Synergistic action of phage phiIPLA-RODI and lytic protein CHAPSH3b : a combination strategy to target Staphylococcus aureus biofilms

Staphylococcus aureus is considered a priority pathogen due to its increasing acquisition of antibiotic resistance determinants. Additionally, this microbe has the ability to form recalcitrant biofilms on different biotic and inert surfaces. In this context, bacteriophages and their derived lytic proteins may be a forward-looking strategy to help combat staphylococcal biofilms. However, these antimicrobials exhibit individual limitations that may be overcome by combining them with other compounds. This work investigates the combination of a phage-derived lytic protein, CHAPSH3b, and the virulent bacteriophage phiIPLA-RODI. The obtained results show the synergy between both antimicrobials for the treatment of 24-h-old S. aureus biofilms, with greater reductions in viable cell counts observed when phage and lysin are applied together compared to the individual treatments. Time-kill curves and confocal microscopy revealed that the fast antibacterial action of CHAPSH3b reduces the population up to 7 hours after initial exposure, which is subsequently followed by phage predation, limiting regrowth of the bacterial population. Moreover, at least 90% of bacteriophage insensitive mutants are susceptible to the lytic protein. Therefore, CHAPSH3b might help curtail the development of phage resistance during treatment. The combination of the lysin and phiIPLA-RODI also showed promising results in an ex vivo pig skin model of wound infection. Overall, the results of this study demonstrate that the combination of phage-derived lytic proteins and bacteriophages can be a viable strategy to develop improved antibiofilm products

Transcriptional activation of the proapoptotic bik gene by E2F proteins in cancer cells

AbstractBH3-only proteins are required for execution of apoptotic cell death. We have found that one of these proteins, Bik, is strongly induced in cancer cells treated with chemotherapeutic agents. Furthermore, we showed that chemotherapy-induced expression of bik is independent of p53. Consistent with its pro-apoptotic activity, blockade of bik expression reduces the adriamycin-mediated apoptotic cell death. We also found that the bik gene is transcriptionally activated by E2F proteins. Consistently, adriamycin induces the E2F-bik pathway. In addition, E2Fs transactivate bik by a p53-independent mechanism. Thus, our data indicate that transcriptional regulation of bik contributes to the efficient apoptotic response to chemotherapeutic agents

Reduction rate by decompression as a treatment of odontogenic cysts

Background: Odontogenic cysts are defined as those cysts that arise from odontogenic epithelium and occur in the tooth-bearing regions of the jaws. Cystectomy, marsupialization or decompression of odontogenic cyst are treatment approach to this pathology. The aim of this study was to evaluate the effectiveness of the decompression as the primary treatment of the cystic lesions of the jaws and them reduction rates involving different factors. Material and Methods: 23 patients with odontogenic cysts of the jaws, previously diagnosed by anatomical histopathology (follicular cysts (7) and radicular cysts (16)) underwent decompression as an initial treatment. Clinical examination and pre and post panoramic radiograph were measured and analyzed. In addition, data as gender, age, time reduction and location of the lesion were collected. Results: Significant results were obtained in relation to the location of lesions and the reduction rate (p<0.01). In a higher initial lesion, a greater reduction rate was observed (p<0.05). Conclusions: Decompression as an initial treatment of cystic lesions of the jaws was effective; it reduces the size of the lesions avoiding a possible damage to adjacent structures. Cystic lesions in the mandible, regardless of the area where they occur will have a higher reduction rate if it is compared with the maxilla. Similar behavior was identified in large lesions compared to smaller. Key words:Decompression, reduction rate, cyst, maxilla, mandible

Transfer of extracellular vesicle-microRNA controls germinal center reaction and antibody production

