Wasteosomes (corpora amylacea) as a hallmark of chronic glymphatic insufficiency.

In different organs and tissues, the lymphatic system serves as a drainage system for interstitial fluid and is useful for removing substances that would otherwise accumulate in the interstitium. In the brain, which lacks lymphatic circulation, the drainage and cleaning function is performed by the glymphatic system, called so for its dependence on glial cells and its similar function to that of the lymphatic system. In the present article, we define glymphatic insufficiency as the inability of the glymphatic system to properly perform the brain cleaning function. Furthermore, we propose that corpora amylacea or wasteosomes, which are protective structures that act as waste containers and accumulate waste products, are, in fact, a manifestation of chronic glymphatic insufficiency. Assuming this premise, we provide an explanation that coherently links the formation, distribution, structure, and function of these bodies in the human brain. Moreover, we open up new perspectives in the study of the glymphatic system since wasteosomes can provide information about which variables have the greatest impact on the glymphatic system and which diseases occur with chronic glymphatic insufficiency. For example, based on the presence of wasteosomes, it seems that aging, sleep disorders, and cerebrovascular pathologies have the highest impact on the glymphatic system, whereas neurodegenerative diseases have a more limited impact. Furthermore, as glymphatic insufficiency is a risk factor for neurodegenerative diseases, information provided by wasteosomes could help to define the strategies and actions that can prevent glymphatic disruptions, thus limiting the risk of developing neurodegenerative diseases.

Corpora amylacea act as containers that remove waste products from the brain.

Corpora amylacea (CA) in the human brain are granular bodies formed by polyglucosan aggregates that amass waste products of different origins. They are generated by astrocytes, mainly during aging and neurodegenerative conditions, and are located predominantly in periventricular and subpial regions. This study shows that CA are released from these regions to the cerebrospinal fluid and are present in the cervical lymph nodes, into which cerebrospinal fluid drains through the meningeal lymphatic system. We also show that CA can be phagocytosed by macrophages. We conclude that CA can act as containers that remove waste products from the brain and may be involved in a mechanism that cleans the brain. Moreover, we postulate that CA may contribute in some autoimmune brain diseases, exporting brain substances that interact with the immune system, and hypothesize that CA may contain brain markers that may aid in the diagnosis of certain brain diseases.

A Machine-Learning Model for Lung Age Forecasting by Analyzing Exhalations.

Spirometers are important devices for following up patients with respiratory diseases. These are mainly located only at hospitals, with all the disadvantages that this can entail. This limits their use and consequently, the supervision of patients. Research efforts focus on providing digital alternatives to spirometers. Although less accurate, the authors claim they are cheaper and usable by many more people worldwide at any given time and place. In order to further popularize the use of spirometers even more, we are interested in also providing user-friendly lung-capacity metrics instead of the traditional-spirometry ones. The main objective, which is also the main contribution of this research, is to obtain a person's lung age by analyzing the properties of their exhalation by means of a machine-learning method. To perform this study, 188 samples of blowing sounds were used. These were taken from 91 males (48.4%) and 97 females (51.6%) aged between 17 and 67. A total of 42 spirometer and frequency-like features, including gender, were used. Traditional machine-learning algorithms used in voice recognition applied to the most significant features were used. We found that the best classification algorithm was the Quadratic Linear Discriminant algorithm when no distinction was made between gender. By splitting the corpus into age groups of 5 consecutive years, accuracy, sensitivity and specificity of, respectively, 94.69%, 94.45% and 99.45% were found. Features in the audio of users' expiration that allowed them to be classified by their corresponding lung age group of 5 years were successfully detected. Our methodology can become a reliable tool for use with mobile devices to detect lung abnormalities or diseases.

Astrocytes and neurons produce distinct types of polyglucosan bodies in Lafora disease.

