Gene expression dynamics during temperature-dependent sex determination in a sea turtle.

Fifty years ago, researchers discovered a link between ambient temperature and the sex of turtle embryos. More recently, significant progress has been made in understanding the influence of temperature on freshwater turtles. However, our understanding of the key genetic factors in other turtle groups, such as sea turtles, remains limited. To address this gap, we conducted RNA-seq analyses on embryonic tissues from the sea olive ridley turtle during the thermosensitive period (stages 21-26) at temperatures known to produce males (26 °C) and females (33 °C). Our findings revealed that incubation temperatures primarily influence genes with broad expression across tissues due to differential cell division rates and later have an effect regulating gonad-specific transcripts. This effect is mostly related to gene activation rather than transcription repression. We performed transcriptome analyses following shifts in incubation temperatures of bi-potential gonads. This approach allowed us to identify genes that respond rapidly and may be closer to the beginning of the temperature-sensing pathway. Notably, we observed swift adaptations in the expression levels of chromatin modifiers JARID2 and KDM6B, as well as the splicing factor SRSF5, and transcription regulators THOC2, DDX3X and CBX3, but little impact in the overall gonad-specific pathways, indicating that temperature-sensing genes may change rapidly but the rewiring of the gonad's developmental fate is complex and resilient. AUTHOR SUMMARY: Sea turtles, one of the most iconic creatures of our oceans, confront a troubling reality of endangerment, a peril magnified by the looming specter of climate change. This climatic shift is gradually increasing the temperature of the nesting beaches thus causing dramatic male/female population biases. Conservation efforts will need genetic and molecular information to reverse the negative effects of climate change on the populations. In this study, we conducted the first transcriptomic analysis of embryonic tissues, including gonads, brain, liver, and mesonephros, in the olive ridley sea turtle during the critical thermosensitive period spanning stages 21-26. We examined both male-producing (26 °C) and female-producing (33 °C) temperatures and found that incubation temperatures influence temperature-sensitive genes that are either expressed globally or specifically associated with the gonads. These findings indicate that incubation temperatures predominantly sway genes with broad expression patterns due to differential cell division rates. This natural process was opted in the gonads to drive sex determination. We also identified genes that are rapidly capable of sensing temperature changes and that could play a role in the activation of the sex determination pathway. Overall, our study sheds light on the intricate interplay between temperature and gene expression during sea turtle development, revealing dynamic changes in the transcriptome and highlighting the involvement of key genetic players in sex determination.

Comparison of Phytochemical Composition and Untargeted Metabolomic Analysis of an Extract from Cnidoscolus aconitifolius (Mill.) I. I. Johnst and Porophyllum ruderale (Jacq.) Cass. and Biological Cytotoxic and Antiproliferative Activity In Vitro.

Cnidoscolus aconitifolius (CA) and Porophyllum ruderale (PR) are representative edible plants that are a traditional food source in Mexico. This research aimed to analyze the phytochemical composition and untargeted metabolomics analysis of CA and PR and evaluate their antiproliferative effect in vitro. The phytochemical composition (UPLC-DAD-QToF/MS-ESI) identified up to 38 polyphenols and selected organic acids that were clustered by the untargeted metabolomics in functional activities linked to indolizidines, pyridines, and organic acids. Compared with PR, CA displayed a higher reduction in the metabolic activity of human SW480 colon adenocarcinoma cells (LC50: 10.65 mg/mL), and both extracts increased the total apoptotic cells and arrested cell cycle at G0/G1 phase. PR increased mRNA Apc gene expression, whereas both extracts reduced mRNA Kras expression. Rutin/epigallocatechin gallate displayed the highest affinity to APC and K-RAS proteins in silico. Further research is needed to experiment on other cell lines. Results suggested that CA and PR are polyphenol-rich plant sources exhibiting antiproliferative effects in vitro.

Analysis of High-Risk Neuroblastoma Transcriptome Reveals Gene Co-Expression Signatures and Functional Features.

