Parkinson's disease is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra, and as a consequence, by decreased dopamine levels in the striatum. ...Currently available therapies are not able to stop or reverse the progression of the disease. A novel therapeutic approach is based on cell therapy with stem cells, in order to replace degenerated neurons. Among stem cells, mesenchymal stem cells seemed the most promising thanks to their capacities to differentiate toward dopaminergic neurons and to release neurotrophic factors. Indeed, mesenchymal stem cells are able to produce different molecules with immunomodulatory, neuroprotective, angiogenic, chemotactic effects and that stimulate differentiation of resident stem cells. Mesenchymal stem cells were isolated for the first time from bone marrow, but can be collected also from adipose tissue, umbilical cord and other tissues. In this review, we focused our attention on mesenchymal stem cells derived from different sources and their application in Parkinson's disease animal models.
BRAF(V600E) mutation is the most frequent genetic alteration in papillary thyroid carcinomas (PTCs) that are 80–90% of all thyroid cancers. We evaluated the relationship between BRAF(V600E) and ...tumor, host, and environmental factors in PTCs from all geographical areas of Sicily. By PCR, BRAF(V600E) was investigated in a series of 323 PTCs diagnosed in 2002–2005. The correlation between clinicopathological tumor, host, and environmental characteristics and the presence of BRAF(V600E) were evaluated by both univariate and multivariate analyses. BRAF(V600E) was found in 38.6% PTCs, with a 52% frequency in the classical PTCs and 26.4% in the tall cell variant. Univariate analysis indicated that BRAF(V600E) was associated with greater tumor size (P=0.0048), extra-thyroid invasion (P<0.0001), and cervical lymph nodal metastases (P=0.0001). Multivariate logistic regression analysis confirmed that BRAF(V600E) was an independent predictor of extra-thyroid invasion (P=0.0001) and cervical lymph nodal metastasis (P=0.0005). The association between BRAF(V600E) and extra-thyroid invasion was also found in micro-PTCs (P=0.006). In 60 classical PTCs, BRAF(V600E) was positively correlated with matrix metalloproteinase-9 expression (P=0.0047), suggesting a possible mechanism for BRAF(V600E) effect on PTC invasiveness. No association was found between BRAF(V600E) and patient age, gender, or iodine intake. In contrast, a strong association was found with residency in Eastern Sicily (P<0.0001 compared with Western Sicily). These results indicate that BRAF(V600E) mutation is a marker of aggressive disease in both micro- and macro-PTCs. Moreover, for the first time, a possible link between BRAF(V600E) mutation and environmental carcinogens is suggested.
The unique deformability and the compliance ability of thin sheets on soft substrates attract much interest for studying the phenomena related to elastic instabilities as well as for sensing very ...weak forces such as those generated by live cells in vitro. However, the techniques used currently for producing such platforms are affected by a high degree of complexity and poor repeatability. Moreover, their deformability is usually used as a passive response to the action of an external force. Herein we propose a novel concept for a reliable and dynamic skin-over-liquid system made of a periodic array of highly compliant microbumps actuated through electrode-free electrohydrodynamic (EHD) pressure. We demonstrate that these structures are highly repeatable and capable of swelling and deflating easily under a simple thermal stimulation driven by the pyroelectric effect, thus providing a challenging platform that can be actively controlled at the microscale. Furthermore, we show the proof of principle by swelling these microbumps for mechanically stimulating live cells in vitro, thus opening the route to more reliable and easy to accomplish assays in the field of mechanobiology.Biomechanics: Giving artificial skin the goosebumpsResearchers curious about how mechanical stress impacts cell growth can now turn to silicone plastics that grow microbumps on demand. Simonetta Grilli from Italy’s Institute of Applied Sciences and Intelligent Systems in Pozzuoli and colleagues created a controllable ‘skin-over-liquid’ system by coating a silicone polymer solution onto lithium niobate, a crystal that contains patterned regions of electric charge. An initial plasma treatment produced a flat stiff film on top of the liquid silicone. Thermal heating caused the skin to swell and form dimpled, air mattress-like patterns, corresponding to the underlying lithium niobate domains. Removal of heat restored the sheet to its initial state. The chemically inert skin supported live fibroblast cells and was used to determine reactions to mechanical stress. Significant variations in nuclei and cytoskeletons were observed between cells grown under flat or swollen-skin conditions.
