Root canal disinfection of immature dog teeth with apical periodontitis: comparison of three different protocols

Objectives: The present in vivo study was designed to assess the efficacy of 3 root canal disinfection protocols in immature dog teeth with apical periodontitis (AP). Material and Methods: Forty immature premolars with pulp necrosis and AP of five Beagle dogs were used. Three experimental disinfection protocols were established. After irrigation with 40 ml 5.25% sodium hypochlorite using the Endovac system, in Group 1 canals were flushed with QMix solution; in Group 2, canals were flushed with QMix solution and 2% chlorhexidine gel dressing was placed for two weeks; and in Group 3, triantibiotic paste dressing was placed for two weeks. Canals were sampled after periapical lesions were radiographically visible (S1), after the first disinfection session (S2) and, in groups 2 and 3, after dressing (S3). Results: After the first session of the disinfection protocol (S2), there was significant (p < 0.05) bacterial reduction in the three experimental groups. Microorganisms were absent in 100% of S2 samples in groups 1 and 2, and in 75% of group 3 (p > 0.05). After dressing, 87.5% of the S3 samples showed increased bacterial count: in group 2, CFU counts (median = 891) were significantly higher than in group 3 (median = 18) (p = 0.03). Conclusions: In immature dog teeth with AP, root canal irrigation using QMix solution, with or without chlorhexi - dine gel dressing, or a triantibiotic paste dressing, provides the same level of disinfection than irrigation with 5.25% sodium hypochlorite alone in only one session

Genetic variation in the NEIL2 DNA glycosylase gene is associated with oxidative DNA damage in BRCA2 mutation carriers

In this report, we have tried to gain molecular insight into a single nucleotide polymorphism (SNP) in the NEIL2 gene previously identified as "cancer risk modifier" for BRCA2 mutation carriers. To that end, we studied the role of this SNP (rs804271) on NEIL2 transcriptional regulation, oxidative DNA damage and genome instability in two independent set of samples: The first one was a series of eighty-six BRCA1 and BRCA2 mutation carriers and eighty non-carrier controls in which we evaluated the effect of the SNP on NEIL2 gene expression and oxidative DNA damage accumulation. The second was a set of twenty lymphoblastoid cell lines (LCLs), thirteen BRCA1 mutation carriers and seven non-carriers control, that were used to analyze the correlation between NEIL2 mRNA and/or protein levels, the oxidative and the double stranded break (DSB) DNA damage levels. Our results suggest that an excessive production of NEIL2 enzyme, associated with the SNP, may have a deleterious effect modifying cancer risk susceptibility in BRCA2 mutation carriers. We hypothesize that due to the SNP impact on NEIL2 transcriptional upregulation, a cascade of events may converge in the accumulation of oxidative DNA damage and its posterior conversion into DSBs for this specific group of patients.We thank Alicia Barroso her technical assistance. Also to Dr. Thomas Helleday, (Karolinska Institutet, Stockholm, Sweden) that kindly provided NEIL2 and UNG purified enzymes. The Genotype-Tissue Expression (GTEx) Project was supported by the Common Fund of the Office of the Director of the National Institutes of Health. Additional funds were provided by the NCI, NHGRI, NHLBI, NIDA, NIMH, and NINDS. Donors were enrolled at Biospecimen Source Sites funded by NCI/SAIC-Frederick, Inc. (SAIC-F) subcontracts to the National Disease Research Interchange (10XS170),Roswell Park Cancer Institute (10XS171), and Science Care, Inc. (X10S172). The Laboratory, Data Analysis, and Coordinating Center (LDACC) were funded through a contract (HHSN268201000029C) to The Broad Institute, Inc. Biorepository operations were funded through an SAIC-F subcontract to the Van Andel Institute (10ST1035). Additional data repository and project management were provided by SAIC-F (HHSN261200800001E). The Brain Bank was supported by supplements to University of Miami grants DA006227 & DA033684 and to contract N01MH000028. Statistical Methods development grants were made to the University of Geneva (MH090941 & MH101814), the University of Chicago (MH090951, MH090937, MH101820, MH101825), the University of North Carolina - Chapel Hill (MH090936 & MH101819), Harvard University (MH090948), Stanford University (MH101782), Washington University St Louis (MH101810), and the University of Pennsylvania (MH101822). The data used for the analyses described in this manuscript were obtained from: [insert, where appropriate] the GTEx Portal on 01/12/2015 and/or dbGaP accession number phs000424.v7.p2 on 01/10/2017.S

