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1.
Artigo em Inglês | MEDLINE | ID: mdl-32833254

RESUMO

Nature adopts complex chemical networks to finely tune biochemical processes. Indeed, small biomolecules play a key role in regulating the flux of metabolic pathways. Chemistry, which was traditionally focused on reactions in simple mixtures, is dedicating an increasing attention to the network reactivity of highly complex synthetic systems, able to display new kinetic phenomena. Here we show that the addition of monophosphate nucleosides to a mixture of amphiphiles and reagents leads to the selective templated formation of self-assembled structures, which can accelerate a reaction between two hydrophobic reactants. The correct matching between nucleotide and amphiphile's head group is fundamental for the selective formation of the assemblies and for the consequent upregulation of the chemical reaction. Transient stability of the nanoreactors is obtained under dissipative conditions, driven by enzymatic dephosphorylation of the templating nucleotides. These results show that small molecules can play a key role in modulating network reactivity, by selectively templating self-assembled structures that are able to upregulate chemical reaction pathways.

2.
Artigo em Inglês | MEDLINE | ID: mdl-32634287

RESUMO

BACKGROUND: Although language difficulties are one of the most distinctive characteristics of the neuropsychological profile of children with sex chromosome trisomies (SCT), the analysis of the maternal input addressed to them is a neglected topic. AIMS: The present study aims to analyse the lexical, morphosyntactic, and functional features of the input addressed to children with SCT comparing them with those of the input directed to typically developing children (TD). METHODS & PROCEDURES: Participants were 38 mothers and their 8-month-old children, 19 with SCT and 19 TD children. Maternal utterances, collected during video-recorded play sessions, have been transcribed and coded. OUTCOMES & RESULTS: No significant differences between groups have been found in the lexical and syntactic characteristics of maternal input. However, considering the input functional features, the proportion of directives and questions was significantly higher in the maternal input addressed to children with SCT than in the input addressed to TD children whereas the opposite pattern was found in the proportion of affect-salient speech. CONCLUSIONS & IMPLICATIONS: The awareness of a possible delay in their children's language development could influence the way the mothers speak to them. In particular, the functional features of maternal input could be affected. Support groups for parents of children with SCT at the preverbal stage could be useful to reassure the mothers about their role in their children's language development. What this paper adds What is already known on the subject Studies in the literature identified a high percentage of language delays or impairments in children with SCT. To date, according to our knowledge, there are no studies that analyse the linguistic input addressed to these children. What this study adds The lexical and syntactic features of maternal input addressed to 8-month-old children with SCT are adequate to the children's communicative skills. However, the mothers of children with SCT seem to provide additional scaffolding in their verbal input, using a lower proportion of affect-salient speech and a higher proportion of questions. In addition, a higher proportion of directives suggests the use of a more demanding style. Clinical implications of this study The awareness of possible language delays could influence the functional features of input leading mothers to use a more supportive and demanding input. Support groups for parents of children with SCT at the preverbal stage could be useful to reassure the mothers about their role in their children's language development.

4.
iScience ; 16: 340-355, 2019 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-31203189

RESUMO

Genetically Encoded Ca2+ Indicators (GECIs) are extensively used to study organelle Ca2+ homeostasis, although some available probes are still plagued by a number of problems, e.g., low fluorescence intensity, partial mistargeting, and pH sensitivity. Furthermore, in the most commonly used mitochondrial Förster Resonance Energy Transfer based-GECIs, the donor protein ECFP is characterized by a double exponential lifetime that complicates the fluorescence lifetime analysis. We have modified the cytosolic and mitochondria-targeted Cameleon GECIs by (1) substituting the donor ECFP with mCerulean3, a brighter and more stable fluorescent protein with a single exponential lifetime; (2) extensively modifying the constructs to improve targeting efficiency and fluorescence changes caused by Ca2+ binding; and (3) inserting the cDNAs into adeno-associated viral vectors for in vivo expression. The probes have been thoroughly characterized in situ by fluorescence microscopy and Fluorescence Lifetime Imaging Microscopy, and examples of their ex vivo and in vivo applications are described.

