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The feasibility of Near Infrared Spectroscopy was assessed for aging traceability of steaks of Angus beef (Biceps femoris) individually vacuum-packaged, as well as for the prediction of the refrigeration storage time (0, 7, and 14 days). For this purpose, a total of 288 steaks homogeneously distributed among the sampling times were used. The model developed by Partial Least Squares-Discriminant Analysis offered high discrimination ability between aged beef vs. non-aged. The accuracy after external validation exceeded 90%. Regarding the predictive capacity of the storage time, it was greater on the set of aged samples, in which the accuracy achieved values higher than 96%, while the accuracy decreased to 75% for the non-aged samples. Results obtained support the ability of NIRS technology to be considered in any digital transformation strategy for traceability across the meat supply chain.
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Espectroscopía Infrarroja Corta , Espectroscopía Infrarroja Corta/métodos , Espectroscopía Infrarroja Corta/instrumentación , Animales , Bovinos , Carne/análisis , Almacenamiento de AlimentosRESUMEN
Aging is an inevitable and gradual decline in several biological functions. Mitochondrial dysfunction is one of the most important hallmarks of aging. In this context, alterations in metabolites associated with mitochondrial dysfunction may serve as a significant biomarker. This study aimed to investigate the existence of a relationship between the key metabolites involved in bioenergetics metabolism and aging. 53 volunteers ranged 20-85 years participated in the study. We tested the association between different tricarboxylic acid (TCA) cycle metabolites, fatty acid metabolism, and amino acid metabolism with age, sex, body composition, and proxy markers of aging such as walking speed, grip strength and chair test. We found that lactic acid negatively correlated with age while several fatty acid metabolites, such as azelaic, sebacic, and linoleic acids, showed positive correlations with age (p < 0.05). Sex-specific trends, such as glycerol, and dodecanoic acid, were also observed for certain metabolites. Furthermore, citric acid levels were found to have a significant association with physical function and body composition measures. Participants with higher citric acid levels displayed improved performance in physical tests and favorable body composition indices. Additionally, fumaric acid and adipic acid showed positive correlations with fat-free body mass, while sebacic acid was negatively associated with measures of fat mass. These findings underscore the importance of understanding the role of circulating bioenergetics metabolites with age, sex variations, and their potential implications in body composition and physical performance.
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Composición Corporal , Metabolismo Energético , Humanos , Masculino , Femenino , Anciano , Persona de Mediana Edad , Adulto , Anciano de 80 o más Años , Adulto Joven , Envejecimiento Saludable , Ácidos Grasos/metabolismo , Ácidos Grasos/sangre , Ciclo del Ácido Cítrico , Biomarcadores/sangre , Aminoácidos/metabolismo , Aminoácidos/sangre , Envejecimiento/metabolismo , Envejecimiento/fisiología , Fuerza de la Mano/fisiologíaRESUMEN
Congenital ichthyoses (CI) comprise a heterogeneous group of monogenic genetic skin diseases characterized by diffuse scaling, often associated with skin inflammation. Diagnosis of the individual form of ichthyosis is complex and is guided by clinical expertise. CI usually has a major impact on quality of life (QOL) and thus requires lifelong treatment. To date, there are no curative therapies, although various symptomatic treatment options exist. The present protocol for the management of CI has been drawn up in accordance with the recommendations published in 2012 by the French National Authority for Health, based on a literature review, with the help and validation of members of the French network for rare skin diseases (FIMARAD). It provides a summary of evidence and expert-based recommendations and is intended to help clinicians with the management of these rare and often complex diseases.
