ABSTRACT
Glioblastoma (GBM) is a highly aggressive brain tumor associated with poor patient survival. The current standard treatment involves invasive surgery, radiotherapy, and chemotherapy employing temozolomide (TMZ). Resistance to TMZ is, however, a major challenge. Previous work from our group has identified candidate genes linked to TMZ resistance, including genes encoding translesion synthesis (TLS) DNA polymerases iota (PolÉ©) and kappa (Polκ). These specialized enzymes are known for bypassing lesions and tolerating DNA damage. Here, we investigated the roles of PolÉ© and Polκ in TMZ resistance, employing MGMT-deficient U251-MG glioblastoma cells, with knockout of either POLI or POLK genes encoding PolÉ© and Polκ, respectively, and assess their viability and genotoxic stress responses upon subsequent TMZ treatment. Cells lacking either of these polymerases exhibited a significant decrease in viability following TMZ treatment compared to parental counterparts. The restoration of the missing polymerase led to a recovery of cell viability. Furthermore, knockout cells displayed increased cell cycle arrest, mainly in late S-phase, and lower levels of genotoxic stress after TMZ treatment, as assessed by a reduction of γH2AX foci and flow cytometry data. This implies that TMZ treatment does not trigger a significant H2AX phosphorylation response in the absence of these proteins. Interestingly, combining TMZ with Mirin (double-strand break repair pathway inhibitor) further reduced the cell viability and increased DNA damage and γH2AX positive cells in TLS KO cells, but not in parental cells. These findings underscore the crucial roles of PolÉ© and Polκ in conferring TMZ resistance and the potential backup role of homologous recombination in the absence of these TLS polymerases. Targeting these TLS enzymes, along with double-strand break DNA repair inhibition, could, therefore, provide a promising strategy to enhance TMZ's effectiveness in treating GBM.
ABSTRACT
El tratamiento con implantes dentales hoy en día es un procedimiento clínico de rutina que permite rehabilitar a los pacientes con prótesis fijas. En este caso presentamos un tratamiento complejo de implantación inmediata del sector anterior con pérdida parcial de la cortical vestibular en el que se realizó una regeneración ósea guiada y provisionalización en un tiempo quirúrgico en un paciente con patología renal. Complementamos el estudio con una revisión de la efectividad de las técnicas utilizadas y las posibles respuestas celular asociadas a la patología renal.
Treatment with dental implants nowadays is a routine clinical procedure that allows patient rehabilitation with fixed prostheses. In this case we present a complex treatment of immediate implantation of the anterior sector with partial loss of the vestibular cortex, in which guided bone regeneration and provisionalization was performed in surgical time in a patient with kidney pathology. The study was complemented with a review of the effectiveness of the techniques used and the possible cellular responses associated with kidney pathology.
ABSTRACT
In the search for alternatives to overcome the challenge imposed by drug resistance development in cancer treatment, the modulation of autophagy has emerged as a promising alternative that has achieved good results in clinical trials. Nevertheless, most of these studies have overlooked a novel and selective type of autophagy: chaperone-mediated autophagy (CMA). Following its discovery, research into CMA's contribution to tumor progression has accelerated rapidly. Therefore, we now understand that stress conditions are the primary signal responsible for modulating CMA in cancer cells. In turn, the degradation of proteins by CMA can offer important advantages for tumorigenesis, since tumor suppressor proteins are CMA targets. Such mutual interaction between the tumor microenvironment and CMA also plays a crucial part in establishing therapy resistance, making this discussion the focus of the present review. Thus, we highlight how suppression of LAMP2A can enhance the sensitivity of cancer cells to several drugs, just as downregulation of CMA activity can lead to resistance in certain cases. Given this panorama, it is important to identify selective modulators of CMA to enhance the therapeutic response.
