Osteogenic potential of apical papilla stem cells mediated by platelet-rich fibrin and low-level laser.

To evaluate the osteogenic potential of platelet-rich fibrin (PRF) and low-level laser therapy (LLLT) on human stem cells from the apical papilla (SCAP) we isolated, characterized, and then cultured in an osteogenic medium cells with PRF and/or LLLT (660 nm, 6 J/m2-irradiation). Osteogenic differentiation was assessed by bone nodule formation and expression of bone morphogenetic proteins (BMP-2 and BMP-4), whereas the molecular mechanisms were achieved by qRT-PCR and RNA-seq analysis. Statistical analysis was performed by ANOVA and Tukey's post hoc tests (p < 0.05* and p < 0.01**). Although PRF and LLLT increased bone nodule formation after 7 days and peaked at 21 days, the combination of PRF + LLLT led to the uppermost nodule formation. This was supported by increased levels of BMP-2 and -4 osteogenic proteins (p < 0.005). Furthermore, the PRF + LLLT relative expression of specific genes involved in osteogenesis, such as osteocalcin, was 2.4- (p = 0.03) and 28.3- (p = 0.001) fold higher compared to the PRF and LLLT groups, and osteopontin was 22.9- and 1.23-fold higher, respectively (p < 0.05), after 7 days of interaction. The transcriptomic profile revealed that the combination of PRF + LLLT induces MSX1, TGFB1, and SMAD1 expression, after 21 days of osteogenic differentiation conditions exposition. More studies are required to understand the complete cellular and molecular mechanisms of PRF plus LLLT on stem cells. Overall, we demonstrated for the first time that the combination of PRF and LLLT would be an excellent therapeutic tool that can be employed for dental, oral, and craniofacial repair and other tissue engineering applications.

Effect of Dentin Conditioning with EDTA and Diode Lasers on Expression of Odontoblast-like Cell Markers of Dental Pulp Stem Cells.

Regenerative endodontic procedures rely on the delivery of mesenchymal stem cells into the root canal and on the effect of local growth factors from the dentin and blood clot. The aim of this study was to assess the effect of dentin conditioning with ethylenediamine tetraacetic acid (EDTA) and diode lasers with different wavelengths (808 nm and 980 nm) on the expression of odontoblast-like cell markers. Forty dentin cylinders were divided into four groups according to the irrigation protocol: EDTA, EDTA + 808 nm diode laser, EDTA + 980 nm diode laser, and phosphate-buffered saline as the control group. Dental pulp stem cells were seeded into the previously conditioned cylinders and incubated for 14 days. The quantitative real-time polymerase chain reaction was used to evaluate the expression of dentin sialophosphoprotein (DSPP), dentin morphoprotein-1 (DMP-1), and transforming growth factor-beta 1 (TGF-ß1). Data analysis was performed using the Kruskal-Wallis test. The activation of EDTA with 980 nm and 808 nm diode lasers resulted in lower DSPP and DMP-1 expression than that for EDTA alone (p < 0.05 and p < 0.01, respectively). The expression of TGF was similar among all groups. The highest level of expression of odontoblast-like differentiation markers was observed with EDTA alone. However, the use of an 808 nm diode laser during EDTA irrigation reduced the expression of odontoblastic differentiation markers.

Sintomatología Depresiva y Bienestar en Personas Mayores de Concepción, Chile: Rol moderador del apoyo social