Neuroblastoma represents a neoplastic expansion of neural crest cells in the developing sympathetic nervous system and is childhood's most common extracranial solid tumor. The heterogeneity of gene expression in different types of cancer is well-documented, and genetic features of neuroblastoma have been described by classification, development stage, malignancy, and progression of tumors. Here, we aim to analyze RNA sequencing datasets, publicly available in the GDC data portal, of neuroblastoma tumor samples from various patients and compare them with normal adrenal gland tissue from the GTEx data portal to elucidate the gene expression profile and regulation networks they share. Our results from the differential expression, weighted correlation network, and functional enrichment analyses that we performed with the count data from neuroblastoma and standard normal gland samples indicate that the analysis of transcriptome data from 58 patients diagnosed with high-risk neuroblastoma shares the expression pattern of 104 genes. More importantly, our analyses identify the co-expression relationship and the role of these genes in multiple biological processes and signaling pathways strongly associated with this disease phenotype. Our approach proposes a group of genes and their biological functions to be further investigated as essential molecules and possible therapeutic targets of neuroblastoma regardless of the etiology of individual tumors.

Urine collection methods in precontinent children treated at the paediatric emergency department.

AIM: To describe the urine collection methods used in precontinent children presenting at the Paediatric Emergency Department (PED) and compare results and contamination rates. METHODS: Retrospective observational cohort study that included 1678 urine cultures collected in infants <24 months of age between January 2016 and December 2019. Urine cultures were compared based on collection technique, sex and patient age. RESULTS: In total, 60.4% of samples were collected by clean-catch urine collection (CCUC), 26.4% by urethral catheterisation (UC) and 13.2% by urine bag (UB). Contamination rates were 2.9% (95% CI 1.3, 4.4) for UC, 11.3% (95% CI 9.3, 13.2) for CCUC and 23.4% (95% CI 17.8, 29.0) for UB. Significant differences in contamination rates were found between UC and CCUC in the 6-12-month age group (1.9% [95% CI 0.0-4.0] versus 12.0% [95% CI 7.2-16.8] [p < 0.0009]), and between UC and UB for all ages. CONCLUSIONS: CCUC is the most common method for urine culture collection in infants <24 months of age at the PED in our centre. UC has the lowest contamination rates, but significant differences were only observed between CCUC and UC in the 6-12-month age group. CCUC is a non-invasive alternative for urine collection in infants.

[Comparison of the outcome of orthodontic dental movement using aligners versus fixed orthodontics: a review]. / Comparación del resultado del movimiento dentario ortodóncico usando alineadores versus ortodoncia fija: una revisión.

Introduction: The aligners have become a preferred alternative in terms of orthodontic treatment, surpassing the choice of conventional brackets, due to the comfort and aesthetics that the aligners represent, however, the final result with this system continues to be a controversial issue. The objective of this review was to verify the effectiveness of the treatment completed with aligners versus conventional brackets. Materials and methods: An exhaustive search was carried out in the PubMed, ScienceDirect, Scopus and Embase databases up to the date of January 5, 2023, including comparative studies that evaluated the final result and treatment time of the aligners compared to conventional braces. Two researchers carefully selected the articles evaluated and analyzed different key topics on the subject. Results: In this study, 8 articles were included, according to the studies, the vast majority did not find significant differences in completion between the compared groups. In addition to this, the treatment time in 3 articles was faster with aligners, in 3 articles it was faster with conventional brackets, and in one article no significant difference was found, so it is not yet established whether a system is faster than the other. Conclusions: Both aligners and conventional fixed orthodontics had good results at the end of orthodontic treatment and presented a similar treatment time, however, the majority of treated cases belonged to malocclusions of medium to low complexity, so these cannot be generalized. findings to all malocclusions.

Vertical and In-Plane Electronic Transport of Graphene Nanoribbon/Nanotube Heterostructures.

All-carbon systems have proven to present interesting transport properties and are often used in electronic devices. Motivated by recent resonant responses measured on graphene/fullerene junction, we propose coupled nanoribbons/carbon-nanotube heterostructures for use as charge filters and to allow tuned transport. These hybrid systems are engineered as a four-terminal device, and we explore multiple combinations of source and collector leads. The armchair-edge configuration results in midgap states when the transport is carried through top/bottom terminals. Such states are robust against the lack of perfect order on the tube and are revealed as sharp steps in the characteristic current curves when a bias potential is turned on. The zigzag-edge systems exhibit differential negative resistance, with features determined by the details of the hybrid structures.

Lack of age-related mosaic loss of W chromosome in long-lived birds.