ABSTRACT
Caspase recruitment domain (CARD)-containing protein 14 (CARD14) is an intracellular protein that mediates nuclear factor-kappa B (NF-ĸB) signaling and proinflammatory gene expression in skin keratinocytes. Several hyperactivating CARD14 mutations have been associated with psoriasis and other inflammatory skin diseases. CARD14-induced NF-ĸB signaling is dependent on the formation of a CARD14-BCL10-MALT1 (CBM) signaling complex, but upstream receptors and molecular mechanisms that activate and regulate CARD14 signaling are still largely unclear. Using unbiased affinity purification and mass spectrometry (AP-MS) screening, we discover polo-like kinase 1 (PLK1) as a novel CARD14-binding protein. CARD14-PLK1 binding is independent of the CARD14 CARD domain but involves a consensus phospho-dependent PLK1-binding motif in the CARD14 linker region (LR). Expression of the psoriasis-associated CARD14(E138A) variant in human keratinocytes induces the recruitment of PLK1 to CARD14-containing signalosomes in interphase cells, but does not affect the specific location of PLK1 in mitotic cells. Finally, disruption of the PLK1-binding motif in CARD14(E138A) increases CARD14-induced proinflammatory signaling and gene expression. Together, our data identify PLK1 as a novel CARD14-binding protein and indicate a negative regulatory role for PLK1 in CARD14 signaling.
ABSTRACT
Self-activated luminescent calcium phosphate (CaP) nanoparticles, including hydroxyapatite (HA) and amorphous calcium phosphate (ACP), are promising for bioimaging and theragnostic applications in nanomedicine, eliminating the need for activator ions or fluorophores. In this study, we developed luminescent and stable citrate-functionalized carbonated ACP nanoparticles for bioimaging purposes. Our findings revealed that both the CO32- content and the posterior heating step at 400 °C significantly influenced the composition and the structural ordering of the chemically precipitated ACP nanoparticles, impacting the intensity, broadness, and position of the defect-related photoluminescence (PL) emission band. The heat-treated samples also exhibited excitation-dependent PL under excitation wavelengths typically used in bioimaging (λexc = 405, 488, 561, and 640 nm). Citrate functionalization improved the PL intensity of the nanoparticles by inhibiting non-radiative deactivation mechanisms in solution. Additionally, it resulted in an increased colloidal stability and reduced aggregation, high stability of the metastable amorphous phase and the PL emission for at least 96 h in water and supplemented culture medium. MTT assay of HepaRG cells, incubated for 24 and 48 h with the nanoparticles in concentrations ranging from 10 to 320 µg mL-1, evidenced their high biocompatibility. Internalization studies using the nanoparticles self-activated luminescence showed that cellular uptake of the nanoparticles is both time (4-24 h) and concentration (160-320 µg mL-1) dependent. Experiments using confocal laser scanning microscopy allowed the successful imaging of the nanoparticles inside cells via their intrinsic PL after 4 h of incubation. Our results highlight the potential use of citrate-functionalized carbonated ACP nanoparticles for use in internalization assays and bioimaging procedures.
Subject(s)
Calcium Phosphates , Nanoparticles , Calcium Phosphates/chemistry , Nanoparticles/chemistry , Humans , Particle Size , Luminescence , Optical Imaging , Cell Survival/drug effects , Carbonates/chemistryABSTRACT
Insects of economic importance such as Leucoptera coffeella can cause high defoliation in plants and reduce crop yields. We aimed to identify changes in the ecological niche and potential zones of the invasion. Occurrence records were obtained from databases and bibliography. WorldClim V2.0 bioclimatic layers were used. For the modeling of the potential distribution, the kuenm R package was used by executing the Maxent algorithm. The potential distribution models suggested greatest environmental suitability extends from Europe, South Asia, and Central and South Africa, showing the "tropical and subtropical moist broadleaf forests" as the ecoregion that presents the greatest probability of the presence of L. coffeella. The potential distribution model projected in the invaded area agrees with the known distribution in the region (America), although the results show that it is occupying environmental spaces not present in the area of origin. This species presented a large proportion of the invaded niche that overlaps the native niche and is colonizing new environmental conditions in the invaded area relative to its native distribution (Africa). This information could be used in the planning of coffee crops on the American continent.
