Metformin and Glucose Concentration as Limiting Factors in Retinal Pigment Epithelial Cell Viability and Proliferation.

Metformin is a well-established drug for the treatment of type 2 diabetes; however, the mechanism of action has not been well described and many aspects of how it truly acts are still unknown. Moreover, regarding in vitro experiments, the glycaemic status when metformin is used is generally not considered, which, added to the suprapharmacological drug concentrations that are commonly employed in research, has resulted in gaps of its mechanism of action. The aim of this study was to determine how glucose and metformin concentrations influence cell culture. Considering that diabetic retinopathy is one of the most common complications of diabetes, a retinal pigment epithelial cell line was selected, and cell viability and proliferation rates were measured at different glucose and metformin concentrations. As expected, glucose concentration by itself positively influenced cell proliferation rates. When the metformin was considered, results were conditioned, as well, by metformin concentration. This conditioning resulted in cell death when high concentrations of metformin were used under physiological concentrations of glucose, while this did not happen when clinically relevant concentrations of metformin were used independently of glucose status. Our study shows the importance of in vitro cell growth conditions when drug effects such as metformin's are being analysed.

Reduced C-reactive protein levels after root canal treatment in clinically healthy young apical periodontitis individuals at cardiovascular risk. A prospective study.

AIM: To determine the systemic inflammatory burden, including hsCRP and its monomeric forms, in patients with apical lesions of endodontic origin treated with root canal treatment (RCT). METHODOLOGY: Prospective pre-/post-study. Apical periodontitis (AP) individuals aged 16-40 were included (N = 29). Individuals received RCT and were followed at 1 and 6 months. Fasting blood samples were obtained. Apical lesions of endodontic origin (ALEO) diameter (mm), and periapical index (PAI), were recorded. The serum concentrations of total hsCRP were determined by turbidimetry. Tumour necrosis factor (TNF)-α, interleukin (IL)-6, IL-10, IL-1ß, and soluble (s) E-selectin were assessed by Multiplex assay. Additionally, mCRP forms were determined in the serum of AP patients with a baseline moderate to high cardiovascular risk based on hsCRP stratification (hsCRP ≥1 mg/L) by immunowestern blot (n = 15). Also, CRP isoforms were explored in ALEOs from AP individuals (n = 4). Data were analysed with StataV16. RESULTS: Periapical index and ALEO sizes were reduced at both follow-up visits after RCT (p < .05). Serum levels of TNF-α, IL-6, IL-10, IL-1ß, and sE-selectin did not show significant differences. CRP was borderline reduced at 1 month (p = .04); however, in AP individuals at cardiovascular risk (hsCRP ≥ 1 mg/L), hsCRP and its monomeric isoform significantly decreased at 1 and 6 months (p < .05). CONCLUSIONS: High-sensitivity CRP and mCRP are reduced after RCT in AP individuals at cardiovascular risk.

A Rare Presentation of Systemic Sclerosis.

Systemic sclerosis (SSc) is a persistent autoimmune disorder. While it commonly impacts the cardiac valves, particularly the mitral valve, involvement of the aortic valve has been seldom documented. We report a case of a 47-year-old male with a history of progressive SSc who displayed complications related to cardiac issues, which were verified through a left atrial appendage biopsy revealing thickening due to fibrosis. This cardiac involvement led to a condition necessitating the replacement of the aortic valve due to aortic regurgitation. This instance underscores the importance of identifying this uncommon association, enabling the delivery of appropriate patient treatment, and reducing complications linked to the underlying condition.

Exodoncia de diente supernumerario geminado en maxilar superior

Introducción: Los dientes supernumerarios constituyen una anomalía de número en la cual se excede la cantidad normal de piezas dentales. La geminación es una anomalía de forma que se caracteriza por la presencia de una corona bífida y una raíz común. Los dientes supernumerarios geminados son de rara aparición; indicar exámenes imagenológicos son importantes para determinar sus características y su relación con las estructuras vecinas. Objetivo: Presentar el caso clínico de un paciente con un diente supernumerario geminado. Caso clínico: Paciente de sexo masculino de 23 años de edad, acude a la consulta odontológica por presentar una elevación del lado izquierdo en el paladar, sin molestias. En el examen clínico se observa un aumento de volumen de mucosa palatina de piezas dentales 24 y 25. Los exámenes de imagen fueron realizados y se diagnosticó un diente supernumerario geminado retenido, se realizó tratamiento quirúrgico y el paciente evolucionó favorablemente. Conclusión: Los dientes supernumerarios geminados son poco frecuentes, una oportuna y correcta evaluación imagenológica, le permitirá al clínico realizar un adecuado diagnóstico y tratamiento.