Females and males often exhibit different survival in nature, and it has been hypothesized that sex chromosomes may play a role in driving differential survival rates. For instance, the Y chromosome in mammals and the W chromosome in birds are often degenerated, with reduced numbers of genes, and loss of the Y chromosome in old men is associated with shorter life expectancy. However, mosaic loss of sex chromosomes has not been investigated in any non-human species. Here, we tested whether mosaic loss of the W chromosome (LOW) occurs with ageing in wild birds as a natural consequence of cellular senescence. Using loci-specific PCR and a target sequencing approach we estimated LOW in both young and adult individuals of two long-lived bird species and showed that the copy number of W chromosomes remains constant across age groups. Our results suggest that LOW is not a consequence of cellular ageing in birds. We concluded that the inheritance of the W chromosome in birds, unlike the Y chromosome in mammals, is more stable.

BAX But Not BCL2 Is Necessary for Apoptosis in Neuroblastoma Cells Treated With Casiopeína® IIIia.

BACKGROUND/AIM: The emerging antineoplastics Casiopeínas® induce uncoupling of the respiratory chain, production of reactive oxygen species (ROS), entry of Bax into mitochondria, and exit of Ca2+ and Bcl-2 from them, leading to apoptosis. This study aimed to elucidate whether BAX and BCL2 are necessary for apoptosis. MATERIALS AND METHODS: We silenced BAX and BCL2 by CRISPR-Cas9, assessed ROS and calcium retention capacity (CRC) by spectrofluorometry, and caspase-3 with inmunoblotting in neuroblastoma (NB) cells and 3T3-L1 fibroblasts treated with cisplatin and Casiopeína IIIia (CasIIIia). RESULTS: We observed an increase in O2•- production only in BCL2KO NB cells treated with cisplatin (three-fold) and CasIIIia (five-fold), whereas the production of H2O2 in BCL2KO NB cells treated with cisplatin and CasIIIia increased five-fold and three-fold, respectively. The baseline calcium-retention capacity (CRC) was 1.7 relative fluorescence units (RFU) in both cell types. In BAXKO, cisplatin and CasIIIia increased CRC to ~2.3 RFU, and in BCL2KO, they decreased CRC to ~1.1 RFU. We did not detect caspase-3 in BAXKO NB cells. CONCLUSION: Only BAX is essential for CasIIIia-induced apoptosis.

Viruses and Mycoplasma pneumoniae are the main etiological agents of community-acquired pneumonia in hospitalized pediatric patients in Spain.

OBJECTIVES: To describe the etiology of community-acquired pneumonia (CAP) in hospitalized children in Spain and analyze the predictors of the etiology. HYPOTHESIS: The different etiological groups of pediatric CAP are associated with different clinical, radiographic, and analytical data. DESIGN: Observational, multicenter, and prospective study. PATIENT SELECTION: This study included children aged 1 month to 17 years with CAP, who were hospitalized between April 2012 and May 2019. METHODS: An extensive microbiological workup was performed. The clinical, radiographic, and analytical parameters were analyzed for three etiological groups. RESULTS: Among the 495 children included, at least one causative pathogen was identified in 262 (52.9%): pathogenic viruses in 155/262 (59.2%); atypical bacteria (AB), mainly Mycoplasma pneumonia, in 84/262 (32.1%); and typical bacteria (TyB) in 40/262 (15.3%). Consolidation was observed in 89/138 (64.5%) patients with viral CAP, 74/84 (88.1%) with CAP caused by AB, and 40/40 (100%) with CAP caused by TyB. Para-pneumonic pleural effusion (PPE) was observed in 112/495 (22.6%) patients, of which 61/112 (54.5%) presented a likely causative pathogen: viruses in 12/61 (19.7%); AB in 23/61 (37.7%); and TyB in 26/61 (42.6%). Viral etiology was significantly frequent in young patients and in those with low oxygen saturation, wheezing, no consolidation, and high lymphocyte counts. CAP patients with AB as the etiological agent had a significantly longer and less serious course as compared to those with other causative pathogens. CONCLUSIONS: Viruses and M. pneumoniae are the main causes of pediatric CAP in Spain. Wheezing, young age, and no consolidation on radiographs are indicative of viral etiology. Viruses and AB can also cause PPE. Since only a few cases can be directly attributed to TyB, the indications for antibiotics must be carefully considered in each patient.