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Ictiosis Lamelar , Ictiosis , Humanos , Calidad de Vida , Ictiosis Lamelar/diagnóstico , Ictiosis Lamelar/genética , Ictiosis Lamelar/terapia , Ictiosis/diagnóstico , Ictiosis/genética , Ictiosis/terapia , Piel , Diagnóstico Diferencial , Literatura de Revisión como AsuntoAsunto(s)
Acné Vulgar , Isotretinoína , Factor de Necrosis Tumoral alfa , Humanos , Acné Vulgar/inducido químicamente , Acné Vulgar/tratamiento farmacológico , Acné Vulgar/patología , Factores Inmunológicos/efectos adversos , Isotretinoína/efectos adversos , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Masculino , AdolescenteRESUMEN
The COVID-19 pandemic has challenged the entire world, and patients with diabetes mellitus (DM) have been particularly affected. We aimed to evaluate predictors of mortality during the first 30 days of hospitalization in critically ill patients with COVID-19 and comorbid DM. This prospective study included 110 critically ill patients admitted with COVID-19 infection. Thirty-two (29%) patients had a previous diagnosis of DM. Clinical variables, laboratory tests, and vascular biomarkers, such as VCAM-1, syndecan-1, ICAM-1, angiopoietin-1, and angiopoeitin-2, were evaluated after intensive care unit (ICU) admission. A comparison was made between patients with and without DM. No difference in mortality was observed between the groups (48.7 vs 46.9%, P=0.861). In the multivariate Cox regression analysis, VCAM-1 levels at ICU admission (HR: 1 [1-1.001], P<0.006) were associated with death in patients with DM. Among patients with DM, advanced age (HR 1.063 [1.031-1.096], P<0.001), increased Ang-2/Ang-1 ratio (HR: 4.515 [1.803-11.308] P=0.001), and need for dialysis (HR: 3.489 [1.409-8.642], P=0.007) were independent predictors of death. Higher levels of VCAM-1 in patients with DM was better at predicting death of patients with severe COVID-19 and comorbid DM, and their cut-off values were useful for stratifying patients with a worse prognosis. Vascular biomarkers VCAM-1 and Ang-2/Ang-1 ratio were predictors of death in patients with severe COVID-19 and comorbid DM and those without DM. Additionally, kidney injury was associated with an increased risk of death.
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COVID-19 , Diabetes Mellitus , Humanos , Enfermedad Crítica , Estudios Prospectivos , Pandemias , Molécula 1 de Adhesión Celular Vascular , Mortalidad Hospitalaria , Unidades de Cuidados Intensivos , Biomarcadores , Estudios RetrospectivosRESUMEN
OBJECTIVES: To evaluate the efficacy of subcutaneous administration of dexmedetomidine/atipamezole at the Governing Vessel 20 (GV20) acupuncture point compared with other administration routes (intramuscular and intravenous) in dogs presented for orthopaedic radiographs. MATERIALS AND METHODS: Prospective, randomised, blinded, controlled clinical study. Sixty-four client-owned dogs were randomly injected with 200 µg/m2 of dexmedetomidine intramuscular (lumbar muscles) (n=20), intravenous (n=23) or subcutaneous at the GV20 point (n=21). Following radiographs, dogs received 2000 µg/m2 of atipamezole intramuscular (n=31), or subcutaneous at the GV20 point (n=27). Degree and time to sedation and recovery were assessed using a sedation scale and a Dynamic and Interactive Visual Analog Scale (DIVAS). Clinical physiological variables and adverse events were used. Statistical linear mixed-effect models (analysis of variance) and Cox models were performed. Significance was set at P-value <0.05. RESULTS: Sedation was insufficient to perform orthopaedic radiographs in six dogs in the intramuscular group. The time to sedation was significantly longer, and sedation scale and DIVAS scores were significantly lower in the intramuscular group. The intravenous group had significantly higher sedation scale and DIVAS scores than the GV20 group. No significant differences were observed between the intramuscular and GV20 recovery groups, although the time effect was significantly more pronounced in the GV20 recovery group. CLINICAL SIGNIFICANCE: Subcutaneous administration of dexmedetomidine and atipamezole at GV20 provided effective sedation and recovery in dogs undergoing orthopaedic radiographic studies. GV20 administration provided a clinically similar level of sedation to the intravenous route, and greater and faster sedation and similar recovery to intramuscular.