Introduction: Supernumerary teeth constitute an anomaly of number where the normal number of teeth is exceeded and gemination is an anomaly of shape characterized by the presence of a bifid crown, a common root. Geminate supernumerary teeth are of rare occurrence; indicating imaging examinations are important to determine their characteristics and their relationship with neighboring structures. Objective: To present the clinical case of a patient with a geminate supernumerary tooth. Clinical case: A 23-year-old male patient came to the dental office due to an elevation on the left side of the palate, without discomfort. In the clinical examination, an increase in the volume of the palatal mucosa of teeth 24 and 25 was observed. The imaging examinations were performed and a retained geminated supernumerary tooth was diagnosed, surgical treatment was performed and the patient progressed favorably. Conclusion: Geminate supernumerary teeth are rare; a timely and correct imaging evaluation will allow the clinician to make an adequate diagnosis and treatment.

Mixing Meds and Milk: Evaluation of a Performance Gap Intervention for Provider Education in Breastfeeding and Maternal Medication Use.

The need for maternal medications is a known barrier to breastfeeding. Though most medications are compatible with lactation, healthcare providers use abundant caution, often viewing medications and breastfeeding as mutually exclusive. A dual intervention of an educational webinar and access to a mobile app for lactation pharmacology was used to enhance provider familiarity, confidence, and access to knowledge in medication use during breastfeeding. Surveys were administered before, one week after, and three months after the webinar to evaluate performance gap improvement. Usage data of the mobile app was collected over twelve months to monitor topic engagement. Results suggested the interventions temporarily increased provider confidence in maternal medication use during lactation; however, the increase was not sustained at three months. Even with one-time training and lactation-specific mobile app access, simply providing an informational resource is insufficient to support evidence-informed care for lactating patients. Longitudinal training on evidence-based medication safety is critical to care for the lactating dyad.

Screening for Adolescent Risk Behaviors: Preliminary Evidence for a Family Functioning Tool.

BACKGROUND: Family functioning is associated with adolescent drug use, alcohol use, cigarette use, and sexual risk behaviors. Assessing adolescents for family functioning, commonly associated with multiple risk behaviors, may help identify adolescents at risk for adverse health outcomes. This study examined whether a latent family functioning construct, encompassing multiple dimensions of family functioning, was associated with adolescents' substance use and sexual risk behaviors. METHOD: This study used data harmonization with three intervention trials, including data from 1451 adolescents (M = 13.6, SD = 1.0), to perform a full-information item bifactor analysis on 46 family functioning items from five pre-existing family functioning measures. Regression analysis was used to examine the association between the identified subset of items and the following outcomes: cigarette use, alcohol use, drug use, and condom use. RESULTS: Bifactor analysis identified a 26-item latent family functioning construct. Regression analysis indicated that a 26-item latent family functioning construct was associated negatively with lifetime and past 90-day cigarette use, alcohol use, and drug use. CONCLUSION: In sum, the multi-dimensional latent family functioning construct may target specific barriers to risk screening in adolescent populations, including time constraint, hesitancy in discussing sensitive health topics, and use culturally appropriate and age-appropriate assessments.

Lipopolysaccharides from Porphyromonas endodontalis and Porphyromonas gingivalis promote angiogenesis via Toll-like-receptors 2 and 4 pathways in vitro.

AIM: Angiogenesis contributes to the development of apical periodontitis, periodontitis, and other oral pathologies; however, it remains unclear how this process is triggered. The aim was to evaluate whether lipopolysaccharide (LPS) from Porphyromonas endodontalis and Porphyromonas gingivalis induced angiogenesis-related effects in vitro via TLR2 and TLR4. METHODOLOGY: Porphyromonas endodontalis LPS (ATCC 35406 and clinical isolate) was purified with TRIzol, whereas P. gingivalis LPS was obtained commercially. The effects of the different LPS (24 h) in endothelial cell migration were analysed by Transwell assays, following quantification in an optical microscope (40×). The effects of LPS on FAK Y397 phosphorylation were assessed by Western blotting. Angiogenesis in vitro was determined in an endothelial tube formation assay (14 h) in Matrigel in the absence or presence of either LPS. IL-6 and VEGF-A levels were determined in cell supernatants, following 24 h treatment with LPS, and measured in multiplex bead immunoassay. The involvement of TLR2 and TLR4 was assessed with blocking antibodies. The statistical analysis was performed using STATA 12® (StataCorp LP). RESULTS: The results revealed that P. endodontalis LPS, but not P. gingivalis LPS, stimulated endothelial cell migration. Pre-treatment with anti-TLR2 and anti-TLR4 antibodies prevented P. endodontalis LPS-induced cell migration. P. endodontalis LPS promoted FAK phosphorylation on Y397, as observed by an increased p-FAK/FAK ratio. Both P. gingivalis and P. endodontalis LPS (ATCC 35406) induced endothelial tube formation in a TLR-2 and -4-dependent manner, as shown by using blocking antibodies, however, only TLR2 blocking decreased tube formation induced by P. endodontalis (clinical isolate). Moreover, all LPS induced IL-6 and VEGF-A synthesis in endothelial cells. TLR2 and TLR4 were required for IL-6 induction by P. endodontalis LPS (ATCC 35406), while only TLR4 was involved in IL-6 secretion by the other LPS. Finally, VEGF-A synthesis did not require TLR signalling. CONCLUSION: Porphyromonas endodontalis and P. gingivalis LPS induced angiogenesis via TLR2 and TLR4. Collectively, these data contribute to understanding the role of LPS from Porphyromonas spp. in angiogenesis and TLR involvement.