Antecedentes: La población sobre 60 años va en aumento y es relevante tener más conocimiento sobre los factores que inciden en su bienestar y salud mental. Objetivo: Este estudio busca comprender la relación entre bienestar, apoyo social y sintomatología depresiva en personas mayores de la provincia de Concepción. Método: 538 adultos mayores fueron seleccionados aleatoriamente de 15 Centros de Salud Primaria de la provincia de Concepción, Chile. Se midieron sus niveles de bienestar (con el Pemberton Happiness Index), de apoyo Social (con la Escala Multidimensional de Percepción de Apoyo Social de Zimet) y de sintomatología depresiva (mediante el Patient Health Questionnaire-9). Resultados: Los adultos mayores mostraron un puntaje elevado (M=8,54; DE = 1,34) en bienestar, sin diferencias por sexo (t(536) = −1,065, p = 0,288 > 0,05). En los análisis de regresión lineal, el apoyo social (β =0,463, p<0,001) y la sintomatología depresiva (β =-1,585, p<0,001) aparecen como influyentes en el bienestar de las personas mayores; y el apoyo social actúa como un factor moderador (β=0,049, p=0,007, IC =0,021; 0,077) en la relación entre sintomatología depresiva y bienestar. Conclusiones: Las personas mayores muestran elevado bienestar. Los resultados refuerzan la relevancia del apoyo social como un factor protector en las personas mayores. Ello corrobora la importancia del fomento de este aspecto en los programas promocionales y preventivos destinados al bienestar y salud mental de adultos mayores.

Background: The population over 60 years of age is increasing. Aim: This research aims to understand the relationship between well-being, social support, and depressive symptoms in older Chilean people. Method: 538 older adults were randomly selected from 15 Primary Health Centers in the province of Concepción, Chile, 341 women and 197 men. Well-being was measured with the Pemberton Happiness Index, Social Support with the Zimet Multidimensional Scale of Perceived Social Support, and Depressive Symptomatology using the Patient Health Questionnaire-9. Results: Older adults showed a high mean score (M=8.54; SD=1.34) in well-being, with no differences by sex (t(536) = −1.065, p = .288 > .05.) In linear regression analyses, social support (β =0,463, p<0,001) and depressive symptomatology (β =-1,585, p<0,001) influence the well-being of the elderly, and social support acts as a moderating factor (β=0.049, p=0.007, CI =0.021; 0.077) in the relationship between social support and well-being. Conclusions: Older people show high well-being. The results reinforce the relevance of social support as a protective factor for the positive mental health of older adults. This corroborates the importance of including this aspect in promotional and preventive mental health programs for older adults.

Life satisfaction, positive affect, depression and anxiety symptoms, and their relationship with sociodemographic, psychosocial, and clinical variables in a general elderly population sample from Chile.

Background: This study aims to describe the relationship between life satisfaction, positive affect, depression and anxiety symptoms with sociodemographic, psychosocial and clinical variables, and to identify the relative importance of these predictor groups. Methods: We evaluated life satisfaction (SWLS), positive affect (PANAS), depressive (PHQ-9), and anxiety (GAI) symptoms and their association with sociodemographic, psychosocial and clinical variables in a multistage, random general population sample of fully functioning individuals aged 60-80 years from the Concepción province and Gran Santiago, Chile (n = 396). We performed weighted multiple regression analysis, considering the complex sample structure with age group, sex, and geographical area, complemented with general and conditional dominance analyses to estimate the relevance of the predictor groups. Results: We found significant associations with the geographical area, sex, age, education level, household members, having a partner, employment status, caregiver status, economic satisfaction, presence of chronic diseases, medication use, and alcohol use. Satisfaction with health was the most important predictor for positive affect (p < 0.001), depressive (p < 0.001), and anxiety (p < 0.001) symptoms, while alcohol use was the most significant predictor for life satisfaction (p < 0.001). Conclusion: Simultaneously studying the positive and negative dimensions of wellbeing and mental health in older adults allows for a more comprehensive perspective on the challenges faced during this stage of life. This study accounts for previously unknown associations and contributes to the identification of common and specific predictors in both dimensions.

Effects of 970 nm diode laser irradiation on morphology, proliferation, and differentiation of gingival mesenchymal stem cells / Efectos de la irradiación con láser de diodo de 970 m sobre la morfología, la proliferación y la diferenciación de las células madre mesenquimales gingivales