6.
Chemistry ; 25(45): 10606-10615, 2019 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-31107567

RESUMO

A comparative study between two novel, highly water soluble, ruthenium(II) polypyridyl complexes, [Ru(phen)2 L'] and [Ru(phen)2 Cu(II)L'] (L and L-CuII ), containing the polyaazamacrocyclic unit 4,4'-(2,5,8,11,14-pentaaza[15])-2,2'-bipyridilophane (L'), is herein reported. L and L-CuII interact with calf-thymus DNA and efficiently cleave DNA plasmid when light-activated. They also possess great penetration abilities and photo-induced biological activities, evaluated on an A375 human melanoma cell line, with L-CuII being the most effective. Our study highlights the key role of the Fenton active CuII center within the macrocycle framework, that would play a synergistic role with light activation in the formation of cytotoxic ROS species. Based on these results, an optimal design of RuII polypyridyl systems featuring specific CuII -chelating polyamine units could represent a suitable strategy for the development of novel and effective photosensitizers in photodynamic therapy.


Assuntos
Complexos de Coordenação/química , Fármacos Fotossensibilizantes/química , Rutênio/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/farmacologia , DNA/química , Clivagem do DNA/efeitos dos fármacos , Humanos , Microscopia Confocal , Fármacos Fotossensibilizantes/farmacologia , Piridinas/química , Oxigênio Singlete/metabolismo
7.
Neurophysiol Clin ; 49(3): 217-226, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31101389

RESUMO

OBJECTIVE: To compare brainstem acoustic evoked potentials (BAEP) and magnetic resonance imaging (MRI) in the differential diagnosis of intracranial hypotension (IH), Chiari malformation (CM) and sensorineural hearing loss (SNHL). METHODS: BAEP were recorded in 18 IH, 18 CM, 20 SNHL patients and 52 controls. MRI were acquired in all IH and CM patients. RESULTS: Abnormal BAEP were observed in 94% of IH patients, in 33% of CM and 70% of SNHL patients. After recovery from IH, BAEP abnormalities disappeared. Internal auditory canal (IAC) MRI abnormalities were described in 88% of IH patients. MRI signs of IH were observed in 33-78% in IH patients, but the most frequent MRI sign was 8th nerve T2 hyperintensity, with contrast enhancement in T1 sequences. This finding, combined with wave I latency, yielded highest specificity and sensitivity for IH diagnosis. CONCLUSIONS: Our study points out how IH can be effectively distinguished from CM and SNHL through the contribution of neurophysiology and MRI; in particular, evaluation of the 8th nerve achieves a high sensitivity and specificity in patients with IH. Further studies are required to examine the combined use of BAEP recordings ad MRI in diagnosis and monitoring of patients affected by IH.


Assuntos
Malformação de Arnold-Chiari/diagnóstico , Perda Auditiva Neurossensorial/diagnóstico , Hipotensão Intracraniana/diagnóstico , Imagem por Ressonância Magnética , Adulto , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Diagnóstico Diferencial , Potenciais Evocados Auditivos do Tronco Encefálico , Feminino , Humanos , Hipotensão Intracraniana/patologia , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade
8.
Epilepsy Behav Case Rep ; 11: 120-121, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31011534

RESUMO

•CAD is one of the most important predisposing factors of cerebral ischaemia in young adults.•Focal motor seizures with the involvement of the neck may represent a risk condition of developing CAD•A heightened awareness for CAD should be recommended in patients affected by epilepsy, carotid dopplers may confirm the clinical suspicion in case of significant neck trauma associated with seizures.

