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Background: Glioblastoma (GBM) is an aggressive brain tumor that exhibits resistance to current treatment, making the identification of novel therapeutic targets essential. In this context, cellular prion protein (PrP C ) stands out as a potential candidate for new therapies. Encoded by the PRNP gene, PrP C can present increased expression levels in GBM, impacting cell proliferation, growth, migration, invasion and stemness. Nevertheless, the exact molecular mechanisms through which PRNP/PrP C modulates key aspects of GBM biology remain elusive., Methods: To elucidate the implications of PRNP/PrP C in the biology of this cancer, we analyzed publicly available RNA sequencing (RNA-seq) data of patient-derived GBMs from four independent studies. First, we ranked samples profiled by bulk RNA-seq as PRNP high and PRNP low and compared their transcriptomic landscape. Then, we analyzed PRNP + and PRNP - GBM cells profiled by single-cell RNA-seq to further understand the molecular context within which PRNP/PrP C might function in this tumor. We explored an additional proteomics dataset, applying similar comparative approaches, to corroborate our findings., Results: Functional profiling revealed that vesicular dynamics signatures are strongly correlated with PRNP/PrP C levels in GBM. We found a panel of 73 genes, enriched in vesicle-related pathways, whose expression levels are increased in PRNP high /PRNP + cells across all RNA-seq datasets. Vesicle-associated genes, ANXA1, RAB31, DSTN and SYPL1, were found to be upregulated in vitro in an in-house collection of patient-derived GBM. Moreover, proteome analysis of patient-derived samples reinforces the findings of enhanced vesicle biogenesis, processing and trafficking in PRNP high /PRNP + GBM cells., Conclusions: Together, our findings shed light on a novel role for PrP C as a potential modulator of vesicle biology in GBM, which is pivotal for intercellular communication and cancer maintenance. We also introduce GBMdiscovery, a novel user-friendly tool that allows the investigation of specific genes in GBM biology., (© 2024. The Author(s).)

Extracellular vesicles (EVs) are membrane-delimited vesicular bodies carrying different molecules, classified according to their size, density, cargo, and origin. Research on this topic has been actively growing through the years, as EVs are associated with critical pathological processes such as neurodegenerative diseases and cancer. Despite that, studies exploring the physiological functions of EVs are sparse, with particular emphasis on their role in organismal development, initial cell differentiation, and morphogenesis. In this review, we explore the topic of EVs from a developmental perspective, discussing their role in the earliest cell-fate decisions and neural tissue morphogenesis. We focus on the function of EVs through development to highlight possible conserved or novel processes that can impact disease progression. Specifically, we take advantage of what was learned about their role in development so far to discuss EVs impact on glioblastoma, a particular brain tumor of stem-cell origin and poor prognosis, and how their function can be hijacked to improve current therapies., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: Thyroid nodules diagnosed as 'atypia of undetermined significance/follicular lesion of undetermined significance' (AUS/FLUS) or 'follicular neoplasm/suspected follicular neoplasm' (FN/SFN), according to Bethesda's classification, represent a challenge in clinical practice. Computerized analysis of nuclear images (CANI) could be a useful tool for these cases. Our aim was to evaluate the ability of CANI to correctly classify AUS/FLUS and FN/SFN thyroid nodules for malignancy., Methods: We studied 101 nodules cytologically classified as AUS/FLUS (n = 68) or FN/SFN (n = 33) from 97 thyroidectomy patients. Slides with cytological material were submitted for manual selection and analysis of the follicular cell nuclei for morphometric and texture parameters using ImageJ software. The histologically benign and malignant lesions were compared for such parameters which were then evaluated for the capacity to predict malignancy using the classification and regression trees gini model. The intraclass coefficient of correlation was used to evaluate method reproducibility., Results: In AUS/FLUS nodule analysis, the benign and malignant nodules differed for entropy (P < 0.05), while the FN/SFN nodules differed for fractal analysis, coefficient of variation (CV) of roughness, and CV-entropy (P < 0.05). Considering the AUS/FLUS and FN/SFN nodules separately, it correctly classified 90.0 and 100.0% malignant nodules, with a correct global classification of 94.1 and 97%, respectively. We observed that reproducibility was substantially or nearly complete (0.61-0.93) in 10 of the 12 nuclear parameters evaluated., Conclusion: CANI demonstrated a high capacity for correctly classifying AUS/FLUS and FN/SFN thyroid nodules for malignancy. This could be a useful method to help increase diagnostic accuracy in the indeterminate thyroid cytology.