Objective: The objective of this study was to investigate the morphology, proliferation, and differentiation of gingival mesenchymal stem cells (GMSCs) irradiated with a 970 nm Diode Laser (LLLT). It is essential to validate the efficacy of treatment, optimize irradiation conditions and guarantee the safety and quality of stem cells for future use in dental applications. Materials and Methods: GMSCs were cultured in standard conditions and irradiated with a Diode laser (970 nm, 0.5W) with an energy density of 9J/cm2. Cell proliferation was assessed with the WST-1 proliferation kit. GMSCs were differentiated into chondrogenic and osteogenic lineages. Cell morphology was performed with Hematoxylin/eosin staining, and quantitative nuclear analysis was done. Cell viability was monitored with trypan blue testing. Results: GMSCs subjected to irradiation demonstrated a significant increase in proliferation at 72 hours compared to the non-irradiated controls (p=0.027). This indicates that the 970 nm diode laser has a stimulatory effect on the proliferation of GMSCs. LLLT-stimulated GMSCs exhibited the ability to differentiate into chondrogenic and osteogenic lineages. A substantial decrease in cell viability was observed 24 hours after irradiation (p=0.024). However, after 48 hours, the cell viability recovered without any significant differences. This indicates that there might be a temporary negative impact on cell viability immediately following irradiation, but the cells were able to recover and regain their viability over time. Conclusions: This study support that irradiation with a 970 nm diode laser could stimulate the proliferation of GMSCs, maintain their ability to differentiate into chondrogenic and osteogenic lineages, and has minimal impact on the mor- phological characteristics of the cells. These results support the potential use of NIR Lasers in combination with GMSCs as a promising strategy for dental treatments.

Objetivo: El objetivo de este estudio fue investigar la morfología, proliferación y diferenciación de las células madre mesenquimatosas (GMSC) irradiadas con un láser de diodo de 970 nm (LLLT). Es fundamental validar la eficacia del tratamiento, optimizar las condiciones de irradiación y garantizar la seguridad y calidad de las células madre para su uso futuro en aplicaciones dentales.Materiales y Métodos: Las GMSC se cultivaron en condiciones estándar y se irradiaron con un láser de diodo (970 nm, 0,5 W) con una densidad de energía de 9 J/cm2. La proliferación celular se evaluó con el kit de proliferación WST-1. Las GMSC se diferenciaron en linajes condrogénicos y osteogénicos. La morfología celular se realizó con tinción de hematoxilina/eosina y se realizó un análisis nuclear cuantitativo. La viabilidad celular se controló con prueba de azul de tripano. Resultados: Las GMSC sometidas a irradiación demostraron un aumento significativo en la proliferación a las 72 horas en comparación con los controles no irradiados (p=0,027). Esto indica que el láser de diodo de 970 nm tiene un efecto estimulante sobre la proliferación de GMSC. Las GMSC estimuladas con LLLT exhibieron la capacidad de diferenciarse en linajes condrogénicos y osteogénicos. Se observó una disminución sustancial de la viabilidad celular 24 horas después de la irradiación (p=0,024). Sin embargo, después de 48 horas, la viabilidad celular se recuperó sin diferencias significativas. Esto indica que podría haber un impacto negativo temporal en la viabilidad de las células inmediatamente después de la irradiación, pero las células pudieron recuperarse y recuperar su viabilidad con el tiempo. Conclusión: En conclusión, este estudio respalda que la irradiación con un láser de diodo de 970 nm podría estimular la proliferación de GMSC, mantener su capacidad para diferenciarse en linajes condrogénicos y osteogénicos y tiene un impacto mínimo en las características morfológicas de las células. Estos resultados respaldan el uso potencial de láseres NIR en combinación con GMSC como una estrategia prometedora para tratamientos dentales.

Sampling from four geographically divergent young female populations demonstrates forensic geolocation potential in microbiomes.

Studies of human microbiomes using new sequencing techniques have increasingly demonstrated that their ecologies are partly determined by the lifestyle and habits of individuals. As such, significant forensic information could be obtained from high throughput sequencing of the human microbiome. This approach, combined with multiple analytical techniques demonstrates that bacterial DNA can be used to uniquely identify an individual and to provide information about their life and behavioral patterns. However, the transformation of these findings into actionable forensic information, including the geolocation of the samples, remains limited by incomplete understanding of the effects of confounding factors and the paucity of diverse sequences. We obtained 16S rRNA sequences of stool and oral microbiomes collected from 206 young and healthy females from four globally diverse populations, in addition to supporting metadata, including dietary and medical information. Analysis of these microbiomes revealed detectable geolocation signals between the populations, even for populations living within the same city. Accounting for other lifestyle variables, such as diet and smoking, lessened but does not remove the geolocation signal.

