Improving the mechanical and biological functions of cell sheet constructs: The interplay of human-derived periodontal ligament stem cells, endothelial cells and plasma rich in growth factors.

OBJECTIVE: The aim of this study was to produce and characterize triple-layered cell sheet constructs with varying cell compositions combined or not with the fibrin membrane scaffold obtained by the technology of Plasma Rich in Growth Factors (mPRGF). MATERIALS AND METHODS: Human primary cultures of periodontal ligament stem cells (hPDLSCs) were isolated, and their stemness nature was evaluated. Three types of triple-layered composite constructs were generated, composed solely of hPDLSCs or combined with human umbilical vein endothelial cells (HUVECs), either as a sandwiched endothelial layer or as coculture sheets of both cell phenotypes. These three triple-layered constructs were also manufactured using mPRGF as cell sheets' support. Necrosis, glucose consumption, secretion of extracellular matrix proteins and synthesis of proangiogenic factors were determined. Histological evaluations and proteomic analyses were also performed. RESULTS: The inclusion of HUVECs did not clearly improve the properties of the multilayered constructs and yet hindered their optimal conformation. The presence of mPRGF prevented the shrinkage of cell sheets, stimulated the metabolic activity and increased the matrix synthesis. At the proteome level, mPRGF conferred a dramatic advantage to the hPDLSC constructs in their ability to provide a suitable environment for tissue regeneration by inducing the expression of proteins necessary for bone morphogenesis and cellular proliferation. CONCLUSIONS: hPDLSCs' triple-layer construct onto mPRGF emerges as the optimal structure for its use in regenerative therapeutics. CLINICAL RELEVANCE: These results suggest the suitability of mPRGF as a promising tool to support cell sheet formation by improving their handling and biological functions.

Pine wilt disease: what do we know from proteomics?

Pine wilt disease (PWD) is a devastating forest disease caused by the pinewood nematode (PWN), Bursaphelenchus xylophilus, a migratory endoparasite that infects several coniferous species. During the last 20 years, advances have been made for understanding the molecular bases of PWN-host trees interactions. Major advances emerged from transcriptomic and genomic studies, which revealed some unique features related to PWN pathogenicity and constituted fundamental data that allowed the development of postgenomic studies. Here we review the proteomic approaches that were applied to study PWD and integrated the current knowledge on the molecular basis of the PWN pathogenicity. Proteomics has been useful for understanding cellular activities and protein functions involved in PWN-host trees interactions, shedding light into the mechanisms associated with PWN pathogenicity and being promising tools to better clarify host trees PWN resistance/susceptibility.

Engaging with the claim of Roma people through twitter: who is behind the hashtags?

Grassroots Roma communities play a pivotal role in organizing at the grassroots level, advocating for their rights, and challenging anti-Roma sentiment across Europe. Nevertheless, there remains a need for a deeper understanding of how these efforts manifest within the digital landscape. Within the overarching framework of the Narratives4Change project (EU Marie Curie Action, Nr. 841,355), this study seeks to examine the use of Twitter as a platform for advocating Roma-related issues. Specifically, it investigates the types of actors engaged in tweeting about Roma-related content and explores potential variations in profiles (organizations vs. individuals) based on the content being tweeted. The analysis encompasses six Roma-related hashtags spanning the years 2017 to 2020: #RomaLivesMatter, #InternationalRomaDay, #OpreRoma, #OpreRomnia, #MujerGitana, and #RomaWomen. The findings reveal that organizations are most active when employing the hashtags #InternationalRomaDay and #MujerGitana, whereas for the other hashtags studied, individual users dominate. Our data underscore the untapped potential of social media spaces and their ecosystems as strategic tools for advocacy and mobilization in support of Roma rights throughout Europe.

Untangling the association between COVID-19 health literacy, trust in the pandemic response, and mental distress, during the COVID-19 pandemic in Ireland: a repeated cross-sectional study.