Subject(s)
Ecosystem , Introduced Species , Animals , Animal Distribution , Lepidoptera , Coffea , MothsABSTRACT
The Y-family DNA polymerases - Pol ι, Pol η, Pol κ and Rev1 - are most well-known for their roles in the DNA damage tolerance pathway of translesion synthesis (TLS). They function to overcome replication barriers by bypassing DNA damage lesions that cannot be normally replicated, allowing replication forks to continue without stalling. In this work, we demonstrate a novel interaction between each Y-family polymerase and the nucleotide excision repair (NER) proteins, RAD23A and RAD23B. We initially focus on the interaction between RAD23A and Pol ι, and through a series of biochemical, cell-based, and structural assays, find that the RAD23A ubiquitin-binding domains (UBA1 and UBA2) interact with separate sites within the Pol ι catalytic domain. While this interaction involves the ubiquitin-binding cleft of UBA2, Pol ι interacts with a distinct surface on UBA1. We further find that mutating or deleting either UBA domain disrupts the RAD23A-Pol ι interaction, demonstrating that both interactions are necessary for stable binding. We also provide evidence that both RAD23 proteins interact with Pol ι in a similar manner, as well as with each of the Y-family polymerases. These results shed light on the interplay between the different functions of the RAD23 proteins and reveal novel binding partners for the Y-family TLS polymerases.
Subject(s)
DNA Repair Enzymes , DNA-Binding Proteins , DNA-Directed DNA Polymerase , DNA Damage , DNA Polymerase iota/chemistry , DNA Repair , DNA Replication , DNA-Directed DNA Polymerase/metabolism , Ubiquitins/chemistry , DNA-Binding Proteins/chemistry , DNA Repair Enzymes/chemistryABSTRACT
Parkinson disease (PD) is caused by the loss of ventral midbrain dopaminergic neurons (mDANs) in the substantia nigra pars compacta (SNpc). These cells are especially vulnerable to stress but can be protected by autophagy enhancement strategies in vitro and in vivo. In our recent study, we focused on the LIM (Lin11, Isl-1, and Mec-3)-domain homeobox transcription factors LMX1A (LIM homeobox transcription factor 1 alpha) and LMX1B (LIM homeobox transcription factor 1 beta), crucial drivers of mDAN differentiation with roles in autophagy gene expression for stress protection in the developed brain. Using human induced pluripotent stem cell (hiPSC)-derived mDANs and transformed human cell lines, we found that these autophagy gene transcription factors are themselves regulated by autophagy-mediated turnover. LMX1B possesses a non-canonical LC3-interacting region (LIR) in its C-terminus through which it interacts with ATG8 family members. The LMX1B LIR-like domain enables binding to ATG8 proteins in the nucleus, where ATG8 proteins act as co-factors for robust transcription of LMX1B target genes. Thus, we propose a novel role for ATG8 proteins as autophagy gene transcriptional co-factors for mDAN stress protection in PD.
ABSTRACT
The BAF (BRG1/BRM-associated factor) chromatin remodelling complex is essential for the regulation of DNA accessibility and gene expression during neuronal differentiation. Mutations of its core subunit SMARCB1 result in a broad spectrum of pathologies, including aggressive rhabdoid tumours or neurodevelopmental disorders. Other mouse models have addressed the influence of a homo- or heterozygous loss of Smarcb1, yet the impact of specific non-truncating mutations remains poorly understood. Here, we have established a new mouse model for the carboxy-terminal Smarcb1 c.1148del point mutation, which leads to the synthesis of elongated SMARCB1 proteins. We have investigated its impact on brain development in mice using magnetic resonance imaging, histology, and single-cell RNA sequencing. During adolescence, Smarcb11148del/1148del mice demonstrated rather slow weight gain and frequently developed hydrocephalus including enlarged lateral ventricles. In embryonic and neonatal stages, mutant brains did not differ anatomically and histologically from wild-type controls. Single-cell RNA sequencing of brains from newborn mutant mice revealed that a complete brain including all cell types of a physiologic mouse brain is formed despite the SMARCB1 mutation. However, neuronal signalling appeared disturbed in newborn mice, since genes of the AP-1 transcription factor family and neurite outgrowth-related transcripts were downregulated. These findings support the important role of SMARCB1 in neurodevelopment and extend the knowledge of different Smarcb1 mutations and their associated phenotypes.
