EXOSC10 is required for RPA assembly and controlled DNA end resection at DNA double-strand breaks

The exosome is a ribonucleolytic complex that plays important roles in RNA metabolism. Here we show that the exosome is necessary for the repair of DNA double-strand breaks (DSBs) in human cells and that RNA clearance is an essential step in homologous recombination. Transcription of DSB-flanking sequences results in the production of damage-induced long non-coding RNAs (dilncRNAs) that engage in DNA-RNA hybrid formation. Depletion of EXOSC10, an exosome catalytic subunit, leads to increased dilncRNA and DNA-RNA hybrid levels. Moreover, the targeting of the ssDNA-binding protein RPA to sites of DNA damage is impaired whereas DNA end resection is hyper-stimulated in EXOSC10-depleted cells. The DNA end resection deregulation is abolished by transcription inhibitors, and RNase H1 overexpression restores the RPA recruitment defect caused by EXOSC10 depletion, which suggests that RNA clearance of newly synthesized dilncRNAs is required for RPA recruitment, controlled DNA end resection and assembly of the homologous recombination machinery.España, Ministerio de Economía y Competitividad R + D + I project grant SAF2016-74855-P to P.

The TRPM8 antagonist RGM8-51 displays analgesic activity in different pain models

TRPM8 channels are overexpressed in sensory neurons after nerve injury or inflammation, resulting in enhanced sensitivity (allodynia and hyperalgesia) to physical stimulation, and have been implicated in migraine, but the interest of TRPM8 antagonists is still a matter of controversy (1,2). The aim of our work was to evaluate the analgesic activity of a TRPM8 antagonist, RGM8-51, in different pain models, looking for similarities and differences with other antagonists. To this end, we used the mouse oxaliplatin-induced peripheral neuropathy, the chronic constriction injury of the rat sciatic nerve (CCI) and mouse NTG-induced migraine-like models. Compound RGM8- 51 reduces the cold allodynia induced by oxaliplatin, from 15 to 60 min after administration (0.1-1 μg, i.pl.), decreases the nocifensive responses to cold, heat and mechanical stimuli in the CCI model (10 μg, i.pl., 30 mg/Kg, i.p.), and relief chronic pain associated to migraine in mouse, in a sex-dependent manner (10 or 30 mg/Kg, i.v.). The β–lactam derivative RGM8-51 not only has analgesic activity in all assayed animal models, but also seems to have a different mode of interaction with the TRPM8 channel than other antagonists, as suggested by docking studies

DD04107-Derived neuronal exocytosis inhibitor peptides: Evidences for synaptotagmin-1 as a putative target

The analgesic peptide DD04107 (Pal-EEMQRR-NH2) and its acetylated analogue inhibit a-calcitonin gene-related peptide (a-CGRP) exocytotic release from primary sensory neurons. Examining the crystal structure of the SNARE-Synaptotagmin-1(Syt1) complex, we hypothesized that these peptides could inhibit neuronal exocytosis by binding to Syt1, hampering at least partially its interaction with the SNARE complex. To address this hypothesis, we first interrogate the role of individual side-chains on the inhibition of a-CGRP release, finding that E1, M3, Q4 and R6 residues were crucial for activity. CD and NMR conformational analysis showed that linear peptides have tendency to adopt a-helical conformations, but the results with cyclic analogues indicated that this secondary structure is not needed for activity. Isothermal titration calorimetry (ITC) measurements demonstrate a direct interaction of some of these peptides with Syt1-C2B domain, but not with Syt7-C2B region, indicating selectivity. As expected for a compound able to inhibit a-CGRP release, cyclic peptide derivative Pal-E-cyclo[EMQK]R-NH2 showed potent in vivo analgesic activity, in a model of inflammatory pain. Molecular dynamics simulations provided a model consistent with KD values for the interaction of peptides with Syt1-C2B domain, and with their biological activity. Altogether, these results identify Syt1 as a potential new analgesic target. © 202

Metabolic strategies for the degradation of the neuromodulator agmatine in mammals

Agmatine (1-amino-4-guanidinobutane), a precursor for polyamine biosynthesis, has been identified as an important neuromodulator with anticonvulsant, antineurotoxic and antidepressant actions in the brain. In this context it has emerged as an important mediator of addiction/satiety pathways associated with alcohol misuse. Consequently, the regulation of the activity of key enzymes in agmatine metabolism is an attractive strategy to combat alcoholism and related addiction disorders. Agmatine results from the decarboxylation of L-arginine in a reaction catalyzed by arginine decarboxylase (ADC), and can be converted to either guanidine butyraldehyde by diamine oxidase (DAO) or putrescine and urea by the enzyme agmatinase (AGM) or the more recently identified AGM-like protein (ALP). In rat brain, agmatine, AGM and ALP are predominantly localised in areas associated with roles in appetitive and craving (drug-reinstatement) behaviors. Thus, inhibitors of AGM or ALP are promising agents for the treatment of addictions. In this review, the properties of DAO, AGM and ALP are discussed with a view to their role in the agmatine metabolism in mammals

Micro-tomographic characterization of the root and canal system morphology of mandibular first premolars in a Chilean population