BACKGROUND: The impact of the COVID-19 pandemic on rates of mental distress is well described. However, the contribution of poor health literacy and low levels of trust in state institutions to mental distress is less well defined. This study aimed to assess the impact of COVID-19 health literacy and trust in the pandemic response (Trust) on mental distress during the COVID-19 pandemic in Ireland. METHODS: We did this nationally representative cross-sectional survey of adult Irish residents during three study periods: from May 26 to June 17, 2020 (n=947); from July 1 to July 23, 2020 (n=995); and from Sept 5 to Sept 28, 2020 (n=972). Participants were contacted using random-digit-dialling and interviewed by a professional market research organisation (Ipsos MRBI' about 80% via mobile phone, 20% via landline). Mental distress was assessed by the Patient Health Questionnaire Anxiety Depression Scale (PHQ-ADS), for which a score of 10 or higher indicated mental distress. Heath literacy and trust were each assessed with three questions, which defined three categories: low, moderate, and high (appendix). Descriptive analysis and multivariate (MVA) Poisson regression were conducted in STATA17, Incidence Rate Ratios (IRR) are reported. FINDINGS: 2914 participants completed the survey across three study periods (median age 46 years, 1510 [51·8%] women, 1401 [48·1%] men, three [0·1%] non-binary). 804 (27·6%) of 2914 participants experienced mental distress (n=804). More women experienced mental distress than men (508 [34%] of women vs 294 [21%] of men). Mental distress was inversely associated with age (from 43% in those aged <30 years [n=232/539] to 19% in those aged >70 years [n=66/349]). Most participants had high health literacy (n=2,530, 86·8%). While health literacy was positively and significantly associated with trust, it was not associated with mental distress and it was therefore excluded from the MVA. Level of trust was captured for 2693 adults; 42·2% participants reported low trust (n=457) or moderate trust (n=679). The prevalence of mental distress was inversely associated with trust; increasing from 24% in those with high trust (n=374/1557), 30% in those with moderate trust (n=202/679), to 36% in those with low trust (n=166/457). In MVA higher rates of mental distress were associated with low trust (IRR 1·45, 95% CI 1·20-1·75; p=0·000) and moderate trust (IRR 1·24, 1·04-1·47, p=0·016) compared with high trust when adjusted for age, sex, employment status, and income,. INTERPRETATION: In Ireland, low levels of trust in the COVID-19 pandemic response were associated with higher levels of mental distress. Although poor health literacy was associated with low levels of trust, it was not independently associated with mental distress. Inference on the nature and direction of causal effects must be cautious given the cross-sectional study design. FUNDING: Health Research Board.

Advances in Platelet Rich Plasma-Derived Extracellular Vesicles for Regenerative Medicine: A Systematic-Narrative Review.

The use of platelet-rich plasma (PRP) has gained increasing interest in recent decades. The platelet secretome contains a multitude of growth factors, cytokines, chemokines, and other biological biomolecules. In recent years, developments in the field of platelets have led to new insights, and attention has been focused on the platelets' released extracellular vesicles (EVs) and their role in intercellular communication. In this context, the aim of this review was to compile the current evidence on PRP-derived extracellular vesicles to identify the advantages and limitations fortheir use in the upcoming clinical applications. A total of 172 articles were identified during the systematic literature search through two databases (PubMed and Web of Science). Twenty publications met the inclusion criteria and were included in this review. According to the results, the use of PRP-EVs in the clinic is an emerging field of great interest that represents a promising therapeutic option, as their efficacy has been demonstrated in the majority of fields of applications included in this review. However, the lack of standardization along the procedures in both the field of PRP and the EVs makes it extremely challenging to compare results among studies. Establishing standardized conditions to ensure optimized and detailed protocols and define parameters such as the dose or the EV origin is therefore urgent. Further studies to elucidate the real contribution of EVs to PRP in terms of composition and functionality should also be performed. Nevertheless, research on the field provides promising results and a novel basis to deal with the regenerative medicine and drug delivery fields in the future.

Cellular composition modifies the biological properties and stability of platelet rich plasma membranes for tissue engineering.