Subject(s)
Hydrocephalus , Transcription Factor AP-1 , Animals , Mice , Hydrocephalus/genetics , Mutation/genetics , Point Mutation/genetics , Signal Transduction , Transcription Factor AP-1/geneticsABSTRACT
Autophagy is an intracellular lysosomal degradation pathway by which cytoplasmic cargoes are removed to maintain cellular homeostasis. Monitoring autophagy flux is crucial to understand the autophagy process and its biological significance. However, assays to measure autophagy flux are either complex, low throughput or not sensitive enough for reliable quantitative results. Recently, ER-phagy has emerged as a physiologically relevant pathway to maintain ER homeostasis but the process is poorly understood, highlighting the need for tools to monitor ER-phagy flux. In this study, we validate the use of the signal-retaining autophagy indicator (SRAI), a fixable fluorescent probe recently generated and described to detect mitophagy, as a versatile, sensitive and convenient probe for monitoring ER-phagy. This includes the study of either general selective degradation of the endoplasmic reticulum (ER-phagy) or individual forms of ER-phagy involving specific cargo receptors (e.g., FAM134B, FAM134C, TEX264 and CCPG1). Crucially, we present a detailed protocol for the quantification of autophagic flux using automated microscopy and high throughput analysis. Overall, this probe provides a reliable and convenient tool for the measurement of ER-phagy.
Subject(s)
Autophagy , Membrane Proteins , Membrane Proteins/metabolism , Endoplasmic Reticulum/metabolism , Endoplasmic Reticulum Stress , Autophagy-Related Proteins/metabolismABSTRACT
Atypical teratoid/rhabdoid tumors (ATRTs) represent a rare, but aggressive pediatric brain tumor entity. They are genetically defined by alterations in the SWI/SNF chromatin remodeling complex members SMARCB1 or SMARCA4. ATRTs can be further classified in different molecular subgroups based on their epigenetic profiles. Although recent studies suggest that the different subgroups have distinct clinical features, subgroup-specific treatment regimens have not been developed thus far. This is hampered by the lack of pre-clinical in vitro models representative of the different molecular subgroups. Here, we describe the establishment of ATRT tumoroid models from the ATRT-MYC and ATRT-SHH subgroups. We demonstrate that ATRT tumoroids retain subgroup-specific epigenetic and gene expression profiles. High throughput drug screens on our ATRT tumoroids revealed distinct drug sensitivities between and within ATRT-MYC and ATRT-SHH subgroups. Whereas ATRT-MYC universally displayed high sensitivity to multi-targeted tyrosine kinase inhibitors, ATRT-SHH showed a more heterogeneous response with a subset showing high sensitivity to NOTCH inhibitors, which corresponded to high expression of NOTCH receptors. Our ATRT tumoroids represent the first pediatric brain tumor organoid model, providing a representative pre-clinical model which enables the development of subgroup-specific therapies.