Indexación ScopusThis study aimed to analyze the root anatomy and root canal system morphology of mandibular first premolars in a Chilean population. 186 teeth were scanned using micro-computed tomography and reconstructed three-dimensionally. The root canal system morphology was classified using both Vertucci’s and Ahmed’s criteria. The radicular grooves were categorized using the ASUDAS system, and the presence of Tomes’ anomalous root was associated with Ahmed’s score. A single root canal was identified in 65.05% of teeth, being configuration type I according to Vertucci’s criteria and code 1MP1 according to Ahmed’s criteria. Radicular grooves were observed in 39.25% of teeth. The ASUDAS scores for radicular grooves were 60.75%, 13.98%, 12.36%, 10.22%, 2.15%, and 0.54%, from grade 0 to grade 5, respectively. The presence of Tomes’ anomalous root was identified only in teeth with multiple root canals, and it was more frequently associated with code 1MP1–2 of Ahmed’s criteria. The root canal system morphology of mandibular first premolars showed a wide range of anatomical variations in the Chilean population. Teeth with multiple root canals had a higher incidence of radicular grooves, which were closely related to more complex internal anatomy. Only teeth with multiple root canals presented Tomes’ anomalous root. © 2021, The Author(s).https://www-nature-com.recursosbiblioteca.unab.cl/articles/s41598-020-80046-

Effect of the presence of the plant growth promoting rhizobacterium (PGPR) Chryseobacterium balustinum Aur9 and salt stress in the pattern of flavonoids exuded by soybean roots

In this work we studied how biotic and abiotic stresses can alter the pattern of flavonoids exuded by Osumi soybean roots. A routine method was developed for the detection and characterization of the flavonoids present in soybean root exudates using HPLC-MS/MS. Then, a systematic screening of the flavonoids exuded under biotic stress, the presence of a plant growth promoting rhizobacterium, and salt stress was carried out. Results obtained indicate that the presence of Chryseobacterium balustinum Aur9 or 50 mM NaCl changes qualitatively the pattern of flavonoids exuded when compared to control conditions. Thus, in the presence of C. balustinum Aur9, soybean roots did not exude quercetin and naringenin and, under salt stress, flavonoids daidzein and naringenin could not be detected. Soybean root exudates obtained under saline conditions showed a diminished capacity to induce the expression of the nodA gene in comparison to the exudates obtained in the absence of salt. Moreover, lipochitooligosaccharides (LCOs) were not detected or weakly detected when Sinorhizobium fredii SMH12 was grown in the exudates obtained under salt stress conditions or under salt stress in the presence of C. balustinum Au9, respectively.Fil: Dardanelli, Marta Susana. Universidad de Sevilla. Facultad de Farmacia; España. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Manyani, Hamid. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: González Barroso, Sergio. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: Rodríguez Carvajal, Miguel A.. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: Gil Serrano, Antonio M.. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: Espuny, Maria R.. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: López Baena, Francisco Javier. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: Bellogín, Ramon A.. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: Megías, Manuel. Universidad de Sevilla. Facultad de Farmacia; EspañaFil: Ollero, Francisco J.. Universidad de Sevilla. Facultad de Farmacia; Españ

Melatonin MT1 and MT2 receptors in the ram reproductive tract

Some melatonin functions in mammals are exerted through MT1 and MT2 receptors. However, there are no reports of their presence in the reproductive tract of the ram, a seasonal species. Thus, we have investigated their existence in the ram testis, epididymis, accessory glands and ductus deferens. Real-time polymerase chain reaction (qPCR) revealed higher levels of m-RNA for both receptors in the testis, ampulla, seminal vesicles, and vas deferens, than in the other organs of the reproductive tract (p < 0.05). Western blot analyses showed protein bands compatible with the MT1 in the testis and cauda epididymis, and for the MT2 in the cauda epididymis and deferent duct. Immunohistochemistry analyses revealed the presence of MT1 receptors in spermatogonias, spermatocytes, and spermatids, and MT2 receptors in the newly-formed spermatozoa in the testis, whereas both receptors were located in the epithelial cells of the ampulla, seminal vesicles, and ductus deferens. Indirect immunofluorescence showed significant differences in the immunolocation of both receptors in spermatozoa during their transit in the epididymis. In conclusion, it was demonstrated that melatonin receptors are present in the ram reproductive tract. These results open the way for new studies on the molecular mechanism of melatonin and the biological significance of its receptors

