Efficacy and limitations of combined A2.CAR Treg and anti-CD154 therapy in a mouse model of haplo-mismatched heart transplantation.

The ability to induce allograft specific tolerance would reduce the need for daily pharmacological immunosuppression, improve post-transplant quality of life and transplant outcomes. Adoptive cell therapy with regulatory T cells expressing donor-specific Chimeric Antigen Receptors (CAR-Tregs) is a promising strategy, but monotherapy has not resulted in prolonged survival of grafts with multiple MHC mismatches. We used a clinically-relevant, haplo-mismatched model of heart transplantation in immune-competent C57BL/6 recipients to test the ability of HLA-A2-specific (A2) CAR Tregs to synergize with CD154 blockade to enhance graft survival. We found that in combination with a single low dose of anti-CD154, A2.CAR Tregs significantly prolonged heart allograft survival. Through use of grafts expressing the 2W-OVA transgene, tetramer tracking of 2W- and OVA-specific cells revealed that in mice with accepted grafts, the effects of A2.CAR Tregs included inhibition of endogenous donor-specific CD4 + and CD8 + T cell expansion, and B cell and antibody responses. Moreover, within the 2W-specific CD4 + T cell population, there was a significant increase in the proportion of FoxP3 pos cells, suggestive of infectious tolerance. In mice where CD154 blockade and A2.CAR Tregs failed to prolong graft survival, there was preferential expansion of FoxP3 neg A2.CAR T cells within the rejecting allograft. Thus, this study provides the first evidence for a synergistic effect between CAR Tregs and CD40-pathway blockade and supports the further refinement of this strategy as a promising future direction towards the goal of transplantation tolerance.

Protocol description and initial experience in kidney graft perfusion using infrared thermography.

PURPOSE: Protocol description for renal perfusion study using thermographic technology and description of the thermographic and clinical behavior of the transplanted kidneys before and after unclamping. METHODS: Infrared thermographic images of renal grafts are obtained before kidney reperfusion, 10 min after and just before closing the surgical wound. Thermographic data is evaluated together with the type of graft and donor, cold ischemia time, hypovascularized areas determined by the surgeon during surgical intervention, alterations in vascular flow in postoperative echo-Doppler, time at the beginning of graft function and serum creatinine monitoring during postoperative follow-up. RESULTS: 17 grafts were studied. The mean temperature of the grafts before reperfusion, 10 min after and at the end of the surgery were 18.7 °C (SD 6.27), 32.36 °C (SD1.47) and 32.07 °C (SD1.78) respectively. 4 grafts presented hypoperfused areas after reperfusion. These areas presented a lower temperature compared to the well perfused parenchyma surface using thermographic images. CONCLUSION: The study of the usefulness and applicability of thermography can allow the development of tools that provide additional objective information on organ perfusion in real time and non-invasive manner. Our protocol and initial results can contribute to provide new evidence. Further analyses should be developed to shed light on the role of this technology.

Azithromycin and moxifloxacin resistance determinants in Mycoplasma genitalium in Lleida, Spain.

OBJECTIVE: Mycoplasma genitalium (MG) is a microorganism related to sexually transmitted infections. Antibiotic resistance of MG leads to an increase in treatment failure rates and the persistence of the infection. The aim of this study was to describe the most frequent mutations associated with azithromycin and moxifloxacin resistance in our geographical area. METHODS: A prospective study from May 2019 to May 2023 was performed. MG-positive samples were collected. Real-time PCRs (AllplexTM MG-AziR Assay and AllplexTM MG-MoxiR Assay, Seegene) were performed in MG positive samples to detect mutations in 23S rRNA V domain and parC gene. RESULTS: A 37.1% of samples presented resistance determinants to azithromycin and the most common mutation detected was A2059G (57.9%). Resistance to moxifloxacin was studied in 72 azithromycin-resistant samples and 36.1% showed mutations, being G248T the most prevalent (73.1%). CONCLUSIONS: The resistance to different lines of treat ment suggests the need for a targeted therapy and the performing of a test of cure afterwards.

Why do physicians go to work when they are sick? Presenteeism at different career stages.

