Characterisation of five new variants of HLA-DPA1.

Genomic sequence of HLA-DPA1*01:03:01:73, -DPA1*01:03:01:80, DPA1*01:03:01:82, -DPA1*01:155:01:02, -DPA1*02:02:02:16 alleles in Spanish individuals.

Genomic sequence of five new variants of HLA-DQA1 in non-coding regions.

Characterisation of HLA-DQA1*01:02:01:30, HLA-DQA1*01:03:01:14, HLA-DQA1*03:03:01:20, HLA-DQA1*04:01:01:13, HLA-DQA1*05:05:01:40 alleles in Spanish individuals.

Technical success, procedural safety, and efficacy of the Silk Vista Baby in the treatment of cerebral aneurysms over a mid-to-long-term follow-up.

Background: Long-term follow-up of cerebral aneurysms treated with the Silk Vista Baby (SVB) flow diverter is lacking. This study aimed to assess the technical success, procedural safety, and efficacy of the SVB (Balt, Montmorency, France) for the treatment of intracranial aneurysms in small cerebral vessels over a mid-to long-term follow-up. Methods: We retrospectively analyzed a prospectively maintained database of patients treated with the SVB between September 2018 and June 2021. Data regarding patient demographics, aneurysm characteristics, and technical procedures were also collected. Angiographic and clinical findings were recorded during the procedure and over a period of at least 12 months. Results: Angiographic and clinical follow-up data were available for 50 patients/50 aneurysms. The procedural complication rate was 8%. At 12 months, the final results showed a technical success rate of 100%, the re rupture rate was 0%, neuromorbidity and mortality rates of 4 and 0%, respectively, and an almost complete occlusion rate of 94%. Conclusion: Treatment of complex intracranial aneurysms with the SVB was safe and effective. Long-term results showed high rates of adequate and stable occlusions.

[Translated article] Compounding training programme for pharmacy technicians using a digital platform and simulation techniques.

OBJECTIVE: To describe a compounding training plan in a tertiary hospital pharmacy service. The project aimed pharmacy assistant technicians to use a digital training platform and simulation techniques. METHODS: Two training programs were designed, one for sterile and the other for non-sterile drugs. Each programme consists of several phases: a basic online training course (digital e-learning platform), a practical simulation workshop, a supervised practice in real conditions, and a final verification by the pharmacist to qualify the technician. Both programs include continuous and accredited evaluation by the hospital's Continued Education Commission. A satisfaction survey on training (e-lerning platform) was designed and conducted for pharmacy technicians (sterile and non-sterile). RESULTS: The project has been running for 3 years. Six specialist pharmacists from different areas involved in compounding are responsible for training and continuous evaluation. Twenty-one technicians have been trained, and nineteen have obtained qualifications. Two of them were found to be unfit. Currently, we employ13 pharmacy technicians who were initially trained. The participation rate in the satisfaction survey on training (e-learning platform) was 61.5% (n=8) out of a total number of current developers (n=13). Overall, 62.5% of technicians reported being satisfied or very satisfied, while the remaining 37.5% were indifferent. CONCLUSIONS: Compounding training program is currently in its third year. It has been crucial in delegating tasks to pharmacy technicians. The use of digital technology is essential in this training. We consider that, specific training in compounding, is indispensable and should be included in the academic plan for pharmacy technicians.

Four new intronic variants of HLA-DQB1 detected by next-generation sequencing.

Characterization of HLA-DQB1*02:02:01:18, -DQB1*02:02:01:19, -DQB1*03:02:01:20 and -DQB1*05:03:01:10 alleles in Spanish individuals.

Exploring large-scale gene coexpression networks in peach (Prunus persica L.): a new tool for predicting gene function.

Peach is a model for Prunus genetics and genomics, however, identifying and validating genes associated to peach breeding traits is a complex task. A gene coexpression network (GCN) capable of capturing stable gene-gene relationships would help researchers overcome the intrinsic limitations of peach genetics and genomics approaches and outline future research opportunities. In this study, we created four GCNs from 604 Illumina RNA-Seq libraries. We evaluated the performance of every GCN in predicting functional annotations using an algorithm based on the 'guilty-by-association' principle. The GCN with the best performance was COO300, encompassing 21 956 genes. To validate its performance predicting gene function, we performed two case studies. In case study 1, we used two genes involved in fruit flesh softening: the endopolygalacturonases PpPG21 and PpPG22. Genes coexpressing with both genes were extracted and referred to as melting flesh (MF) network. Finally, we performed an enrichment analysis of MF network and compared the results with the current knowledge regarding peach fruit softening. The MF network mostly included genes involved in cell wall expansion and remodeling, and with expressions triggered by ripening-related phytohormones, such as ethylene, auxin, and methyl jasmonate. In case study 2, we explored potential targets of the anthocyanin regulator PpMYB10.1 by comparing its gene-centered coexpression network with that of its grapevine orthologues, identifying a common regulatory network. These results validated COO300 as a powerful tool for peach and Prunus research. This network, renamed as PeachGCN v1.0, and the scripts required to perform a function prediction analysis are available at https://github.com/felipecobos/PeachGCN.