Lafora disease (LD), the most devastating adolescence-onset epilepsy, is caused by mutations in the EPM2A or EPM2B genes, which encode the proteins laforin and malin, respectively. Loss of function of one of these proteins, which are involved in the regulation of glycogen synthesis, induces the accumulation of polyglucosan bodies (PGBs)-known as Lafora bodies (LBs) and associated with neurons-in the brain. Ageing and some neurodegenerative conditions lead to the appearance of another type of PGB called corpora amylacea, which are associated with astrocytes and contain neo-epitopes that can be recognized by natural antibodies. Here we studied the PGBs in the cerebral cortex and hippocampus of malin knockout mice, a mouse model of LD. These animals presented not only LBs associated with neurons but also a significant number of PGBs associated with astrocytes. These astrocytic PGBs were also increased in mice from senescence-accelerated mouse-prone 8 (SAMP8) strain and mice with overexpression of Protein Targeting to Glycogen (PTGOE ), indicating that they are not exclusive of LD. The astrocytic PGBs, but not neuronal LBs, contained neo-epitopes that are recognized by natural antibodies. The astrocytic PGBs appeared predominantly in the hippocampus but were also present in some cortical brain regions, while neuronal LBs were found mainly in the brain cortex and the pyramidal layer of hippocampal regions CA2 and CA3. Our results indicate that astrocytes, contrary to current belief, are involved in the etiopathogenesis of LD.

Serial block-face scanning electron microscopy applied to study the trafficking of 8D3-coated gold nanoparticles at the blood-brain barrier.

Due to the physical and physiological properties of the blood-brain barrier (BBB), the transport of neurotherapeutics from blood to brain is still a pharmaceutical challenge. We previously conducted a series of experiments to explore the potential of the anti-transferrin receptor 8D3 monoclonal antibody (mAb) to transport neurotherapeutics across the BBB. In that study, gold nanoparticles (AuNPs) were coated with the 8D3 antibody and administered intravenously to mice. Transmission electron microscopy was used and a two-dimensional (2D) image analysis was performed to detect the AuNPs in the brain capillary endothelial cells (BCECs) and brain parenchyma. In the present work, we determined that serial block-face scanning electron microscopy (SBF-SEM) is a useful tool to study the transcytosis of these AuNPs across the BBB in three dimensions and we, therefore, applied it to gain more knowledge of their transcellular trafficking. The resulting 3D reconstructions provided additional information on the endocytic vesicles containing AuNPs and the endosomal processing that occurs inside BCECs. The passage from 2D to 3D analysis reinforced the trafficking model proposed in the 2D study, and revealed that the vesicles containing AuNPs are significantly larger and more complex than described in our 2D study. We also discuss tradeoffs of using this technique for our application, and conclude that together with other volume electron microscopy imaging techniques, SBF-SEM is a powerful approach that is worth of considering for studies of drug transport across the BBB.

Trafficking of Gold Nanoparticles Coated with the 8D3 Anti-Transferrin Receptor Antibody at the Mouse Blood-Brain Barrier.

Receptor-mediated transcytosis has been widely studied as a possible strategy to transport neurotherapeutics across the blood-brain barrier (BBB). Monoclonal antibodies directed against the transferrin receptor (TfR) have been proposed as potential carrier candidates. A better understanding of the mechanisms involved in their cellular uptake and intracellular trafficking is required and could critically contribute to the improvement of delivery methods. Accordingly, we studied here the trafficking of gold nanoparticles (AuNPs) coated with the 8D3 anti-transferrin receptor antibody at the mouse BBB. 8D3-AuNPs were intravenously administered to mice and allowed to recirculate for a range of times, from 10 min to 24 h, before brain extraction and analysis by transmission electron microscope techniques. Our results indicated a TfR-mediated and clathrin-dependent internalization process by which 8D3-AuNPs internalize individually in vesicles. These vesicles then follow at least two different routes. On one hand, most vesicles enter intracellular processes of vesicular fusion and rearrangement in which the AuNPs end up accumulating in late endosomes, multivesicular bodies or lysosomes, which present a high AuNP content. On the other hand, a small percentage of the vesicles follow a different route in which they fuse with the abluminal membrane and open to the basal membrane. In these cases, the 8D3-AuNPs remain attached to the abluminal membrane, which suggests an endosomal escape, but not dissociation from TfR. Altogether, although receptor-mediated transport continues to be one of the most promising strategies to overcome the BBB, different optimization approaches need to be developed for efficient drug delivery.

Neo-epitopes emerging in the degenerative hippocampal granules of aged mice can be recognized by natural IgM auto-antibodies.