INTRODUCTION AND OBJECTIVE: Physicians find it difficult to take on the role of the patient and they show unusual behaviors when ill. One of these behaviors is presenteeism, which is working while sick. The objective of this research is to analyze the factors that contribute to the phenomenon of presenteeism in Spanish physicians. MATERIAL AND METHODS: Mixed methodology study: one national survey through the General Council of Medical Associations website (quantitative part), 22 semistructured interviews with sick residents and practicing physicians, and three focus groups involving professionals from the occupational health services (qualitative). A bivariate analysis using parametric and non-parametric tests. The significance level was p<0.05 (95% confidence interval). Qualitative analysis using the comparative-constant method until saturation of information. RESULTS: Presenteeism is reported by 89.4% of doctors who responded to the survey, and it is more common among women. Contributing factors include fear of overburdening colleagues (the main reason and more common among women 58.14% vs 48.35%), self-perception of doing one's duty (the second reason and more common among men, 44.63% vs 33.14%) and economic impact and difficulty in accepting the role of a sick person. This behavior has an impact on patient safety, and is part of the hidden curriculum that also affects the training of medical professionals. CONCLUSIONS: Presenteeism is a widespread and accepted practice among medical professionals. Although normalized, and even appreciated as a way to avoid overburdening colleagues, presenteeism has important implications for clinical ethics and patient safety.

Prevalence of subclinical infectious agents in a blood donor population tested on every donation.

OBJECTIVES: This study aimed to assess the prevalence of blood-borne infectious agents in healthy, client-owned dogs from a blood donor population in Portugal and Spain, and to address the importance of a screening protocol on every donation. MATERIALS AND METHODS: Client-owned healthy dogs were tested before each donation on a veterinary blood bank. Blood samples from new potential donors, and from regular donors, were tested by real-time polymerase chain reaction for Leishmania species, Ehrlichia species, Brucella species, Babesia species and Anaplasma species Serological tests were also performed for Leishmania species, Ehrlichia species and Dirofilaria immitis. All donors were tested for every infectious agent in each donation. RESULTS: The study found that out of a total of 8036 donors and 35,120 samples tested, 3.9% of blood donors tested positive for at least one of the agents, with the most prevalent being Anaplasma species (2.1%). Serological tests also revealed positive results in 14.0% of donors, with the highest percentage for Leishmania species (7.7%). Moreover, the study found that 28.2% of positive results were from dogs with negative results in donations performed 3 to 12 months before, and 18.0% of positive results were recent infections. CLINICAL SIGNIFICANCE: These findings indicate a high prevalence of infectious agents in seemingly healthy, selected dogs eligible to become blood donors in the Iberian Peninsula, highlighting the importance of regular testing on every donation. This study emphasises the importance of a regular screening protocol for every donation instead of annual testing, as is commonly performed in veterinary medicine.

[Translated article] Clinical and Epidemiological Characteristics of Patients Being Treated for Scabies in Spain: Opportunities for Improvement - A Multicenter Cross-Sectional CLINI-AEDV Study.

BACKGROUND: Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. METHODS: We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). RESULTS: Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had clinical features of classic scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. CONCLUSIONS: In a high proportion of scabies cases, the patient has already received treatment. In those cases, we observe several remediable shortcomings that could explain why some of these treatments fail. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides.

Clinical and Epidemiological Characteristics of Patients Being Treated for Scabies in Spain: Opportunities for Improvement - A Multicenter Cross-Sectional CLINI-AEDV Study. / Características clínicas y epidemiológicas de pacientes diagnosticados de escabiosis en España: oportunidades de mejora. Estudio transversal multicéntrico CLINI-AEDV.