Intercellular communication orchestrates effective immune responses against disease-causing agents. Extracellular vesicles (EVs) are potent mediators of cell-cell communication. EVs carry bioactive molecules, including microRNAs, which modulate gene expression and function in the recipient cell. Here, we show that formation of cognate primary T-B lymphocyte immune contacts promotes transfer of a very restricted set of T-cell EV-microRNAs (mmu-miR20-a-5p, mmu-miR-25-3p, and mmu-miR-155-3p) to the B cell. Transferred EV-microRNAs target key genes that control B-cell function, including pro-apoptotic BIM and the cell cycle regulator PTEN. EV-microRNAs transferred during T-B cognate interactions also promote survival, proliferation, and antibody class switching. Using mouse chimeras with Rab27KO EV-deficient T cells, we demonstrate that the transfer of small EVs is required for germinal center reaction and antibody production in vivo, revealing a mechanism that controls B-cell responses via the transfer of EV-microRNAs of T-cell origin. These findings also provide mechanistic insight into the Griscelli syndrome, associated with a mutation in the Rab27a gene, and might explain antibody defects observed in this pathogenesis and other immune-related and inflammatory disorders.This manuscript was funded by grants SAF2017-82886-R (FS-M) from the Spanish Ministry of Economy and Competitiveness; CAM (S2017/BMD-3671-INFLAMUNE-CM) from the Comunidad de Madrid (FS-M); CIBERCV (CB16/11/00272), BIOIMID PIE13/041 from the Instituto de Salud Carlos III and from the Fundación La MaratóTV3(grant122/C/2015). The current research has received funding from “la Caixa” Foundation under the project code HR17-00016. VGY is supported by the AECC foundation. A.R.R. is supported by CNIC funding. This project was funded by the Spanish Ministerio de Ciencia, Innovacion y Universidades SAF2016-75511-R, and La Caixa Health Research Program HR17-00247 grant to A.R.R. Grants from Ramón Areces Foundation “Ciencias de la Vida y de la Salud” (XIX Concurso-2018) and from Ayuda Fundación BBVA y Equipo de Investigación Científica (BIOMEDICINA-2018) (to FSM). The CNIC is supported by the Ministerio de Ciencia, Innovacion y Universidades and the Pro-CNIC Foundation, and is a Severo Ochoa Center of Excellence (SEV-2015-0505).S

miRNA profiling during antigen-dependent T cell activation: A role for miR-132-3p

microRNAs (miRNAs) are tightly regulated during T lymphocyte activation to enable the establishment of precise immune responses. Here, we analyzed the changes of the miRNA profiles of T cells in response to activation by cognate interaction with dendritic cells. We also studied mRNA targets common to miRNAs regulated in T cell activation. pik3r1 gene, which encodes the regulatory subunits of PI3K p50, p55 and p85, was identified as target of miRNAs upregulated after T cell activation. Using 3'UTR luciferase reporter-based and biochemical assays, we showed the inhibitory relationship between miR-132-3p upregulation and expression of the pik3r1 gene. Our results indicate that specific miRNAs whose expression is modulated during T cell activation might regulate PI3K signaling in T cells.We thank Miguel Vicente-Manzanares for help with English editing and Almudena R. Ramiro for helpful discussions. We appreciate help from Gloria Martinez del Hoyo on DCs experiments set up. We also thank the CNIC Genomics, Bioinformatics and Cellomics Units for technical support. This work was supported by grants SAF2014-55579R from Ministerio de Economia y Competitividad-Spain, ERC-2011-AdG 294340-GENTRIS, CIBER CARDIOVASCULAR (FEDER and Instituto de Salud Carlos III), PIE-13-00041 and INDISNET S2011-BMD-2332 (F.S.M.). The Centro Nacional de Investigaciones Cardiovasculares (CNIC, Spain) is supported by the Ministerio de Economia y Competitividad-Spain and the Pro-CNIC Foundation.S

Toxic metals in toenails as biomarkers of exposure: A review

Toenails have been used as biomarkers of exposure to toxic metals, but their validity for this purpose is not yet clear and might differ depending on the specific agent. To evaluate this issue, we reviewed the literature on: a) the time-window of exposure reflected by toenails; b) the reproducibility of toenail toxic-metal levels in repeated measures over time; c) their relationship with other biomarkers of exposure, and; d) their association with potential determinants (i.e. sociodemographic, anthropometric, or lifestyle characteristics) or with sources of exposure like diet or environmental pollution. Thus, we performed a systematic review, searching for articles that provided original data for levels of any of the following toxic metals in toenails: aluminum, beryllium, cadmium, chromium, mercury, nickel, lead, thallium and uranium. We identified 88 articles, reporting data from 67 different research projects, which were quite heterogeneous with regard to population profile, sample size and analytical technique. The most commonly studied metal was mercury. Concerning the time-window of exposure explored by toenails, some reports indicate that toenail cadmium, nickel and lead may reflect exposures that occurred 7-12 months before sampling. For repeated samples obtained 1-6 years apart, the range of intraindividual correlation coefficients of aluminum, chromium and mercury was 0.33-0.56. The correlation of toxic metal concentrations between toenails and other matrices was higher for hair and fingernails than for urine or blood. Mercury levels were consistently associated with fish intake, while other toxic metals were occasionally associated with specific sources (e.g. drinking water, place of residence, environmental pollution, and occupation). The most frequently evaluated health endpoints were cardiovascular diseases, cancer, and central nervous system diseases. Available data suggest that toenail mercury levels reflected long-term exposures and showed positive associations with fish intake. The lack of standardization in sample collection, quality control, analytical techniques and procedures - along with the heterogeneity and conflicting results among studies - mean it is still difficult to conclude that toenails are a good biomarker of exposure to toxic metals. Further studies are needed to draw solid conclusions about the suitability of toenails as biomarkers of exposure to toxic metals.This work was supported by FIS grants PI12/00150, PI17CIII/00034, PI18/00287 (Instituto de Salud Carlos III, State Secretary of R + D + I and European Union (ERDF/ESF, “Investing in your future")), P42ES010349 and P30ES009089 (National Institute of Environmental Health Sciences).S