BACKGROUND: Degenerative granular structures appear progressively with age in the hippocampus of most mouse strains. We recently reported that these granules contain a neo-epitope that is recognised by IgM antibodies present as contaminants in many commercial antibodies obtained from mouse ascites and mouse or rabbit serum. We hypothesise that these anti-neo-epitope IgMs are in fact natural auto-antibodies that are generated spontaneously during the foetal stage without previous contact with external antigens and whose repertoire and reactivity pattern have been determined through evolution, being remarkably stable within species and even between species. FINDINGS: In the present work we found that mice from the ICR-CD1, BALB/C and SAMP8 strains have anti-neo-epitope IgM antibodies in their plasma at all ages tested and even when maintained under specific opportunistic pathogen-free conditions. Moreover, we determined that these anti-neo-epitope IgMs are also present in rabbit, goat and rat serum. We also found that, in each mouse that presented hippocampal granules, the anti-neo-epitope IgMs contained in its plasma recognised the neo-epitopes in its own granules. CONCLUSIONS: This study led to the conclusion that anti-neo-epitope IgMs are widespread natural auto-antibodies contained in the plasma of mice and other species. The presence of these natural auto-antibodies not only explains why they are frequently found as contaminants in commercial antibodies, but also paves the way for a new approach to a treatment and diagnosis of pathological brain processes based on natural IgMs and neo-epitopes.

A green strategy for federated and heterogeneous clouds with communicating workloads.

Providers of cloud environments must tackle the challenge of configuring their system to provide maximal performance while minimizing the cost of resources used. However, at the same time, they must guarantee an SLA (service-level agreement) to the users. The SLA is usually associated with a certain level of QoS (quality of service). As response time is perhaps the most widely used QoS metric, it was also the one chosen in this work. This paper presents a green strategy (GS) model for heterogeneous cloud systems. We provide a solution for heterogeneous job-communicating tasks and heterogeneous VMs that make up the nodes of the cloud. In addition to guaranteeing the SLA, the main goal is to optimize energy savings. The solution results in an equation that must be solved by a solver with nonlinear capabilities. The results obtained from modelling the policies to be executed by a solver demonstrate the applicability of our proposal for saving energy and guaranteeing the SLA.

Database constraints applied to metabolic pathway reconstruction tools.

Our group developed two biological applications, Biblio-MetReS and Homol-MetReS, accessing the same database of organisms with annotated genes. Biblio-MetReS is a data-mining application that facilitates the reconstruction of molecular networks based on automated text-mining analysis of published scientific literature. Homol-MetReS allows functional (re)annotation of proteomes, to properly identify both the individual proteins involved in the process(es) of interest and their function. It also enables the sets of proteins involved in the process(es) in different organisms to be compared directly. The efficiency of these biological applications is directly related to the design of the shared database. We classified and analyzed the different kinds of access to the database. Based on this study, we tried to adjust and tune the configurable parameters of the database server to reach the best performance of the communication data link to/from the database system. Different database technologies were analyzed. We started the study with a public relational SQL database, MySQL. Then, the same database was implemented by a MapReduce-based database named HBase. The results indicated that the standard configuration of MySQL gives an acceptable performance for low or medium size databases. Nevertheless, tuning database parameters can greatly improve the performance and lead to very competitive runtimes.

Increase in wasteosomes (corpora amylacea) in frontotemporal lobar degeneration with specific detection of tau, TDP-43 and FUS pathology.