9.
Clin Neurophysiol Pract ; 4: 27-29, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30859143

RESUMO

Objective: Insular lobe seizures generally represent a misconceived ictal phenomenon characterized by specific neurological signs. Aphasia can be a rare presenting sign associated with insular lobe epilepsy which could be easily mistaken for a manifestation of other acute brain diseases. Method: We describe an insular status epilepticus (SE) characterized by sudden onset of language disturbance associated with hypersalivation and paraesthesia. A concomitant EEG recording showed the presence of Lateralized Periodic Discharges plus superimposed fast activity (LPDs + F). After an adequate acute endovenous anti-seizure treatment, a normalization of the EEG abnormalities with a complete resolution of all the neurological symptoms was achieved. Discussion: Language disturbances can be usually found in various pathological acute pictures involving the dominant frontal and temporal lobes. The presence of certain EEG pattern, could rise the suspect of aphasia as a critical manifestation. LPDs pattern is usually correlated with structural lesions. The association between LPDs and seizure is controversial but it seems to be more consistent when they are associated with "Plus modifiers" and with an high periodic frequency. Conclusion: Our case underlines the importance of considering focal SE in the differential diagnosis of patients presenting aphasia, even in the absence of previous history of epilepsy. We describe how LPDs can be associated with SE in a patient affected by a brain tumour, supporting the idea that some characteristic periodic patterns could be associated with seizure occurrence.

10.
Aging Cell ; 18(3): e12924, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30793475

RESUMO

Mitochondrial dysfunction is implicated in most neurodegenerative diseases, including Alzheimer's disease (AD). We here combined experimental and computational approaches to investigate mitochondrial health and bioenergetic function in neurons from a double transgenic animal model of AD (PS2APP/B6.152H). Experiments in primary cortical neurons demonstrated that AD neurons had reduced mitochondrial respiratory capacity. Interestingly, the computational model predicted that this mitochondrial bioenergetic phenotype could not be explained by any defect in the mitochondrial respiratory chain (RC), but could be closely resembled by a simulated impairment in the mitochondrial NADH flux. Further computational analysis predicted that such an impairment would reduce levels of mitochondrial NADH, both in the resting state and following pharmacological manipulation of the RC. To validate these predictions, we utilized fluorescence lifetime imaging microscopy (FLIM) and autofluorescence imaging and confirmed that transgenic AD neurons had reduced mitochondrial NAD(P)H levels at rest, and impaired power of mitochondrial NAD(P)H production. Of note, FLIM measurements also highlighted reduced cytosolic NAD(P)H in these cells, and extracellular acidification experiments showed an impaired glycolytic flux. The impaired glycolytic flux was identified to be responsible for the observed mitochondrial hypometabolism, since bypassing glycolysis with pyruvate restored mitochondrial health. This study highlights the benefits of a systems biology approach when investigating complex, nonintuitive molecular processes such as mitochondrial bioenergetics, and indicates that primary cortical neurons from a transgenic AD model have reduced glycolytic flux, leading to reduced cytosolic and mitochondrial NAD(P)H and reduced mitochondrial respiratory capacity.

11.
BMJ Case Rep ; 11(1)2018 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-30567263

RESUMO

Dropped head syndrome (DHS) is a rare condition, characterised by weakness of the cervical paraspinal muscles with sagging of the head. It is usually seen in association with neurological disorders and rarely can follow radiotherapy. We report a case of a 54-year-old man survivor of Hodgkin lymphoma (HL), who developed DHS 28 years after radiotherapy. He was referred to our department due to progressive weakness and atrophy of cervical paraspinal and shoulder girdle musculature. Physical and neurophysiological examination, electromyography and MRI confirmed the diagnosis of DHS. In the following years, there was no progression of symptoms.


Assuntos
Neuropatias do Plexo Braquial/fisiopatologia , Terapia por Exercício/métodos , Doença de Hodgkin/radioterapia , Atrofia Muscular Espinal/fisiopatologia , Músculos do Pescoço/efeitos da radiação , Radioterapia/efeitos adversos , Neuropatias do Plexo Braquial/etiologia , Neuropatias do Plexo Braquial/reabilitação , Sobreviventes de Câncer , Humanos , Masculino , Pessoa de Meia-Idade , Atrofia Muscular Espinal/etiologia , Atrofia Muscular Espinal/reabilitação , Músculos do Pescoço/fisiopatologia , Aparelhos Ortopédicos , Síndrome , Fatores de Tempo , Resultado do Tratamento
12.
Neurol Sci ; 39(11): 1887-1894, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30088164

