References Values of Soluble alpha-Klotho Serum Levels Using an Enzyme-Linked Immunosorbent Assay in Healthy Adults Aged 18-85 Years

Hector Vazquez-Lorente (FPU18/03655) and Lucas Jurado-Fasoli (FPU19/01609) are under an FPU fellowship from the Spanish Ministry of Education. This study is part of a Ph.D. thesis conducted in the Biomedicine Doctoral Studies of the University of Granada, Spain. The APC was funded by San Cecilio University Hospital, FIBAO, IBS-Granada, Spain.alpha-Klotho protein is a powerful predictor of the aging process and lifespan. Although lowered circulating soluble alpha-Klotho levels have been observed in aged non-healthy individuals, no specific reference values across a wide range of ages and sex using an enzyme-linked immunosorbent assay (ELISA) are available for larger cohorts of healthy individuals. The present analytical cross-sectional study was aimed to establish the reference values of soluble alpha-Klotho serum levels in healthy adults by age and sex groups. A total of 346 (59% women) healthy individuals aged from 18 to 85 years were recruited. Subjects were divided by sex and age as: (i) young (18-34.9 years), (ii) middle-aged (35-54.9 years), and (iii) senior (55-85 years) individuals. The soluble alpha-Klotho levels were measured in serum using ELISA. Senior adults were the age-group that presented the lowest soluble alpha-Klotho serum levels (p < 0.01), with age showing a negative association with soluble alpha-Klotho serum levels (p < 0.001). No differences between sexes were observed. Therefore, soluble alpha-Klotho levels were especially decreased-regardless of sex-in our cohort of healthy individuals because of the physiological decline derived from the aging process. We recommend routine assessments of soluble alpha-Klotho levels using ELISA as a simple and cheap detectable marker of aging that improves quality of life in the elderly.Spanish Government FPU18/03655 FPU19/01609San Cecilio University Hospital, FIBAO, IBS-Granada, Spai

Mild hypophosphatasia may be twice as prevalent as previously estimated: an effective clinical algorithm to detect undiagnosed cases

Objectives: Since the prevalence of hypophosphatasia (HPP), a rare genetic disease, seems to be underestimated in clinical practice, in this study, a new diagnostic algorithm to identify missed cases of HPP was developed and implemented. Methods: Analytical determinations recorded in the Clinical Analysis Unit of the Hospital Universitario Clínico San Cecilio in the period June 2018 – December 2020 were reviewed. A new clinical algorithm to detect HPP-misdiagnosed cases was used including the following steps: confirmation of persistent hypophosphatasemia, exclusion of secondary causes of hypophosphatasemia, determination of serum pyridoxal- 5′-phosphate (PLP) and genetic study of ALPL gene. Results: Twenty-four subjects were selected to participate in the study and genetic testing was carried out in 20 of them following clinical algorithm criteria. Eighty percent of patients was misdiagnosed with HPP following the current standard clinical practice. Extrapolating these results to the current Spanish population means that there could be up to 27,177 cases of undiagnosed HPP in Spain. In addition, we found a substantial proportion of HPP patients affected by other comorbidities, such as autoimmune diseases (∼40 %). Conclusions: This new algorithm was effective in detecting previously undiagnosed cases ofHPP, which appears to be twice as prevalent as previously estimated for the European population. In the near future, our algorithm could be globally applied routinely in clinical practice to minimize the underdiagnosis of HPP. Additionally, some relevant findings, such as the high prevalence of autoimmune diseases in HPP-affected patients, should be investigated to better characterize this disorder.Instituto de Salud Carlos III grants PI21-01069 co-funded by the European Regional Development Fund (FEDER) and by Junta de Andalucía grant PI-0268-2019Operational Programme for Youth Employment of the Junta de Andalucía with Ref: POEJ_04/2022-12Instituto de Salud Carlos III with co-funding by FEDER (CD20/00022)(FI19/00118 and CM21/00221) from Instituto de Salud CarlosPostdoctoral fellowship from the Junta de Andalucía (RH-0141-2020

Identification of hereditary breast and ovarian cancer germline variants in Granada (Spain): NGS perspective