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Puntos de Acupuntura , Enfermedades de los Perros , Hipnóticos y Sedantes , Ortopedia , Animales , Perros , Dexmedetomidina/administración & dosificación , Dexmedetomidina/efectos adversos , Método Doble Ciego , Hipnóticos y Sedantes/administración & dosificación , Hipnóticos y Sedantes/efectos adversos , Estudios Prospectivos , Enfermedades de los Perros/diagnóstico por imagen , Inyecciones Subcutáneas/métodosRESUMEN
PURPOSE: To investigate the prevalence of negative behaviour in preschool children attending dental clinics and its association with sociodemographic, oral health-related and parental psychosocial factors. METHODS: A cross-sectional study was carried out with 145 parents/guardians and their children aged 4-6 years attending paediatric dentistry training programmes in a capital city of Midwest Brazil. Data were obtained from children's dental records, interviews, and questionnaires for parents/guardians. The outcome was negative child behaviour, based on the dentists' use or indication of behavioural control measures during the dental appointments, as registered in the children's dental records. Covariates were sociodemographic, clinical and parent/guardian psychosocial factors religiosity (DUREL index) and Sense of Coherence (SOC-13 scale). Bivariate analyses and Poisson regression with robust variance were performed. RESULTS: The prevalence of negative behaviour was 24.1% (95% CI = 17.9-31.7). In the bivariate analyses, the variables initially selected for the regression models (p < 0.25) were the parent/guardians' number of children and religiosity, and the children's dental pain and caries status in deciduous teeth. After adjustment, the prevalence of negative behaviour was 2.12 higher in children with teeth extracted due to caries. CONCLUSION: The prevalence of negative behaviour was high and associated with the presence of missing teeth due to caries, regardless of sociodemographic, psychosocial, and other oral health factors.
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Caries Dental , Salud Bucal , Humanos , Preescolar , Brasil/epidemiología , Prevalencia , Estudios Transversales , Caries Dental/epidemiologíaRESUMEN
Anchored Hybrid Enrichment (AHE) is a tool for capturing orthologous regions of the nuclear genome shared in low or single copy across lineages. Despite the increasing number of studies using this method, its usefulness to estimate relationships at deeper taxonomic levels in plants has not been fully explored. Here we present a proof of concept about the performance of nuclear loci obtained with AHE to infer phylogenetic relationships and explore the use of gene sampling schemes to estimate divergence times in Asterales. We recovered low-copy nuclear loci using the AHE method from herbarium material and silica-preserved samples. Maximum likelihood, Bayesian inference, and coalescence approaches were used to reconstruct phylogenomic relationships. Dating analyses were conducted under a multispecies coalescent approach by jointly inferring species tree and divergence times with random gene sampling schemes and multiple calibrations. We recovered 403 low-copy nuclear loci for 63 species representing nine out of eleven families of Asterales. Phylogenetic hypotheses were congruent among the applied methods and previously published results. Analyses with concatenated datasets were strongly supported, but coalescence-based analyses showed low support for the phylogenetic position of families Argophyllaceae and Alseuosmiaceae. Estimated family ages were congruent among gene sampling schemes, with the mean age for Asterales around 130 Myr. Our study documents the usefulness of AHE for resolving phylogenetic relationships at deep phylogenetic levels in Asterales. Observed phylogenetic inconsistencies were possibly due to the non-inclusion of families Phellinceae and Pentaphragmataceae. Random gene sampling schemes produced consistent age estimates with coalescence and species tree relaxed clock approaches.