Dentin slice model of dental stem cells in a fibrin-agarose construct for dental pulp regeneration / Modelo de trozo de dentina de células madre dentales en una construcción de fibrina-agarosa para la regeneración de la pulpa dental

Objectives: To implement a dentin slice model of mesenchymal stem cells derived from dental tissues in a fibrin-agarose construct for dental pulp regeneration. Material and Methods: MSCs derived from different oral cavity tissues were combined with a fibrin-agarose construct at standard culture conditions. Cell viability and proliferation tests were assayed using a fluorescent cell dye Calcein/Am and WST-1 kit. The proliferation assay was evaluated at 24, 48, 72, and 96 hours. Also, we assessed the dental pulp stem cells (DPSCs) cell morphology inside the construct with histological stains such as Hematoxylin and Eosin, Masson's trichrome, and Periodic acid­Schiff. In addition, we elaborated a tooth dentin slice model using a culture of DPSC in the fibrin­agarose constructs co-adhered to dentin walls. Results: The fibrin-agarose construct was a biocompatible material for MSCs derived from dental tissues. It provided good conditions for MSCs' viability and proliferation. DPSCs proliferated better than the other MSCs, but the data did not show significant differences. The morphology of DPSCs inside the construct was like free cells. The dentin slice model was suitable for DPSCs in the fibrin-agarose construct. Conclusion: Our findings support the dentin slice model for future biological use of fibrin-agarose matrix in combination with DPSCs and their potential use in dental regeneration. The multipotency, high proliferation rates, and easy obtaining of the DPSCs make them an attractive source of MSCs for tissue regeneration.

Objetivos: Implementar un modelo de dentina con células madre mesenquimales derivadas de tejidos dentales en una constructo de fibrina-agarosa para la regeneración de la pulpa dental. Material y Métodos: Las MSC derivadas de diferentes tejidos de la cavidad oral se combinaron con una construcción de fibrina-agarosa en condiciones de cultivo estándar. Las pruebas de viabilidad y proliferación celular se ensayaron utilizando un kit de colorante celular fluorescente Calcein/Am y WST-1. El ensayo de proliferación se evaluó a las 24, 48, 72 y 96 horas. Además, evaluamos la morfología celular de las células madre de la pulpa dental (DPSC) dentro de la construcción con tinciones histológicas como hematoxilina y eosina, tricrómico de Masson y ácido peryódico de Schiff. Además, elaboramos un modelo de rebanadas de dentina dental utilizando un cultivo de DPSC en las construcciones de fibrina-agarosa coadheridas a las paredes de la dentina. Resultados: La construcción de fibrina-agarosa fue un material biocompatible para las MSC derivadas de tejidos dentales. Proporcionó buenas condiciones para la viabilidad y proliferación de las MSC. Las DPSC proliferaron mejor que las otras MSC, pero los datos no mostraron diferencias significativas. La morfología de las DPSC dentro de la construcción era como la de las células libres. El modelo de corte de dentina fue adecuado para DPSC en la construcción de fibrina-agarosa.Conclusión: Nuestros hallazgos respaldan el modelo de corte de dentina para el futuro uso biológico de la matriz de fibrina-agarosa en combinación con DPSC y su uso potencial en la regeneración dental. El multipotencial, las altas tasas de proliferación y la fácil obtención de las DPSC las convierten en una fuente atractiva de MSC para la regeneración de tejidos.

[Suicidal behavior among Chilean young people self-identified as cisgender, lesbian, gay, and bisexual]. / Ideación e intento suicida en jóvenes lesbianas, gays y bisexuales de Chile: estudio comunitario online.