Alta prevalencia de malnutrición por déficit e inseguridad alimentaria en pm institucionalizadas en establecimientos de larga estadía (ELEAM) de las comunas de Chillán y Chillán Viejo, Chile

El acceso y disponibilidad a alimentos seguros y nutritivos, es prioritario para obtener una buena nutrición, con el fin de mejorar la salud y calidad de vida de las personas mayores (PM). El objetivo del estudio fue determinar la prevalencia de malnutrición por déficit e inseguridad alimentaria en PM institucionalizadas en ELEAM, de las comunas de Chillán y Chillán Viejo, Región de Ñuble. Estudio descriptivo que incluyó a 63 personas de 65-96 años, residentes en 7 ELEAM de las comunas de Chillán y Chillán Viejo que aceptaron participar del estudio. Se evaluó el estado nutricional mediante Índice de Masa Corporal (IMC), circunferencia braquial (CB), perímetro de pantorrilla (PP); y se aplicó Escala de Seguridad Alimentaria adaptada y validada. La información se analizó con estadística descriptiva uni y bivariada según tipo de variables. El grupo estuvo conformado mayoritariamente por mujeres (74,6%). La situación nutricional más prevalente fue la malnutrición por déficit (42,9%), seguida por el estado nutricional normal (31,7%). La CB mostró valores promedios de 26,9±3,5 cm en las mujeres y de 26,8±4,5 cm en los hombres; según PP el 39,6% presentó déficit de masa muscular. La seguridad alimentaria estuvo presente en el 20,6% de los encuestados y la inseguridad alimentaria en el 79,3%. Se concluye que las PM pertenecientes a los ELEAM estudiados, presentaron mayoritariamente malnutrición por déficit, e inseguridad alimentaria, lo que no contribuye a una mejor salud del grupo estudiado.

Access to and availability of sufficient, safe, and nutritious food is a priority for quality nutrition for the health and quality of life of the elderly. The objective of this study was to determine the prevalence of undernutrition due to food deficit and insecurity in institutionalized elderly individuals in long-term care facilities (ELEAM) in the communes of Chillán and Chillán Viejo, Ñuble Region, Chile. This descriptive study included 63 individuals aged 65-96 residing in seven ELEAM in the communes of Chillán and Chillán Viejo that accepted to participate. Nutritional status was evaluated by Body Mass Index (IMC), mid-arm circumference (MAC), and calf perimeter (CP); and an adapted and validated Food Security Scale was applied. The information was analyzed using univariate and bivariate descriptive statistics by type of variable. The group consisted mainly of women (74.6%). The most prevalent nutritional status was deficit malnutrition (42.9%), followed by normal nutritional status (31.7%). CB showed mean values of 26.9 ± 3.5 cm in women and 26.8± 4.5 cm in men; according to CP, 39.6% presented a deficit of muscular mass. Food security and insecurity were present in 20.6% and 79.3% of the respondents, respectively. It was concluded that the elderly belonging to the ELEAMs studied presented mostly deficit malnutrition, and food insecurity, which does not contribute to the better health of the group studied.

Preclinical evaluation of Insulin-like growth factor receptor 1 (IGF1R) and Insulin Receptor (IR) as a therapeutic targets in triple negative breast cancer.

Triple Negative Breast Cancer (TNBC), a subtype of breast cancer, has fewer successful therapeutic therapies than other types of breast cancer. Insulin-like growth factor receptor 1 (IGF1R) and the Insulin receptor (IR) are associated with poor outcomes in TNBC. Targeting IGF1R has failed clinically. We aimed to test if inhibiting both IR/IGF1R was a rationale therapeutic approach to treat TNBC. We showed that despite IGF1R and IR being expressed in TNBC, their expression is not associated with a negative survival outcome. Furthermore, targeting both IR/IGF1R with inhibitors in multiple TNBC cell lines did not inhibit cell growth. Linsitinib, a small molecule inhibitor of both IGF1R and IR, did not block tumour formation and had no effect on tumour growth in vivo. Cumulatively these data suggest that while IGF1R and IR are expressed in TNBC, they are not good therapeutic targets. A potential reason for the limited anti-cancer impact when IR/IGF1R was targeted may be because multiple signalling pathways are altered in TNBC. Therefore, targeting individual signalling pathways may not be sufficient to inhibit cancer growth.