A predominant involvement of the triple seropositive patients and others with rheumatoid factor in the association of smoking with rheumatoid arthritis

The major environmental risk factor for rheumatoid arthritis (RA) is smoking, which according to a widely accepted model induces protein citrullination in the lungs, triggering the production of anticitrullinated protein antibodies (ACPA) and RA development. Nevertheless, some research findings do not fit this model. Therefore, we obtained six independent cohorts with 2253 RA patients for a detailed analysis of the association between smoking and RA autoantibodies. Our results showed a predominant association of smoking with the concurrent presence of the three antibodies: rheumatoid factor (RF), ACPA and anti-carbamylated protein antibodies (ACarPA) (3 Ab vs. 0 Ab: OR = 1.99, p = 2.5 × 10?8). Meta-analysis with previous data (4491 patients) confirmed the predominant association with the concurrent presence of the three antibodies (3 Ab vs. 0 Ab: OR = 2.00, p = 4.4 ×10?16) and revealed that smoking was exclusively associated with the presence of RF in patients with one or two antibodies (RF+ 1+2 vs. RF? 0+1+2: OR = 1.32, p = 0.0002). In contrast, no specific association with ACPA or ACarPA was found. Therefore, these results showed the need to understand how smoking favors the concordance of RA specific antibodies and RF triggering, perhaps involving smoking-induced epitope spreading and other hypothesized mechanisms

Staphylococcus aureus Nasal Colonization in Spanish Children. The COSACO Nationwide Surveillance Study

Objective: To assess the prevalence and risk factors for S. aureus and methicillin-resistant S. aureus (MRSA) nasal colonization in Spanish children. Methods: Cross-sectional study of patients <14 years from primary care centers all over Spain. Clinical data and nasal aspirates were collected from March to July 2018. Results: A total of 1876 patients were enrolled. Prevalence of S. aureus and MRSA colonization were 33% (95% CI 30.9-35.1) and 1.44% (95% CI 0.9-2), respectively. Thirty-three percent of the children (633/1876) presented chronic conditions, mainly atopic dermatitis, asthma and/or allergy (524/633). Factors associated with S. aureus colonization were age ≥5 years (OR 1.10, 95% CI 1.07-1.12), male sex (OR 1.43, 95% CI 1.17-1.76), urban setting (OR 1.46, 95% CI 1.08-1.97) and the presence of asthma, atopic dermatitis or allergies (OR 1.25; 95% CI: 1.093-1.43). Rural residence was the only factor associated with MRSA colonization (OR 3.62, 95% CI 1.57-8.36). MRSA was more frequently resistant than methicillin-susceptible S. aureus to ciprofloxacin [41.2% vs 2.6%; p<0.0001], clindamycin [26% vs 16.9%; p=0.39], and mupirocin [14.3% vs 6.7%; p=0.18]. None of the MRSA strains was resistant to tetracycline, fosfomycin, vancomycin or daptomycin. Conclusions: The main risk factors for S. aureus colonization in Spanish children are being above five years of age, male gender, atopic dermatitis, asthma or allergy, and residence in urban areas. MRSA colonization is low, but higher than in other European countries and is associated with rural settings.This study has been supported by The Spanish Ministry of Science and Innovation – Instituto de Salud Carlos III, and Fondos FEDER of the EU, Grant Nº PI18CIII/00372 [Fondo de Investigaciones Sanitarias-Spanish Health Research Fund (ISCIII)]; Grant Award “Jose María Corretger” from the Spanish Society for Pediatric Infectious Diseases; Grant Research Award from the Spanish Association of Pediatric Primary Care; and a Small Grant Award from the European Society for Pediatric Infectious Diseases.S