BACKGROUND: Several studies support the hypothesis that scabies is on the rise in Spain. There are also concerns about the possible development of resistance to treatment and an increase in atypical presentations. The aims of this study were to describe the demographic and clinical characteristics of patients with scabies seen by dermatologists in Spain, to identify the possible emergence of atypical forms of scabies, and to explore the frequency of treatment failures and associated risk factors. METHODS: We conducted an observational, cross-sectional, multicenter study of data collected prospectively in April and May 2023 using the CLINI-AEDVp platform created by the Spanish Academy of Dermatology and Venereology (AEDV). RESULTS: Participating dermatologists from 31 hospitals in 15 of Spain's autonomous communities recorded 186 cases of active scabies (51% in women) during the study period. A diagnostic certainty level of A, B or C as per the International Alliance for the Control of Scabies Consensus Criteria was required for diagnosis. Overall, 92% of patients had typical scabies and 66% had already been treated with a scabicide for the current episode. Of the treated patients, only 36% had received and completed adequate treatment (including the simultaneous treatment of all household members) and 50% had not received clear written recommendations. CONCLUSIONS: In a high proportion of the cases of scabies studied, the patient had already received treatment. In those cases, we observed several remediable shortcomings that could explain why some of these treatments had failed. Remedying these deficiencies should lead to better control of scabies and an improved assessment of the actual effectiveness of currently available scabicides.

Reduction of NrF2 as coadjuvant during the development of persistent periapical lesions.

BACKGROUND: Persistent periapical lesions (PPL) are the result of pulpar necrosis induced by bacterial infection resulting in bone degradation and culminating with the loss of dental piece. Pathological changes in the peripapice are associated with the presence of free radicals. The transcription factor Nrf2 is the main regulator of the endogenous antioxidant response against oxidative stress and has been implicated in the regulation of osteoclastogenesis.The aim is to determine the oxidative condition in samples from patients with Persistent Periapical Injuries as a detonating factor of tissue damage. MATERIAL AND METHODS: An observational, descriptive, cross-sectional study was carried out in samples with PPL (cases) and samples by removal of third molars (controls) obtained in the clinic of the specialty in endodontics, University of Guadalajara. Samples were submitted to histological staining with Hematoxylin-Eosin, lipoperoxide analysis, Superoxide Dismutase (SOD), Glutathione-Peroxidase (GPx) and Catalase (CAT) activities were determined by immunoenzymatic assays and NrF2 by Western Blot analysis. RESULTS: Samples from PPL patients histologically showed an increased presence of lymphocytes, plasma cells, and eosinophils, as well as a decrease in extracellular matrix proteins and fibroblast cells. There was a rise in lipid peroxidation, GPx and SOD activities, but an important decline (36%) in Catalase activity was observed (p<0.005); finally, NrF2-protein was diminished at 10.41%. All comparisons were between cases vs controls. CONCLUSIONS: The alterations in antioxidants endogenous NrF2-controlled are related to osseous destruction in patients with PPL.

Balloon-Expandable Pulmonary Valves for Patched or Native Right Ventricular Outflow Tracts.

The implantation of percutaneous balloon expandable valves in native or patched right ventricular outflow tracts (nRVOT) is a challenging technique due to the diversity of anatomies and shapes, the large sizes, and the distensibility of the nRVOT, for which specific techniques have been developed. We present a single center experience with balloon expandable percutaneous pulmonary valves in nRVOT, describing the techniques used, complications observed, and a short-mid term follow-up.. This is a single center descriptive study of patients who underwent a percutaneous pulmonary valve implantation in a nRVOT with a balloon expandable pulmonary valve in our center between September 2012 and June 2022.. We implanted successfully 45 valves in 46 patients (20 Sapien and 25 Melody). Tetralogy of Fallot or pulmonary atresia with VSD were the main congenital heart disease (n = 32). All were pre-stented, 18 in a one step procedure. We used a Dryseal sheath in 13/21 Sapien. In 6 patients we used the anchoring technique, 5 with a very large nRVOT and one pyramidal nRVOT. In the 3.5 year follow-up 7 patients developed endocarditis and 3 required a valve redilation, no fractures were observed. PPVI of native RVOT with balloon expandable valves is feasible in a number of selected anatomies, including large or pyramidal nRVOT, using specific techniques, (presenting, LPA anchoring).

Improving growth of Cupriavidus necator H16 on formate using adaptive laboratory evolution-informed engineering.