Combination of palbociclib with navitoclax based-therapies enhances in vivo antitumoral activity in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is a very aggressive subtype of breast cancer with a poor prognosis and limited effective therapeutic options. Induction of senescence, arrest of cell proliferation, has been explored as an effective method to limit tumor progression in metastatic breast cancer. However, relapses occur in some patients, possibly as a result of the accumulation of senescent tumor cells in the body after treatment, which promote metastasis. In this study, we explored the combination of senescence induction and the subsequent removal of senescent cells (senolysis) as an alternative approach to improve outcomes in TNBC patients. We demonstrate that a combination treatment, using the senescence-inducer palbociclib and the senolytic agent navitoclax, delays tumor growth and reduces metastases in a mouse xenograft model of aggressive human TNBC (hTNBC). Furthermore, considering the off-target effects and toxicity derived from the use of navitoclax, we propose a strategy aimed at minimizing the associated side effects. We use a galacto-conjugated navitoclax (nav-Gal) as a senolytic prodrug that can preferentially be activated by ß-galactosidase overexpressed in senescent cells. Concomitant treatment with palbociclib and nav-Gal in vivo results in the eradication of senescent hTNBC cells with consequent reduction of tumor growth, while reducing the cytotoxicity of navitoclax. Taken together, our results support the efficacy of combination therapy of senescence-induction with senolysis for hTNBC, as well as the development of a targeted approach as an effective and safer therapeutic opportunity.

MMP-9 Levels in the Gingival Crevicular Fluid of Chilean Rosacea Patients.

Rosacea is a chronic inflammatory skin disease whose prevalence rates remain unknown in Chile. Laboratory benchmark testing for this disease is not useful, therefore, we aimed to evaluate the gingival crevicular fluid (GCF) levels of extracellular metalloproteinases (MMP)-2 and MMP-9 as novel rosacea biomarkers. We designed a cross-sectional study with a control group. Participants were systemically healthy adults (n = 20) and persons with rosacea (n = 18). We performed a periodontal evaluation and collected gingival crevicular fluid to measure MMP-2 and MMP-9 levels. Analysis showed mean and standard deviation of MMP-9 concentrations in the GCF for patients with rosacea was 764.52 ± 569.83 pg/mL; for healthy patients, it was 260.69 ± 170.43 pg/mL (p < 0.05). The diagnosis of rosacea was responsible for the levels of MMP-9 in the GCF (p < 0.05), as opposed to periodontitis, smoking, and age (p > 0.05). The Area under ROC for MMP-9 was 0.869 (95%, C.I: 0.719−0.956), with a sensitivity of 72.22% and specificity of 81.58% for the diagnosis of rosacea. We conclude that the quantification of MMP-9 in the GCF could be used as a biomarker of rosacea. Also, rosacea was responsible for increasing the levels of MMP-9 in the GCF independent of periodontal status.

Lipopolysaccharide from Porphyromonas gingivalis, but Not from Porphyromonas endodontalis, Induces Macrophage M1 Profile.

Apical Lesions of Endodontic Origin (ALEO) are initiated by polymicrobial endodontic canal infection. Porphyromonas gingivalis (Pg) and Porphyromonas endodontalis (Pe) lipopolysaccharides (LPS) can induce a pro-inflammatory macrophage response through their recognition by TLR2 and TLR4. However, polarization responses induced by Pg and/or Pe LPS in macrophages are not fully understood. We aimed to characterize the polarization profiles of macrophages differentiated from THP-1 cells following Pg and/or Pe LPS stimulation from reference strain and clinical isolates. A modified LPS purification protocol was implemented and the electrophoretic LPS profiles were characterized. THP-1 human monocytes differentiated to macrophages were stimulated with Pg and Pe LPS. Polarization profiles were characterized through cell surface markers and secreted cytokines levels after 24 h of stimulation. TLR2 and TLR4 cell surfaces and transcriptional levels were determined after 24 or 2 h of LPS stimulation, respectively. LPS from Pg induced a predominant M1 profile in macrophages evidenced by changes in the expression of the surface marker CD64 and pro-inflammatory cytokine profiles, TNF-α, IL-1ß, IL-6, and IL-12. Pe LPS was unable to induce a significant response. TLR2 and TLR4 expressions were neither modified by Pg or Pe LPS. Pg LPS, but not Pe LPS, induced a macrophage M1 Profile.