Sustained blood glutamate scavenging enhances protection in ischemic stroke

Stroke is a major cause of morbidity, mortality, and disability. During ischemic stroke, a marked and prolonged rise of glutamate concentration in the brain causes neuronal cell death. This study explores the protective effect of a bioconjugate form of glutamate oxaloacetate transaminase (hrGOT), which catalyzes the depletion of blood glutamate in the bloodstream for ~6 days following a single administration. When treated with this bioconjugate, a significant reduction of the infarct volume and a better retention of sensorimotor function was observed for ischemic rats compared to those treated with saline. Moreover, the equivalent dose of native hrGOT yielded similar results to the saline treated group for some tests. Targeting the bioconjugate to the blood-brain-barrier did not improve its performance. The data suggest that the bioconjugates draw glutamate out of the brain by displacing homeostasis between the different glutamate pools of the body

Workshop on mitigation measures to reduce bycatch of short-beaked common dolphins in the Bay of Biscay (WKEMBYC2)

Following the special request from the DG MARE, the Workshop on mitigation measures to minimize bycatch of short-beaked common dolphins in the Bay of Biscay (WKEMBYC2) was established by ICES. WKEMBYC2 was tasked with updating and revaluating the scenarios previously proposed in the ICES special request advice in 2020. The group was asked to consider recent data on bycatch of common dolphins in commercial fisheries and total fishing effort in the Bay of Biscay and Iberian Coast ecosystem, as well as taking into account results from any mitigation trials carried out since the meeting in 2020. In section 2 of this report the efficacy of the scenarios provided during WKEMBYC in 2020 were reassessed using updated bycatch estimates calculated from at-sea monitoring and stranding data collected between 2019 and 2021. Scenarios and methods remained unchanged, to ensure comparability between both evaluations. Similarly, PBR values considered in the 2020 scenarios were again considered here, with the addition of the mPBR which was developed by OSPAR since the last workshop. The annual mortality due to bycatch inferred from French strandings in the Bay of Biscay and along the Western Channel was estimated at about 9,040 (95%CI [6,640 - 13,300]) common dolphins between 2019 and 2021. In the Bay of Biscay and Iberian Coast (areas 8 and 9), the mean annual bycatch estimated from at-sea observations between 2019-2021 across all métiers was 5938 (95% CI 3081-9700) common dolphins. The abundance estimate and PBR values used in this report were the same as those used in 2020; with common dolphin abundance estimated to be 634 286 (CV=0.307) for the European Atlantic Assessment Unit, and PBR for the species calcu-lated as 4926 individuals per year. The management objective of PBR is to ensure that “a population will remain at, or recover to, its maximum net productivity level (typically 50% of the populations carrying capacity), with 95% probability, within a 100-year period”. A modified PBR (mPBR) value of 985 was also used, with the management objective of ensuring “a population should be able to recover to or be maintained at 80% of carrying capacity, with probability 0.8, within a 100-year period”. Considering bycatch estimates calculated from at-sea monitoring, scenarios with a combination of pingers on OTM/PTB and at least 4-week closure in winter can reach the management objective of bycatch remaining below the PBR. The removal bycatch over a three-month period between the January and March winter period, and an addi-tional month closure in July/August potentially reduces bycatch below the mPBR level for at-sea monitoring bycatch estimates alone. Considering estimates inferred from strandings, a mini-mum of 6-week closure combined with pingers can achieve the objective of reducing bycatch below PBR. None of the scenarios can reduce bycatch below mPBR for either monitoring and stranding bycatch estimates. The narrower the fishery closure, the higher the risk of not achieving the management objective, as the peak of mortality can be missed. In section 3 of the report the workshop participants chose to further explore the scenarios with bycatch rates and estimate bycatch at a finer spatial and temporal scale and to consider additional mitigation measures based on results of newly conducted preliminary trials. This exploratory analysis allowed particular areas, métiers or periods with evidence of elevated bycatch rates to be identified at higher resolution, however, this approach requires significant at-sea monitoring in all strata. If such data were available, this method could be used to highlight specific areas, métiers and periods where particular effort in vessel observation should be deployed or mitigation measures implemented. WKEMBYC2 also recommended a series of monitoring actions to improve bycatch estimates, monitoring to data analysis, mitigation and the assessment of the northeast Atlantic common dolphin