Dynamics of Reverse Transcription-Polymerase Chain Reaction and Serologic Test Results in Children with SARS-CoV-2 Infection.

OBJECTIVES: To determine the time to reverse transcription-polymerase chain reaction (RT-PCR) negativity after the first positive RT-PCR test, factors associated with longer time to RT-PCR negativity, proportion of children seroconverting after proven severe acute respiratory syndrome coronavirus 2 infection, and factors associated with the lack of seroconversion. STUDY DESIGN: The Epidemiological Study of Coronavirus in Children of the Spanish Society of Pediatrics is a multicenter study conducted in Spanish children to assess the characteristics of coronavirus disease 2019. In a subset of patients, 3 serial RT-PCR tests on nasopharyngeal swab specimens were performed after the first RT-PCR test, and immunoglobulin G serology for severe acute respiratory syndrome coronavirus 2 antibodies was performed in the acute and follow-up (<14 and ≥14 days after diagnosis) phase. RESULTS: In total, 324 patients were included in the study. The median time to RT-PCR negativity was 17 days (IQR, 8-29 days), and 35% of patients remained positive more than 4 weeks after the first RT-PCR test. The probability of RT-PCR negativity did not differ across groups defined by sex, disease severity, immunosuppressive drugs, or clinical phenotype. Globally, 24% of children failed to seroconvert after infection. Seroconversion was associated with hospitalization, persistence of RT-PCR positivity, and days of fever. CONCLUSIONS: Time to RT-PCR negativity was long, regardless of the severity of symptoms or other patient features. This finding should be considered when interpreting RT-PCR results in a child with symptoms, especially those with mild symptoms. Seroprevalence and postimmunization studies should consider that 11 in 4 infected children fail to seroconvert.

Success rate of infrazygomatic miniscrews considering their design and insertion techniques. A review.

Miniscrews offer the possibility of performing dental movements, minimizing unwanted side effects and enhancing effectiveness. Extra-alveolar miniscrews are ideal as they provide excellent primary stability and avoid anatomical structures. However, in some cases the primary stability is lost before achieving the success of the mechanics used and thus, the most likely causes of failure should be determined. The purpose of this review was to analyze the success rate of infrazygomatic miniscrews, considering their design and the insertion techniques used. Data collection of this literature review was carried out by searching PubMed, Wiley, Google Scholar sites, SCIELO, Elsevier and Dialnet for publication made from 2003 to June 2022. The search was carried out on June 10th, 2022 and the following keywords were used; infrazygomatic crest, miniscrews, anchorage and stability. Different topics were analyzed and discussed highlighting their clinical relevance. After analyzing the 798 articles, 566 were excluded. The remaining articles were re-analyzed and 153 articles were excluded for the title or abstract and 33 articles were excluded for the methodology. Finally, 46 items remained. After thoroughly analyzing all the articles included, this study concluded that the alloy of the miniscrew (stainless steel or titanium), perforation of the maxillary sinus and the placement area (adhered mucosa or mobile mucosa) do not influence the survival of the miniscrew. The evidence also indicates that the percentage of failure is lower in infrazygomatic compared to intraradicular miniscrews. Orthodontists can confidently and safely include infrazygomatic miniscrew in different orthodontic procedures.

Los minitornillos ofrecen la posibilidad de realizar movimientos dentales, lo que minimiza los efectos secundarios no deseados y potencia la eficacia. Los minitornillos extraalveolares son ideales ya que proporcionan una excelente estabilidad primaria y evitan estructuras anatómicas. Sin embargo, en algunos casos se pierde la estabilidad primaria antes de lograr el éxito de la mecánica utilizada y, por lo tanto, se deben determinar las causas más probables de la falla. El propósito de esta revisión fue analizar la tasa de éxito de los minitornillos infracigomáticos, considerando su diseño y las técnicas de inserción utilizadas. La recolección de datos de esta revisión bibliográfica se realizó mediante la búsqueda en PubMed, Wiley, sitios de Google Scholar, SciELO, Elsevier y Dialnet para publicaciones realizadas desde 2003 hasta junio de 2022. La búsqueda se realizó el 10 de junio de 2022 y se utilizaron las siguientes palabras clave: cresta infracigomática, minitornillos, anclaje y estabilidad. Se analizaron y discutieron diferentes temas destacando su relevancia clínica. Tras analizar los 798 artículos, se excluyeron 566. Los restantes fueron reanalizados y 153 resultaron excluidos por el título o resumen, y 33 por la metodología. Finalmente, quedaron 46 artículos. Tras analizar exhaustivamente todos los artículos incluidos, este estudio concluye que la aleación del minitornillo (acero inoxidable o titanio), la perforación del seno maxilar y la zona de colocación (mucosa adherida o mucosa móvil) no influyen en la supervivencia del minitornillo. La evidencia también indica que el porcentaje de fracaso es menor en los minitornillos infracigomáticos en comparación con los intrarradiculares. Los ortodoncistas pueden incluir con confianza y seguridad minitornillos infracigomáticos en diferentes procedimientos de ortodoncia.