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Magnoliopsida , Filogenia , Magnoliopsida/genética , Teorema de Bayes , Genoma , Núcleo Celular/genéticaRESUMEN
The COVID-19 pandemic has challenged the entire world, and patients with diabetes mellitus (DM) have been particularly affected. We aimed to evaluate predictors of mortality during the first 30 days of hospitalization in critically ill patients with COVID-19 and comorbid DM. This prospective study included 110 critically ill patients admitted with COVID-19 infection. Thirty-two (29%) patients had a previous diagnosis of DM. Clinical variables, laboratory tests, and vascular biomarkers, such as VCAM-1, syndecan-1, ICAM-1, angiopoietin-1, and angiopoeitin-2, were evaluated after intensive care unit (ICU) admission. A comparison was made between patients with and without DM. No difference in mortality was observed between the groups (48.7 vs 46.9%, P=0.861). In the multivariate Cox regression analysis, VCAM-1 levels at ICU admission (HR: 1 [1-1.001], P<0.006) were associated with death in patients with DM. Among patients with DM, advanced age (HR 1.063 [1.031-1.096], P<0.001), increased Ang-2/Ang-1 ratio (HR: 4.515 [1.803-11.308] P=0.001), and need for dialysis (HR: 3.489 [1.409-8.642], P=0.007) were independent predictors of death. Higher levels of VCAM-1 in patients with DM was better at predicting death of patients with severe COVID-19 and comorbid DM, and their cut-off values were useful for stratifying patients with a worse prognosis. Vascular biomarkers VCAM-1 and Ang-2/Ang-1 ratio were predictors of death in patients with severe COVID-19 and comorbid DM and those without DM. Additionally, kidney injury was associated with an increased risk of death.
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Objective. The goal of this work is to experimentally compare the 3D spatial and energy resolution of a semi-monolithic detector suitable for total-body positron emission tomography (TB-PET) scanners using different surface crystal treatments and silicon photomultiplier (SiPM) models.Approach. An array of 1 × 8 lutetium yttrium oxyorthosilicate (LYSO) slabs of 25.8 × 3.1 × 20 mm3separated with Enhanced Specular Reflector (ESR) was coupled to an array of 8 × 8 SiPMs. Three different treatments for the crystal were evaluated: ESR + RR + B,with lateral faces black (B) painted and a retroreflector (RR) layer added to the top face; ESR +RR, with lateral faces covered with ESR and a RR layer on the top face and; All ESR, with lateral and top sides with ESR. Additionally, two SiPM array models from Hamamatsu Photonics belonging to the series S13361-3050AE-08 (S13) and S14161-3050AS-08 (S14) have been compared. Coincidence data was experimentally acquired using a22Na point source, a pinhole collimator, a reference detector and moving the detector under study in 1 mm steps in thex- andDOI- directions. The spatial performance was evaluated by implementing a neural network (NN) technique for the impact position estimation in thex- (monolithic) andDOIdirections.Results. Energy resolution values of 16 ± 1%, 11 ± 1%, 16 ± 1%, 15 ± 1%, and 13 ± 1% were obtained for theS13-ESR + B + RR,S13-AllESR,S14-ESR + B + RR,S14-ESR + RR,andS14-AllESR, respectively. Regarding positioning accuracy, mean average error of 1.1 ± 0.5, 1.3 ± 0.5 and 1.3 ± 0.5 were estimated for thex- direction and 1.7 ± 0.8, 2.0 ± 0.9 and 2.2 ± 1.0 for theDOI- direction, for the ESR + B + RR, ESR + RR and All ESR cases, respectively, regardless of the SiPM model.Significance. Overall, the obtained results show that the proposed semi-monolithic detectors are good candidates for building TB-PET scanners.
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Redes Neurales de la Computación , Tomografía de Emisión de PositronesRESUMEN
PURPOSE: To study Oral Health related Quality of Life (OHRQoL) of pre-school children and its associated factors. METHODS: Cross-sectional study with sample data collected through an on-line questionnaire to a non-probabilistic sample. The questionnaire included the Portuguese version of the Early Childhood Oral Health Impact Scale (ECOHIS) and questions about sociodemographic characteristics, oral health behaviours, and reported oral health of the child. Data analyses included descriptive statistics, Mann-Whitney and Kruskal-Wallis tests and Linear regression (α = 0.05). RESULTS: The sample consisted of 1475 parents of pre-school children (3-5 years-old), residing in Portugal. The mean ECOHIS total score was 1.5 (sd = 3.4). In the bivariate analysis the child's age, starting toothbrushing after one year of age, frequent intake of sugary food and drinks, previous appointment with an oral health professional, all reported of oral health problems, and a negative perception of oral health were negatively associated with OHRQoL (p < 0,05). In the regression model the factors that most contributed to a worse quality of life were dental caries and abscess, contributing to an ECOHIS average increase of 2.56 and 3.34, respectively. CONCLUSIONS: The studied population presented a good OHRQoL, with the item related to pain being the most relevant for the ECOHIS score. Worst OHRQoL was found in older children, whose parents negatively rated the children's oral health and with reported dental caries, trauma, and abscess.