BACKGROUND: Suicidal behavior is more common among young people and those who identify as lesbian, gay, or bisexual. AIM: To assess suicidal behavior among young people self-identified as cisgender lesbian, gay, and bisexual. MATERIAL AND METHODS: The Okasha Suicide Scale was answered online by1,683 Chilean people aged 18 to 30 years, self-identified as cisgender lesbian, gay, or bisexual. RESULTS: Thirty four percent of respondents reported suicidal ideation in some moment of their lives and 16% reported repeated suicidal thoughts. Thirty five percent reported at least one suicide attempt. Women had a higher risk of suicide attempts than men. Bisexuals had a higher risk of suicide attempts than lesbians and homosexuals. Those with a non-conforming or non-binary gender identity had a higher risk of suicide attempts than those with a female and male gender identity. CONCLUSIONS: These results confirm the higher suicide risk among young sexual minority groups.

International Validation of the Canadian Syncope Risk Score : A Cohort Study.

BACKGROUND: The Canadian Syncope Risk Score (CSRS) was developed to predict 30-day serious outcomes not evident during emergency department (ED) evaluation. OBJECTIVE: To externally validate the CSRS and compare it with another validated score, the Osservatorio Epidemiologico della Sincope nel Lazio (OESIL) score. DESIGN: Prospective cohort study. SETTING: Large, international, multicenter study recruiting patients in EDs in 8 countries on 3 continents. PARTICIPANTS: Patients with syncope aged 40 years or older presenting to the ED within 12 hours of syncope. MEASUREMENTS: Composite outcome of serious clinical plus procedural events (primary outcome) and the primary composite outcome excluding procedural interventions (secondary outcome). RESULTS: Among 2283 patients with a mean age of 68 years, the primary composite outcome occurred in 7.2%, and the composite outcome excluding procedural interventions occurred in 3.1% at 30 days. Prognostic performance of the CSRS was good for both 30-day composite outcomes and better compared with the OESIL score (area under the receiver-operating characteristic curve [AUC], 0.85 [95% CI, 0.83 to 0.88] vs. 0.74 [CI, 0.71 to 0.78] and 0.80 [CI, 0.75 to 0.84] vs. 0.69 [CI, 0.64 to 0.75], respectively). Safety of triage, as measured by the frequency of the primary composite outcome in the low-risk group, was higher using the CSRS (19 of 1388 [0.6%]) versus the OESIL score (17 of 1104 [1.5%]). A simplified model including only the clinician classification of syncope (cardiac syncope, vasovagal syncope, or other) variable at ED discharge-a component of the CSRS-achieved similar discrimination as the CSRS (AUC, 0.83 [CI, 0.80 to 0.87] for the primary composite outcome). LIMITATION: Unable to disentangle the influence of other CSRS components on clinician classification of syncope at ED discharge. CONCLUSION: This international external validation of the CSRS showed good performance in identifying patients at low risk for serious outcomes outside of Canada and superior performance compared with the OESIL score. However, clinician classification of syncope at ED discharge seems to explain much of the performance of the CSRS in this study. The clinical utility of the CSRS remains uncertain. PRIMARY FUNDING SOURCE: Swiss National Science Foundation & Swiss Heart Foundation.

Ideación e intento suicida en jóvenes lesbianas, gays y bisexuales de Chile: estudio comunitario online

Background: Suicidal behavior is more common among young people and those who identify as lesbian, gay, or bisexual. Aim: To assess suicidal behavior among young people self-identified as cisgender lesbian, gay, and bisexual. Material and Methods: The Okasha Suicide Scale was answered online by1,683 Chilean people aged 18 to 30 years, self-identified as cisgender lesbian, gay, or bisexual. Results: Thirty four percent of respondents reported suicidal ideation in some moment of their lives and 16% reported repeated suicidal thoughts. Thirty five percent reported at least one suicide attempt. Women had a higher risk of suicide attempts than men. Bisexuals had a higher risk of suicide attempts than lesbians and homosexuals. Those with a non-conforming or non-binary gender identity had a higher risk of suicide attempts than those with a female and male gender identity. Conclusions: These results confirm the higher suicide risk among young sexual minority groups.

Infusing the trauma-informed approach in youth suicide research: Lessons from the field.