Rats exhibit age-related mosaic loss of chromosome Y.

Mosaic loss of the Y chromosome (LOY) is the most frequent chromosomal aberration in aging men and is strongly correlated with mortality and disease. To date, studies of LOY have only been performed in humans, and so it is unclear whether LOY is a natural consequence of our relatively long lifespan or due to exposure to human-specific external stressors. Here, we explored whether LOY could be detected in rats. We applied a locus-specific PCR and target sequencing approach that we used as a proxy to estimate LOY in 339 samples covering eleven tissues from young and old individuals. We detected LOY in four tissues of older rats. To confirm the results from the PCR screening, we re-sequenced 60 full genomes from old rats, which revealed that the Y chromosome is the sole chromosome with low copy numbers. Finally, our results suggest that LOY is associated with other structural aberrations on the Y chromosome and possibly linked to the mosaic loss of the X chromosome. This is the first report, to our knowledge, demonstrating that the patterns of LOY observed in aging men are also present in a rodent, and conclude that LOY may be a natural process in placental mammals.

Evaluación de la resistencia de genotipos de Passifloras a Fusarium solani f.sp. passiflorae en granadilla / Evaluation of the resistance of Passifloras genotypes to Fusarium solani f.sp. passiflorae in granadilla / Avaliação da resistência de genótipos de Passifloras a Fusarium solani f.sp. passiflorae em granadilla

RESUMEN En Colombia la enfermedad más limitante en el cultivo de la granadilla es la pudrición del cuello causada por Fusarium solani f.sp. passiflorae. Este patógeno afecta a las plantas en todos los estados de desarrollo en la base de los tallos y causa la muerte en un corto tiempo. El objetivo de esta investigación fue evaluar la resistencia en dieciocho especies de la familia Passifloraceae a la pudrición del cuello. Para este experimento se utilizó un aislamiento altamente virulento según los resultados de las pruebas de patogenicidad con plantas de granadilla. Según los resultados después de la inoculación con el patógeno once especies de Passifloras fueron susceptibles, en contraste cinco genotipos de maracuyás (PE10, PE11, PE13, PE15 y PE17) y dos cholupas (PM02 y PM04), fueron resistentes no manifestaron síntomas. Se recomienda utilizar las especies resistentes para el mejoramiento genético de esta especie o emplearlos como porta injertos.

ABSTRACT In Colombia the most limiting disease in the cultivation of granadilla is neck rot caused by Fusarium solani f.sp, passiflorae. This pathogen affects plants in all stages of development at the base of the stems and causes death in a short time. The objective of this research was to evaluate the resistance in eighteen species of the Passifloraceae family to neck rot. For this experiment, a highly virulent isolate was used according to the results of pathogenicity tests with passion fruit plants. According to the results after inoculation with the pathogen, eleven Passifloras species were susceptible, in contrast, five passion fruit genotypes (PE10, PE11, PE13, PE15 and PE17) and two cholupas (PM02 and PM04), were resistant and did not show symptoms. It is recommended to use resistant species for the genetic improvement of this species or to use them as rootstock.