RESUMO

Transcranial sonography (TCS) is a noninvasive, easily performed, and commonly available neuroimaging technique useful for the study of brain parenchyma in movement disorders. This tool has been increasingly used in the diagnosis of Parkinson's disease and atypical parkinsonism. The aim of the study was to evaluate the applicability of this technique as supportive tool in the early diagnosis of movement disorders. We performed TCS on 315 individuals which were diagnosed as healthy controls or affected by idiopathic Parkinson's disease, monogenetic subtypes of Parkinson's disease, atypical parkinsonism, and Dementia with Lewy bodies. Five TCS diagnostic patterns were defined on the basis of substantia nigra's and lenticular nuclei's echogenicity. TCS evaluations were performed by two blinded neuro-sonographers. Clinical diagnosis on all individuals was performed at baseline and at 4-year follow-up. The concordance rate between TCS patterns and clinical diagnosis and the specificity of TCS pattern to discriminate each group of individuals were compared at baseline and at follow-up. The concordance rate between TCS patterns and clinical diagnosis of all individuals was 84% at baseline and increased at follow-up (91%) significantly (p = 0.01). The specificity of TCS pattern in the comparison between patients diagnosed as affected by idiopathic Parkinson's disease and atypical parkinsonism showed a significant increase at follow-up (p = 0.03). Our study strongly confirms the role of TCS as a noninvasive and cost-effective tool in early diagnosis of movement disorders.


Assuntos
Transtornos dos Movimentos/diagnóstico por imagem , Ultrassonografia Doppler Transcraniana/métodos , Idoso , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes
13.
Nanoscale ; 10(25): 11913-11922, 2018 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-29901055

RESUMO

Here we present a comprehensive study on the photophysics of QDs-fullerene blends, aiming to elucidate the impact of ligands on the extraction of carriers from QDs. We investigated how three different ligands (oleylamine, octadecanethiol and propanethiol) influence the dynamics of charge generation, separation, and recombination in blends of CdSe/CdS core/shell QDs and PCBM. We accessed each relevant process directly by combining the results from both optical and magnetic spectroscopies. Transient absorption measurements revealed a faster interaction dynamics in thiol-capped ligands. Through phenomenological modeling of the interaction processes, i.e., energy transfer and electron transfer, we estimated the suppression of exciton migration and the enhancement of electron transfer processes when alkyl-thiols are employed as ligands. Contextually, we report the profound impact of the ligands' alkyl chain length, leading to strengthened interactions with PCBM acceptors. Quantitatively, we measured a 10-fold increase in the electron transfer rate when oleylamine ligands were exchanged with propanethiol ligands. EPR spectroscopy gave access to subtle details regarding both the enhanced charge generation and lower binding energy of charge-transfer states in blends compared to PCBM alone. Moreover, through pulsed EPR techniques, we inferred the localization of deep electron traps in localized sites close to QDs in the blends. Therefore, our thorough characterization evidenced the essential role of ligands in determining QD interactions. We believe that these discoveries will contribute to the efficient incorporation of QDs in existing organic PV technologies.

14.
J Nanopart Res ; 19(9): 316, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28959137

RESUMO

Screening nanoparticle toxicity directly on cell culture can be a fast and cheap technique. Nevertheless, to obtain results in accordance with those observed in live animals, the conditions in which cells are cultivated should resemble the one encountered in live systems. Microfluidic devices offer the possibility to satisfy this requirement, in particular with endothelial cell lines, because they are capable to reproduce the flowing media and shear stress experienced by these cell lines in vivo. In this work, we exploit a microfluidic device to observe how human umbilical vein endothelial cells (HUVEC) viability changes when subject to a continuous flow of culture medium, in which spherical citrate-stabilized gold nanoparticles of different sizes and at varying doses are investigated. For comparison, the same experiments are also run in multiwells where the cells do not experience the shear stress induced by the flowing medium. We discuss the results considering the influence of mode of exposure and nanoparticle size (24 and 13 nm). We observed that gold nanoparticles show a lower toxicity under flow conditions with respect to static and the HUVEC viability decreases as the nanoparticle surface area per unit volume increases, regardless of size.