Conversion of CO2 to value-added products presents an opportunity to reduce GHG emissions while generating revenue. Formate, which can be generated by the electrochemical reduction of CO2, has been proposed as a promising intermediate compound for microbial upgrading. Here we present progress towards improving the soil bacterium Cupriavidus necator H16, which is capable of growing on formate as its sole source of carbon and energy using the Calvin-Benson-Bassham (CBB) cycle, as a host for formate utilization. Using adaptive laboratory evolution, we generated several isolates that exhibited faster growth rates on formate. The genomes of these isolates were sequenced, and resulting mutations were systematically reintroduced by metabolic engineering, to identify those that improved growth. The metabolic impact of several mutations was investigated further using RNA-seq transcriptomics. We found that deletion of a transcriptional regulator implicated in quorum sensing, PhcA, reduced expression of several operons and led to improved growth on formate. Growth was also improved by deleting large genomic regions present on the extrachromosomal megaplasmid pHG1, particularly two hydrogenase operons and the megaplasmid CBB operon, one of two copies present in the genome. Based on these findings, we generated a rationally engineered ΔphcA and megaplasmid-deficient strain that exhibited a 24% faster maximum growth rate on formate. Moreover, this strain achieved a 7% growth rate improvement on succinate and a 19% increase on fructose, demonstrating the broad utility of microbial genome reduction. This strain has the potential to serve as an improved microbial chassis for biological conversion of formate to value-added products.

A French community-based intervention for parents of French-Canadian children with behavior problems: The EQUIPE program.

OBJECTIVES: Behavior problems are one of the most common reasons for seeking mental health services in pediatric populations. The objectives are to evaluate the effects of the EQUIPE program (Étude Québécoise d'Intervention pour les Parents d'Enfants avec des problèmes de comportement) and to analyze the impact of the severity of behavior problems and of parental characteristics. METHODS: This program was translated from the Community Parent Education Program. The effects of EQUIPE, as compared to a control group, were evaluated by using Child Behavior Checklist and Parent Stress Index questionnaires before (T0) and after the intervention program (T1), and at 6 (T2) and 12months (T3) follow-up visits. RESULTS: In total, 533 participants were enrolled in intervention (n=465) (with "severe" or "mild" subgroups according to CBCL-T score) and a control group (n=68). In the two groups, the results showed a statistically significant decrease in Child Behavior Checklist and Parent Stress Index total scores at T1, T2 and T3, with the exception of Child Behavior Checklist total scores in the control group at T2. In the intervention group Child Behavior Checklist total scores were significantly higher in the "severe"; which was not the case for Parent Stress Index total scores at T2 and T3. DISCUSSION: Socioeconomic characteristics, family details, parental medical history and the age of the children appeared to influence changes in Child Behavior Checklist and Parent Stress Index total scores. CONCLUSION: The EQUIPE program is an effective intervention for reducing behavior problems and parents' stress in a French-Canadian population.

Gender analysis of Spanish National Questionnaire on behaviours and attitudes of doctors towards their own illness (CAMAPE).

INTRODUCTION AND OBJECTIVES: Physicians' health is a key element for quality healthcare. Medical professionals have difficulty accepting their role as patients and it might be different among sexes. The aim was to describe behaviours and attitudes of doctors towards their own illness. MATERIALS AND METHODS: An online survey was launched through the General Council of Medical Associations webpage for all Spanish registered doctors. A bivariate analysis by sex was performed for all the questionnaire variables using parametric and non-parametric tests. The significance level was p<0.05 (95% confidence interval). RESULTS: A total of 4,308 registered doctors (1,858 men and 2,450 women) answered. Women were younger, single, and worked mainly in non-surgical specialities in the public sector. Men were older, married, and worked more frequently in public-private practice. Women had less chronic conditions, except for anxiety disorders (11.52% vs 15.18%). Both sexes, especially women, primarily self-treated (94.29% vs 95.02%), went to work while ill (88.16% vs 90.29%), visited their GP (56% vs 70%), and half of them underwent annual occupational health checks (40% vs 48%). Women self-prescribed more analgesics (93.43% vs 95.63%), more presenteeism (88% vs 90%) and felt more insecure when treating sick fellows (9.96% vs 20.12%) and requested training for it. More women agreed to make deontological recommendations about doctors' health (91.55% vs 96.16%) and considered revalidation may contribute to improve doctors' health (65.29% vs 66.16%). CONCLUSIONS: Male and female doctors show illness-health behaviours and attitudes at work to improve. There are differences among male and female doctors. Regarding, medical feminization, ethical recommendations may be of benefit regarding doctors' health-illness issues and considering gender perspective.