RESUMO Na Colômbia, a doença mais limitante no cultivo do granadilla é a podridão do pescoço causada pelo Fusarium solani f.sp, passiflorae. Este patógeno afeta as plantas em todos os estágios de desenvolvimento na base dos caules e causa a morte em um curto espaço de tempo. O objetivo desta pesquisa era avaliar a resistência de dezoito espécies da família Passifloraceae à podridão do colar Para esta experiência, foi utilizado um isolado altamente virulento de acordo com os resultados dos testes de patogenicidade em plantas de maracujá. De acordo com os resultados após a inoculação com o patógeno onze espécies de Passifloraceae eram suscetíveis, em contraste, cinco genótipos de maracujá (PE10, PE11, PE13, PE15 e PE17) e dois cholupas (PM02 e PM04) eram resistentes e não apresentavam sintomas. Recomenda-se usar as espécies resistentes para o melhoramento genético desta espécie ou usá-las como porta-enxertos.

Urine collection methods for infants under 3 months of age in clinical practice.

BACKGROUND: Methods of urine collection used in precontinent children are a controversial issue. Definitive diagnosis of urinary tract infection (UTI) requires an uncontaminated urine culture. We aimed to describe methods used to collect urine for culture in infants under 3 months of age and compare results and contamination rates. METHODS: This retrospective observational cohort study included 721 urine cultures collected from infants <3 months of age at the Hospital Universitario Infanta Sofía, Madrid, between January 2016 and December 2019. Urine cultures were compared based on collection technique, sex, and patient age. RESULTS: Median patient age was 36 days and 54.6% were male. In total, 592 (82.1%) samples were collected using clean-catch urine stimulation technique (CCUST), 77 (10.7%) by urethral catheterization (UC) and 52 (7.2%) by urine bag (UB). Positive cultures were obtained in 11.7% (95% confidence interval [CI] 9.1, 14.3) of CCUST samples and in 28.6% (95% CI 18.5, 38.7) of UC samples (p<0.001). The contamination rate was 13.7% (95% CI 10.9, 16.4] for CCUST, 23.1% (95% CI 11.6, 34.6) for UB and 5.2% (95% CI 0.2, 10.2) for UC, with statistically significant differences (p=0.007) between UB and UC collection. CONCLUSIONS: CCUST is the most commonly used method in our hospital for collecting urine in infants younger than 3 months. The contamination rate of UC is lower but not significantly different to that of CCUST. Urine collection by CCUST serves as a non-invasive alternative to UC for diagnosis of UTI in infants under 3 months of age in routine clinical practice. Graphical abstract.

Expression Evolution of Ancestral XY Gametologs across All Major Groups of Placental Mammals.

Placental mammals present 180 million-year-old Y chromosomes that have retained a handful of dosage-sensitive genes. However, the expression evolution of Y-linked genes across placental groups has remained largely unexplored. Here, we expanded the number of Y gametolog sequences by analyzing ten additional species from previously unexplored groups. We detected seven remarkably conserved genes across 25 placental species with known Y repertoires. We then used RNA-seq data from 17 placental mammals to unveil the expression evolution of XY gametologs. We found that Y gametologs followed, on average, a 3-fold expression loss and that X gametologs also experienced some expression reduction, particularly in primates. Y gametologs gained testis specificity through an accelerated expression decay in somatic tissues. Moreover, despite the substantial expression decay of Y genes, the combined expression of XY gametologs in males is higher than that of both X gametologs in females. Finally, our work describes several features of the Y chromosome in the last common mammalian ancestor.

Viviparous Reptile Regarded to Have Temperature-Dependent Sex Determination Has Old XY Chromosomes.

The water skinks Eulamprus tympanum and Eulamprus heatwolei show thermally induced sex determination where elevated temperatures give rise to male offspring. Paradoxically, Eulamprus species reproduce in temperatures of 12-15 °C making them outliers when compared with reptiles that use temperature as a cue for sex determination. Moreover, these two species are among the very few viviparous reptiles reported to have thermally induced sex determination. Thus, we tested whether these skinks possess undetected sex chromosomes with thermal override. We produced transcriptome and genome data for E. heatwolei. We found that E. heatwolei presents XY chromosomes that include 14 gametologs with regulatory functions. The Y chromosomal region is 79-116 Myr old and shared between water and spotted skinks. Our work provides clear evidence that climate could be useful to predict the type of sex determination systems in reptiles and it also indicates that viviparity is strictly associated with sex chromosomes.

Post-transcriptional regulation of Rad51c by miR-222 contributes cellular transformation.