Wasteosomes (or corpora amylacea) are polyglucosan bodies that appear in the human brain with aging and in some neurodegenerative diseases, and have been suggested to have a potential role in a nervous system cleaning mechanism. Despite previous studies in several neurodegenerative disorders, their status in frontotemporal lobar degeneration (FTLD) remains unexplored. Our study aims to characterize wasteosomes in the three primary FTLD proteinopathies, assessing frequency, distribution, protein detection, and association with aging or disease duration. Wasteosome scores were obtained in various brain regions from 124 post-mortem diagnosed sporadic FTLD patients, including 75 participants with tau (FTLD-tau), 42 with TAR DNA-binding protein 43 (FTLD-TDP), and 7 with Fused in Sarcoma (FTLD-FUS) proteinopathies, along with 29 control subjects. The wasteosome amount in each brain region for the different FLTD patients was assessed with a permutation test with age at death and sex as covariables, and multiple regressions explored associations with age at death and disease duration. Double immunofluorescence studies examined altered proteins linked to FTLD in wasteosomes. FTLD patients showed a higher accumulation of wasteosomes than control subjects, especially those with FTLD-FUS. Unlike FTLD-TDP and control subjects, wasteosome accumulation did not increase with age in FTLD-tau and FTLD-FUS. Cases with shorter disease duration in FTLD-tau and FTLD-FUS seemed to exhibit higher wasteosome quantities, whereas FTLD-TDP appeared to show an increase with disease progression. Immunofluorescence studies revealed the presence of tau and phosphorylated-TDP-43 in the periphery of isolated wasteosomes in some patients with FTLD-tau and FTLD-TDP, respectively. Central inclusions of FUS were observed in a higher number of wasteosomes in FTLD-FUS patients. These findings suggest a role of wasteosomes in FTLD, especially in the more aggressive forms of FLTD-FUS. Detecting these proteins, particularly FUS, in wasteosomes from cerebrospinal fluid could be a potential biomarker for FTLD.

Quitline nurses' experiences in providing telephone-based smoking cessation help to mental health patients: A mixed methods study.

WHAT IS KNOWN ON THE SUBJECT?: Quitlines are known to be effective in helping people quit smoking, including those with mental health conditions. It is particularly important to address smoking in this population as the prevalence of smoking ranges from 40% to 75%. However, professionals working in quitlines often face barriers due to their limited training and resources to effectively support these smokers quit, especially if they are not mental health professionals. Therefore, training programmes should be developed to enhance their knowledge and skills in providing smoking cessation support to this vulnerable population. WHAT THE PAPER ADDS TO EXISTING KNOWLEDGE?: The '061 QUIT-MENTAL study' evaluated the efficacy of a proactive telephone-based intervention for smoking cessation among smokers with severe mental health disorders. Conducted through a quitline service in Catalonia, Spain, the study focused on training non-mental health specialized nurses and other health professionals to provide evidence-based interventions for promoting smoking cessation among individuals with mental health disorders. The objective of this study is to assess the changes in nurses' knowledge and readiness to treat smokers with mental health conditions, while also capturing their insights and perceptions regarding the facilitators and barriers to providing smoking cessation interventions. The training and insights of the nurses were integral to conducting this research and providing valuable information for the future sustainability of such interventions. This is particularly important as quitlines hold the potential to offer cessation support to these patients at the community level. WHAT ARE THE IMPLICATIONS FOR PRACTICE?: While the training programme was successful in improving non-mental health specialized nurses' knowledge and motivation skills to help patients with mental health disorders quit smoking, they encountered obstacles in delivering this intervention over the phone. These difficulties were mainly due to challenges in reaching participants and delivering the intervention as detailed in the protocol. The study highlights the need of reducing barriers for providers in attending to these patients, particularly if they are non-mental health specialized professionals. By minimizing the stigmatization associated with caring for mentally ill individuals and promoting coordination with specialists, innovative approaches may be introduced to alleviate the burden of tobacco-related diseases among this population. ABSTRACT: Introduction The viewpoint of those who implement a programme for the first time is crucial for understanding its impact and ensuring its long-term viability. The 061 QUIT-MENTAL study was a pragmatic randomized controlled trial evaluating a proactive telephone-based intervention addressed to mental health patients conducted by non-psychiatric specialized nurses. Aim We assessed nurses' knowledge of smoking cessation interventions addressed to this population before and after receiving training and their insights after delivering the intervention. Method Mixed methods study: (1) Pre-post evaluation to assess self-reported knowledge, self-efficacy and opinions about smoking cessation. (2) In-depth interviews with key nurses to ascertain their perceptions regarding the impact of the training received in delivering the study intervention. Results The training enhanced nurses' knowledge of psychological and pharmacological resources to aid these patients, as well as their ability to increase their motivation to quit. However, nurses reported difficulties in delivering population-based interventions to individuals with mental health disorders. These challenges primarily arose from participants being hard to reach, exhibiting low motivation to quit, struggling to comprehend instructions or follow recommendations, and nurses feeling unsure about their capacity to assist individuals with mental illnesses in quitting, despite the training they received. Discussion Despite the training and protocol designed to facilitate the delivery of the intervention, nurses faced difficulties in providing population-based interventions to individuals with mental health disorders. Implications for Practice Future quitline programmes aimed at the population with mental health disorders should strive to reduce barriers for providers in attending to these patients, particularly if they are non-mental health specialized professionals. By minimizing the stigmatization associated with caring for mentally ill individuals and promoting coordination with specialists, innovative approaches may be introduced to alleviate the burden of tobacco-related diseases among this population.