Adolescent suicide is a serious public health problem in Chile. Given the high prevalence of suicidal behaviors and suicide, youth suicide researchers must approach participants, families, and communities with care. Special attention must be given to suicide-related trauma among survivors of suicidal behaviors and of suicide. In this article, we discuss how investigators can infuse the trauma-informed model in their work. Our discussion is organized around research stages and tasks: study design, recruitment, data collection and analysis, and care for the research team. We illustrate the integration of the trauma-informed (TI) model key elements and principles with examples from our work in youth suicide research. We posit that infusing the TI model in research aligns with the ethical mandate of beneficence. Our goal is to help other researchers reflect on how to design and implement TI informed research that is attuned to participants, staff, and communities. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Low-Level Laser Irradiation Promotes Proliferation and Differentiation on Apical Papilla Stem Cells.

Introduction: Low-level laser therapy (LLLT) has been reported to improve cell proliferation and differentiation. The stem cells derived from dental apical papilla (SCAPs) are a promising therapy because they are easily obtained from immature human teeth. The effect of LLLT over SCAPs is still unknown. This study aimed to evaluate the proliferation and osteogenic potential of the SCAPs stimulated with LLLT. Methods: SCAPs were isolated from the third molars of a healthy donor and characterized according to the minimum established criteria. SCAPs were cultured for 24 hours before being exposed to LLLT. Cells were exposed to different doses, energy, and wavelengths for selecting the irradiation parameters. SCAPs proliferation was evaluated with the MTT assay at 24 hours and 7-day post-laser exposure. VEGF and TGFß2 expression were assessed with a specific enzyme-linked immunosorbent assay (ELISA). The osteogenic differentiation potential was analyzed with alizarin red staining, and the nodule quantification was performed by the relative optical density (ROD) analysis using ImageJ software. Results: The cells isolated from the apical papilla showed phenotype and stem cell properties. SCAPs irradiated with one dose at 6 J/m2 and 650 nm exhibited significantly higher proliferation (P>0.05) than the controls nonirradiated. LLLT stimulated SCAPs' expression of factors VEGF and TGFß2. Also, SCAPs irradiated showed higher osteogenic activity (P<0.05). Conclusion: LLLT promotes proliferation, osteogenic differentiation, and VEGF and TGFß2 expression on SCAPs. LLLT is a practical approach for the preconditioning of SCAPs in vitro for future regenerative therapies. More studies are needed to determine the underlying molecular processes that determine the mechanism of the LLLT.

Psychometric Properties of the Patient Health Questionnaire-9 in Elderly Chilean Primary Care Users.

Background: This study aimed to assess the measurement properties (reliability, factor structure, and criterion validity) of the Patient Health Questionnaire (PHQ-9) as an instrument for screening major depressive disorder (MDD) in elderly primary care users in Chile. Method: About 582 participants aged between 65 and 80 years were enrolled from primary care centers. They completed the Composite International Diagnostic Interview (CIDI), a survey with sociodemographic characteristics and the PHQ-9. Results: The PHQ-9 revealed an acceptable internal consistency (ω = 0.79 [95% CI: 0.75-0.80] and α = 0.78 [95% CI: 0.75-0.81]); confirmatory factor analysis demonstrated a good fit for both 1- and 2-factor solutions. The chi-square difference test (χ2 = 0.61, gl = 1, p = 0.43) and correlation between the somatic and the cognitive-effective latent factors were very high (r = 0.97, p < 0.001), indicating that the 1 factor model was more parsimonious. Utilizing the CIDI as the gold standard, the area under the curve (AUC) was 0.88 (SE = 0.04, 95% CI: 0.84-0.90). The optimal cut-off score of ≥ 6 yielded good sensitivity and specificity for detecting MDD (0.95 and 0.76, respectively). However, considering the clinical utility index, the cut-off score of ≥9 proved to be a more effective marker for discarding cases of MDD. Conclusion: The PHQ-9 has adequate psychometric properties for elderly primary care users. In clinical settings, it showed its greatest utility in ruling out the presence of an MDD, however, its clinical value for identifying possible cases of MDD is limited. In cases above the cut-off point, it is recommended to perform a more thorough evaluation.

Dental Forensic Kit (DFK®) and Quick Extract™ FFPE DNA extraction kit, a new workflow for obtaining dental DNA for human genetic identity.