Tailoring two-photon fluorescent probes for pH bioimaging in living cells

Fluorescence biosensors are indispensable basic tools in modern biology. These type of molecules allow real-time visualization of biological events inside living cells. Especially important in many of these processes (proliferation, apoptosis or defense tasks) is the control of the cellular pH. In consequence, a great variety of structural models have been developed for pH bioimaging in fluorescence microscopy. Nonetheless, these efforts have been mainly focused on the development of one-photon (OP) probes. Recently, we described a biosensor with excellent photophysical properties and appropriate two-photon absorption (TPA) behavior. This sensor allows selective and specific detection of hydroxyl radicals solely inside lysosomes.Based on this scaffold, we have synthesized and characterized new TPA fluorescent probes. These molecules have an “off-on” response to different pH environments with a strong selectivity and sensitivity toward H+. These naphthalene-indolenine derivatives have a high synthetic versatility through affordable and efficient synthesis. The synthetic modification of this model allows tuning subcellular targets through minor modifications and without affecting their emission properties. The effectiveness of these probes and their structural modifications for different pH-related applications has been probed in mouse embrionary fibroblast (MEF) cells.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Soluble platelet-endothelial cell adhesion molecule-1, a biomarker of ventilator-induced lung injury

Introduction: Endothelial cell injury is an important component of acute lung injury. Platelet-endothelial cell adhesion molecule-1 (PECAM1) is a transmembrane protein that connects endothelial cells to one another and can be detected as a soluble, truncated protein (sPECAM1) in serum. We hypothesized that injurious mechanical ventilation (MV) leads to shedding of PECAM1 from lung endothelial cells resulting in increasing sPECAM1 levels in the systemic circulation. Methods: We studied 36 Sprague–Dawley rats in two prospective, randomized, controlled studies (healthy and septic) using established animal models of ventilator-induced lung injury. Animals (n = 6 in each group) were randomized to spontaneous breathing or two MV strategies: low tidal volume (VT) (6 ml/kg) and high-VT (20 ml/kg) on 2 cmH2O of positive end-expiratory pressure (PEEP). In low-VT septic animals, 10 cmH2O of PEEP was applied. We performed pulmonary histological and physiological evaluation and measured lung PECAM1 protein content and serum sPECAM1 levels after four hours ventilation period. Results: High-VT MV caused severe lung injury in healthy and septic animals, and decreased lung PECAM1 protein content (P < 0.001). Animals on high-VT had a four- to six-fold increase of mean sPECAM1 serum levels than the unventilated counterpart (35.4 ± 10.4 versus 5.6 ± 1.7 ng/ml in healthy rats; 156.8 ± 47.6 versus 35.6 ± 12.6 ng/ml in septic rats) (P < 0.0001). Low-VT MV prevented these changes. Levels of sPECAM1 in healthy animals on high-VT MV paralleled the sPECAM1 levels of non-ventilated septic animals. Conclusions: Our findings suggest that circulating sPECAM1 may represent a promising biomarker for the detection and monitoring of ventilator-induced lung injury

Castillejo del Bonete (Terrinches, Ciudad Real): un complejo tumular prehistórico de la Cultura de las Motillas en el Alto Guadalquivir

Situado en las estribaciones orientales de Sierra Morena, dentro de la cuenca hidrológica del Guadalquivir, Castillejo del Bonete es un gran complejo arquitectónico que consta de una cueva monumentalizada mediante estructuras varias, entre las que destacan varios corredores megalíticos y túmulos, todos ellos asociados a contextos funerarios y depósitos de ofrendas. El presente artículo se centra en la explicación detallada de los elementos que integran este complejo constructivo para posteriormente discutir el avance que supone para la investigación de la Prehistoria Reciente en La Mancha. Castillejo del Bonete tiene el potencial de convertirse en un yacimiento clave para la comprensión de las prácticas funerarias y la creciente jerarquización social durante el tránsito del III al II milenios cal ANE

Efficacy and safety of universal (TCRKO) ARI-0001 CAR-T cells for the treatment of B-cell lymphoma

Autologous T cells expressing the Chimeric Antigen Receptor (CAR) have been approved as advanced therapy medicinal products (ATMPs) against several hematological malignancies. However, the generation of patient-specific CART products delays treatment and precludes standardization. Allogeneic off-theshelf CAR-T cells are an alternative to simplify this complex and timeconsuming process. Here we investigated safety and efficacy of knocking out the TCR molecule in ARI-0001 CAR-T cells, a second generation aCD19 CAR approved by the Spanish Agency of Medicines and Medical Devices (AEMPS) under the Hospital Exemption for treatment of patients older than 25 years with Relapsed/Refractory acute B cell lymphoblastic leukemia (B-ALL). We first analyzed the efficacy and safety issues that arise during disruption of the TCR gene using CRISPR/Cas9. We have shown that edition of TRAC locus in T cells using CRISPR as ribonuleorproteins allows a highly efficient TCR disruption (over 80%) without significant alterations on T cells phenotype and with an increased percentage of energetic mitochondria. However, we also found that efficient TCRKO can lead to on-target large and medium size deletions, indicating a potential safety risk of this procedure that needs monitoring. Importantly, TCR edition of ARI-0001 efficiently prevented allogeneic responses and did not detectably alter their phenotype, while maintaining a similar anti-tumor activity ex vivo and in vivo compared to unedited ARI-0001 CAR-T cells. In summary, we showed here that, although there are still some risks of genotoxicity due to genome editing, disruption of the TCR is a feasible strategy for the generation of functional allogeneic ARI-0001 CAR-T cells. We propose to further validate this protocol for the treatment of patients that do not fit the requirements for standard autologous CAR-T cells administration.Spanish ISCIII Health Research FundEuropean Commission PI15/02015 PI18/00337 PI21/00298Red TerAv RD21/ 0017/0004 PI18/ 00330 PI17/00672CECEyU and CSyF of the Junta de Andalucia FEDER/European Cohesion Fund (FSE) for Andalusia 2016000073391-TRA 2016000073332-TRA PI-57069 PAIDIBio326 CARTPI-0001- 201 PECART-0031-2020 PI0014-2016 PEER-0286-2019Spanish Government 00123009/SNEO-20191072 PLEC2021-008094regional Ministry of Health 0006/2018 C2-0002-2019Spanish Government FPU16/05467 FPU17/02268 FPU17/04327Junta de Andalucia PECART-00312020Consejeria de Salud y Familias PECART-0027-2020 MCI DIN2018-010180 DIN2020-01155