Genetic Variation and Evolutionary Analysis of Eggplant Mottled Dwarf Virus Isolates from Spain.

The genetic variation and population structure of gene N (nucleocapsid) and part of gene L (replicase) from 13 eggplant mottle dwarf virus (EMDV) isolates from Spain were evaluated and compared with sequences of EMDV isolates from other countries retrieved from GenBank. Phylogenetic inference of part of gene L showed three main clades, one containing an EMDV isolate from Australia and the other two containing isolates from Iran and Europe, as well as four subclades. EMDV isolates from Spain were genetically very similar and grouped in a subclade together with one isolate from Germany and one from the UK. No new recombination events were detected in addition to one recombination previously reported, suggesting that recombination is rare for EMDV. The comparison of synonymous and non-synonymous rates showed that negative selection played an important role, and only two codons were under positive selection. Genetic differentiation (Fst test), phylogenetic and nucleotide diversity analyses suggest a unique introduction of EMDV to Spain and low gene flow with other countries. In contrast, Greece and Italy showed diverse populations with high gene flow between both.

Genomic sequence of the HLA-B*40:01:02:61, -B*48:01:01:14, and -C*01:02:01:69 alleles.

Characterization of HLA-B*40:01:02:61, -B*48:01:01:14, and -C*01:02:01:69 alleles in Spanish individuals.

BCAS1 defines a heterogeneous cell population in diffuse gliomas.

Oligodendrocyte precursor markers have become of great interest to identify new diagnostic and therapeutic targets for diffuse gliomas, since state-of-the-art studies point towards immature oligodendrocytes as a possible source of gliomagenesis. Brain enriched myelin associated protein 1 (BCAS1) is a novel marker of immature oligodendrocytes and was proposed to contribute to tumorigenesis in non-central nervous system tumors. However, BCAS1 role in diffuse glioma is still underexplored. This study analyzes the expression of BCAS1 in different tumor samples from patients with diffuse gliomas (17 oligodendrogliomas; 8 astrocytomas; 60 glioblastomas) and uncovers the molecular and ultrastructural features of BCAS1+ cells by immunostaining and electron microscopy. Our results show that BCAS1+ cells exhibit stellate or spherical morphology with similar ultrastructural features. Stellate and spherical cells were detected as isolated cells in all studied gliomas. Nevertheless, only stellate cells were found to be proliferative and formed tightly packed nodules with a highly proliferative rate in oligodendrogliomas. Our findings provide a comprehensive characterization of the BCAS1+ cell population within diffuse gliomas. The observed proliferative capacity and distribution of BCAS1+ stellate cells, particularly in oligodendrogliomas, highlight BCAS1 as an interesting marker, warranting further investigation into its role in tumor malignancy.

Dietary Milk or Isolated Legume Proteins Modulate Intestinal Microbiota Composition in Rats.

Shifts toward increased proteolytic fermentation, such as, for example, in athlete and high-protein weight loss diets, may alter the relative abundance of microbial species in the gut and generate bioactive, potentially deleterious metabolic products. In the current investigation, intestinal (caecal) microbiota composition was studied in rats fed diets differing only in their constituent proteins: milk (casein (CAS), lactalbumin (LA)) or legume (Cicer arietinum, Lupinus angustifolius) protein isolates (chickpea protein isolate (CPI), lupin protein isolate (LI)). ANOSIM and Discriminant Analysis showed significant (p < 0.05) differences at both family and genus levels in both microbiota composition and functionality as a consequence of feeding the different proteins. Differences were also significant (p < 0.05) for predicted functionality parameters as determined by PICRUSt analysis. LA induced a generally healthier microbiota composition than CAS, and higher amounts of Methanobrevibacter spp. and Methanogenic_PWY were found in the LI group. LEfSe analysis of bacterial composition and functional activities revealed a number of groups/functions able to explain the different effects found with milk and legume protein isolates. In conclusion, the mostly beneficial modulation of intestinal microbiota generally found with legume-based diets is likely to be due, at least in part, to their constituent proteins.

Three novel variants of HLA-DQB1*06 identified by next-generation sequencing.

Characterization of HLA-DQB1*06:472, -DQB1*06:02:01:34 and -DQB1*06:04:01:06 alleles in Spanish individuals.

Nitrogenase cofactor biosynthesis using proteins produced in mitochondria of Saccharomyces cerevisiae.