Objective: Follicular lesions of the thyroid with papillary carcinoma nuclear characteristics are classified as infiltrative follicular variant of papillary thyroid carcinoma-FVPTC (IFVPTC), encapsulated/well demarcated FVPTC with tumour capsular invasion (IEFVPTC), and the newly described category "non-invasive follicular thyroid neoplasm with papillary-like nuclear features" (NIFTP) formerly known as non-invasive encapsulated FVPTC. This study evaluated whether computerized image analysis can detect nuclear differences between these three tumour subtypes., Methods: Slides with histological material from 15 cases of NIFTP and 33 cases of FVPTC subtypes (22 IEFVPTC, and 11 IFVPTC) were analyzed using the Image J image processing program. Tumour cells were compared for both nuclear morphometry and chromatin textural characteristics., Results: Nuclei from NIFTP and IFVPTC tumours differed in terms of chromatin textural features (grey intensity): mean (92.37 ± 21.01 vs 72.99 ± 14.73, p = 0.02), median (84.93 ± 21.17 vs 65.18 ± 17.08, p = 0.02), standard deviation (47.77 ± 9.55 vs 39.39 ± 7.18; p = 0.02), and coefficient of variation of standard deviation (19.96 ± 4.01 vs 24.75 ± 3.31; p = 0.003). No differences were found in relation to IEFVPTC., Conclusion: Computerized image analysis revealed differences in nuclear texture between NIFTP and IFVPTC, but not for IEFVPTC.

Objective: The aim of this study was to evaluate the influence of the incorporation of chlorhexidine-hexametaphosphate nanoparticles (CHX-HMP NPs) on antibacterial, cytotoxic and physicochemical properties of AH Plus (AH), MTA Fillapex (MTA) and Pulp Canal Sealer (PCS)., Methods: The NPs were synthesized and characterized by Scanning Electron Microscopy (SEM), Dynamic Light Scattering (DLS), zeta potential, Atomic Force Microscopy (AFM) and Energy-Dispersive X-ray Spectroscopy (EDS). The incorporation was made by weight, 2% and 5% of NPs. The antimicrobial activity, cytotoxicity, flow, radiopacity, setting time, solubility and pH were evaluated. The statistical analysis was performed by two-way analysis of variance test and Tukey post hoc test (P < 0.05)., Results: SEM analysis showed the tendency for CHX-HMP NPs to cluster, the effective mean diameter measured by DLS: 169.39 nm and the zeta potential: -10.18 mV. The NPs were individually measured by AFM: 22.99-52.75 nm. EDS analysis identified the presence of C, N, O, Na, P, Cl. After incorporation: The Direct Contact Test showed an increase in the antimicrobial action of AH, PCS and MTA; the sealers showed a decrease in flow and at 24 h of immersion also an increase in solubility, but did not affect the radiopacity of the samples; AH setting time increased and MTA did not reach setting under any of the conditions tested. All samples showed a decrease in pH value as the immersion time progressed., Significance: The incorporation of NPs can improve the antimicrobial performance of endodontic sealers without impairing other biological and physicochemical properties., (Copyright © 2021 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

Despite great advances in our knowledge of the consequences of Zika virus to human health, many questions remain unanswered, and results are often inconsistent. The small sample size of individual studies has limited inference about the spectrum of congenital Zika manifestations and the prognosis of affected children. The Brazilian Zika Cohorts Consortium addresses these limitations by bringing together and harmonizing epidemiological data from a series of prospective cohort studies of pregnant women with rash and of children with microcephaly and/or other manifestations of congenital Zika. The objective is to estimate the absolute risk of congenital Zika manifestations and to characterize the full spectrum and natural history of the manifestations of congenital Zika in children with and without microcephaly. This protocol describes the assembly of the Consortium and protocol for the Individual Participant Data Meta-analyses (IPD Meta-analyses). The findings will address knowledge gaps and inform public policies related to Zika virus. The large harmonized dataset and joint analyses will facilitate more precise estimates of the absolute risk of congenital Zika manifestations among Zika virus-infected pregnancies and more complete descriptions of its full spectrum, including rare manifestations. It will enable sensitivity analyses using different definitions of exposure and outcomes, and the investigation of the sources of heterogeneity between studies and regions.