BACKGROUND: Most of the protocols described for obtaining DNA from dental tissues are methods that involve major laboratory equipment and many hours of work. They are also methodologies that almost destroy the tooth. AIM: Our aim was to develop an innovative workflow for optimizing dental DNA extraction from teeth. Our methodological proposal is a new workflow for obtaining dental DNA for human genetic identity using Dental Forensic Kit (DFK®) and Formalin-Fixed Paraffin-Embedded (FFPE™) DNA extraction kit. METHODS: Two different dental samples groups were assayed with DFK® and FFPE™. The first group corresponded to extracted teeth from living donors and the second group was considered in real conditions with challenging teeth from corpses. Genomic Dental DNA was amplified and genotyped with platforms Identifiler Plus™, Power Plex 21™ and Global Filer ™ kits. RESULTS: Our workflow was useful in obtaining dental DNA and partial and complete genetic profiles, from teeth of both study groups. DFK® kit worked in a conservative treatment of teeth generating dental tissues (pulp and cement) and FFPE™ for DNA extraction was a very cheap, quick and easy method for obtaining genomic dental DNA. CONCLUSIONS: The innovative method and the workflow proposed herein allows obtaining robust and reliable genetic profiles, from dental tissues. DFK® kit works optimizing the treatment of dental tissues and FFPE™ demonstrates a new use in forensic genetics.

Mesenchymal Stem Cells Derived from Human Inflamed Dental Pulp Exhibit Impaired Immunomodulatory Capacity In Vitro.

INTRODUCTION: Dental pulp stem cells (DPSC) are very attractive in regenerative medicine. In this study, we focused on the characterization of the functional properties of mesenchymal stem cells derived from DPSCs. Currently, it is unknown whether inflammatory conditions present in an inflamed dental pulp tissue could alter the immunomodulatory properties of DPSCs. This study aimed to evaluate the immunomodulatory capacity in vitro of DPSCs derived from healthy and inflamed dental pulp. METHODS: DPSCs from 10 healthy and inflamed dental pulps (irreversible pulpitis) were characterized according to the minimal criteria of the International Society for Cell Therapy, proliferation, differential potential, and colony-forming units. Furthermore, the immunomodulatory capacity of DPSCs was tested on the proliferation of T lymphocytes by flow cytometry and the in vitro enzyme activity of indoleamine 2, 3-dioxygenase. RESULTS: There were no significant differences in the DPSC characteristics and properties such as immunophenotype, tridifferentiation, colony-forming units, and proliferation of the DPSCs derived from normal and inflamed pulp tissue. Furthermore, there were significant differences in the immunomodulatory capacity of DPSCs obtained from human healthy dental pulp and with the diagnosis of irreversible pulpitis. CONCLUSIONS: Our results showed that DPSCs isolated from inflamed dental pulp showed typical characteristics of MSCs and diminished immunosuppressive capacity in vitro in comparison with MSCs derived from healthy dental pulp. Further investigation in vivo is needed to clarify the mechanism of this diminished immunosuppressive capacity.

Optimizing DNA recovery and forensic typing of degraded blood and dental remains using a specialized extraction method, comprehensive qPCR sample characterization, and massively parallel sequencing.

Human dental remains encountered in criminal casework evidence, missing person cases, or mass disaster tragedies provide a valuable sample source for DNA typing when suitable soft tissue is unavailable. Using traditional methods, teeth samples can be challenging to process, resulting in low-quantity and/or quality nuclear DNA and insufficient profiles for comparisons. This study examines the performance of a three-part nuclear DNA analysis workflow for teeth samples based on (1) improved dental tissue recovery using the Dental Forensic Kit (DFKMR) (Universidad de los Andes) and DNA extraction with QuickExtract™ FFPE DNA Extraction Kit (Lucigen®), (2) quantification with InnoQuant® HY (InnoGenomics Technologies) for sensitive assessment of total human and male DNA quantity/quality, and (3) massively parallel sequencing for simultaneous genotyping of 231 short tandem repeat (STR) and single-nucleotide polymorphism (SNP) markers with the ForenSeq® DNA Signature Prep Kit (Verogen, Inc.). Initial evaluation of artificially degraded blood samples (n = 10) achieved highly sensitive and informative quantification results with InnoQuant® HY, enabling successful first pass genotyping with the MiSeq FGx® System. Twenty-three STR alleles (out of 85) and 70 identity informative SNP loci (out of 94) were recovered from two pg total long target DNA input (0.86 ng total short target input) and an InnoQuant degradation index (DI) of 460 (severely degraded). The three-part workflow was subsequently applied to teeth samples (dental pulp, root cement tissues; n = 13) with postmortem intervals (PMI) of the teeth ranging from 8 days to approximately 6 months. Informative SNP and STR DNA profiles were obtained, to include 78 STR alleles and 85 identity informative SNP loci typed (of 94 total SNP targets) in a 1 month, four-day PMI root cement sample with one pg total long target DNA input and a DI of 76. These data indicate successful performance of the proposed workflow from degraded DNA from teeth samples.