Subject(s)
Brain Neoplasms , Rhabdoid Tumor , Teratoma , Child , Humans , Teratoma/drug therapy , Teratoma/genetics , SMARCB1 Protein/genetics , Brain Neoplasms/drug therapy , Brain Neoplasms/genetics , Rhabdoid Tumor/drug therapy , Rhabdoid Tumor/genetics , Rhabdoid Tumor/metabolism , Receptors, Notch , Epigenomics , DNA Helicases , Nuclear Proteins , Transcription Factors/geneticsABSTRACT
The LIM homeodomain transcription factors LMX1A and LMX1B are essential mediators of midbrain dopaminergic neuronal (mDAN) differentiation and survival. Here we show that LMX1A and LMX1B are autophagy transcription factors that provide cellular stress protection. Their suppression dampens the autophagy response, lowers mitochondrial respiration, and elevates mitochondrial ROS, and their inducible overexpression protects against rotenone toxicity in human iPSC-derived mDANs in vitro. Significantly, we show that LMX1A and LMX1B stability is in part regulated by autophagy, and that these transcription factors bind to multiple ATG8 proteins. Binding is dependent on subcellular localization and nutrient status, with LMX1B interacting with LC3B in the nucleus under basal conditions and associating with both cytosolic and nuclear LC3B during nutrient starvation. Crucially, ATG8 binding stimulates LMX1B-mediated transcription for efficient autophagy and cell stress protection, thereby establishing a novel LMX1B-autophagy regulatory axis that contributes to mDAN maintenance and survival in the adult brain.
Subject(s)
Autophagy-Related Protein 8 Family , LIM-Homeodomain Proteins , Mesencephalon , Neurons , Transcription Factors , Humans , Autophagy , Brain/cytology , Brain/metabolism , LIM-Homeodomain Proteins/genetics , LIM-Homeodomain Proteins/metabolism , Mesencephalon/metabolism , Transcription Factors/metabolism , Autophagy-Related Protein 8 Family/genetics , Neurons/cytologyABSTRACT
Introduction: Cell membrane-covered biomimetic nanosystems have allowed the development of homologous nanostructures to bestow nanoparticles with enhanced biointerfacing capabilities. The stability of these structures, however, still represents a challenge for the scientific community. This study is aimed at developing and optimizing cell derived membrane-coated nanostructures upon applying design of experiments (DoE) to improve the therapeutic index by homotypic targeting in cancer cells. Methods: Important physicochemical features of the extracted cell membrane from tumoral cells were assessed by mass spectrometry-based proteomics. PLGA-based nanoparticles encapsulating temozolomide (TMZ NPs) were successfully developed. The coating technology applying the isolated U251 cell membrane (MB) was optimized using a fractional two-level three-factor factorial design. All the formulation runs were systematically characterized regarding their diameter, polydispersity index (PDI), and zeta potential (ZP). Experimental conditions generated by DoE were also subjected to morphological studies using negative-staining transmission electron microscopy (TEM). Its short-time stability was also assessed. MicroRaman and Fourier-Transform Infrared (FTIR) spectroscopies and Confocal microscopy were used as characterization techniques for evaluating the NP-MB nanostructures. Internalization studies were carried out to evaluate the homotypic targeting ability. Results and Discussion: The results have shown that nearly 80% of plasma membrane proteins were retained in the cell membrane vesicles after the isolation process, including key proteins to the homotypic binding. DoE analysis considering acquired TEM images reveals that condition run five should be the best-optimized procedure to produce the biomimetic cell-derived membrane-coated nanostructure (NP-MB). Storage stability for at least two weeks of the biomimetic system is expected once the original characteristics of diameter, PDI, and ZP, were maintained. Raman, FTIR, and confocal characterization results have shown the successful encapsulation of TMZ drug and provided evidence of the effective coating applying the MB. Cell internalization studies corroborate the proteomic data indicating that the optimized NP-MB achieved specific targeting of homotypic tumor cells. The structure should retain the complex biological functions of U251 natural cell membranes while exhibiting physicochemical properties suitable for effective homotypic recognition. Conclusion: Together, these findings provide coverage and a deeper understanding regarding the dynamics around extracted cell membrane and polymeric nanostructures interactions and an in-depth insight into the cell membrane coating technology and the development of optimized biomimetic and bioinspired nanostructured systems.