Malnutrición por exceso en niños de 5 a 10 años y su asociación con el estado nutricional pre y gestacional, lactancia materna y patología materna

La malnutrición por exceso es un problema de salud pública relevante, con origen multifactorial y factores asociados como el estado nutricional pregestacional (ENP), durante y post gestación, ganancia excesiva de peso (EGP) gestacional y duración de la lactancia materna. El objetivo del estudio fue evaluar la malnutrición por exceso en niños de 5 a 10 años y su asociación con el estado nutricional pre y gestacional, lactancia materna exclusiva (LME) y patologías maternas. Se realizó un estudio con 213 niños y niñas estudiantes de 5 a 10 años. Los datos biosociodemográficos se obtuvieron mediante entrevistas con las madres y datos maternos previos y el embarazo con el carné de control. El estimó estado nutricional se estimó con z-score del peso para la talla e IMC/E, según sexo. Para los análisis estadísticos se usó el software STATA 15.0. El 56% de los estudiantes tuvo malnutrición por exceso. Un 53,1% de las madres presentó malnutrición por exceso previo al embarazo que aumentó a 74,6% post gestación. El 71,4% de los niños tuvo una LME ≥6 meses. Se relacionó estadísticamente la edad materna ≥35 años (p= 0,044) y la malnutrición por exceso pregestacional (p= 0,014). La edad materna ≥35 años aumentó el riesgo de malnutrición por exceso casi dos veces (OR= 1,78; IC: 1,029-3,046), al igual que el ENP (OR= 2,11; IC: 1,193-3,693) y en patologías maternas (OR= 1,41; IC: 1, 073-2,694). En conclusión, los niños preescolares de 5 a 10 años con factores de edad materna ≥35, ENP y patologías maternas tuvieron entre 1,4 y 2,11 veces más riesgo de presentar malnutrición por exceso comparado con aquellos niños sin estos factores.

Overnutrition is a relevant public health problem with a multifactorial origin. Associated factors include maternal nutritional status before, during, and after gestation, excessive gestational weight gain, and breastfeeding duration. The objective of the study was to evaluate the association between overnutrition in children aged 5-10 years and pre- and gestational nutritional status, exclusive breastfeeding, and maternal pathologies. The study consisted of 213 schoolchildren aged 5-10 years. Bio-socio-demographic data were obtained through interviews with mothers, previous maternal data and pregnancy follow-up records. Nutritional status was measured using weight-for-height and BMI z-score by age and sex. The STATA 15.0 software was used for statistical analysis. A total of 56% of the schoolchildren had overnutrition. Meanwhile, 53.1% of the mothers demonstrated pre-pregnancy overnutrition, which increased to 74.6% post-pregnancy. It was found that 71.4% of schoolchildren had experienced exclusive breastfeeding ≥6 months. Maternal age ≥35 years (p= 0.044) and pre-gestational nutritional status (p= 0.014) were statistically related. Maternal age ≥35 years increased overnutrition by almost two-fold (OR= 1.78; IC: 1.029-3.046), as did pre-gestational nutritional status (OR= 2.11; IC: 1.193-3.693) and maternal pathologies (OR= 1.41; IC: 1.073-2.694). In conclusion, schoolchildren aged 5-10 years with mothers ≥35 years of age, who had overnutrition in the pre-gestational period, and a pathology were 1.4 to 2.44 times more at risk of developing overnutrition than children without these factors.

Pharmacological senolysis reduces doxorubicin-induced cardiotoxicity and improves cardiac function in mice.

Many anticancer agents used in clinics induce premature senescence in healthy tissues generating accelerated aging processes and adverse side-effects in patients. Cardiotoxicity is a well-known limiting factor of anticancer treatment with doxorubicin (DOX), a very effective anthracycline widely used as antitumoral therapy in clinical practice, that leads to long-term morbidity and mortality. DOX exposure severely affects the population of cardiac cells in both mice and human hearts by inducing premature senescence, which may represent the molecular basis of DOX-induced cardiomyopathy. Here, we demonstrate that senescence induction in the heart contributes to impaired cardiac function in mice upon DOX treatment. Concomitant elimination of senescent cells with the senolytic Navitoclax in different formulations produces a significant decrease in senescence and cardiotoxicity markers together with the restoration of the cardiac function in mice followed by echocardiography. These results evidence the potential clinical use of senolytic therapies to alleviate cardiotoxicities induced in chemotherapy-treated patients.

Role of TLR9 methylation on its active transcription in apical inflammation.