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Caries Dental , Salud Bucal , Calidad de Vida , Niño , Preescolar , Humanos , Absceso , Estudios Transversales , Caries Dental/epidemiología , Padres , Portugal , Encuestas y CuestionariosRESUMEN
PURPOSE: To study the results of surgical treatment of acquired concomitant esotropia and its posible causes. METHODS: Retrospective study of the patients with acute acquired esotropia that were operated on since 2017. Distance and near horizontal deviation, spherical equivalent and the excessive use of the tablets and smartphones were analyzed. A good sensorial outcome was considered when stereopsis was recovered and good motor outcome if horizontal deviation was ≤10 prism diopters at the end of the follow-up. RESULTS: A total of 15 cases were included (60% women), mean age: 29.07 years (86.66% ≥16 years). Mean time since the beginning of the symptoms to the surgery was 23.3 months. Mean horizontal preoperative deviation was 19.73 ± 7.2 pd at near and 22.93 ± 7.3 dp at distance decreasing to 3.33 ± 3.8 pd at near and 3.86 ± 3.9 pd at distance (P < .001). The 66.6% had diplopía. The 46.66% had decompensated esophoria or microesotropia, 86,66 % myopia and the 33.33% referred excesive use of the tablets and smartphones. The medial rectus recession were done in 10 cases. A good motor outcome was obtained in the 93.33% of the cases and sensorial outcome in the 53.3% without diplopía. CONCLUSION: Surgery was an excellent treatment to resolve the deviation and diplopia. The esophorias and microesotropias represented near the half of the cases and the excesive use of the tablets and smartphones were the third part of the sample. Most of the patients had myopia.
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Esotropía , Miopía , Enfermedad Aguda , Adulto , Diplopía/cirugía , Esotropía/diagnóstico , Esotropía/etiología , Esotropía/cirugía , Femenino , Humanos , Masculino , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
The economically important cotton and cacao family (Malvaceae sensu lato) have long been recognized as a monophyletic group. However, the relationships among some subfamilies are still unclear as discordant phylogenetic hypotheses keep arising when different sources of molecular data are analyzed. Phylogenetic discordance has previously been hypothesized to be the result of both introgression and incomplete lineage sorting (ILS), but the extent and source of discordance have not yet been evaluated in the context of loci derived from massive sequencing strategies and for a wide representation of the family. Furthermore, no formal methods have been applied to evaluate if the detected phylogenetic discordance among phylogenomic datasets influences phylogenetic dating estimates of the concordant relationships. The objective of this research was to generate a phylogenetic hypothesis of Malvaceae from nuclear genes, specifically we aimed to (1) investigate the presence of major discordance among hundreds of nuclear gene histories of Malvaceae; (2) evaluate the potential source of discordance; and (3) examine whether discordance and loci heterogeneity influence on time estimates of the origin and diversification of subfamilies. Our study is based on a comprehensive dataset representing 96 genera of the nine subfamilies and 268 nuclear loci. Both concatenated and coalescence-based approaches were followed for phylogenetic inference. Using branch lengths and topology, we located the placement of introgression events to directly evaluate whether discordance is due to introgression rather than ILS. To estimate divergence times, concordance and molecular rate were considered. We filtered loci based on congruence with the species tree and then obtained the molecular rate of each locus to distribute them into three different sets corresponding to shared molecular rate ranges. Bayesian dating was performed for each of the different sets of loci with the same parameters and calibrations. Phylogenomic discordance was detected between methods, as well as gene histories. At deep coalescent times, we found discordance in the position of five subclades probably due to ILS and a relatively small proportion of introgression. Divergence time estimation with each set of loci generated overlapping clade ages, indicating that, even with different molecular rate and gene histories, calibrations generally provide a strong prior.