The phytoalexin Resveratrol (3,5,4'-trihydroxystilbene; RSV) has been related to numerous beneficial effects on health by its cytoprotection and chemoprevention activities. Liver fibrosis is characterized by the extracellular matrix accumulation after hepatic injury and can lead to cirrhosis. Hepatic stellate cells (HSC) play a crucial role during fibrogenesis and liver wound healing by changing their quiescent phenotype to an activated phenotype for protecting healthy areas from damaged areas. Strategies on promoting the activated HSC death, the quiescence return or the cellular activation stimuli decrease play an important role on reducing liver fibrosis. Here, we evaluated the RSV effects on some markers of activation in GRX, an HSC model. We further evaluated the RSV influence in the ability of GRX on releasing inflammatory mediators. RSV at 1 and 10 µM did not alter the protein content of α-SMA, collagen I and GFAP; but 50 µM increased the content of these activation-related proteins. Also, RSV did not change the myofibroblast-like morphology of GRX. Interestingly, RSV at 10 and 50 µM decreased the GRX migration and collagen-I gel contraction. Finally, we showed that RSV triggered the increase in the TNF-α and IL-10 content in culture media of GRX while the opposite occurred for the IL-6 content. Altogether, these results suggested that RSV did not decrease the activation state of GRX and oppositely, triggered a pro-activation effect at the 50 µM concentration. However, despite the increase of TNF- α in culture media, these results on IL-6 and IL-10 secretion were in accordance with the anti-inflammatory role of RSV in our model.

Aim: To assess the impact of conservative endodontic access cavities (CEC) and truss access cavities (TAC) during root canal treatment performed on mandibular molars in terms of: ability to shape and fill root canals, microbial reduction in canals, and cleaning of the pulp chamber. In addition, the fracture resistance of the teeth after coronal restoration was assessed. Traditional endodontic cavities (TEC) were used as a reference technique for comparison., Methodology: Thirty extracted intact mandibular molars were scanned in a microcomputed tomography device (micro-CT), matched based on similar anatomical features and assigned to TEC, CEC or TAC groups (n = 10). The specimens were accessed accordingly, and root canals were contaminated with bacterial suspensions of Enterococcus faecalis (21 days). Subsequently, the first microbial sample was collected from root canals (S1). The canals were initially prepared with Reciproc Blue R25 instrument followed by a second instrumentation using Reciproc Blue R40. Eight mL of 0.5% NaOCl were used as an irrigant for each instrument. A final irrigation protocol was performed with 2 mL of 0.5% NaOCl, 2 mL of 17% EDTA and another 2 mL of 0.5% NaOCl. Microbial samples were collected from root canals after R25 (S2), R40 (S3) and final irrigation (S4). The teeth were rescanned after S4. Then, root canals were filled, rescanned, restored and the teeth subjected to fracture resistance tests. The statistical analysis was performed with type I negative binomial and beta 0-1 inflation regression models for microbiological analysis. Instrumentation, filling and resistance to fracture results were subjected to anova and Tukey tests (P < 0.05)., Results: S4 revealed no significant variations in microbial reduction amongst the groups (P > 0.05). TEC had a significantly lower percentage of unprepared surface area than CEC (P < 0.05). No differences were found regarding the percentage of dentine removed, transportation, centring ability and filling voids amongst the groups (P > 0.05). The TEC group had a significantly lower volume of remaining root filling material within the pulp chamber than CEC and TAC groups (P < 0.05). There was no difference regarding fracture resistance amongst the groups (P > 0.05)., Conclusion: Conservative access cavities did not offer any advantage in comparison with the traditional endodontic cavities in any of the parameters considered. Furthermore, conservative methods were associated with larger percentages of unprepared canal surface area and larger volumes of remaining root filling material within the pulp chamber., (© 2020 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Tumor cells can employ epithelial-mesenchymal transition (EMT) or autophagy in reaction to microenvironmental stress. Importantly, EMT and autophagy negatively regulate each other, are able to interconvert, and both have been shown to contribute to drug-resistance in glioblastoma (GBM). EMT has been considered one of the mechanisms that confer invasive properties to GBM cells. Autophagy, on the other hand, may show dual roles as either a GBM-promoter or GBM-suppressor, depending on microenvironmental cues. The Wingless (WNT) signaling pathway regulates a plethora of developmental and biological processes such as cellular proliferation, adhesion and motility. As such, GBM demonstrates deregulation of WNT signaling in favor of tumor initiation, proliferation and invasion. In EMT, WNT signaling promotes induction and stabilization of different EMT activators. WNT activity also represses autophagy, while nutrient deprivation induces β-catenin degradation via autophagic machinery. Due to the importance of the WNT pathway to GBM, and the role of WNT signaling in EMT and autophagy, in this review we highlight the effects of the WNT signaling in the regulation of both processes in GBM, and discuss how the crosstalk between EMT and autophagy may ultimately affect tumor biology., (Copyright © 2020 Coelho, Fernandes, Boccacino, Souza, Melo-Escobar, Alves, Prado, Iglesia, Cangiano, Mazzaro and Lopes.)