15.
J Phys Chem Lett ; 8(10): 2236-2242, 2017 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-28471190

RESUMO

The controversial nature of the fluorescent properties of carbon dots (CDs), ascribed either to surface states or to small molecules adsorbed onto the carbon nanostructures, is an unresolved issue. To date, an accurate picture of CDs and an exhaustive structure-property correlation are still lacking. Using two unconventional spectroscopic techniques, fluorescence correlation spectroscopy (FCS) and time-resolved electron paramagnetic resonance (TREPR), we contribute to fill this gap. Although electron micrographs indicate the presence of carbon cores, FCS reveals that the emission properties of CDs are based neither on those cores nor on molecular species linked to them, but rather on free molecules. TREPR provides deeper insights into the structure of carbon cores, where C sp2 domains are embedded within C sp3 scaffolds. FCS and TREPR prove to be powerful techniques, characterizing CDs as inherently heterogeneous systems, providing insights into the nature of such systems and paving the way to standardization of these nanomaterials.

16.
Chemistry ; 23(48): 11549-11559, 2017 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-28544114

RESUMO

The next generation of adaptive, intelligent chemical systems will rely on a continuous supply of energy to maintain the functional state. Such systems will require chemical methodology that provides precise control over the energy dissipation process, and thus, the lifetime of the transiently activated function. This manuscript reports on the use of structurally diverse chemical fuels to control the lifetime of two different systems under dissipative conditions: transient signal generation and the transient formation of self-assembled aggregates. The energy stored in the fuels is dissipated at different rates by an enzyme, which installs a dependence of the lifetime of the active system on the chemical structure of the fuel. In the case of transient signal generation, it is shown that different chemical fuels can be used to generate a vast range of signal profiles, allowing temporal control over two orders of magnitude. Regarding self-assembly under dissipative conditions, the ability to control the lifetime using different fuels turns out to be particularly important as stable aggregates are formed only at well-defined surfactant/fuel ratios, meaning that temporal control cannot be achieved by simply changing the fuel concentration.

17.
Phys Chem Chem Phys ; 18(45): 31286-31295, 2016 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-27824179

RESUMO

The design of novel nanostructures with tailored opto-electronic properties is a crucial step for third-generation photovoltaics, and the development of cheap and environmentally compatible materials is still a challenge. Carbon quantum dots (CQDs) emerged as promising candidates but usually a low processability and poor electron-donor properties hampered their photovoltaic applications. We tackle these issues through the synthesis and photophysical characterization of N-doped CQDs functionalized with different thiophene-containing groups. Functionalization was aimed at enhancing the electron donating properties of the carbon dots and improving the solubility in nonpolar solvents. The increased solubility in organic solvents allowed us to investigate the photoinduced interactions of the functionalized carbon dots with the fullerene derivative PCBM in solution and in solid blends. The investigation was carried out by cyclic voltammetry, photoluminescence spectroscopy and electron paramagnetic resonance (EPR). The remarkable oxidation potential shift of the functionalized carbon dots with respect to the pristine materials and the HOMO-LUMO energies strongly suggest them as good electron donors towards PCBM. The electron transfer process between CQDs and PCBM resulted in efficient fluorescence quenching in solution and in total quenching in solid blends. By using EPR spectroscopy in the solid blends, we demonstrated the efficient electron transfer by observing the photoinduced formation of a PCBM radical anion in the presence of functionalized CQDs. Time-resolved EPR allowed us to identify differences in the charge transport efficiency for different CQD:PCBM blends. The enhanced processability of CQDs with PCBM and the promising charge-generation and separation properties pave the way to the development of "all-carbon" photovoltaic devices.