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PURPOSE: The school environment may positively influence student health behaviours and learning. This study aimed to investigate the association between cities' quality of education and adolescent students' oral health-related behaviours. METHODS: Cross-sectional study using data of the 2015 Brazilian National Adolescent School-Based Health Survey and other public databases. The sample was composed of adolescents (N = 23,674) from public schools of the 27 Brazilian state capitals. Outcomes were four oral health-related behaviours: toothbrushing, sweets and soft-drink consumption, and dental visits. The explanatory variable was the cities' quality of education, measured by the Brazilian Basic Education Development Index. Covariates were individual (sociodemographic) and contextual (socioeconomic, oral health coverage and oral health-promoting schools). Multilevel logistic regression was performed considering two levels: individual (adolescents) and contextual (city). RESULTS: The prevalence of the outcomes were: low daily toothbrushing frequency 6.7% (95% CI 6.0-7.4); high weekly sweets consumption 41.5% (95% CI 40.3-42.7); high weekly soft-drink consumption 28.5% (95% CI 27.2-29.9); and low frequency of annual dental visits 31.2% (95% CI 30.1-32.3). In the adjusted models, cities whose schools had higher scores of education quality were more likely to have students with low toothbrushing frequency and high frequency of sweets consumption. CONCLUSION: The cities' quality of education was associated with unhealthy oral health-related behaviours, particularly the frequency of toothbrushing and sweets consumption. Therefore, appropriate health promotion strategies as well as high-quality education are needed in schools.
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Conductas Relacionadas con la Salud , Conocimientos, Actitudes y Práctica en Salud , Adolescente , Brasil/epidemiología , Estudios Transversales , Evaluación Educacional , Escolaridad , Humanos , Análisis Multinivel , Factores SocioeconómicosRESUMEN
Oral tissue regeneration following chronic diseases and injuries is limited by the natural endogenous wound-healing process. Current regenerative approaches implement exogenous systems, including stem cells, scaffolds, growth factors, and plasmid DNA/viral vectors, that induce variable clinical outcomes. An innovative approach that is safe, effective, and inexpensive is needed. The lipid nanoparticle-encapsulated nucleoside-modified messenger RNA (mRNA) platform has proven to be a successful vaccine modality against coronavirus disease 2019, demonstrating safety and high efficacy in humans. The same fundamental technology platform could be applied to facilitate the development of mRNA-based regenerative therapy. While the platform has not yet been studied in the field of oral tissue regeneration, mRNA therapeutics encoding growth factors have been evaluated and demonstrated promising findings in various models of soft and hard tissue regeneration such as myocardial infarction, diabetic wound healing, and calvarial and femoral bone defects. Because restoration of both soft and hard tissues is crucial to oral tissue physiology, this new therapeutic modality may help to overcome challenges associated with the reconstruction of the unique and complex architecture of oral tissues. This review discusses mRNA therapeutics with an emphasis on findings and lessons in different regenerative animal models, and it speculates how we can apply mRNA-based platforms for oral tissue regeneration.
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COVID-19 , Ingeniería de Tejidos , Animales , Regeneración Ósea/genética , Humanos , Péptidos y Proteínas de Señalización Intercelular , Liposomas , Nanopartículas , ARN Mensajero , Tecnología , Cicatrización de Heridas/genéticaRESUMEN
BACKGROUND: As COVID-19 became a pandemic, the urgent need to find an effective treatment vaccine has been a major objective. Vaccines contain adjuvants which are not exempt from adverse effects and can trigger the autoimmune/inflammatory syndrome induced by adjuvants (ASIA). There is very little information about autoimmune endocrine disease and the ASIA after the use of mRNA-based SARS-CoV2 vaccination. CASE SERIES: We report three cases and also review the literature showing that the thyroid gland can be involved in the ASIA induced by the mRNA-based SARS-CoV2 vaccination. We present the first case to date of silent thyroiditis described in the context of SARS-CoV2 vaccination with Pfizer/BioNTech. Also, we discuss the first subacute thyroiditis in the context of SARS-CoV2 vaccination with the Moderna's vaccine. Finally, we provide another case to be added to existing evidence on Graves' disease occurring post-vaccination with the Pfizer/BioNTech vaccine. DISCUSSION: Adjuvants play an important role in vaccines. Their ability to increase the immunogenicity of the active ingredient is necessary to achieve the desired immune response. Both the Moderna and the Pfizer/BioNTech vaccines use mRNA coding for the SARS-CoV2 S protein enhanced by adjuvants. In addition, the cross-reactivity between SARS-CoV2 and thyroid antigens has been reported. This would explain, at least, some of the autoimmune/inflammatory reactions produced during and after SARS-CoV2 infection and vaccination. CONCLUSION: The autoimmune/inflammatory syndrome induced by adjuvants involving the thyroid could be an adverse effect of SARS-CoV2 vaccination and could be underdiagnosed.