Biological nitrogen fixation, the conversion of inert N2 to metabolically tractable NH3, is only performed by certain microorganisms called diazotrophs and is catalyzed by the nitrogenases. A [7Fe-9S-C-Mo-R-homocitrate]-cofactor, designated FeMo-co, provides the catalytic site for N2 reduction in the Mo-dependent nitrogenase. Thus, achieving FeMo-co formation in model eukaryotic organisms, such as Saccharomyces cerevisiae, represents an important milestone toward endowing them with a capacity for Mo-dependent biological nitrogen fixation. A central player in FeMo-co assembly is the scaffold protein NifEN upon which processing of NifB-co, an [8Fe-9S-C] precursor produced by NifB, occurs. Prior work established that NifB-co can be produced in S. cerevisiae mitochondria. In the present work, a library of nifEN genes from diverse diazotrophs was expressed in S. cerevisiae, targeted to mitochondria, and surveyed for their ability to produce soluble NifEN protein complexes. Many such NifEN variants supported FeMo-co formation when heterologously produced in the diazotroph A. vinelandii. However, only three of them accumulated in soluble forms in mitochondria of aerobically cultured S. cerevisiae. Of these, two variants were active in the in vitro FeMo-co synthesis assay. NifEN, NifB, and NifH proteins from different species, all of them produced in and purified from S. cerevisiae mitochondria, were combined to establish successful FeMo-co biosynthetic pathways. These findings demonstrate that combining diverse interspecies nitrogenase FeMo-co assembly components could be an effective and, perhaps, the only approach to achieve and optimize nitrogen fixation in a eukaryotic organism.IMPORTANCEBiological nitrogen fixation, the conversion of inert N2 to metabolically usable NH3, is a process exclusive to diazotrophic microorganisms and relies on the activity of nitrogenases. The assembly of the nitrogenase [7Fe-9S-C-Mo-R-homocitrate]-cofactor (FeMo-co) in a eukaryotic cell is a pivotal milestone that will pave the way to engineer cereals with nitrogen fixing capabilities and therefore independent of nitrogen fertilizers. In this study, we identified NifEN protein complexes that were functional in the model eukaryotic organism Saccharomyces cerevisiae. NifEN is an essential component of the FeMo-co biosynthesis pathway. Furthermore, the FeMo-co biosynthetic pathway was recapitulated in vitro using only proteins expressed in S. cerevisiae. FeMo-co biosynthesis was achieved by combining nitrogenase FeMo-co assembly components from different species, a promising strategy to engineer nitrogen fixation in eukaryotic organisms.

Iron Homeostasis in Azotobacter vinelandii.

Iron is an essential nutrient for all life forms. Specialized mechanisms exist in bacteria to ensure iron uptake and its delivery to key enzymes within the cell, while preventing toxicity. Iron uptake and exchange networks must adapt to the different environmental conditions, particularly those that require the biosynthesis of multiple iron proteins, such as nitrogen fixation. In this review, we outline the mechanisms that the model diazotrophic bacterium Azotobacter vinelandii uses to ensure iron nutrition and how it adapts Fe metabolism to diazotrophic growth.

Three novel alleles HLA-A*02:1092, -A*29:01:01:11 and -B*44:03:01:58 identified by next-generation sequencing.

Three novel HLA Class I alleles, -A*02:1092, -A*29:01:01:11 and -B*44:03:01:58, characterized in Spanish individuals.

Four novel HLA alleles: HLA-DPA1*01:03:01:68, -DPA1*01:03:01:71, -DQA1*02:01:01:06, and -DQB1*06:03:01:19.

Four novel HLA Class II alleles, HLA-DPA1*01:03:01:68, -DPA1*01:03:01:71, -DQA1*02:01:01:06, and -DQB1*06:03:01:19, characterized in Spanish individuals.

Manifestaciones laríngeas de COVID-19 / Laryngeal manifestations of COVID-19

Introducción: la infección por COVID-19 afecta el tracto aerodigestivo superior a través de la enzima convertidora de angiotensina 2 (ECA2) y/o la proteasa transmembrana serina 2 (TMPRSS2). Sus manifestaciones agudas y secuelas han sido muy variadas y no todas están relacionadas con la intubación orotraqueal. El objetivo es describir las características sociodemográficas, clínicas y los hallazgos endoscópicos de los pacientes con síntomas laringofaríngeos posteriores a una infección por SARS-CoV-2 evaluados en el Hospital Militar Central y Hospital Universitario Clínica San Rafael entre marzo de 2020 y marzo de 2022. Materiales y métodos: estudio observacional de corte transversal con datos sociodemográficos, comorbilidades, necesidad de intubación orotraqueal, variedad de síntomas y sus hallazgos endoscópicos. Resultados: se recolectaron datos de 118 pacientes; la edad media fue de 51 años ± 14,4. El síntoma más frecuente fue la disfonía (69,5 %), seguido de la disnea (39,8 %). El 58,9 % requirió intubación orotraqueal y, de estos, la manifestación más frecuente fue disfonía por tensión muscular (DTM) y estenosis subglótica-traqueal. En el 41,1 % restante su hallazgo más frecuente fue la laringitis irritativa. Conclusiones: la COVID-19 tiene múltiples manifestaciones laringofaríngeas en relación con su mecanismo de infección e invasión en los tejidos de esta zona, de tipo inflamatorio y estructural, y no todos están relacionados con la intubación.