18.
Nat Chem ; 8(7): 725-31, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27325101

RESUMO

Dissipative self-assembly is exploited by nature to control important biological functions, such as cell division, motility and signal transduction. The ability to construct synthetic supramolecular assemblies that require the continuous consumption of energy to remain in the functional state is an essential premise for the design of synthetic systems with lifelike properties. Here, we show a new strategy for the dissipative self-assembly of functional supramolecular structures with high structural complexity. It relies on the transient stabilization of vesicles through noncovalent interactions between the surfactants and adenosine triphosphate (ATP), which acts as the chemical fuel. It is shown that the lifetime of the vesicles can be regulated by controlling the hydrolysis rate of ATP. The vesicles sustain a chemical reaction but only as long as chemical fuel is present to keep the system in the out-of-equilibrium state. The lifetime of the vesicles determines the amount of reaction product produced by the system.


Assuntos
Vesículas Citoplasmáticas/química , Complexos Multiproteicos/química , Trifosfato de Adenosina/química , Compostos Aza/química , Modelos Teóricos , Piperidinas/química , Fatores de Tempo
19.
Dalton Trans ; 44(35): 15712-20, 2015 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-26266469

RESUMO

The new complexes cis-[Ir(CO)2Cl(4-(para-di-n-butylaminostyryl)pyridine)] () and [Ir(cyclometallated-2-phenylpyridine)2(4,4'-(para-di-n-butylaminostyryl)-2,2'-bipyridine)][PF6] () were synthesized and fully characterized along with the known complex Ir(cyclometallated-2-phenylpyridine)2(5-Me-1,10-phenanthroline)][PF6] (). Remarkably, complex , with an Ir(i) centre, displays fluorescence - as opposed to the phosphorescence typical of many Ir(iii) complexes - with a modestly high quantum yield in solution, opening a new route for the design of iridium-based emitters which should not be limited to the +3 oxidation state. It is also characterized by an unexpectedly large two-photon absorption (TPA) cross section, an order of magnitude higher than that previously reported for Ir(iii) or Pt(ii) complexes. The great potential of cyclometallated Ir(iii) complexes for photodynamic therapy was confirmed, with and showing a good singlet oxygen generation ability, coupled with a modest TPA activity for .

20.
Microvasc Res ; 97: 147-55, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25446009

RESUMO

A new in vitro model system, adding advection and shear stress associated with a flowing medium, is proposed for the investigation of nanoparticles uptake and toxicity towards endothelial cells, since these processes are normally present when nanoparticles formulations are intravenously administered. In this model system, mechanical forces normally present in vivo, such as advection and shear stress were applied and carefully controlled by growing human umbilical vein endothelial cells inside a microfluidic device and continuously infusing gold nanoparticle (Au NPs) solution in the device. The tests performed in the microfluidic device were also run in multiwells, where no flow is present, so as to compare the two model systems and evaluate if gold nanoparticles toxicity differs under static and flow culture conditions. Full characterization of Au NPs in water and in culture medium was accomplished by standard methods. Two-photon fluorescence correlation spectroscopy was also employed to map the flow speed of Au NPs in the microfluidic device and characterize Au NPs before and after interactions with the cells. Au NPs uptake in both in vitro systems was investigated through electron and fluorescence microscopy and ICP-AES, and NPs toxicity measured through standard bio-analytical tests. Comparison between experiments run in multiwells and in microfluidic device plays a pivotal role for the investigation of nanoparticle-cell interaction and toxicity assessment: our work showed that administration of equal concentrations of Au NPs under flow conditions resulted in a reduced sedimentation of nanoparticle aggregates onto the cells and lower cytotoxicity with respect to experiments run in ordinary static conditions (multiwells).


Assuntos
Compostos de Ouro/toxicidade , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Nanopartículas Metálicas/toxicidade , Velocidade do Fluxo Sanguíneo , Técnicas de Cultura de Células , Células Cultivadas , Compostos de Ouro/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Células Endoteliais da Veia Umbilical Humana/ultraestrutura , Humanos , Técnicas Analíticas Microfluídicas , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Fluxo Sanguíneo Regional , Espectrometria de Fluorescência/métodos , Estresse Mecânico , Fatores de Tempo
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