Funding for open access charge: Universidad de Granada/CBUA. Maria Molina-Zayas has been a recipient of the 2018 AEFA Post-residency Grant (Spanish Association of Clinical Laboratory) and Dr. Carmen Garrido-Navas holds a postdoctoral fellowship from the Ministry of Economy, Competitiveness, Enterprises and Universities (DOC_01682).The aim of this study was to assess the prevalence of germline variants in cancer-predisposing genes by either targeted (BRCA1/2) or multigene NGS panel in a high-risk Hereditary Breast and Ovarian Cancer (HBOC) cohort. Samples from 824 Caucasian probands were retrospectively collected and the impact of genetic diagnosis and genetic variants epidemiology in this cohort was evaluated. Performance of risk-reducing prophylactic measures, such as prophylactic mastectomy and/or prophylactic oophorectomy, was assessed through clinical follow-up of patients with a positive genetic result. Pathogenic variants predisposing to HBOC were identified in 11.9% (98/824) individuals at BRCA2 (47/98), BRCA1 (24/98), PALB2 (8/51), ATM (7/51), CHEK2 (6/51) MSH6, (2/51), RAD51C (2/51) and TP53 (2/386). Of them, 11 novel pathogenic variants and 12 VUS were identified, characterized, and submitted to ClinVar. Regarding clinical impact, the risk of developing basal or Her2 breast cancer was increased 15.7 times or 37.5 times for BRCA1 and MSH6 pathogenic variants respectively. On the contrary, the risk of developing basal or luminal A breast cancer was reduced to 81% or 77% for BRCA2 and BRCA1 pathogenic variants, respectively. Finally, 53.2% of individuals testing positive for class IV/V variants underwent prophylactic surgery (mastectomy, oophorectomy or both) being significantly younger at the cancer diagnosis than those undertaking prophylactic measures (p = 0.008). Of them, 8 carried a pathogenic/likely pathogenic variant in other genes different from BRCA1 and BRCA2, and the remaining (46.7%) decided to continue with clinical follow-up. No differences in pathogenicity or risk of developing cancer were found for BRCA1/2 between targeted and multigene sequencing strategies; however, NGS was able to resolve a greater proportion of high-risk patients.Universidad de Granada/CBUA2018 AEFA Post-residency Grant (Spanish Association of Clinical Laboratory)Ministry of Economy, Competitiveness, Enterprises and Universities DOC_0168

Role of age and comorbidities in mortality of patients with infective endocarditis

[Purpose]: The aim of this study was to analyse the characteristics of patients with IE in three groups of age and to assess the ability of age and the Charlson Comorbidity Index (CCI) to predict mortality. [Methods]: Prospective cohort study of all patients with IE included in the GAMES Spanish database between 2008 and 2015.Patients were stratified into three age groups:<65 years,65 to 80 years,and ≥ 80 years.The area under the receiver-operating characteristic (AUROC) curve was calculated to quantify the diagnostic accuracy of the CCI to predict mortality risk. [Results]: A total of 3120 patients with IE (1327 < 65 years;1291 65-80 years;502 ≥ 80 years) were enrolled.Fever and heart failure were the most common presentations of IE, with no differences among age groups.Patients ≥80 years who underwent surgery were significantly lower compared with other age groups (14.3%,65 years; 20.5%,65-79 years; 31.3%,≥80 years). In-hospital mortality was lower in the <65-year group (20.3%,<65 years;30.1%,65-79 years;34.7%,≥80 years;p < 0.001) as well as 1-year mortality (3.2%, <65 years; 5.5%, 65-80 years;7.6%,≥80 years; p = 0.003).Independent predictors of mortality were age ≥ 80 years (hazard ratio [HR]:2.78;95% confidence interval [CI]:2.32–3.34), CCI ≥ 3 (HR:1.62; 95% CI:1.39–1.88),and non-performed surgery (HR:1.64;95% CI:11.16–1.58).When the three age groups were compared,the AUROC curve for CCI was significantly larger for patients aged <65 years(p < 0.001) for both in-hospital and 1-year mortality. [Conclusion]: There were no differences in the clinical presentation of IE between the groups. Age ≥ 80 years, high comorbidity (measured by CCI),and non-performance of surgery were independent predictors of mortality in patients with IE.CCI could help to identify those patients with IE and surgical indication who present a lower risk of in-hospital and 1-year mortality after surgery, especially in the <65-year group

Desarrollo de un método para la detección simultánea de las mutaciones más frecuentes de la β-Talasemia en España

Tesis Univ. Granada. Departamento de Bioquímica, Biología Molecular e Inmunología III. Leída el 18 de abril de 200

Actuación comercial de las cooperativas vitivinícolas: análisis crítico del caso Montilla-Moriles

El articulo analiza la problemática situación en que se encuentran las cooperativas vitivinícolas, centrándose en el caso de la zona Montilla-Moriles. Se exponen los resultados más significativos de una encuesta realizada a las veinte cooperativas existentes en dicha zona: excedentes crónicos escaso valor añadido al producto, bajo nivel gerencial, etc. A continuación, se realiza un análisis critico de la situación, que concluye con una clasificación de las cooperativas en cuatro grandes campos atendiendo a su volumen de producción (variable cuantitativa), y nivel empresarial (variable cualitativa). Finalmente se exponen unas recomendaciones sobre actuaciones futuras, elaborándose una propuesta de actuaciones colectivas que tiene en cuenta la actitud de las distintas cooperativas frente a las posibles acciones a realizar

Marketing the inputs agrarios: caracterización y estrategias de las empresas suministradoras