DNA repair inhibition has been described as an essential event leading to the initiation of carcinogenesis. In a previous study, we observed that the exposure to metal mixture induces changes in the miR-nome of the cells that was correlated with the sub-expression of mRNA involved in processes and diseases associated with metal exposure. From this analysis, one of the miRNAs that shows changes in its expression is miR-222, which is overexpressed in various cancers associated with exposure to metals. In silico studies showed that a possible target for the microRNA-222 could be Rad 51c, a gene involved in the double-stranded DNA repair. We could appreciate that up-regulation of miR-222 reduces the expression both gene and as a protein expression of Rad51c by RT-PCR and immunoblot, respectively. A luciferase assay was performed to validate Rad51c as miR-222 target. Neutral comet assay was performed in order to evaluate DNA double-strand breaks under experimental conditions. Here, we demonstrate that miR-222 up-regulation, directly regulates Rad51c expression negatively, and impairs homologous recombination of double-strand break DNA repair during the initiation stage of cell transformation. This inhibition triggers morphological transformation in a two-stage Balb/c 3T3 cell assay, suggesting that this small RNA acts as an initiator of the carcinogenesis process.

As-Cd-Pb Mixture Induces Cellular Transformation via Post-Transcriptional Regulation of Rad51c by miR-222.

BACKGROUND/AIMS: Exposure to heavy metals is today a threat to society. The understanding of the molecular processes related to diseases related to exposure to metals mixture involve changes in the expression of microRNAs. Changes on microRNAs expression may alter several cellular processes, among them, DNA repair inhibition has been described as an essential event leading to the initiation of metal-induced carcinogenesis. METHODS: We evaluate the miR-222 expression in the two-stage transformation Balb/c 3T3 cell assay treated with As-Cd-Pb mixture. RESULTS: We could appreciate that up-regulation of miR-222 reduces the expression both gene and as a protein expression of Rad51c by RT-PCR and immunoblot, respectively. CONCLUSION: Here, we demonstrate that the mixture of As-Cd-Pb at epidemiologically relevant concentrations induces miR-222 up-regulation, which directly negatively regulates Rad51c expression and impairs homologous recombination of DNA during the initiation stage of cell transformation. This inhibition triggers morphological transformation in a murine two-stage Balb/c 3T3 cell assay, suggesting that this small RNA acts as an initiator of the carcinogenesis process.

Deciphering Ancestral Sex Chromosome Turnovers Based on Analysis of Male Mutation Bias.

The age of sex chromosomes is commonly obtained by comparing the substitution rates of XY gametologs. Coupled with phylogenetic reconstructions, one can refine the origin of a sex chromosome system relative to specific speciation events. However, these approaches are insufficient to determine the presence and duration of ancestral sex chromosome systems that were lost in some species. In this study, we worked with genomic and transcriptomic data from mammals and squamates and analyzed the effect of male mutation bias on X-linked sequences in these groups. We searched for signatures indicating whether monotremes shared the same sex chromosomes with placental mammals or whether pleurodonts and acrodonts had a common ancestral sex chromosome system. Our analyses indicate that platypus did not share the XY chromosomes with placental mammals, in agreement with previous work. In contrast, analyses of agamids showed that this lineage maintained the pleurodont XY chromosomes for several million years. We performed multiple simulations using different strengths of male mutation bias to confirm the results. Overall, our work shows that variations in substitution rates due to male mutation bias could be applied to uncover signatures of ancestral sex chromosome systems.

Symmetry-protected metallic and topological phases in penta-materials.

We analyze the symmetry and topological features of a family of materials closely related to penta-graphene, derived from it by adsorption or substitution of different atoms. Our description is based on a novel approach, called topological quantum chemistry, that allows to characterize the topology of the electronic bands, based on the mapping between real and reciprocal space. In particular, by adsorption of alkaline (Li or Na) atoms we obtain a nodal line metal at room temperature, with a continuum of Dirac points around the perimeter of the Brillouin zone. This behavior is also observed in some substitutional derivatives of penta-graphene, such as penta-PC2. Breaking of time-reversal symmetry can be achieved by the use of magnetic atoms; we study penta-MnC2, which also presents spin-orbit coupling and reveals a Chern insulator phase. We find that for this family of materials, symmetry is the source of protection for metallic and nontrivial topological phases that can be associated to the presence of fractional band filling, spin-orbit coupling and time-reversal symmetry breaking.