ABSTRACT
DNA-targeting agents have a significant clinical use, although toxicity remains an issue that plays against their widespread application. Understanding the mechanism of action and DNA damage response elicited by such compounds might contribute to the improvement of their use in anticancer chemotherapy. In a previous study, our research group characterized a new DNA-targeting agent - pradimicin-IRD. Since DNA-targeting agents and DNA repair are close-related subjects, the present study used in silico-modelling and a transcriptomic approach seeking to characterize the DNA repair pathways activated in HCT 116 cells following pradimicin-IRD treatment. Molecular docking analysis showed pradimicin-IRD as a DNA intercalating agent and a potential inhibitor of DNA-binding proteins. Furthermore, the transcriptomic study highlighted DNA repair functions related to genes modulated by pradimicin-IRD, such as nucleotide excision repair, telomeres maintenance and double-strand break repair. When validating these functions, PCNA protein levels decreased after exposure to pradimicin. Furthermore, molecular docking analysis suggested DNA-pradimicin-PCNA interaction. In addition, hTERT and POLH showed reduced mRNA levels after 6 h of treatment with pradimicin-IRD. Moreover, POLH-deficient cells displayed higher resistance to pradimicin-IRD than POLH-proficient cells and the compound prevented formation of the POLH/DNA complex (molecular docking). Since the modulation of DNA repair genes by pradimicin-IRD is TP53-independent, unlike doxorubicin, dissimilarities between the mechanism of action and the DNA damage response of pradimicin-IRD and doxorubicin open new insights for further studies of pradimicin-IRD as a new antineoplastic compound.
Subject(s)
Antineoplastic Agents , Humans , Molecular Docking Simulation , Proliferating Cell Nuclear Antigen , Antineoplastic Agents/pharmacology , DNA Repair , DNA , Doxorubicin/pharmacology , DNA DamageABSTRACT
Human DNA polymerases can bypass DNA lesions performing translesion synthesis (TLS), a mechanism of DNA damage tolerance. Tumor cells use this mechanism to survive lesions caused by specific chemotherapeutic agents, resulting in treatment relapse. Moreover, TLS polymerases are error-prone and, thus, can lead to mutagenesis, increasing the resistance potential of tumor cells. DNA polymerase eta (pol eta) - a key protein from this group - is responsible for protecting against sunlight-induced tumors. Xeroderma Pigmentosum Variant (XP-V) patients are deficient in pol eta activity, which leads to symptoms related to higher sensitivity and increased incidence of skin cancer. Temozolomide (TMZ) is a chemotherapeutic agent used in glioblastoma and melanoma treatment. TMZ damages cells' genomes, but little is known about the role of TLS in TMZ-induced DNA lesions. This work investigates the effects of TMZ treatment in human XP-V cells, which lack pol eta, and in its complemented counterpart (XP-V comp). Interestingly, TMZ reduces the viability of XP-V cells compared to TLS proficient control cells. Furthermore, XP-V cells treated with TMZ presented increased phosphorylation of H2AX, forming γH2AX, compared to control cells. However, cell cycle assays indicate that XP-V cells treated with TMZ replicate damaged DNA and pass-through S-phase, arresting in the G2/M-phase. DNA fiber assay also fails to show any specific effect of TMZ-induced DNA damage blocking DNA elongation in pol eta deficient cells. These results show that pol eta plays a role in protecting human cells from TMZ-induced DNA damage, but this can be different from its canonical TLS mechanism. The new role opens novel therapeutic possibilities of using pol eta as a target to improve the efficacy of TMZ-based therapies against cancer.
Subject(s)
Antineoplastic Agents , Xeroderma Pigmentosum , Antineoplastic Agents/pharmacology , DNA , DNA Damage , DNA-Directed DNA Polymerase/genetics , DNA-Directed DNA Polymerase/metabolism , Humans , Temozolomide/pharmacology , Xeroderma Pigmentosum/geneticsABSTRACT
Introduction: There are multiple techniques for vertical bone augmentation. Guided bone regeneration is one of them; however, the literature is diverse and includes different study designs, which makes it difficult to synthesize results. Objective: To analyze the general technical characteristics, clinical results, and complications of vertical bone augmentation performed with guided bone regeneration in humans. Material and Methods: This scoping review was based on the PRISMA-ScR guidelines. A search was performed in the Pubmed, Scielo, and Worldcat databases. Papers published from 1990 to April 2020 were included in the study. Research articles not conducted in humans or published in languages other than English and Spanish were excluded. Title and abstract were screened by two reviewers, then full studies were extracted, and data tabulated. Results: 89 studies were included. The highest percentage reported having obtained a vertical bone increase of less than 5 mm and having used non-resorbable membranes. The most frequent type of graft is autogenous and combinations of grafts, the most common being autogenous with xenograft. All studies that reported bone stability of implants in regenerated bone were favorable, as was implant survival, reporting values between 83.8% and 100%. Membrane exposure is the most frequently reported complication, followed by infection or abscesses, and tissue dehiscence. Conclusion: Vertical bone regeneration is a reliable technique, with high predictability and low incidence of complications compared to other vertical bone augmentation techniques.