PI3 k/akt inhibition induces apoptosis through p38 activation in neurons.

Accumulating evidence suggests that the PI3K/AKT pathway is a pro-survival signalling system in neurons. Therefore, the inhibition of this pathway may be implicated in the degeneration of neurons in Parkinson's disease (PD), Alzheimer's disease (AD), and other neurological disorders. Here we study the participation of the mitogen-activated protein kinase (MAPK) pathway on apoptosis induced by PI3K/AKT inhibition in cultured cerebellar granule cells (CGCs). LY294002, a specific PI3K/AKT inhibitor, selectively activated the p38 MAPK kinase pathway and enhanced c-Jun phosphorylation, but did not activate JNK. The pharmacological inhibitors SB203580 (p38 inhibitor) and SP600125 (a JNK inhibitor) protected primary cultures of rat CGCs from LY294002-induced apoptosis. Furthermore, both compounds decreased the phosphorylation of c-Jun and lowered mRNA levels of the pro-apoptotic gene dp5, a direct target of c-Jun. Taken together, our data demonstrate that PI3K/AKT inhibition induces neuronal apoptosis, a process that is mediated by the activation of p38 MAPK/c-Jun/dp5.

The cloud paradigm applied to e-Health.

BACKGROUND: Cloud computing is a new paradigm that is changing how enterprises, institutions and people understand, perceive and use current software systems. With this paradigm, the organizations have no need to maintain their own servers, nor host their own software. Instead, everything is moved to the cloud and provided on demand, saving energy, physical space and technical staff. Cloud-based system architectures provide many advantages in terms of scalability, maintainability and massive data processing. METHODS: We present the design of an e-health cloud system, modelled by an M/M/m queue with QoS capabilities, i.e. maximum waiting time of requests. RESULTS: Detailed results for the model formed by a Jackson network of two M/M/m queues from the queueing theory perspective are presented. These results show a significant performance improvement when the number of servers increases. CONCLUSIONS: Platform scalability becomes a critical issue since we aim to provide the system with high Quality of Service (QoS). In this paper we define an architecture capable of adapting itself to different diseases and growing numbers of patients. This platform could be applied to the medical field to greatly enhance the results of those therapies that have an important psychological component, such as addictions and chronic diseases.

Uncovering tau in wasteosomes (corpora amylacea) of Alzheimer's disease patients.

Brain corpora amylacea, recently renamed as wasteosomes, are polyglucosan bodies that appear during aging and some neurodegenerative conditions. They collect waste substances and are part of a brain cleaning mechanism. For decades, studies on their composition have produced inconsistent results and the presence of tau protein in them has been controversial. In this work, we reanalyzed the presence of this protein in wasteosomes and we pointed out a methodological problem when immunolabeling. It is well known that to detect tau it is necessary to perform an antigen retrieval. However, in the case of wasteosomes, an excessive antigen retrieval with boiling dissolves their polyglucosan structure, releases the entrapped proteins and, thus, prevents their detection. After performing an adequate pre-treatment, with an intermediate time of boiling, we observed that some brain wasteosomes from patients with Alzheimer's disease (AD) contained tau, while we did not detect tau protein in those from non-AD patients. These observations pointed the different composition of wasteosomes depending on the neuropathological condition and reinforce the role of wasteosomes as waste containers.

Analyzing the Virchow pioneering report on brain corpora amylacea: shedding light on recurrent controversies.