miR-2861 endorsing osteoblast differentiation through the overexpression of Runt-related transcription factor 2 (RUNX2) protein has been recently described. In this study we evaluated: the ...performance of living construct, composed by human Periodontal Ligament Stem Cells (hPDLSCs) and 3D scaffold (EXg), and the behaviour of miR-2861/RUNX2 expression pathway on the osteogenic commitment. Human PDLSCs were seeded with and without EXg scaffold and cultured under basal and osteogenic conditions. Morphological features, adhesiveness and differentiation abilities were analysed using scanning electron and confocal laser scanning microscopy. Time-course of RUNX2, ALP, OPN and miR-2861 were evaluated through RT-PCR analysis. Our results highlighted that the osteogenic differentiation was mostly obvious in the hPDLSCs, grown onto 3D scaffold in presence of osteoinductive medium. Moreover, the overexpression of miR-2861 and RUNX2 in hPDLSCs cultured in presence of EXg under osteogenic and standard conditions was demonstrated. In synthesis, the increased expression of miR-2861/RUNX2 provides new insights regarding miRNA signaling network in the presence of scaffold providing an additional method to evaluate the performance of biomaterial in bone regeneration.
Biofilms are detrimental to human life and industrial processes due to potential infections, contaminations, and deterioration. Therefore, the evaluation of microbial capability to form biofilms is ...of fundamental importance for assessing how different environmental factors may affect their vitality. Nowadays, the approaches used for biofilm evaluation are still poor in reliability and rapidity and often provide contradictory results. Here, we present what we call biofilm electrostatic test (BET) as a simple, rapid, and highly reproducible tool for evaluating in vitro the ability of bacteria to form biofilms through electrostatic interaction with a pyroelectrified carrier. The results show how the BET is able to produce viable biofilms with a density 6-fold higher than that on the control, after just 2 h incubation. The BET could pave the way to a rapid standardization of the evaluation of bacterial resistance among biofilm-producing microorganisms. In fact, due to its simplicity and cost-effectiveness, it is well suited for a rapid and easy implementation in a microbiology laboratory.
Multiple Sclerosis (MS) is a global concern disease leading to a progressive, chronic and demyelinating condition, affecting the central nervous system (CNS). The pathology has an ...inflammatory/autoimmune origin; nevertheless, neuronal cell death mechanisms are not to be underestimated. The present study was designed to test the effects of intraperitoneal administration of cannabidiol (CBD), the main non-psychotropic cannabinoid of Cannabis sativa (CS), in an experimental model of MS. The aim is to evaluate the capability of CBD administration to thwart the cascade of mediators involved in MS-induced apoptosis.
Experimental Autoimmune Encephalomyelitis (EAE) was induced by immunization with myelin oligodendroglial glycoprotein (MOG)35-55 peptide in mice. After immunization, mice were observed daily for signs of EAE and weight loss. Disease signs were evaluated using a standardized scoring system.
Immunohistochemical and Western blot assessments of key apoptotic markers reveal that CBD treatment is able to avoid Fas pathway activation, phospho-ERK p42/44 and cleaved caspase-3 triggering as well as alterations in mitochondrial permeability due to Bax/Bcl-2 unbalance. Moreover, CBD interferes with p53-p21 axis activation. As results, the absence of tissue apobody formation in spinal cord tissues of EAE-mice treated with CBD was established. Most of therapeutic properties of CS are currently ascribed to the psychotropic effects of phenylterpenoid delta-9 tetrahydrocannabinol.
We have demonstrated that, alone, purified CBD possesses an anti-apoptotic power against the neurodegenerative processes underlying MS development. This represents an interesting new profile of CBD that could lead to its introduction in the clinical management of MS.
Easter Island (Rapa Nui), Chile, is remote, located in the Polynesian Triangle in Oceania. The closest continental point is Chile, 3,512 km east. It has a population of 7,750 inhabitants, who are ...Chilean citizens, and receives more than 60,000 tourists a year. For this entire population, there is a medium complexity hospital without a neurology specialist. In 2019, local professionals were trained in a Telestroke program with remote clinical support conducted by neurologists located on mainland Chile. We present a 50-year-old native male, with unknown medical history, who suddenly presented right-half-body weakness and aphasia. He was evaluated via Telestroke consultation, and thrombolysis with tenecteplase was indicated. The patient improved rapidly and 45 min later the NIHSS score was 0 points. To our knowledge, this is the first reported case of Telestroke treatment in such a remote area, highlighting the importance of telemedicine to overcome geographical and technological stroke care barriers and to improve patients’ outcome, no matter where they live.
We aimed to characterize spontaneous cervical artery dissection (CeAD) patients with and without stroke and describe risk factors for cerebrovascular complications in a Chilean prospective cohort.
...Methods: Consecutive CeAD patients admitted to a Chilean center confirmed by neuroimaging. Logistic regression was used.
