The extracellular wall-bound β-N-acetylglucosaminidase from Lactobacillus casei is involved in the metabolism of the human milk oligosaccharide lacto-N-triose

Human milk oligosaccharides (HMOs) are considered to play a key role in establishing and maintaining the infant gut microbiota. Lacto-N-triose forms part of both type 1 and type 2 HMOs and also of the glycan moieties of glycoproteins. Upstream of the previously characterized gene cluster involved in lacto-N-biose and galacto-N-biose metabolism from Lactobacillus casei BL23, there are two genes, bnaG and manA, encoding a β-N-acetylglucosaminidase precursor and a mannose-6-phosphate isomerase, respectively. In this work, we show that L. casei is able to grow in the presence of lacto-N-triose as a carbon source. Inactivation of bnaG abolished the growth of L. casei on this oligosaccharide, demonstrating that BnaG is involved in its metabolism. Interestingly, whole cells of a bnaG mutant were totally devoid of β-N-acetylglucosaminidase activity, suggesting that BnaG is an extracellular wall-attached enzyme. In addition to hydrolyzing lacto-N-triose into N-acetylglucosamine and lactose, the purified BnaG enzyme also catalyzed the hydrolysis of 3′-N-acetylglucosaminyl-mannose and 3′-N-acetylgalactosaminyl-galactose. L. casei can be cultured in the presence of 3′-N-acetylglucosaminyl-mannose as a carbon source, but, curiously, the bnaG mutant strain was not impaired in its utilization. These results indicate that the assimilation of 3′-N-acetylglucosaminyl-mannose is independent of BnaG. Enzyme activity and growth analysis with a manA-knockout mutant showed that ManA is involved in the utilization of the mannose moiety of 3′-N-acetylglucosaminyl-mannose. Here we describe the physiological role of a β-N-acetylglucosaminidase in lactobacilli, and it supports the metabolic adaptation of L. casei to the N-acetylglucosaminide-rich gut niche.This work was financed by funds of the Spanish Ministry for Economy and Competitiveness (MINECO)/FEDER through projects AGL2014-52996-C2 (1-R and 2-R) and of the Valencian government through project ACOMP/2012/030. G.N.B. was supported by a predoctoral fellowship from the Carolina Foundation and Argentinian Ministry of Education.Peer reviewe

Nanostructured Chitosan-Based Biomaterials for Sustained and Colon-Specific Resveratrol Release

In the present work, we demonstrate the preparation of chitosan-based composites as vehicles of the natural occurring multi-drug resveratrol (RES). Such systems are endowed with potential therapeutic effects on inflammatory bowel diseases (IBD), such as Crohn’s disease (CD) and ulcerative colitis, through the sustained colonic release of RES from long-lasting mucoadhesive drug depots. The loading of RES into nanoparticles (NPs) was optimized regarding two independent variables: RES/polymer ratio, and temperature. Twenty experiments were carried out and a Box–Behnken experimental design was used to evaluate the significance of these independent variables related to encapsulation efficiency (EE). The enhanced RES EE values were achieved in 24 h at 39 °C and at RES/polymer ratio of 0.75:1 w/w. Sizes and polydispersities of the optimized NPs were studied by dynamic light scattering (DLS). Chitosan (CTS) dispersions containing the RES-loaded NPs were ionically gelled with tricarballylic acid to yield CTS-NPs composites. Macro- and microscopic features (morphology and porosity studied by SEM and spreadability), thermal stability (studied by TGA), and release kinetics of the RES-loaded CTS-NPs were investigated. Release patterns in simulated colon conditions for 48 h displayed significant differences between the NPs (final cumulative drug release: 79–81%), and the CTS-NPs composites (29–34%)

Dyadic analysis of emotional intimate partner violence: an estimation of dyadic patterns and influencing individual, family, and couple factors