AIM: To explore the methylation pattern, its role in transcriptional regulation and potential modifiers of methylation of the TLR9 gene in chronic periapical inflammation. METHODOLOGY: In this cross-sectional study, apical lesions of endodontic origin (ALEO, n = 61) and healthy periodontal ligaments (HPL, n = 15) were included. Products from bisulfited and PCR-amplified DNA were analysed for their methylation profiles in the promoter region and at each CpG island. Additionally, TLR9 mRNA levels were quantified by qPCR and bivariate and multiple modelling were performed to better understand the influence of methylation on gene transcription. RESULTS: TLR9 mRNA levels were upregulated in ALEO compared to HPL (p < .001). TLR9 promoter CpG sites and CpG +2086 in the intragenic island 1 were demethylated in ALEO compared to HPL (p < .05). Multivariate analysis, adjusted by smoking and gender, revealed that demethylation of TLR9 promoter sites enhanced transcriptional activity, specifically demethylated CpGs at positions -736 and -683, (p = .02), which are close to CRE binding. Although ALEO reduced the global methylation of the gene promoter and intragenic-island 2 (p < .05) by -42.5 and -9.5 percentage points, respectively, age reduced the global methylation of intragenic-island 3 within the exon 2. CONCLUSIONS: Demethylations of TLR9 promoter CpG sites, along with the intragenic DNA methylation status, were involved in higher transcription in ALEO. Hence, chronic periapical inflammation and ageing modify the methylation status both in the gene promoter and in intragenic CpG islands.

Tannat Grape Skin: A Feasible Ingredient for the Formulation of Snacks with Potential for Reducing the Risk of Diabetes.

In the present work the feasibility of Tannat grape skin (TGS) as a functional ingredient in the formulation of two snacks (yogurt and biscuits) was studied. The research provided novel information on the effects of the food matrix and digestion process, under simulated human oral gastrointestinal conditions, in the bioaccessibility of TGS bioactive compounds composing of the snacks with health promoting properties (antioxidant, anti-inflammatory, and antidiabetic). TGS polyphenolic profile was analyzed by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) finding mainly flavonoids, phenolic acids, and anthocyanins, which may exert antioxidant, anti-inflammatory, and carbohydrase inhibition capacities. TGS digest showed antioxidant and antidiabetic potential compared to the undigested sample (p < 0.05). Yogurt and biscuits with TGS were developed with the nutrition claims "no-added sugars" and "source of fiber" and were digested in vitro to evaluate the bioaccessibility of compounds with health promoting properties after food processing and digestion. After in vitro simulation of digestion, bioactive properties were enhanced for control and TGS snacks which may be attributed to the formation/release of compounds with health-promoting properties. Biscuits showed significant increase in ABTS antioxidant capacity and yogurt showed increased α-glucosidase inhibition capacity by the addition of TGS (p < 0.05). Polyphenols from TGS and bioactive peptides from snacks which may be released during digestion might be responsible for the observed bioactivities. Consumer's acceptance of TGS yogurt and biscuits showed scores of 6.3 and 5.1 (scale 1−9), respectively, showing TGS yogurt had higher overall acceptance. Sensory profile assessed by check-all-that-apply + just-about-right (CATA+JAR) showed most of the attributes were evaluated as "just about right", supporting good food quality. The developed yogurt presented adequate shelf-life parameters for 28 days. TGS yogurt with higher acceptability showed reduced ROS formation (p < 0.05) induced by tert-butyl hydroperoxide (1 mM) in CCD-18Co colon cells and RAW264.7 macrophages when pre-treated with concentrations 500−1000 and 100−500 µg/mL of the digests, respectively. Moreover, TGS yogurt digest pre-treatment reduced nitric oxide (NO) production (p < 0.05) in lipopolysaccharide (LPS)-induced RAW264.7 macrophages, showing anti-inflammatory potential. Bioactive peptides generated during lactic fermentation and digestion process may be contributors to intracellular effects. In conclusion, yogurt and biscuits with Tannat grape skin addition were obtained with nutrition claims "no-added sugars" and "source of fiber" with the potential to modulate key biochemical events associated with diabetes pathogenesis.

CpG Single-Site Methylation Regulates TLR2 Expression in Proinflammatory PBMCs From Apical Periodontitis Individuals.