The mobility of cellular prion protein (PrP C ) in specific cell membrane domains and among distinct cell compartments dictates its molecular interactions and directs its cell function. PrP C works in concert with several partners to organize signaling platforms implicated in various cellular processes. The scaffold property of PrP C is able to gather a molecular repertoire to create heterogeneous membrane domains that favor endocytic events. Dynamic trafficking of PrP C through multiple pathways, in a well-orchestrated mechanism of intra and extracellular vesicular transport, defines its functional plasticity, and also assists the conversion and spreading of its infectious isoform associated with neurodegenerative diseases. In this review, we highlight how PrP C traffics across intra- and extracellular compartments and the consequences of this dynamic transport in governing cell functions and contributing to prion disease pathogenesis.

Cell motility is a central process involved in fundamental biological phenomena during embryonic development, wound healing, immune surveillance, and cancer spreading. Cell movement is complex and dynamic and requires the coordinated activity of cytoskeletal, membrane, adhesion and extracellular proteins. Cellular prion protein (PrP C ) has been implicated in distinct aspects of cell motility, including axonal growth, transendothelial migration, epithelial-mesenchymal transition, formation of lamellipodia , and tumor migration and invasion. The preferential location of PrP C on cell membrane favors its function as a pivotal molecule in cell motile phenotype, being able to serve as a scaffold protein for extracellular matrix proteins, cell surface receptors, and cytoskeletal multiprotein complexes to modulate their activities in cellular movement. Evidence points to PrP C mediating interactions of multiple key elements of cell motility at the intra- and extracellular levels, such as integrins and matrix proteins, also regulating cell adhesion molecule stability and cell adhesion cytoskeleton dynamics. Understanding the molecular mechanisms that govern cell motility is critical for tissue homeostasis, since uncontrolled cell movement results in pathological conditions such as developmental diseases and tumor dissemination. In this review, we discuss the relevant contribution of PrP C in several aspects of cell motility, unveiling new insights into both PrP C function and mechanism in a multifaceted manner either in physiological or pathological contexts.

Alzheimer's disease is a neurodegenerative disorder characterized by extracellular deposition of amyloid-β (Aβ) peptide and hyperphosphorylation of Tau protein, which ultimately leads to the formation of intracellular neurofibrillary tangles and cell death. Increasing evidence indicates that genistein, a soy isoflavone, has neuroprotective effects against Aβ-induced toxicity. However, the molecular mechanisms involved in its neuroprotection are not well understood. In this study, we have established a neuronal damage model using retinoic-acid differentiated SH-SY5Y cells treated with different concentrations of Aβ 25-35 to investigate the effect of genistein against Aβ-induced cell death and the possible involvement of protein kinase B (PKB, also termed Akt), glycogen synthase kinase 3β (GSK-3β), and Tau as an underlying mechanism to this neuroprotection. Differentiated SH-SY5Y cells were pre-treated for 24 hr with genistein (1 and 10 nM) and exposed to Aβ 25-35 (25 μM), and we found that genistein partially inhibited Aβ induced cell death, primarily apoptosis. Furthermore, the protective effect of genistein was associated with the inhibition of Aβ-induced Akt inactivation and Tau hyperphosphorylation. These findings reinforce the neuroprotective effects of genistein against Aβ toxicity and provide evidence that its mechanism may involve regulation of Akt and Tau proteins., (© 2019 John Wiley & Sons, Ltd.)

Heat shock proteins (HSPs) are evolutionary conserved proteins that work as molecular chaperones and perform broad and crucial roles in proteostasis, an important process to preserve the integrity of proteins in different cell types, in health and disease. Their function in cancer is an important aspect to be considered for a better understanding of disease development and progression. Glioblastoma (GBM) is the most frequent and lethal brain cancer, with no effective therapies. In recent years, HSPs have been considered as possible targets for GBM therapy due their importance in different mechanisms that govern GBM malignance. In this review, we address current evidence on the role of several HSPs in the biology of GBMs, and how these molecules have been considered in different treatments in the context of this disease, including their activities in glioblastoma stem-like cells (GSCs), a small subpopulation able to drive GBM growth. Additionally, we highlight recent works that approach other classes of chaperones, such as histone and mitochondrial chaperones, as important molecules for GBM aggressiveness. Herein, we provide new insights into how HSPs and their partners play pivotal roles in GBM biology and may open new therapeutic avenues for GBM based on proteostasis machinery.