Scaffolds should provide structural support for tissue regeneration, allowing their gradual biodegradation and interacting with cells and bioactive molecules to promote remodeling. Thus, the scaffold's intrinsic properties affect cellular processes involved in tissue regeneration, including migration, proliferation, differentiation, and protein synthesis. In this sense, due to its biological effect and clinical potential, Platelet Rich Plasma (PRP) fibrin could be considered a successful scaffold. Given the high variability in commercial PRPs formulations, this research focused on assessing the influence of cellular composition on fibrin membrane stability and remodeling cell activity. The stability and biological effect were evaluated at different time points via D-dimer, type I collagen and elastase quantification in culture media conditioned by Plasma Rich in Growth Factors - Fraction 1 (PRGF-F1), Plasma Rich in Growth Factors - Whole Plasma (PRGF-WP) and Leukocyte-rich Platelet Rich Plasma (L-PRP) membranes, and by gingival fibroblast cells seeded on them, respectively. Ultrastructure of PRP membranes was also evaluated. Histological analyses were performed after 5 and 18 days. Additionally, the effect of fibrin membranes on cell proliferation was determined. According to the results, L-PRP fibrin membranes degradation was complete at the end of the study, while PRGF membranes remained practically unchanged. Considering fibroblast behavior, PRGF membranes, in contrast to L-PRP ones, promoted extracellular matrix biosynthesis at the same time as fibrinolysis and enhanced cell proliferation. In conclusion, leukocytes in PRP fibrin membranes drastically reduce scaffold stability and induce behavioral changes in fibroblasts by reducing their proliferation rate and remodeling ability.

Evolution of the COVID-19 Pandemic after the Introduction of School Referral Nurses in the Province of Granada-A Descriptive Study.

The aim of this research is to describe the evolution of the pandemic in a school context, following the introduction of school nurses into the educational setting. BACKGROUND: The first wave of COVID-19 in Spain prevented social interaction by imposing lockdowns on the population. All non-essential activities, including face-to-face education, were interrupted, affecting the school-aged population during the second and third trimester of the 2019/2020 school year. Schools are places where prevention, identification and case management measures should be rapidly implemented. METHODS: This is a prospective and descriptive study using a quantitative method to study the school population of Granada and its province during the school year 2020/2021, from September 2020 to May 2021. The study participants were all schools under the jurisdiction of the Territorial Delegation of Education of Granada, whether public, private, state-subsidised or charter schools, including all educational stages. RESULTS: The confirmation rate in Granada city (11.2%), in contrast to the Andalusian average (6.9%), places Granada as the province with the highest incidence rate. The infection rates among teachers show the same confirmation rate as the general population of Granada (9%); however, among students this rate is lower (7%). There is a higher incidence of outbreaks taking place outside school and a lower incidence of outbreaks occurring within the school environment. Both partial and total outbreaks are more frequent in early childhood education. CONCLUSION: The early identification and management of reported suspected cases of COVID-19 in schools is proving effective in preventing infection in the school population, achieving good pandemic control in schools, and reducing the number of outbreaks and people affected. Schools have been confirmed to be safe. Establishing screening for asymptomatic schoolchildren could be a tool to improve control in schools.

Quality of Life in the Postpartum Recovery of Women Who Practice Hypopressive Exercise: Randomized Clinical Trial.

Background: Practicing physical exercise is a way of maintaining a healthy lifestyle and it has been demonstrated to provide benefits and improve quality of life after labour. Aim: The objective of this study is to determine if performing a moderate hypopressive exercise program after childbirth improves the HRQoL of women. Materials and Methods: A 12-week randomized clinical trial was carried out. The women in the exercise group followed a training program based on the hypopressive method, which comprises postural and respiratory exercises of moderate intensity. We used the SF-36v2 questionnaire to assess the Health-Related Quality of Life in women. Results: We found statistically significant differences (p < 0.05) in general health components, vitality, emotional role, and mental health in the sample group of exercise group of women, who obtained a higher score. We found statistically significant differences in all aspects between the groups. The interaction between time and group affect in general health components, vitality, emotional role and the score of mental components. In all of them, the score was higher at the end of the programme and in the exercise group. Conclusions: The training program with hypopressive physical exercise improves the quality of life after childbirth. According to the guidelines recommended by the ACOG, the hypopressive method is valid for the improvement of HRQoL in women who perform this method at 16 weeks of delivery. Further studies are recommended to start such exercises one week after delivery in postpartum women.

