partially different dielectric models, a continuum counterion fluctuation model proposed by Mandel and a discrete charged site model, proposed by Minakata. The influence of the quality of the solvent on the dielectric parameters has been investigated in water-methanol and water-glycerol mixtures at different composition, by varying the permittivity (m) and the viscosity eta of the solvent phase. The analysis of the dielectric spectra in solvents where electrostatic and hydrodynamic interactions vary with the solvent composition suggests that both the two models are able, in principle, to account for the observed high-frequency dielectric behavior. However, while some certain assumptions are necessary about the polyion structure within the Mandel model, no structural prerequisite is needed within the Minakata model, where the polarization mechanism invoked considers a radial counterion exchange with the outer medium, which is largely independent of the local polyion conformation.


Assuntos
DNA/química , Eletroquímica , Glicerol/química , Íons/química , Metanol/química , Água/química , Condutividade Elétrica , Modelos Químicos , Modelos Teóricos , Termodinâmica , Viscosidade
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