Randomized cluster trial of a parenting program in Chile: Key mediators in the decrease in behavior problems in preschool children.

BACKGROUND: Parenting training is a proven strategy for the promotion of positive parenting practices and for the prevention and treatment of behavior problems in children. The processes that explain this efficacy are less clear. The aim of this study was to assess the mediating role of parenting practice modification, encouraged through the implementation of a universal parenting training program, for the decrease of behavior problems in 3- to 6-year-old children. METHOD: A cluster randomized trial was carried out in 19 educational centers in low and middle socioeconomic areas. A total of 178 families received the program and 154 of them were the control group. The following parenting practices were assessed: positive reinforcement, involvement, inconsistency, unsuitable treatment behaviors and physical punishment, as well as hostility and humiliation behaviors. Parent-child interaction was also assessed using an observational instrument. A multiple mediation analysis was carried out, identifying the indirect effects. RESULTS: Reduction of harsh discipline and physical punishment, and parental inconsistency mediated the effects observed in the reduction of child behavior problems during the program. CONCLUSION: Within Chilean families, harsh discipline, physical punishment, and parental inconsistency are important aspects to be considered in the implementation of universal parenting training programs.

Adaptive Reflection on Negative Emotional Experiences: Convergences and Divergence Between the Processing-Mode Theory and the Theory of Self-Distancing Reflection.

Reflecting on negative emotional experiences can be adaptive but it can also maintain or intensify detrimental emotional states. Which factors determine whether reflection can have one consequence or another is unclear. This study focused on two research programs that have concentrated on this topic in the last decades: processing-mode theory (PMT) and self-distancing theory (SDT). The article described and contrasted both programs and their findings. The promising results that PMT and SDT have achieved in identifying the differences between the forms of adaptive and maladaptive reflection are highlighted. Likewise, the disconcerting contradictions observed between both programs that make integrating the findings difficult are indicated. The PMT states that adaptive reflection is concrete, and it is focused on the how of the experience. The SDT states that adaptive reflection is self-distanced and focused on the global meaning of the experience. The article finishes by indicating possible explanations for these apparent contradictions and outlines the challenges to be solved to improve comprehension of the topic.

Effectiveness of a group-based psychosocial program to prevent depression and anxiety in older people attending primary health care centres: a randomised controlled trial.

BACKGROUND: Evidence about the effectiveness of psychosocial interventions to reduce the incidence of depression and anxiety and promote subjective well-being in older people is limited, particularly in Latin-American countries. This study thus aims to assess a program specifically designed to address this issue in persons aged 65 to 80 and attending primary health care centres. METHOD: Older people who use primary care centres are to be randomly assigned to the program or to a control group. Only independent users will be included; those having had a major depressive disorder or an anxiety disorder in the last 6 months will be excluded. The program is group based; it includes cognitive stimulation, expansion of social support networks and cognitive behaviour strategies. Depressive and anxiety symptoms and disorders, as well as psychological well-being, will be assessed using standardised instruments, once before implementing the program and later, after 18 and 36 weeks. DISCUSSION: Primary care is a setting where interventions to improve mental health can be beneficial. Providing evidence-based programs that work with older people is a priority for public mental health. TRIAL REGISTRATION: A protocol for this study has been registered prospectively at ISRCTN registry on 25 July 2018. Identifier: ISRCTN32235611 .