Chorological news for Carex (Cyperaceae) in the Iberian Peninsula

Se presentan diversas notas corológicas del género Carex para la península ibérica, que incluyen novedades provinciales para cuatro especies (C. elata subsp. elata, C. demissa subsp. demissa, C. oedipostyla, C. sylvatica subsp. sylvatica) así como la confirmación provincial (no indicada en Flora iberica) de la presencia de tres taxónes. Proporcionamos asimismo la segunda cita nacional para Portugal de C. reuteriana subsp. mauritanica. Otras tres citas poseen interés corológico al constituir novedades comarcales y/o de sector biogeográfico, o bien segunda cita provincial. Cinco de las novedades revisten interés conservacionista por tratarse de especies o poblaciones amenazadas, incluidas en Listas Rojas y/o protegidas a nivel legal. Por último, hacemos referencia a la posible naturalización de C. pendula.Various chorological notes for the genus Carex in the Iberian Peninsula are presented, including new provincial records for four species (C. elata subsp. elata, C. demissa subsp. demissa, C. oedipostyla, C. sylvatica subsp. sylvatica), as well as provincial confirmations (not indicated in Flora iberica) for the presence of three taxa. We also provide the second national report for Portugal of C. reuteriana subsp. mauritanica. Three additional reports have chorological interest by constituting new regional and/or biogeographic records, or the second provincial report. Five of the novelties are of conservation concern because they imply endangered species or populations, included in Red Lists and/or legally protected. Finally, we refer to the possible naturalization of C. pendula

Acceleration of the DNA methylation clock among lynch syndrome‑associated mutation carriers

The research leading to these results has received funding from "la Caixa" Foundation (Ref: CAIXA2017/1) for library preparation, sequencing, and employment of research personnel, from The Fundacion Progreso y Salud, Junta de Andalucia, Spain and from DPI2017-84439-R of MINECO, Madrid and FEDER for sequencing and employment of research personnel. Finally, grant ref. A-BIO-470-UGR20 from University of Granada and FEDER has funded article processing charges (APC) and sample processing expenses.Background: DNA methylation (DNAm) age metrics have been widely accepted as an epigenetic biomarker for biological aging and disease. The purpose of this study is to assess whether or not individuals carrying Lynch Syndromeassociated mutations are affected in their rate of biological aging, as measured by the epigenetic clock. Methods: Genome-wide bisulfite DNA sequencing data were generated using DNA from CD4 + T-cells obtained from peripheral blood using 27 patient samples from Lynch syndrome families. Horvath’s DNAm age model based on penalized linear regression was applied to estimate DNAm age from patient samples with distinct clinical and genetic characteristics to investigate cancer mutation-related aging effects. Results: Both Lynch mutation carriers and controls exhibited high variability in their estimated DNAm age, but regression analysis showed steeper slope for the Lynch mutation carriers. Remarkably, six Lynch Syndrome-associated mutation carriers showed a strong correlation to the control group, and two sisters carrying Lynch Syndrome-associated mutations, with no significant difference in lifestyle and similar chronological age, were assigned very different DNAm age. Conclusions: Future studies will be required to explore, in larger patient populations, whether specific epigenetic age acceleration is predictive of time-to-cancer development, treatment response, and survival. Epigenetic clock DNAm metrics may be affected by the presence of cancer mutations in the germline, and thus show promise of potential clinical utility for stratified surveillance strategies based on the relative risk for imminent emergence of tumor lesions in otherwise healthy Lynch Syndrome-associated mutation carriers.La Caixa Foundation CAIXA2017/1Junta de AndaluciaSpanish Government DPI2017-84439-REuropean CommissionUniversity of Granada A-BIO-470-UGR2