Introduction: Apical periodontitis (AP) is a common oral disease caused by the inflammatory destruction of the periapical tissues due to the infection of the root canal system of the tooth. It also contributes to systemic bacterial translocation, where peripheric mononuclear blood cells (PBMCs) can act as carriers. Toll-like receptor (TLR) 2 mediates the response to infection and activates inflammatory responses. DNA methylation can be induced by bacteria and contributes to the modulation of this response. Despite the evidence that supports the participation of PBMCs in immune-inflammatory disorders, the inflammatory profile and epigenetic regulatory mechanisms of PBMCs in AP individuals are unknown. Aim: To determine TLR2 gene methylation and inflammatory profiles of PBMCs in AP. Methods: Cross-sectional exploratory study. Otherwise, healthy individuals with AP (n=27) and controls (n=30) were included. PMBCs were isolated by a Ficoll gradient, cultured for 24 hours, and both RNA and DNA were extracted. DNA was bisulfite-treated, and specific sites at the promoter region of the TLR2 gene were amplified by qPCR using validated primers. To verify its amplification, agarose gels were performed. Then, the PCR product was sequenced. mRNA expression of TLR2 was determined by qPCR. The soluble levels of 105 inflammatory mediators were first explored with Proteome Profiler Human Cytokine Array Kit. Consequently, tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10, IL-6Rα, IL-1ß, and IL-12p70 levels were measured by Multiplex assay. Results: PBMCs from individuals with AP demonstrated a proinflammatory profile showing higher soluble levels of TNF-α, IL-6, and IL-1ß compared to controls (p<0.05). Higher TLR2 expression and higher global methylation pattern of the promoter region of the gene were found in AP compared to controls (p<0.05). The CpGs single-sites at positions -166 and -146 were completely methylated, while the site -102 was totally unmethylated, independently of the presence of AP. DNA methylation of CpG single-sites in positions -77 and +24 was positively associated with TLR2 expression. Conclusions: PBMCs from AP subjects show a hyperinflammatory phenotype and TLR2 upregulation in association with single CpG-sites' methylation from the TLR2 gene promoter, thereby contributing to a sustained systemic inflammatory load in individuals with periapical endodontic diseases.

Acceso a los dispositivos autoajustables para el tratamiento de apneas del sueño durante la pandemia por COVID-19 / Access to auto-adjustable positive airway pressure devices for the treatment of obstructive sleep apnea during the COVID-19 pandemic

Resumen El empleo de presión positiva en la vía aérea es el tratamiento de elección en apneas obstructIvas del sueño. Desde el inicio de la pandemia por el virus SARS- CoV-2 las recomendaciones fueron evitar la administración en el laboratorio de sueño de estas terapias e iniciar tratamiento con equipos autoajus tables. El objetivo fue evaluar el acceso a los equipos autoajustables en el contexto epidemiológico actual. Se incluyeron 66 pacientes, la edad media fue de 50.7 ± 12.8, 44% hombres, con un IAH de 35.5 (22.7-64.2) y un IMC de 38.3 (32.8-46.1). El 45% tenía cobertura médica. De los 50 (74%) que tramitaron el equipo, 19 (29%) accedió al dispositivo. La cobertura médica fue la única variable significativa entre los que accedieron y los que no: 54% vs. 21% (p = 0.016). La demora entre el inicio del trámite y el del tratamiento fue 31 ± 21 días en los que tenían cobertura médica y 65 ± 35 días en aquellos sin cobertura (p = 0.008). El acceso a terapias autoajustables durante la pandemia por COVID estuvo por debajo del de nuestra experiencia previa y de lo descripto en otros trabajos con terapias fijas. Cabe plantearse si esta disminución es producto de la pandemia e independiente del tipo de terapia, o está en relación directa a los mayores costos de los equipos autoajustables o falta de convenio de los mismos por parte de los organismos proveedores.

Abstract The use of positive airway pressure is the treatment of choice for obstructive sleep apnea. Since the beginning of SARS-CoV-2 virus pandemic the recommenda tions were to avoid the administration of this therapy in the sleep laboratory and to start treatment with auto-adjustable continuous positive airway pressure devices. The objective was to evaluate access to these devices in the current epidemiological context. Sixty-six patients were included, the mean age was 50.7 ± 12.8, 44% men with an AHI of 35.5 (22.7-64.2) and a BMI of 38.3 (32.8-46.1). Forty-five percent had health insurance. Of the 50 (74%) who processed the device, 19 (29%) had access. Health insurance was the only significant variable between those who had access to the device and those who did not: 54% vs. 21% (p=0.016). The delay between the beginning of the procedure to access the device and the start of treatment was 31 ± 21 in patients who had medical insurance and 65 ± 35.2 in those who did not (p = 0.008). Access to auto-adjustable therapies during the COVID-19 pandemic was below our former experience and that described in other studies with fixed therapies. It is worth considering whether this decrease is a product of the pandemic and indepen dent of the type of therapy or is directly related to the higher costs of self-adjustable devices or to a lack of coverage from the healthcare providers.

Effects of IFIH1 rs1990760 variants on systemic inflammation and outcome in critically ill COVID-19 patients in an observational translational study.