The first report of corpora amylacea (CA) is attributed to Morgagni, who described them in the prostate in the eighteenth century. Nearly a hundred years later, and following the lead started by Purkinje, Virchow described them in the brain. He made a detailed description of the most useful techniques to visualize them, but he failed to describe the cause of why CA do appear, why they are mainly linked with the elderly, and which is their clinical significance. Although in the last two centuries CA have received little attention, recent data have been able to describe that CA accumulate waste products and that some of them can be found in the cerebrospinal fluid and lymphatic nodes, after being released from the brain. Indeed, CA have been renamed to wasteosomes to underline the waste products they gather and to avoid confusion with the term amyloid used by Virchow, now widely related to certain protein deposits found in the brain. Here, after providing a commented English translation of Virchow's findings, we provide a recent update on these structures and their connection with the glymphatic system insufficiency, for which wasteosomes should be considered a hallmark, and how these bodies could serve as diagnostic or prognostic markers of various brain conditions.

CPT1A in AgRP neurons is required for sex-dependent regulation of feeding and thirst.

BACKGROUND: Fatty acid metabolism in the hypothalamus has an important role in food intake, but its specific role in AgRP neurons is poorly understood. Here, we examined whether carnitinea palmitoyltransferase 1A (CPT1A), a key enzyme in mitochondrial fatty acid oxidation, affects energy balance. METHODS: To obtain Cpt1aKO mice and their control littermates, Cpt1a(flox/flox) mice were crossed with tamoxifen-inducible AgRPCreERT2 mice. Food intake and body weight were analyzed weekly in both males and females. At 12 weeks of age, metabolic flexibility was determined by ghrelin-induced food intake and fasting-refeeding satiety tests. Energy expenditure was analyzed by calorimetric system and thermogenic activity of brown adipose tissue. To study fluid balance the analysis of urine and water intake volumes; osmolality of urine and plasma; as well as serum levels of angiotensin and components of RAAS (renin-angiotensin-aldosterone system) were measured. At the central level, changes in AgRP neurons were determined by: (1) analyzing specific AgRP gene expression in RiboTag-Cpt1aKO mice obtained by crossing Cpt1aKO mice with RiboTag mice; (2) measuring presynaptic terminal formation in the AgRP neurons with the injection of the AAV1-EF1a-DIO-synaptophysin-GFP in the arcuate nucleus of the hypothalamus; (3) analyzing AgRP neuronal viability and spine formations by the injection AAV9-EF1a-DIO-mCherry in the arcuate nucleus of the hypothalamus; (4) analyzing in situ the specific AgRP mitochondria in the ZsGreen-Cpt1aKO obtained by breeding ZsGreen mice with Cpt1aKO mice. Two-way ANOVA analyses were performed to determine the contributions of the effect of lack of CPT1A in AgRP neurons in the sex. RESULTS: Changes in food intake were just seen in male Cpt1aKO mice while only female Cpt1aKO mice increased energy expenditure. The lack of Cpt1a in the AgRP neurons enhanced brown adipose tissue activity, mainly in females, and induced a substantial reduction in fat deposits and body weight. Strikingly, both male and female Cpt1aKO mice showed polydipsia and polyuria, with more reduced serum vasopressin levels in females and without osmolality alterations, indicating a direct involvement of Cpt1a in AgRP neurons in fluid balance. AgRP neurons from Cpt1aKO mice showed a sex-dependent gene expression pattern, reduced mitochondria and decreased presynaptic innervation to the paraventricular nucleus, without neuronal viability alterations. CONCLUSIONS: Our results highlight that fatty acid metabolism and CPT1A in AgRP neurons show marked sex differences and play a relevant role in the neuronal processes necessary for the maintenance of whole-body fluid and energy balance.

Adaptation, implementation, and evaluation of an online health sciences training program for brief smoking intervention: A pre-post study in four European countries.