Introduction: COVID 19 infection affects the upper aerodigestive tract through angiotensin-converting enzyme 2 (ACE2) and/or Transmembrane serine protease 2 (TMPRSS2). Its acute manifestations and sequelae have been very varied, and not all of them are related to orotracheal intubation. The objective is to describe the sociodemographic and clinical characteristics and the endoscopic findings of patients with laryngopharyngeal symptoms after SARS-CoV-2 infection evaluated at the Hospital Militar Central and Hospital Universitario Clínica San Rafael between March 2020 and March 2022. Methods: Cross-sectional observational study, obtaining sociodemographic data, comorbidities, need for orotracheal intubation, variety of symptoms and their endoscopic findings. Results: 118 patients were collected; the mean age was 51 years ± 14.4. The most frequent symptom was dysphonia (69.5%), followed by dyspnea (39.8%). 58.9% required orotracheal intubation and of these the most frequent manifestation was muscular tension dysphonia (MTD) and subglottictracheal stenosis. In the remaining 41.1%, the most frequent finding was irritative laryngitis. Conclusions: COVID-19 has multiple laryngopharyngeal manifestations in relation to its mechanism of infection and invasion in the tissues of this area, as an inflammatory and structural type, and not all of them are related to intubation.

Identification of four novel HLA-DQA1 alleles: HLA-DQA1*01:01:02:04, 01:04:01:05, 03:03:01:19 and 05:01:01:08.

Four novel HLA-DQA1 alleles, -DQA1*01:01:02:04, -DQA1*01:04:01:05, -DQA1*03:03:01:19 and -DQA1*05:01:01:08, characterized in Spanish individuals.

Borrelia miyamotoi Infection in Immunocompromised Man, California, USA, 2021.

Infection with Borrelia miyamotoi in California, USA, has been suggested by serologic studies. We diagnosed B. miyamotoi infection in an immunocompromised man in California. Diagnosis was aided by plasma microbial cell-free DNA sequencing. We conclude that the infection was acquired in California.

Dislocation and survival rate of dual mobility cups in revision total hip arthroplasty in patients with high risk of instability.

PURPOSE: Instability remains as an unsolved complication after revision total hip arthroplasty (rTHA). Dual mobility (DM) cups have decreased dislocation rate in rTHA; however, there are few clinical studies that evaluate the risk of dislocation after rTHA in patients with high risk of instability. METHODS: Between 2007 and 2019, 95 consecutive rTHA using a DM cup, in 93 patients with instability risk factors were retrospectively reviewed. Instability risk factors were defined as follows: deficient abductor mechanism (22.1%), previous dislocation (68.4%), Paprosky 3 acetabular bone defect (21.1%), neuromuscular disease (4.2%), lumbar spine arthrodesis (6.3%), and more than one previous surgery (33.6%). Mean follow-up was 4.3 ± 2.4 years. Clinical and radiographic parameters were reported. DM cup survival was determined using a Kaplan-Meier analysis with re-revision for any cause as the end-point. Surgical complications and re-dislocation rate was recorded. RESULTS: Seven DM cups were re-revised; three due to aseptic loosening, three due to peri-prosthetic joint infection (PJI), and one due to instability. At seven years follow-up, the survival rate of the DM cup using re-revision for any reason as the end-point was 82.4% (95% confidence interval, 66.4-98.4%). There were 12 (12.6%) complications: two (2.1%) dislocations, six (6.3%) PJI, four (4.2%) peri-prosthetic fractures. CONCLUSION: Our findings suggest DM cups are effective reducing dislocation rate in rTHA in patients with a high risk of instability, without increasing early aseptic loosening and with a low rate of dislocation. Nevertheless, longer follow-up is needed to confirm implant stability and dislocation rates in the long term.

Identification of three novel HLA class I alleles: HLA-B*15:640, -B*18:01:01:71 and -C*05:275.

Three novel HLA Class I alleles, -B*15:640, -B*18:01:01:71 and -C*05:275, characterized in Spanish individuals.