Introducción: Existen múltiples técnicas para el aumento óseo vertical siendo una opción la regeneración ósea guiada, sin embargo, la literatura es diversa y con distintos diseños que dificultan la síntesis de resultados. Objetivo: Analizar las características generales técnicas, resultados clínicos y complicaciones del aumento óseo vertical realizado con regeneración ósea guiada en humanos. Material y Métodos: Esta revisión de alcance se basó en la guía PRISMA-ScR. Se realizó una búsqueda en las bases de datos Pubmed, Scielo y Worldcat. Fueron incluidos aquellos publicados desde el año 1990 hasta abril de 2020. Se excluyeron los estudios no realizados en humanos o publicados en idiomas distintos al inglés y español. Dos revisores examinaron título y resumen, luego los estudios completos se extrajeron y se ordenaron los datos en tablas. Resultados: 89 estudios fueron incluidos. El mayor porcentaje reportó haber obtenido un aumento óseo vertical menor a 5 mm y haber utilizado membranas no reabsorbibles. El tipo de injerto que más frecuente es el autógeno y las combinaciones de injertos, siendo el más común autógeno con xenoinjerto. Todos los estudios que reportaron estabilidad ósea de implantes en hueso regenerado fueron favorables, al igual que la supervivencia de implantes, reportando valores entre 83,8% y 100%. La exposición de membrana es la complicación que más se repite en los estudios, seguido por infección o abscesos y dehiscencia de tejidos. Conclusión: La regeneración ósea vertical es una técnica confiable, con alta predictibilidad y baja incidencia de complicaciones en comparación a otras técnicas de aumento óseo vertical.
Subject(s)
Humans , Bone Regeneration , Dental Implants , Guided Tissue Regeneration, Periodontal , Alveolar Ridge Augmentation/methods , Alveolar Bone Loss , Transplants , Alveolar ProcessABSTRACT
The murine esBAF complex plays a major role in the regulation of gene expression during stem cell development and differentiation. As one of its core subunits, Smarcb1 is indispensable for its function and its loss is connected to neurodevelopmental disorders and participates in the carcinogenesis of entities such as rhabdoid tumours. We explored how Smarcb1 regulates gene programs in murine embryonic stem cells (mESC) and in this way orchestrates differentiation. Our data underline the importance of Smarcb1 expression and function for the development of the nervous system along with basic cellular functions, such as cell adhesion and cell organisation. Using ChIP-seq, we were able to portray the consequences of Smarcb1 knockdown (kd) for the binding of esBAF and PRC2 as well as its influence on histone marks H3K27me3, H3K4me3 and H3K27ac. Their signals are changed in gene and enhancer regions of genes connected to nervous system development and offers a plausible explanation for changes in gene expression. Further, we describe a group of genes that are, despite increased BAF binding, suppressed after Smarcb1 kd by mechanisms independent of PRC2 function.