Este trabajo aborda el estudio de la actividad comercial de las empresas suministradoras de medios de producción al agricultor, incidiendo fundamentalmente en la aplicación de técnicas y estrategias de marketing. En primer lugar se realiza una revisión sintética de las características que definirían y diferenciarían los mercados de productores y de consumidores. Posteriormente, se procede a caracterizar para el caso de los inputs agrarios fungibles -semillas, fertilizantes y fitosanitarios- el mercado de productores agrícolas y las estrategias de las empresas que tienen al agricultor como destinatario de sus productos. Dicha caracterización, basada en criterios tales como: características de la demanda, segmentación del mercado y estrategias de producto, precio, distribución y promoción, permite deducir en qué aspectos el mercado de los agricultores se asemeja realmente al de productores y en cuáles al de consumidores, constatándose así su carácter híbrido.This paper addresses the study of commercial activities in companies that supply means of production to the farmer, stressing mainly marketing technique and strategy applicability. Firstly, the features that typically define and differentiate producer and consumer markets are briefly reviewed. Then, the farm producers market and the strategies employed by companies whose products target the farmer are typed for fungible farm inputs -seeds, fertilizers and pesticides. From the above characterization, based on criteria such as demand characteristics, market segmentation and product strategias, pricing, distribution and promotion, it is possible to deduce which features make the farmer' market similar to a producer and which to a consumer market, thus revealing its hybridness

Hypophosphatasia: A Unique Disorder of Bone Mineralization

This research was funded by the Institute of Health Carlos III grants (PI18-00803 and PI18-01235), co-funded by the European Regional Development Fund (FEDER). In addition, JM.V-S and C.G-F are funded by predoctoral and postdoctoral fellowships, respectively, from the Institute of Health Carlos III (CM19/00188; CD20/00022). The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.Hypophosphatasia (HPP) is a rare genetic disease characterized by a decrease in the activity of tissue non-specific alkaline phosphatase (TNSALP). TNSALP is encoded by the ALPL gene, which is abundantly expressed in the skeleton, liver, kidney, and developing teeth. HPP exhibits high clinical variability largely due to the high allelic heterogeneity of the ALPL gene. HPP is characterized by multisystemic complications, although the most common clinical manifestations are those that occur in the skeleton, muscles, and teeth. These complications are mainly due to the accumulation of inorganic pyrophosphate (PPi) and pyridoxal-5 '-phosphate (PLP). It has been observed that the prevalence of mild forms of the disease is more than 40 times the prevalence of severe forms. Patients with HPP present at least one mutation in the ALPL gene. However, it is known that there are other causes that lead to decreased alkaline phosphatase (ALP) levels without mutations in the ALPL gene. Although the phenotype can be correlated with the genotype in HPP, the prediction of the phenotype from the genotype cannot be made with complete certainty. The availability of a specific enzyme replacement therapy for HPP undoubtedly represents an advance in therapeutic strategy, especially in severe forms of the disease in pediatric patients.Instituto de Salud Carlos III PI18-00803 PI18-01235 CM19/00188 CD20/00022European Commissio

Melatonin Treatment Reduces Oxidative Damage and Normalizes Plasma Pro-Inflammatory Cytokines in Patients Suffering from Charcot-Marie-Tooth Neuropathy: A Pilot Study in Three Children

Charcot-Marie-Tooth neuropathy (CMT) is a motor and sensory neuropathy comprising a heterogeneous group of inherited diseases. The CMT1A phenotype is predominant in the 70% of CMT patients, with nerve conduction velocity reduction and hypertrophic demyelination. These patients have elevated oxidative stress and chronic inflammation. Currently, there is no effective cure for CMT; herein, we investigated whether melatonin treatment may reduce the inflammatory and oxidative damage in CMT1A patients. Three patients, aged 8–10 years, were treated with melatonin (60 mg at 21:00 h plus 10 mg at 09:00 h), and plasma levels of lipid peroxidation (LPO), nitrites (NOx), IL-1β, IL-2, IL-6, TNF-α, INF-γ, oxidized to reduced glutathione (GSSG/GSH) ratio, and the activities of superoxide dismutase (SOD), glutathione-S transferase (GST), glutathione peroxidase (GPx), and reductase (GRd), were determined in erythrocytes at 3 and 6 months of treatment. Healthy age- and sex-matched subjects were used as controls. The results showed increased activities of SOD, GST, GPx, and GRd in CMT1A patients, which were reduced at 3 and 6 months of treatment. The GSSG/GSH ratio significantly increased in the patients, returning to control values after melatonin treatment. The inflammatory process was confirmed by the elevation of all proinflammatory cytokines measured, which were also normalized by melatonin. LPO and NOx, which also were elevated in the patients, were normalized by melatonin. The results document beneficial effects of the use of melatonin in CMT1A patients to reduce the hyperoxidative and inflammatory condition, which may correlate with a reduction of the degenerative process