The ALMA Frontier Fields Survey

CONTEXT: Dusty star-forming galaxies are among the most prodigious systems at high redshift (z > 1), characterized by high star-formation rates and huge dust reservoirs. The bright end of this population has been well characterized in recent years, but considerable uncertainties remain for fainter dusty star-forming galaxies, which are responsible for the bulk of star formation at high redshift and thus play a key role in galaxy growth and evolution. AIMS: In this first paper of our series, we describe our methods for finding high redshift faint dusty galaxies using millimeter observations with ALMA. METHODS: We obtained ALMA 1.1 mm mosaic images for three strong-lensing galaxy clusters from the Frontier Fields Survey, which constitute some of the best studied gravitational lenses to date. The ≈2′ × 2′ mosaics overlap with the deep HST WFC3/IR footprints and encompass the high magnification regions of each cluster for maximum intrinsic source sensitivity. The combination of extremely high ALMA sensitivity and the magnification power of these clusters allows us to systematically probe the sub-mJy population of dusty star-forming galaxies over a large surveyed area. RESULTS: We present a description of the reduction and analysis of the ALMA continuum observations for the galaxy clusters Abell 2744 (z = 0.308), MACS J0416.1-2403 (z = 0.396) and MACS J1149.5+2223 (z = 0.543), for which we reach observed rms sensitivities of 55, 59 and 71 μJy beam-1 respectively. We detect 12 dusty star-forming galaxies at S/N ≥ 5.0 across the three clusters, all of them presenting coincidence with near-infrared detected counterparts in the HST images. None of the sources fall close to the lensing caustics, thus they are not strongly lensed. The observed 1.1 mm flux densities for the total sample of galaxies range from 0.41 to 2.82 mJy, with observed effective radii spanning ≲0.̋05 to 0.̋37 ± 0.̋21 . The lensing-corrected sizes of the detected sources appear to be in the same range as those measured in brighter samples, albeit with possibly larger dispersion

Domestic violence. A look from adolescence

Introducción: la violencia intrafamiliar es la acción o la omisión por algún miembro de la familia en relación de poder que incluye el abuso físico, emocional, por descuido y sexual. Objetivo: describir el comportamiento de la violencia intrafamiliar según la percepción de adolescentes provenientes de familias disfuncionales. Métodos: se realizó un estudio descriptivo interpretativo de corte transversal con una estrategia mixta: análisis cuantitativo de variables y cualitativo de casos de familias disfuncionales pertenecientes a dos consultorios médicos de la familia del Policlínico “José Ramón León Acosta” de Santa Clara. Resultados: de las 164 familias registradas el 58,6% resultaron disfuncionales. Los adolescentes víctimas de violencia presentaron condiciones socio-económico-culturales regulares o malas (70,7%) y pertenecían a familias extensas (80,2%). Como formas represivas predominó el método de agresión verbal (100%): críticas, amenazas, gritos, insultos, peleas y burlas. Conclusiones: la violencia intrafamiliar constituye un serio problema de salud que afecta a las familias disfuncionales, su caracterización permitió reconocer como agresores más frecuentes a la madre, al padre y a los hermanos. Se deben realizar acciones de promoción de salud en la comunidad y de tipo preventivas con las familias que constituyen riesgo de violencia.Introduction: domestic violence is the act or omission by a member of the family in a power relationship that includes physical, emotional, carelessness and sexual abuse. Objective: to describe the behavior of intrafamily violence according to the perception of adolescents from dysfunctional families. Methods: a descriptive cross-sectional study with a mixed strategy was carried out: quantitative analysis of variables and qualitative of cases of dysfunctional families that are taken care by two family medical offices of the José Ramón León Acosta Polyclinic in Santa Clara. Results: of 164 registered families, 58.6% were dysfunctional. The adolescents’ victims of violence presented regular or bad socio-economic-cultural conditions (70.7%) and belonging to extended families (80.2%). As a repression forms, the method of verbal aggression prevailed (100%): criticism, threats, shouting, insults, fights and ridicule. Conclusions: intrafamily violence constitutes a serious health problem that affects dysfunctional families, its characterization allowed recognizing as more frequent aggressors the mother, father and siblings. Health promotion and preventive actions should be carried out in the community with families that constitute a risk of violence

Altered myogenesis and premature senescence underlie human TRIM32-related myopathy

TRIM32 is a E3 ubiquitin -ligase containing RING, B-box, coiled-coil and six C-terminal NHL domains. Mutations involving NHL and coiled-coil domains result in a pure myopathy (LGMD2H/STM) while the only described mutation in the B-box domain is associated with a multisystemic disorder without myopathy (Bardet-Biedl syndrome type11), suggesting that these domains are involved in distinct processes. Knock-out (T32KO) and knockin mice carrying the c.1465G > A (p.D489N) involving the NHL domain (T32KI) show alterations in muscle regrowth after atrophy and satellite cells senescence. Here, we present phenotypical description and functional characterization of mutations in the RING, coiled-coil and NHL domains of TRIM32 causing a muscle dystrophy. Reduced levels of TRIM32 protein was observed in all patient muscle studied, regardless of the type of mutation (missense, single amino acid deletion, and frameshift) or the mutated domain. The affected patients presented with variable phenotypes but predominantly proximal weakness. Two patients had symptoms of both muscular dystrophy and Bardet-Biedl syndrome. The muscle magnetic resonance imaging (MRI) pattern is highly variable among patients and families. Primary myoblast culture from these patients demonstrated common findings consistent with reduced proliferation and differentiation, diminished satellite cell pool, accelerated senescence of muscle, and signs of autophagy activation.Health Institute Carlos III PI16-01843 JR15/00042FEDER PI16-01843 JR15/00042Fundación Progreso y Salud, Junta de Andalucía PI-0085-2016Australian National Health and Medical Research Council (NHMRC) APP1122952 APP111751