Several studies have demonstrated the antitumor effect of doxazosin, an α1-adrenergic blocker, against glioma and breast, bladder and prostate cancers. Doxazosin is also being evaluated as a treatment for posttraumatic stress disorder (PTSD) and alcoholism, and α1-adrenergic blockers have been linked to neuroprotection in neurodegenerative disorders, such as Alzheimer's Disease (AD). Cancer and AD have an inverse relationship in many aspects, with several factors that contribute to apoptosis inhibition and proliferation being increased in cancers but decreased in AD. Neuroblastoma (NB) is a pediatric tumor derived from embryonic neural-crest cells, with an overall cure rate of 40%, despite aggressive treatment. Thus, due to the need of new therapeutic strategies against NB and neurodegenerative disorders and the inverse relationship between these diseases, we investigated whether doxazosin may serve as an antitumor and neuroprotective agent. We analyzed the drug's effects on undifferentiated and retinoic acid-differentiated SH-SY5Y human NB cells and on an in vitro model of organotypic hippocampal cultures exposed to amyloid-β. Doxazosin showed antitumor effect on undifferentiated NB cells by induction of apoptosis, necrosis, cell cycle arrest and decrease of p-EGFR Tyr1048 levels. On differentiated cells, doxazosin was less cytotoxic and increased p-EGFR Tyr1048 , p-Akt Ser473 and p-GSK-3β Ser9 levels. Moreover, the drug was able to protect hippocampal slices from amyloid-β toxicity through prevention of GSK-3β activation and of Tau hyperphosphorylation. Therefore, our results show that doxazosin has antitumor activity against undifferentiated NB and is neuroprotective on an in vitro model of Alzheimer's disease., (Copyright © 2019 IBRO. Published by Elsevier Ltd. All rights reserved.)

Background: Computerized image analysis seems to represent a promising diagnostic possibility for thyroid tumors. Our aim was to evaluate the discriminatory diagnostic efficiency of computerized image analysis of cell nuclei from histological materials of follicular tumors., Methods: We studied paraffin-embedded materials from 42 follicular adenomas (FA), 47 follicular variants of papillary carcinomas (FVPC) and 20 follicular carcinomas (FC) by the software ImageJ. Based on the nuclear morphometry and chromatin texture, the samples were classified as FA, FC or FVPC using the Classification and Regression Trees method., Results: We observed high diagnostic sensitivity and specificity rates (FVPC: 89.4% and 100%; FC: 95.0% and 92.1%; FA: 90.5 and 95.5%, respectively). When the tumors were compared by pairs (FC vs FA, FVPC vs FA), 100% of the cases were classified correctly., Conclusion: The computerized image analysis of nuclear features showed to be a useful diagnostic support tool for the histological differentiation between follicular adenomas, follicular variants of papillary carcinomas and follicular carcinomas., (© 2018 The authors.)

Purpose: To investigate the efficacy of allogeneic mesenchymal stem-cells and autologous mononuclear cells to promote sensorimotor recovery and tissue rescue., Methods: Female rabbits were submitted to the epidural balloon inflation method and the intravenous cells administrations were made after 8 hours or seven days after injury induction. Sensorimotor evaluation of the hindlimbs was performed, and the euthanasia was made thirty days after the treatment. Spinal cords were stained with hematoxylin and eosin., Results: Both therapies given 8 hours after the injury promoted the sensorimotor recovery after a week. Only the group treated after a week with mononuclear cells showed no significant recovery at post-injury day 14. In the days 21 and 28, all treatments promoted significant recovery. Histopathological analysis showed no difference among the experimental groups. Our results showed that both bone marrow-derived cell types promoted significant sensorimotor recovery after injury, and the treatment made at least a week after injury is efficient., Conclusion: The possibilities of therapy with bone marrow-derived cells are large, increasing the therapeutic arsenal for the treatment of spinal cord injury.

Glioblastoma (GBM) is the most common and aggressive primary malignant brain tumor in adults. Hypoxia is a distinct feature in GBM and plays a significant role in tumor progression, resistance to treatment, and poor outcome. However, there is lack of studies relating type of cell death, status of Akt phosphorylation on Ser473, mitochondrial membrane potential, and morphological changes of tumor cells after hypoxia and reoxygenation. The rat glioma C6 cell line was exposed to oxygen deprivation (OD) in 5 % fetal bovine serum (FBS) or serum-free media followed by reoxygenation (RO). OD induced apoptosis on both 5 % FBS and serum-free groups. Overall, cells on serum-free media showed more profound morphological changes than cells on 5 % FBS. Moreover, our results suggest that OD combined with absence of serum provided a favorable environment for glioblastoma dedifferentiation to cancer stem cells, since nestin, and CD133 levels increased. Reoxygenation is present in hypoxic tumors through microvessel formation and cell migration to oxygenated areas. However, few studies approach these phenomena when analyzing hypoxia. We show that RO caused morphological alterations characteristic of cells undergoing a differentiation process due to increased GFAP. In the present study, we characterized an in vitro hypoxic microenvironment associated with GBM tumors, therefore contributing with new insights for the development of therapeutics for resistant glioblastoma.