Autologous Platelet Rich Plasma (PRGF) Preserves Genomic Stability of Gingival Fibroblasts and Alveolar Osteoblasts after Long-Term Cell Culture.

Plasma rich in growth factors (PRGF) has several applications in dentistry that may require repeated applications of PRGF. Furthermore, it has been used for ex vivo expansion of human origin cells for their clinical application. One of the most relevant issues in these applications is to guarantee the genetic stability of cells. In this study, the chromosomal stability of gingival fibroblasts and alveolar osteoblasts after long-term culture was evaluated. Cells were expanded with PRGF or foetal bovine serum (FBS) as a culture medium supplement until passage 7 or 8 for gingival fibroblast or alveolar osteoblasts, respectively. A comparative genomic hybridization (CGH) array was used for the genetic stability study. This analysis was performed at passage 3 and after long-term culture with the corresponding culture medium supplements. The cell proliferative rate was superior after PRGF culture. Array CGH analysis of cells maintained with all the three supplements did not reveal the existence of alterations in copy number or genetic instability. The autologous PRGF technology preserves the genomic stability of cells and emerges as a safe substitute for FBS as a culture medium supplement for the clinical translation of cell therapy.

Deletion Syndrome 22q11.2: A Systematic Review.

22q11.2 deletion syndrome (DS 22q11.2) is a rare disease of genetic origin, caused by the loss of the q11.2 region of chromosome 22. It affects one in 4000 live newborns, and among the clinical manifestations that can occur in this syndrome are abnormalities in the parathyroid glands (producing calcium deficits), the palate, the heart and the thymus. It is also known as DiGeorge syndrome or velocardiofacial syndrome, among other names, depending on the clinical presentation of each individual. The main objective of the review was to update information on DS 22q11.2 from publications in the scientific literature. The daily activities of these patients are seriously impaired, due to the impact of the clinical manifestations. Interventions can be performed to improve their social, cognitive and emotional skills, thus increasing their ability to perform different daily activities.

Platelet-Rich Plasma as an Alternative to Xenogeneic Sera in Cell-Based Therapies: A Need for Standardization.

There has been an explosion in scientific interest in using human-platelet-rich plasma (PRP) as a substitute of xenogeneic sera in cell-based therapies. However, there is a need to create standardization in this field. This systematic review is based on literature searches in PubMed and Web of Science databases until June 2021. Forty-one studies completed the selection criteria. The composition of PRP was completely reported in less than 30% of the studies. PRP has been used as PRP-derived supernatant or non-activated PRP. Two ranges could be identified for platelet concentration, the first between 0.14 × 106 and 0.80 × 106 platelets/µL and the second between 1.086 × 106 and 10 × 106 platelets/µL. Several studies have pooled PRP with a pool size varying from four to nine donors. The optimal dose for the PRP or PRP supernatant is 10%. PRP or PRP-derived supernatants a have positive effect on MSC colony number and size, cell proliferation, cell differentiation and genetic stability. The use of leukocyte-depleted PRP has been demonstrated to be a feasible alternative to xenogeneic sera. However, there is a need to improve the description of the PRP preparation methodology as well as its composition. Several items are identified and reported to create guidelines for future research.

Educational Intervention in the Postural Hygiene of School-Age Children.

INTRODUCTION: Healthy lifestyle habits formed in childhood provide the foundation for a healthy adult life; therefore, it is important to encourage healthy habits and to correct poor habits from an early age. AIM: In this study, we aim to evaluate the influence of educational intervention in order to modify postural hygiene habits in school-age children. MATERIALS AND METHODS: We randomly selected three public primary education schools in the Galician provinces of Orense and Pontevedra based on stratified multistage sampling. A sample of 479 students was obtained, representing 2% of all 6- to 12-year-old children registered during the 2015/2016 academic year in those provinces. RESULTS: Following the intervention, the students' postural hygiene improved. We found statistically significant differences regarding a reduction in the number of hours spent watching television (0.531 h/day); the way school books and supplies were carried, with an increase in the use of backpacks with wheels (from 58.5% to 64.1%); and an improvement in postural hygiene when watching television, with an increase from 63.7% to 80.8% of those surveyed opting to watch television whilst seated on a chair instead of lying down. CONCLUSIONS: Educational intervention by inculcating healthy postural hygiene habits in children at an early age can improve and correct unhealthy behaviours.