Background: The co-occurrence of intimate partner violence (IPV) has been usually determined by co-occurring reports of perpetration and victimization, but still not considering the interdependence of both members'' reports. The current research aimed to analyze the dyadic patterns of IPV perpetration and victimization, considering the influence of sexism toward women, family functioning, and relationship duration. Method: The sample involved 242 couples heterosexual couples. The ActorPartner Interdependence Model was calculated to analyze the violent dynamics in the couple. For males, a couple-oriented pattern was identified, but for females the pattern was between couple-oriented and actor-only pattern. Results: These patterns remained stable when personal, family, and relational variables were included in the model, which present different effects on perpetration and victimization: males'' hostile sexism predicted higher levels of male IPV perpetration and victimization as well as higher levels of male and female victimization via male perpetration. Regarding family functioning, higher family good functioning predicted lower IPV perpetration in males and lower victimization in females. Finally, relationship length predicted higher levels of IPV perpetration of both couple members and higher victimization via perpetration. Conclusions: These results are discussed in light of the interdependence of couple members'' IPV perpetration and victimization. Antecedentes: La coocurrencia de la violencia en la pareja (IPV) suele establecerse con el informe de ambos miembros de la pareja, pero rara vez se tiene en cuenta la interdependencia entre sus respuestas. El objetivo del trabajo ha sido analizar los patrones diádicos de la IPV teniendo en cuenta el sexismo hacia las mujeres, la funcionalidad familiar y la duración de la relación. Método: La muestra estaba compuesta por 242 parejas heterosexuales, utilizándose el Actor-Partner Interdependence Model para analizar los patrones diádicos. En el caso de los hombres se identificó el patrón orientado a la díada, mientras que el patrón de las mujeres parecía encontrarse entre el orientado a la díada y al actor. Resultados: Estos patrones se mantuvieron tras haber incluido el sexismo, el funcionamiento familiar y la duración de la relación: el sexismo hostil de los hombres predecía mayores niveles de perpetración en ellos y de victimización en ambos a través de la perpetración de los hombres. El buen funcionamiento familiar predecía una menor perpetración en los hombres y victimización en las mujeres. Finalmente la duración de la relación predijo mayores niveles de perpetración y victimización a través de la perpetración en ambos miembros de la pareja. Conclusiones: Se discuten los resultados a partir de la interdependencia de la IPV de ambos miembros de la pareja

Aplicación de la Dignity Therapy en pacientes oncológicos en situación avanzada.

The Dignity Therapy was designed to cope with suffering, psychosocial and existential distress among patients with advanced or terminal disease. This paper is a cuasi experimental study with sixteen cancer patients in advanced stages of the disease. The aim of the study is to apply, in our context, the Dignity Therapy as a proposal for intervention for the relief of suffering such patients, in order to assess whether there is pre and post intervention changes. The results of Wilcoxon’s test, show statistically significant differences comparing the pre-intervention and the post-intervention for the variables Anxiety and Well-being. Nevertheless, statistically significant differences are not observed for the variables Depression and Serenity. The subjects agreed to participate voluntarily, and all the subjects felt satisfied after the application of the Dignity Therapy and noted that they found it useful enough or very useful. These results suggest that it is an appropriate therapeutic approach to these patients.La Dignity Therapy fue diseñada para hacer frente al sufrimiento, la angustia psicosocial y existencial entre los pacientes con enfermedad avanzada o terminal. El presente trabajo es un estudio cuasi experimental con dieciséis pacientes oncológicos en estado avanzado de la enfermedad. El objetivo del estudio fue aplicar, en nuestro contexto, la Dignity Therapy como propuesta de intervención para el alivio del sufrimiento a dichos pacientes, evaluando si existen cambios pre y post intervención. Tras obtener resultados mediante la prueba de Wilcoxon, se obtienen diferencias estadísticamente significativas comparando la pre-intervención y la post-intervención para las variables Ansiedad y Bienestar. Sin embargo, no se observa diferencias estadísticamente significativas para las variables Depresión y Serenidad. Los sujetos accedieron a participar de forma voluntaria y todos se sintieron satisfechos tras la aplicación de la Dignity Therapy, destacando que les había resultado bastante útil o muy útil. Estos resultados apuntan a que se trata de un acercamiento terapéutic