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Vacunas contra la COVID-19/efectos adversos , Enfermedad de Graves/etiología , Glándula Tiroides/inmunología , Tiroiditis/etiología , Vacunación/efectos adversos , Adulto , Vacunas contra la COVID-19/inmunología , Femenino , Enfermedad de Graves/inmunología , Humanos , Masculino , Tiroiditis/inmunologíaRESUMEN
SARS-CoV-2 is the causative agent of COVID-19. The dimeric form of the viral Mpro is responsible for the cleavage of the viral polyprotein in 11 sites, including its own N and C-terminus. The lack of structural information for intermediary forms of Mpro is a setback for the understanding its self-maturation process. Herein, we used X-ray crystallography combined with biochemical data to characterize multiple forms of SARS-CoV-2 Mpro. For the immature form, we show that extra N-terminal residues caused conformational changes in the positioning of domain-three over the active site, hampering the dimerization and diminishing its activity. We propose that this form preludes the cis and trans-cleavage of N-terminal residues. Using fragment screening, we probe new cavities in this form which can be used to guide therapeutic development. Furthermore, we characterized a serine site-directed mutant of the Mpro bound to its endogenous N and C-terminal residues during dimeric association stage of the maturation process. We suggest this form is a transitional state during the C-terminal trans-cleavage. This data sheds light in the structural modifications of the SARS-CoV-2 main protease during its self-maturation process.
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Péptido Hidrolasas/química , Péptido Hidrolasas/metabolismo , SARS-CoV-2/metabolismo , Proteínas Virales/química , Proteínas Virales/metabolismo , Dominio Catalítico/fisiología , Cristalografía por Rayos X/métodos , Dimerización , HumanosRESUMEN
BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) infection after kidney transplantation (KT) is associated with high mortality. AIM: To analyse an outbreak of infection/colonization with IMP-1-producing CRPA on a KT ward. METHODS: A case-control study was conducted. Cases were identified through routine surveillance culture and real-time polymerase chain reaction for carbapenemase performed directly from rectal swab samples. Controls were randomly selected from patients hospitalized on the same ward during the same period, at a ratio of 3:1. Strain clonality was analysed through pulsed-field gel electrophoresis (PFGE), and whole-genome sequencing was performed for additional strain characterization. FINDINGS: CRPA was identified in 37 patients, in 51.4% through surveillance cultures and in 49.6% through clinical cultures. The median persistence of culture positivity was 42.5 days. Thirteen patients (35.1%) presented a total of 15 infections, of which seven (46.7%) were in the urinary tract; among those, 30-day mortality rate was 46.2%. PFGE analysis showed that all of the strains shared the same pulsotype. Multilocus sequence typing analysis identified the sequence type as ST446. Risk factors for CRPA acquisition were hospital stay >10 days, retransplantation, urological surgical reintervention after KT, use of carbapenem or ciprofloxacin in the last three months and low median lymphocyte count in the last three months. CONCLUSION: KT recipients remain colonized by CRPA for long periods and could be a source of nosocomial outbreaks. In addition, a high proportion of such patients develop infection. During an outbreak, urine culture should be added to the screening protocol for KT recipients.