Composite alginate-gelatin hydrogels incorporating PRGF enhance human dental pulp cell adhesion, chemotaxis and proliferation.

The increasing prevalence of tissue injuries is fueling the development of autologous biological treatments for regenerative medicine. Here, we investigated the potential of three different bioinks based on the combination of gelatin and alginate (GA), enriched in either hydroxyapatite (GAHA) or hydroxyapatite and PRGF (GAHAP), as a favorable microenvironment for human dental pulp stem cells (DPSCs). Swelling behaviour, in vitro degradation and mechanical properties of the matrices were evaluated. Morphological and elemental analysis of the scaffolds were also performed along with cytocompatibility studies. The in vitro cell response to the different scaffolds was also assessed. Results showed that all scaffolds presented high swelling capacity, and those that contained HA showed higher Young's modulus. GAHAP had the lowest degradation rate and the highest values of cytocompatibility. Cell adhesion and chemotaxis were significantly increased when PRGF was incorporated to the matrices. GAHA and GAHAP compositions promoted the highest proliferative rate as well as significantly stimulated osteogenic differentiation. In conclusion, the enrichment with PRGF improves the regenerative properties of the composites favouring the development of personalized constructs.

Cause of Death in Heart Failure Based on Etiology: Long-Term Cohort Study of All-Cause and Cardiovascular Mortality.

We assessed differences in long-term all-cause and cardiovascular (CV) mortality in heart failure (HF) outpatients based on the etiology of HF. Consecutive patients admitted to the HF Clinic from August 2001 to September 2019 (N = 2587) were considered for inclusion. HF etiology was divided into ischemic heart disease (IHD), dilated cardiomyopathy (DCM), hypertensive heart disease, alcoholic cardiomyopathy, drug-induced cardiomyopathy (DICM), valvular heart disease, and hypertrophic cardiomyopathy. All-cause death and CV death were the primary end points. Among 2387 patients included in the analysis (mean age 66.5 ± 12.5 years, 71.3% men), 1317 deaths were recorded (731 from CV cause) over a maximum follow-up of 18 years (median 4.1 years, interquartile range (IQR) 2-7.8). Considering IHD as the reference, only DCM had a lower risk of all-cause death (adjusted hazard ratio (aHR) 0.68, 95% confidence interval (CI) 0.56-0.83, p < 0.001), and only DICM had a higher risk of all-cause death (aHR 1.47, 95% CI 1.02-2.11, p = 0.04). However, almost all etiologies had a significantly lower risk of CV death than IHD. Among the studied HF etiologies, DCM and DICM have the lowest and highest risk of all-cause death, respectively, whereas IHD has the highest adjusted risk of CV death.

The inclusion of leukocytes into platelet rich plasma reduces scaffold stability and hinders extracellular matrix remodelling.

BACKGROUND: Scaffolds should have controllable degradation rate and allow cells to produce their own extracellular matrix. Platelet rich plasma (PRP) is a source of autologous growth factors and proteins embedded in a 3D fibrin scaffold. There is no consensus regarding the obtaining conditions and composition of PRPs. The aim of this study was to evaluate how the inclusion of leukocytes (L-PRP) in plasma rich in growth factors (PRGF) may alter the process of fibrinolysis. The effect of different combinations of cellular phenotypes with PRGF and L-PRP clots on both the fibrinolysis and matrix deposition process was also determined. METHODS: PRGF and L-PRP clots were incubated for 14 days and D-dimer and type I collagen were determined in their conditioned media to evaluate clots' stability. For remodelling assays, gingival fibroblasts, alveolar osteoblasts and human umbilical vein endothelial cells (HUVEC) were seeded onto the two types of clots for 14 days. D-dimer, type I collagen, and laminin α4 were measured by ELISA kits in their conditioned media. Morphological and histological analysis were also performed. Cell proliferation was additionally determined RESULTS: PRGF clots preserved their stability as shown by the low levels of both D-dimer and collagen type I compared to those obtained for L-PRP clots. The inclusion of both gingival fibroblasts and alveolar osteoblasts stimulated a higher fibrinolysis in the PRGF clots. In contrast to this, the degradation rates of both PRGF and L-PRP clots remained unchanged after culturing with the endothelial cells. In all cases, type I collagen and laminin α4 levels were in line with the degree of clots' degradation. In all phenotypes, cell proliferation was significantly higher in PRGF than in L-PRP clots. CONCLUSION: The inclusion of leukocytes in PRGF scaffolds reduced their stability, decreased cell number and slowed down cell remodelling.