Design of an X-ray irradiator based on a standard imaging X-ray tube with FLASH dose-rate capabilities for preclinical research

This work was funded by Comunidad de Madrid under project B2017/BMD-3888 PRONTO-CM "Protontherapy and nuclear techniques for oncology". Support by the Spanish Government (RTI 2018-098868-B-I00, RTC-2015-3772-1, XPHASE-LASER, CPP 2021-008751 NEW-MBI) , as well as European Regional and Resilience Funds, and the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 793576 (CAP-PERAM) is acknowledged. This is a contribution for the Moncloa Campus of International Excellence, "Grupo de Física Nuclear-UCM", Ref. 910059. Part of the calculations of this work were performed in the "Cluster de Calculo para Técnicas Físicas", funded in part by UCM and in part by EU Regional Funds.We propose a new concept of small animal X-ray irradiator based on a conventional imaging X-ray tube for preclinical research. In this work we assessed its feasibility to deliver FLASH dose rates. Our design puts the imaging X-ray tube into a shielded cabinet, which makes the system affordable and suitable to use without disruption in existing laboratories and with minimum regulatory burden. Two conventional 150 kVp X-ray tubes were characterized with Gafchromic films for dose rates and dose uniformity. Monte Carlo simulations were also performed to model the irradiator, and the efficiencies of the tube and dose rates (with and without additional filtration) were calculated and compared with measurements. The feasibility of achieving ultra-high dose rates was determined from the rating charts provided by the manufacturer and measurements. The small animal irradiator proposed in this work was able to deliver conventional dose rate irradiation (0.5-1 Gy/min) at 150 kVp at 20 cm distance with minimum amount of filtration. FLASH irradiations (a 10 Gy dose delivered at >40 Gy/s) were also possible at the maximum capabilities of the tubes by placing the samples at the closest possible distances from the sources. A first prototype has already been built and characterized.Depto. de Estructura de la Materia, Física Térmica y ElectrónicaFac. de Ciencias FísicasTRUEComunidad de MadridGobierno de EspañaEuropean Regional and Resilience FundsEuropean Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grantMoncloa Campus of International Excellence, "Grupo de Fisica Nuclear-UCM"Universidad Complutense de Madrid (UCM)EU Regional Fundspu

Design of beams and slabs of self-compacting concrete fittings working to flexion

Numerous experimental studies indicate that addition of fibers substantially increases the shear strength of concrete and mortar beams, and that stirrups and reinforcing fibers combination can be really effective. When steel fibers are used as a supplement or substitute for vertical stirrups or folded bars, we obtain the following characteristics: The fibers are distributed randomly by the volume of the concrete with a closer separation than can be achieved with the Armor; The tensile strength at first crack, the ultimate tensile strength and the shear friction resistance are increased

The beginning of high mountain occupations in the Pyrenees. Human settlements and mobility from 10,500 cal BP to 4500 cal BP

During the last two decades, the archaeological research carried out in the Pyrenees challenged the traditional images of the past in this mountain area. The archaeological sequence of the range goes back and sites like Balma Margineda, treated until recently as an exception, now are seen as part of more global process. Actual data suggest that main valleys of the Pyrenean frequented by humans at the end of the last glacial period, with sites slightly over 1000 o.s.l. After the Younger Dryas, the human presence ascended to alpine and subalpine areas, in accordance with current archaeological data. The Neolisitation process was early in some hillsides, with intense remains of farming and pastoralism in many sites from dated in the second half of the 6th millennia cal BC. Human settlements like Coro Tracito, Els Trocs and El Sardo confirm the full introduction of agrarian activity in the central part of the Pyrenees between 5300 and 4600 cal BC. After 3500/3300 cal BC the indices oh sheepherding rises to alpine areas, with an abrupt increase of known archaeological sites in alpine areas, above the current timberline. This phenomena, as well as the signs of anthropic disturbance of the alpine environment in sedimentary sequences, suggests a more stable and ubiquitous human presence, probably largely associated with the development of mobile herding practices