Results: 168 patients were included, median follow-up time was 157 days. Stroke occurred in 49 (29.2%) cases, 4 (2%) patients died, all of whom had a stroke, and 10 (6%) presented CeAD recurrence. In univariate analyses, men (odds ratio OR 3.97, 95% confidence interval CI 1.97–8.00, P < 0.001), internal carotid artery CeAD (OR 2.82, 95% CI 1.38–5.78, P = 0.005) and vessel occlusion (OR 4.45, 95% CI 1.38–14.38, P = 0.035) increased stroke risk. Conversely, vertebral artery dissection (OR 0.35, 95% CI 0.16–0.74, P = 0.006) and longer symptom onset to admission (O–A) time (OR 0.79, 95% CI 0.70–0.90, P < 0.001) were associated to decreased stroke risk. After multivariate analysis, men (OR 2.88, 95% CI 1.32-6.27, P = 0.008) and O–A time (OR 0.80, 95% CI 0.69–0.92, P = 0.002) remained independently associated with stroke.
Conclusion: CeAD presented commonly as a non-stroke entity, with favorable prognosis. Albeit to a higher frequency of CeAD in women, stroke occurred predominantly in men, who were admitted earlier.
Background and purpose:
5‐lipoxygenase (5‐LO) is the key enzyme in leukotriene (LT) biosynthesis from arachidonic acid (AA). Here, we examined the role of the 5‐LO‐product, cysteinyl‐LT (Cys‐LT), ...with a 5‐LO inhibitor (zileuton) and a Cys‐LT, receptor antagonist (montelukast), in the inflammatory response and tissue injury associated with spinal cord injury (SCI).
Experimental approach:
SCI was induced in mice by the application of vascular clips to the dura via a two‐level T6 to T7 laminectomy for 1 min. Cord inflammation was assessed histologically and by measuring inflammatory mediators (ELISA) and apoptosis by annexin V, TUNEL, Fas ligand staining and Bax and Bcl‐2 expression (immunohistochemistry and western blots). Motor function in hindlimbs was assessed by a locomotor rating scale, for 10 days after cord injury.
Key results:
SCI in mice resulted in tissue damage, oedema, neutrophil infiltration, apoptosis, tumour necrosis‐α (TNF‐α) and cyclooxygenase‐2 (COX‐2) expression, prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) production, and extracellular signal‐regulated kinase 1/2 (ERK1/2) phosphorylation in injured tissue. Treatment of the mice with zileuton or montelukast reduced the spinal cord inflammation and tissue injury, neutrophil infiltration, TNF‐α, COX‐2 and pERK1/2 expression, PGE2 and LTB4 production, and apoptosis. In separate experiments, zileuton or montelukast significantly improved the recovery of limb function over 10 days.
Conclusions and implications:
Zileuton and montelukast produced a substantial reduction of inflammatory events associated with experimental SCI. Our data underline the important role of 5‐LO and Cys‐LT in neurotrauma.
British Journal of Pharmacology (2008) 153, 568–582; doi:10.1038/sj.bjp.0707577; published online 3 December 2007
Nitrite is protective against renal ischemia/reperfusion injury (IRI); an effect due to its reduction to nitric oxide (NO). In addition to other reductase pathways, endothelial NO synthase (eNOS) may ...also facilitate nitrite reduction in ischemic environments. We investigated the role of eNOS in sodium nitrite (60
μM, 10
ml/kg applied topically 1
min before reperfusion)-induced protection against renal IRI in C57/BL6 wild-type (WT) and eNOS knockout (eNOS KO) mice subjected to bilateral renal ischemia (30
min) and reperfusion (24
h). Markers of renal dysfunction (plasma creatinine and urea), damage (tubular histology) and inflammation (cell recruitment) were elevated following IRI in WT mice; effects significantly reduced following nitrite treatment. Chemiluminescence analysis of cortical and medullary sections of the kidney demonstrated rapid (within 1
min) distribution of nitrite following application. Whilst IRI caused a significant (albeit substantially reduced compared to WT mice) elevation of markers of renal dysfunction and damage in eNOS KO mice, the beneficial effects of nitrite were absent or reduced, respectively. Moreover, nitrite treatment enhanced renal dysfunction in the form of increased plasma creatinine in eNOS KO mice. Confirmation of nitrite reductase activity of eNOS was provided by demonstration of nitrite (100
μM)-derived NO production by kidney homogenates of WT mice, that was significantly reduced by
l-NMMA.
l-NMMA was without effect using kidney homogenates of eNOS KO mice. These results support a role for eNOS in the pathways activated during renal IRI and also identify eNOS as a nitrite reductase in ischemic conditions; activity which in part underlies the protective effects of nitrite.
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