Chemical coding of stomatogastric nervous system (STNS) and enteric nervous system (ENS) of midgut and hindgut in the snail Megalobulimus abbreviatus was investigated using histochemistry, histofluorescence, and immunohistochemistry. The gastrointestinal plexuses, constituted by intrinsic neurons and fibers originating from the subesophageal ganglia and/or STNS, showed intense acetylcholinesterase (AChE) and nicotinamide adenine dinucleotide diaphorase (NADPHd) activity. The enteric neurons and fibers with AChE activity are scattered in the submucosa and between both muscular layers of gastrointestinal tract, whereas NADPHd neurons and fibers are more abundant between muscular layers than in the submucosa. Catecholaminergic nerve fibers and varicosities are found mainly within the submucosa across the mid- and hindgut. Serotoninand FMRFamide-immunoreactive neurons and fibers originating from the STNS are distributed in the submucosa of the intestine and rectum. FMRFamide-immunoreactive neurons and fibers are present in the mucosa, submucosa, and muscular layers of mid- and hindgut. The neuron-like intraepithelial cells exhibited AChE activity, a few NADPHd activity, and immunoreactivity for serotonin and FMRFamide. Intense glial fibrillary acidic protein (GFAP) immunoreaction is found throughout the intestine plexuses and in the STNS ganglia. The GFAP immunoreaction in intramural plexuses suggests the presence of glial cells as an important component of ENS in this pulmonate snail.

The most common cause of spinal cord injury are high impact trauma, which often result in some motor impairment, sensory or autonomic a greater or lesser extent in the distal areas the level of trauma. In terms of survival and complications due to sequelae, veterinary patients have a poor prognosis unfavorable. Therefore justified the study of experimental models of spinal cord injury production that could provide more support to research potential treatments for spinal cord injuries in medicine and veterinary medicine. Preclinical studies of acute spinal cord injury require an experimental animal model easily reproducible. The most common experimental animal model is the rat, and several techniques for producing a spinal cord injury. The objective of this study was to describe and evaluate the effectiveness of acute spinal cord injury production technique through inflation of Fogarty(r) catheter using rabbits as an experimental model because it is a species that has fewer conclusive publications and contemplating. The main requirements of a model as low cost, handling convenience, reproducibility and uniformity. The technique was adequate for performing preclinical studies in neuro-traumatology area, effectively leading to degeneration and necrosis of the nervous tissue fostering the emergence of acute paraplegia.

Glioblastoma is the most frequent and malignant brain tumor. Treatment includes chemotherapy with temozolomide concomitant with surgical resection and/or irradiation. However, a number of cases are resistant to temozolomide, as well as the human glioblastoma cell line U138-MG. We investigated doxazosin's (an antihypertensive drug) activity against glioblastoma cells (C6 and U138-MG) and its neurotoxicity on primary astrocytes and organoptypic hippocampal cultures. For this study, the following methods were used: citotoxicity assays, flow cytometry, western-blotting and confocal microscopy. We showed that doxazosin induces cell death on C6 and U138-MG cells. We observed that doxazosin's effects on the PI3K/Akt pathway were similar as LY294002 (PI3K specific inhibitor). In glioblastoma cells treated with doxasozin, Akt levels were greatly reduced. Upon examination of activities of proteins downstream of Akt we observed upregulation of GSK-3β and p53. This led to cell proliferation inhibition, cell death induction via caspase-3 activation and cell cycle arrest at G0/G1 phase in glioblastoma cells. We used in this study Lapatinib, a tyrosine kinase inhibitor, as a comparison with doxazosin because they present similar chemical structure. We also tested the neurocitotoxicity of doxazosin in primary astrocytes and organotypic cultures and observed that doxazosin induced cell death on a small percentage of non-tumor cells. Aggressiveness of glioblastoma tumors and dismal prognosis require development of new treatment agents. This includes less toxic drugs, more selective towards tumor cells, causing less damage to the patient. Therefore, our results confirm the potential of doxazosin as an attractive therapeutic antiglioma agent.