Current trends in waste valorization.

This paper presents the scientific breakthroughs made in bioprocess engineering and microbial biotechnology for the conversion of wastes into products with added value and/or biofuels. The significant results obtained in the emerging fields of hybrid electrosynthesis, the role of enzymes in the degradation of plastics, polyhydroxyalkanoate and 5-aminolevulinic acid production, fermentation technology and the application of molecular engineering tools to bioprocess technology are highlighted.

Screening and evaluation of biomass upgrading strategies for sustainable transportation fuel production with biomass-derived volatile fatty acids.

Biomass conversion to fuels and chemicals is crucial to decarbonization, but choosing an advantageous upgrading pathway out of many options is challenging. Rigorously evaluating all candidate pathways (process simulation, product property testing) requires a prohibitive amount of research effort; even simple upgrading schemes have hundreds of possible permutations. We present a method enabling high-throughput screening by approximating upgrading unit operations and drop-in compatibility of products (e.g., fuel properties) and apply it to volatile fatty acid (VFA) conversion to liquid transportation fuels via a MATLAB script, VFA Upgrading to Liquid Transportation fUels Refinery Estimation (VULTURE). VULTURE selects upgrading configurations that maximize fuel blend bio-derived content. We validate VULTURE's approximations through surrogate fuel property testing and process simulation. Techno-economic and life cycle analyses suggest that VFA upgrading processes down-selected by VULTURE are profitable and have low carbon intensities, demonstrating the potential for the strategy to accelerate process development timelines at decreased costs.

Five years' experience of the clinical exome sequencing in a Spanish single center.

Nowadays, exome sequencing is a robust and cost-efficient genetic diagnostic tool already implemented in many clinical laboratories. Despite it has undoubtedly improved our diagnostic capacity and has allowed the discovery of many new Mendelian-disease genes, it only provides a molecular diagnosis in up to 25-30% of cases. Here, we comprehensively evaluate the results of a large sample set of 4974 clinical exomes performed in our laboratory over a period of 5 years, showing a global diagnostic rate of 24.62% (1391/4974). For the evaluation we establish different groups of diseases and demonstrate how the diagnostic rate is not only dependent on the analyzed group of diseases (43.12% in ophthalmological cases vs 16.61% in neurological cases) but on the specific disorder (47.49% in retinal dystrophies vs 24.02% in optic atrophy; 18.88% in neuropathies/paraparesias vs 11.43% in dementias). We also detail the most frequent mutated genes within each group of disorders and discuss, on our experience, further investigations and directions needed for the benefit of patients.

[Design and development of a dietary intervention in children with prediabetes]. / Diseño y desarrollo de una intervención dietética en niños con prediabetes.

INTRODUCTION: Prediabetes is a pathological condition where the blood glucose concentration is higher than normal concentrations, but lower than those considered in the diagnosis of type 2 diabetes mellitus (DM2). Until a few years ago, the prediabetes condition occurred in adults with associated risk factors such as overweight or obesity, sedentary lifestyle, poor eating habits, cardiovascular problems, etc. In recent years it has begun to be detected in children, with the family habits that these children have established becoming increasingly important. MATERIAL AND METHODS: In our study, 29 pre-diabetic children from Pedro Abad, Córdoba, have been evaluated. For this, we have carried out a randomized crossover trial with an intervention group (IG) and a control group (CG), using a dietary intervention with nutritional reinforcement. The main objective of the present study was to determine if the eating habits of pre-diabetic children improved with a dietary intervention based on nutritional education. RESULTS: Through a nutritional evaluation through different tests and visits made to the children, the results corresponding to the first phase of the study, relate an improvement in eating habits and adherence to the Mediterranean Diet by the intervened children. CONCLUSIONS: The eating habits of pre-diabetic children improve with a dietary intervention based on nutritional education, since by providing them with knowledge of the different foods and the contribution of nutrients that they provide us, they can make a better selection of foods.