Current Understanding of the Physiopathology, Diagnosis and Therapeutic Approach to Alzheimer's Disease.

Alzheimer's disease (AD) is the most common cause of dementia. It is characterized by cognitive decline and progressive memory loss. The aim of this review was to update the state of knowledge on the pathophysiological mechanisms, diagnostic methods and therapeutic approach to AD. Currently, the amyloid cascade hypothesis remains the leading theory in the pathophysiology of AD. This hypothesis states that amyloid-ß (Aß) deposition triggers a chemical cascade of events leading to the development of AD dementia. The antemortem diagnosis of AD is still based on clinical parameters. Diagnostic procedures in AD include fluid-based biomarkers such as those present in cerebrospinal fluid and plasma or diagnostic imaging methods. Currently, the therapeutic armory available focuses on symptom control and is based on four pillars: pharmacological treatment where acetylcholinesterase inhibitors stand out; pharmacological treatment under investigation which includes drugs focused on the control of Aß pathology and tau hyperphosphorylation; treatment focusing on risk factors such as diabetes; or nonpharmacological treatments aimed at preventing development of the disease or treating symptoms through occupational therapy or psychological help. AD remains a largely unknown disease. Further research is needed to identify new biomarkers and therapies that can prevent progression of the pathology.

The Effect of Therapeutic Hypothermia after Cardiac Arrest on the Neurological Outcome and Survival-A Systematic Review of RCTs Published between 2016 and 2020.

Therapeutic hypothermia is a treatment used for patients who have suffered cardiorespiratory arrest and remain conscious after the recovery of spontaneous circulation. However, its effectiveness is controversial. The objective of this systematic review is to summarize the scientific evidence available about the effect of therapeutic hypothermia on neurological status and survival in this type of patients. METHODOLOGY: A primary search in CINAHL, CUIDEN, Pubmed, Web of Science, and Scopus databases was carried out. Randomized clinical trials (RCT) published from 2016 to 2020 were selected. RESULTS: 17 studies were selected for inclusion and most relevant data were extracted. Methodological quality was assessed by the RoB tool. CONCLUSIONS: Although therapeutic hypothermia is a safe technique with few adverse and manageable effects, it has not shown to improve survival rate and neurological status of adult nor pediatric patients. It is possible that its positive effect on neuroprotection could be achieved only by preventing hyperthermia although further investigation is needed.

Pharmacological Treatments against COVID-19 in Pregnant Women.

The recent respiratory virus known as SARS-CoV-2 has caused millions of deaths worldwide, causing great uncertainty due to the lack of a specific treatment, which has been mitigated by the use of various drugs traditionally used against other types of pathologies. Pregnancy presents special physiological conditions that expose the pregnant woman and the foetus to greater risk. Pregnant women are often excluded from trials due to possible risk of toxicity or side effects, resulting in a lack of knowledge about the use of drugs and treatments during pregnancy. The main objective of this review was to compile existing knowledge about currently available drug treatments for COVID-19 in pregnant women. The review report met the criteria of the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) review protocol and was registered with the registration number CRD42021251036. The electronic databases searched were Scopus, PubMed, CINAHL and SciELO. Finally, 22 articles were included, resulting in an analysis of drugs with an acceptable safety profile in the treatment of pregnant women with COVID-19.