Objective: Surgical treatment of Scheuermann's kyphosis (SK) remains a subject under discussion. In view of the controversy over the best form of surgical tr otomy., Methods: This was a descriptive case-control study with cross-sectional analysis. Twenty-eight patients, split into two groups conducted at different times, were evaluated., Results: The first group comprised patients treated using the double approach, with an average age of 19 years, preoperative kyphosis of 77.6°, postoperative kyphosis of 35.8° and average correction of 53.2%. The second group comprised patients treated using the posterior route associated with Smith-Petersen osteotomy, with a mean age of 27.3 years, preoperative kyphosis of 72.9°, postoperative kyphosis of 44.3° and average correction of 39.3%. Analysis between the two groups showed statistically significant differences in the following variables: age (p = 0.02), postoperative kyphosis (p = 0.04) and degree and percentage of kyphosis correction (p = 0.001). There was no difference concerning preoperative kyphosis (p = 0.33). In the assessment of postoperative pain (VAS), the first group presented an average of 0.6, versus 0.5 in the second group. There were only minor complications: seven in the first group and two in the second., Conclusion: The two surgical techniques studied proved to be adequate for treating SK. In the present study, the deformity correction was greater in the first group, while the pain VAS results were better in the second group, with lower incidence of complications.

Increased efficacy of the multipotent mesenchymal stromal cells (MSCs) for the treatment of CNS injuries has been shown when they are administrated within a collagen scaffold, an environment in three dimensions (3D), when compared to the cultivation over a plastic surface (2D). We evaluated the MSC therapeutic effect in the 2D and 3D conditions using the model of focal cortical ablation. Male Wistar rats were submitted to the ablation by aspiration. Intravenous injection (IV) of MSC cultured in 2D, and the intralesional administration (IL) of MSC cultured in 2D or 3D were tested. Administrations were made 24h after ablation. Unskilled and skilled forelimb movements were evaluated by sensorimotor tests. The level of cytokines was measured two days after ablation in the 2D IV groups. Only the MSC 3D IL promoted recovery of the skilled movements. MSC 2D IV promoted recovery of the unskilled movements in all tests, and the MSC 3D IL promoted it only in the adhesive test. MSC 2D IL was unable to promote any recovery. DAPI-stained MSC was found in the perilesional parenchyma at the third post-ablation day after 2D and 3D IL. A significant reduction in the levels of cytokines by the MSC 2D IV was observed in the plasma. Our study strengthens the evidences of the MSC as a prospective therapeutic approach for the CNS injuries., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Stroke is a disease of the elderly. However, most of the preclinical studies about the treatment of stroke with bone marrow-derived cells have used young animals. Here, it was assessed whether the sensorimotor recovery promoted by the treatment of the brain ischemia with the bone marrow mononuclear cells (BMMCs) is influenced by age and/or gender. Unilateral cortical ischemia by thermocoagulation was made in the primary motor and sensorimotor cortices in young and middle-aged rats of both genders. Twenty four hours after ischemia, animals received intravenous injection of BMMCs or vehicle. Each combination of age and gender received BMMCs from donor with the same combination. Survival rate and ischemic lesion size were quantified. Sensorimotor recovery was evaluated by the cylinder and adhesive tests. The results showed that the treatment with BMMCs resulted in sensorimotor recovery of both young and middle-aged ischemic rats. No important effect of gender was found, but age was a significant factor. Middle-aged animals had increased mortality and lesion sizes. In the adhesive test, middle-aged animals had lower BMMCs-induced sensorimotor recovery. The results suggest that the treatment of stroke with the BMMCs should be beneficial for males and females in the elderly., (Copyright © 2014 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.)

Digestive disorders caused by sudden changes in diet or inappropriate diet are among the most common disorders of the digestive system. Cecal or intestinal tympany, one consequence of inappropriate diet, is characterized by the accumulation of gases, marked distension of the cecum and colon and the induction of inflammatory processes. To know the effects of intestinal tympany on the enteric plexuses, we developed a method of experimental tympanic colic (TC) in the Chinchilla lanigera. This species was used in view of its susceptibility to TC. TC was induced with a diet rich in alfalfa associated with grain overload for two weeks. Physical and clinical examination including the von Frey test confirmed the diagnosis. The chinchillas with acute abdomen were treated with 1% ketoprofen and resumption of a balanced diet. Necropsy and histopathological analysis showed tympany-induced alterations mainly in the cecum and colon. After treatment, the control conditions were restored. The TC protocol is proposed as an experimental approach designed to aid the study of the effects of acute intestinal inflammation and obstruction caused by an inappropriate diet.