Subject(s)
Rhabdoid Tumor , Animals , Carcinogenesis , Cell Differentiation/genetics , Embryonic Stem Cells/metabolism , Mice , Rhabdoid Tumor/genetics , Rhabdoid Tumor/metabolismABSTRACT
Rhabdoid tumors (RT) are rare and highly aggressive pediatric neoplasms. Their epigenetically-driven intertumoral heterogeneity is well described; however, the cellular origin of RT remains an enigma. Here, we establish and characterize different genetically engineered mouse models driven under the control of distinct promoters and being active in early progenitor cell types with diverse embryonic onsets. From all models only Sox2-positive progenitor cells give rise to murine RT. Using single-cell analyses, we identify distinct cells of origin for the SHH and MYC subgroups of RT, rooting in early stages of embryogenesis. Intra- and extracranial MYC tumors harbor common genetic programs and potentially originate from fetal primordial germ cells (PGCs). Using PGC specific Smarcb1 knockout mouse models we validate that MYC RT originate from these progenitor cells. We uncover an epigenetic imbalance in MYC tumors compared to PGCs being sustained by epigenetically-driven subpopulations. Importantly, treatments with the DNA demethylating agent decitabine successfully impair tumor growth in vitro and in vivo. In summary, our work sheds light on the origin of RT and supports the clinical relevance of DNA methyltransferase inhibitors against this disease.
Subject(s)
Rhabdoid Tumor , Animals , Germ Cells/pathology , Humans , Mice , Rhabdoid Tumor/genetics , Rhabdoid Tumor/pathology , SMARCB1 Protein/genetics , Single-Cell Analysis , TranscriptomeABSTRACT
Measurements of CO2 and counting of occupants were carried out in 37 public bus trips during commuting rush hours in Barcelona (NE Spain) with the aim of evaluating parameters governing ventilation inside the vehicles and proposing actions to improve it. The results show that CO2 concentrations (1039 and 934 ± 386 ppm, as average and median, during rush hours but with average reduced occupancy due to the fair to be infected by SARS-CoV-2 during the measurement period, and measured in the middle of the busses) are in the lower range of values recorded in the literature for public buses, however an improvement in ventilation is required in a significant proportion of the journeys. Thus, we found better ventilation in the older Euro 3+ (retrofitted with filter traps and selective catalytic reduction) and Euro 5 buses (average 918 ± 257 ppm) than in the hermetically closed new Euro 6 ones (1111 ± 432 ppm). The opening of the windows in the older buses yielded higher ventilation rates (778 ± 432 ppm). The opening of all doors at all stops increases the ventilation by causing a fall in concentrations of 200-350 ppm below inter-stop concentrations, with this effect typically lasting 40-50 s in the hermetically closed new Euro 6 hybrid buses. Based on these results a number of recommendations are offered in order to improve ventilation, including measurement of CO2 and occupancy, and installation of ventilation fans on the top of the hermetically closed new buses, introducing outdoor air when a given concentration threshold is exceeded. In these cases, a CO2 sensor installed in the outdoor air intake is also recommended to take into account external CO2 contributions.
Subject(s)
Air Pollution, Indoor , COVID-19 , Air Pollution, Indoor/analysis , Carbon Dioxide/analysis , Humans , Motor Vehicles , SARS-CoV-2 , Transportation , VentilationABSTRACT
Nucleotide excision repair (NER) is one of the main pathways for genome protection against structural DNA damage caused by sunlight, which in turn is extensively related to skin cancer development. The mutation spectra induced by UVB were investigated by whole-exome sequencing of randomly selected clones of NER-proficient and XP-C-deficient human skin fibroblasts. As a model, a cell line unable to recognize and remove lesions (XP-C) was used and compared to the complemented isogenic control (COMP). As expected, a significant increase of mutagenesis was observed in irradiated XP-C cells, mainly C>T transitions, but also CC>TT and C>A base substitutions. Remarkably, the C>T mutations occur mainly at the second base of dipyrimidine sites in pyrimidine-rich sequence contexts, with 5'TC sequence the most mutated. Although T>N mutations were also significantly increased, they were not directly related to pyrimidine dimers. Moreover, the large-scale study of a single UVB irradiation on XP-C cells allowed recovering the typical mutation spectrum found in human skin cancer tumors. Eventually, the data may be used for comparison with the mutational profiles of skin tumors obtained from XP-C patients and may help to understand the mutational process in nonaffected individuals.