BACKGROUND: Tobacco cessation intervention has a positive impact on quality of care. For health professionals, limited competency in this area may be associated with poor training during their academic programs. There is a clear need to further develop and implement training programs to improve tobacco cessation knowledge, skills, and attitudes among healthcare students. OBJECTIVES: The aim of this study was to assess the effectiveness of the innovative online training program "Brief Intervention in Smoking Cessation" for healthcare students to improve their knowledge, skills, and attitudes. DESIGN: A pre-post evaluation study with a satisfaction assessment tool was used. SETTING: Seven universities from four European countries, including Belgium, Portugal, Spain, and the United Kingdom, participated. PARTICIPANTS: One thousand and seventy-two (1072) undergraduate students participated, with 851 completing the online program. METHODS: All participants completed the "Brief Intervention in Smoking Cessation" online program, which consisted of five theoretical modules, five videos, and three virtual simulation cases between January 2020 and June 2022. Knowledge was assessed by a multiple-choice test, and practical skills were assessed by a simulation algorithm, both of which were developed by education and smoking cessation experts. Competency was achieved when students successfully completed both assessments. Satisfaction was measured using an ad hoc 16-item questionnaire. Pre-post changes in knowledge were assessed using a paired Student's t-test. RESULTS: Eighty-six percent of the students achieved smoking cessation competency. Students significantly improved their knowledge score on a scale of 0 to 10 points, with a mean pre-program score of 3.79 vs a mean post-program score of 7.33 ([-3.7 - -3.4] p < 0.001), acquiring sufficient attitudes and skills (simulation mean of 7.4 out of 10 points). Students were highly satisfied with the program (8.2 out of 10) and recommended it to other students (8.4 out of 10). CONCLUSIONS: The "Brief Intervention in Smoking Cessation" online training program is effective for the acquisition of smoking cessation competencies among European health profession students.

Expression pattern of ataxia telangiectasia mutated (ATM), p53, Akt, and glycogen synthase kinase-3ß in the striatum of rats treated with 3-nitropropionic acid.

3-Nitropropionic acid (3-NPA) is a mitochondrial toxin used in the laboratory to replicate neurodegenerative conditions that are accompanied by degeneration of the caudate-putamen. 3-NPA induces depletion in ATP production, reactive oxygen species production, and secondary excitotoxicity mediated by activation of N-methyl-D-aspartate receptors that culminates in the triggering of cell death mechanisms, including apoptosis. We here examined by immunohistochemical methods whether cellular expression of phospho(Ser1981) -ataxia telangiectasia mutated (ATM), phospho(Ser15) -p53, phospho(Ser473) -Akt, and phospho(Ser9) -glycogen synthase kinase-3ß (GSK3ß), which are key signal molecules that play a critical role in regulating cellular processes related to cell survival and demise, were involved in the striatal neurodegeneration in the brains of rats treated with 3-NPA. Our results indicate that the toxin induced the activation of ATM and p53 only in astrocytes, and a role for these proteins in neuronal degeneration was ruled out. On the other hand, striatal neurons lost the active form of Akt as soon as they began to appear pyknotic, indicating impairment of the PI3K/Akt/GSK3 pathway in their degenerative process. The inactive form of GSK3ß was detected extensively, mainly in the rim of the striatal lesions around degenerating neurons, which could be attributed to a cell death or cell survival response.

GSK3ß inhibition is involved in the neuroprotective effects of cyclin-dependent kinase inhibitors in neurons.

In the present study, we evaluated the effects of roscovitine (Rosco) and flavopiridol (Flavo), both of which are classified as cyclin-dependent kinase (CDK) inhibitors, on apoptosis induced by the inhibition of PI3K/AKT pathway in cerebellar granule neurons (CGNs). Our results demonstrate that both CDK inhibitors prevented apoptosis induced by LY294002 (LY), as also occurs with SB415286 (SB4), a selective GSK3ß inhibitor. Our findings also indicate that these CDK inhibitors inhibit GSK3ß, representing a potential pharmacological mechanism involved in their neuroprotective properties. Thus, the increased activity of GSK3ß induced by LY294002 and detected by dephosphorylation at Ser9 was prevented by both compounds. Likewise, GSK3ß activity was measured by a radioactivity assay, revealing that CDK inhibitors and SB415286 prevented the increase in GSK3ß activity induced by PI3K inhibition. In addition, we analysed c-Jun, which is also a mediator of PI3K inhibition-induced apoptosis. However, neither of the CDK inhibitors nor SB415286 prevented the increase in c-Jun phosphorylation induced by PI3K inhibition. Therefore, our data identify GSK3ß as a crucial mediator of CGN apoptosis induced by PI3K inhibition and indicate that the antiapoptotic effects of CDKs are mediated by the inhibition of this pharmacological target.