Resveratrol (RSV) is known for its antioxidant properties; however, this compound has been proposed to have cytotoxic and pro-oxidant effects depending on its concentration and time of exposure. We previously reported the cell cycle arrest effect of low doses of RSV in GRX cells, an activated hepatic stellate cell model. Here, we evaluated the effects of RSV treatment (0.1-50 μM) for 24 and 120 h on GRX viability and oxidative status. Only treatment with 50 μM of RSV reduced the amount of live cells. However, even low doses of RSV induced an increased reactive species production at both treatment times. While being diminished within 24 h, RSV induced an increase in the SOD activity in 120 h. The cellular damage was substantially increased at 24 h in the 50 μM RSV-treated group, as indicated by the high lipoperoxidation, which may be related to the significant cell death and low proliferation. Paradoxically, this cellular damage and lipoperoxidation were considerably reduced in this group after 120 h of treatment while the surviving cells proliferated. In conclusion, RSV induced a dose-dependent pro-oxidant effect in GRX cells. The highest RSV dose induced oxidative-related damage, drastically reducing cell viability; but this cytotoxicity seems to be attenuated during 120 h of treatment.

We describe the morphology and innervation of the midgut and hindgut of the giant land snail Megalobulimus abbreviatus for the first time. The midgut (stomach and intestine) and hindgut (rectum and anus) are innervated by the subesophageal ganglia, through the gastrointestinal branch (originated from the visceral nerve) and the rectum-anal nerve, respectively. Backfilling through these nerves revealed neuronal bodies, mainly in the right parietal and visceral ganglia. The enteric plexuses of the midgut and hindgut are formed by extensive axonal networks and several neuronal somata arranged in clusters or as isolated cells. The gastrointestinal branch and the rectum-anal nerve directly innervate the enteric plexuses of the intestine and the hindgut, respectively. However, the outer wall of the stomach has a stomatogastric nervous system, which consists of four ganglia: stomatogastric, gastric, cardic, and pyloric. Fibers of the gastrointestinal branch project to these ganglia. Anterograde tracing from stomatogastric system ganglia revealed that the enteric plexus of the stomach is innervated only by these peripheral ganglia. Anterograde tracing of the gastrointestinal branch did not result in labeling in the enteric plexus of the stomach. Therefore, the midgut and hindgut of M. abbreviatus is controlled by an intrinsic innervation, constituted by the submucous and myenteric plexuses, which are innervated directly by neurons from the subesophageal ganglia or indirectly via the stomatogastric nervous system (for the stomach).

Thirty-seven isolates from 10 HIV-negative and 26 HIV-positive patients in Mato Grosso were evaluated. Direct examination, culturing and chemotyping of species were performed. Ketoconazole, itraconazole, voriconazole, fluconazole and amphotericin B were evaluated. Thirty-seven yeasts of Cryptococcus spp were identified, of which 26 were from HIV-positive patients (25 Cryptococcus neoformans and one Cryptococcus gattii) and 10 from HIV-negative patients (five Cryptococcus neoformans and five Cryptococcus gattii). The Cryptococcus neoformans clinical isolates from HIV-positive patients showed resistance (8% and 8.7%) and dose-dependent susceptibility (20% and 17.4%) to fluconazole and itraconazole, respectively. Among the Cryptococcus neoformans isolates from HIV-negative patients, there was dose-dependent susceptibility (40%) to fluconazole. Cryptococcus gattii isolates from HIV-negative patients were shown to be susceptible to all antifungal agents, except for one isolate of Cryptococcus gattii that showed dose-dependent susceptibility to fluconazole (20%). The Cryptococcus gattii isolate from an HIV-positive patient showed resistance to fluconazole (MIC > or = 256 (1/4)g/ml) and itraconazole (MIC = 3 (1/4)microg/ml).

Resveratrol (RSV) exerts anti-proliferative and pro-apoptotic actions in different cell lines. Hepatic stellate cells (HSCs) are major fibrogenic cell types that contribute to collagen accumulation during chronic liver disease. In the present study, the inhibitory effects of RSV on cell proliferation, cell cycle, and apoptosis were evaluated in the mouse hepatic stellate cell line GRX. Cells treated with 1 nM-1 muM of RSV demonstrated a decrease in cell growth of about 35% after 5 days. GRX cells, treated with RSV (100 nM or 1 muM), were analyzed by flow cytometry; RSV induced an increase in the number of GRX cells in the S- and sub-G1 phases. The increase in sub-G1 phase cells and the nuclear condensation and fragmentation shown by DAPI staining identified a possible pro-apoptotic effect of RSV on GRX cells. Furthermore, the RSV anti-proliferative effects could be explained by an S-phase accumulation caused by a decrease in the progression through the cell cycle or an inhibition of S or G2 phase transition. It is notable that these RSV actions are mediated at nanomolar levels, compatible with the concentrations of free RSV in biological fluids after ingestion of polyphenol-rich foods, suggesting a possible effect of these foods as an adjuvant treatment in chronic liver diseases.