Background: Variants in IFIH1, a gene coding the cytoplasmatic RNA sensor MDA5, regulate the response to viral infections. We hypothesized that IFIH1 rs199076 variants would modulate host response and outcome after severe COVID-19. Methods: Patients admitted to an intensive care unit (ICU) with confirmed COVID-19 were prospectively studied and rs1990760 variants determined. Peripheral blood gene expression, cell populations, and immune mediators were measured. Peripheral blood mononuclear cells from healthy volunteers were exposed to an MDA5 agonist and dexamethasone ex-vivo, and changes in gene expression assessed. ICU discharge and hospital death were modeled using rs1990760 variants and dexamethasone as factors in this cohort and in-silico clinical trials. Results: About 227 patients were studied. Patients with the IFIH1 rs1990760 TT variant showed a lower expression of inflammation-related pathways, an anti-inflammatory cell profile, and lower concentrations of pro-inflammatory mediators. Cells with TT variant exposed to an MDA5 agonist showed an increase in IL6 expression after dexamethasone treatment. All patients with the TT variant not treated with steroids survived their ICU stay (hazard ratio [HR]: 2.49, 95% confidence interval [CI]: 1.29-4.79). Patients with a TT variant treated with dexamethasone showed an increased hospital mortality (HR: 2.19, 95% CI: 1.01-4.87) and serum IL-6. In-silico clinical trials supported these findings. Conclusions: COVID-19 patients with the IFIH1 rs1990760 TT variant show an attenuated inflammatory response and better outcomes. Dexamethasone may reverse this anti-inflammatory phenotype. Funding: Centro de Investigación Biomédica en Red (CB17/06/00021), Instituto de Salud Carlos III (PI19/00184 and PI20/01360), and Fundació La Marató de TV3 (413/C/2021).

Patients with severe COVID-19 often need mechanical ventilation to help them breathe and other types of intensive care. The outcome for many of these patients depends on how their immune system reacts to the infection. If the inflammatory response triggered by the immune system is too strong, this can cause further harm to the patient. One gene that plays an important role in inflammation is IFIH1 which encodes a protein that helps the body to recognize viruses. There are multiple versions of this gene which each produce a slightly different protein. It is possible that this variation impacts how the immune system responds to the virus that causes COVID-19. To investigate, Amado-Rodríguez, Salgado del Riego et al. analyzed the IFIH1 gene in 227 patients admitted to an intensive care unit in Spain for severe COVID-19 between March and December 2020. They found that patients with a specific version of the gene called TT experienced less inflammation and were more likely to survive the infection. Physicians typically treat patients with moderate to severe COVID-19 with corticosteroid drugs that reduce the inflammatory response. However, Amado-Rodríguez, Salgado del Riego et al. found that patients with the TT version of the IFIH1 gene were at greater risk of dying if they received corticosteroids. The team then applied the distribution of IFIH1 variants among different ethnic ancestries to data from a previous clinical trial, and simulated the effects of corticosteroid treatment. This 'mock' clinical trial supported their findings from the patient-derived data, which were also validated by laboratory experiments on immune cells from individuals with the TT gene. The work by Amado-Rodríguez, Salgado del Riego et al. suggests that while corticosteroids benefit some patients, they may cause harm to others. However, a real-world clinical trial is needed to determine whether patients with the TT version of the IFIH1 gene would do better without steroids.

A Healthy Lifestyle Intervention for Hispanic Families: Moderating Effects of Education, Income, Nativity.

OBJECTIVE: To explore the moderation effect of parental social determinant factors-educational attainment, income, and nativity-on intervention effects of a family-based lifestyle intervention, Familias Unidas for Health and Wellness (FUHW). DESIGN: Longitudinal analysis across 6- and 24-months postbaseline of a randomized controlled trial. PARTICIPANTS: Two-hundred and eighty primary caregivers. ANALYSIS: Moderation analysis was used to examine the interactive effects between intervention effects and social determinant factors. MAIN OUTCOME MEASURES: Parental body mass index, added sugar intake, sugar-sweetened beverage intake, and fruits and vegetable intake. RESULTS: Findings indicated that FUHW effects on added sugar intake were moderated by educational attainment (P = 0.006), income (P = 0.023), and nativity (P = 0.024). In addition, FUHW effects on sugar-sweetened beverage intake were moderated by parent education (P = 0.042). CONCLUSIONS AND IMPLICATIONS: This study suggests that FUHW may be helpful in reducing added sugar and sugar-sweetened beverage intake for Hispanic parents who face social determinant factors-educational attainment, income, and foreign-born status-that may influence their dietary behaviors. By including demographic information in intervention development, effects may be enhanced by ensuring that considerations related to participants' socioeconomic and cultural status are incorporated into intervention components and help reduce overweight and obesity.