Factors related to mortality of patients with COVID-19 who are admitted to the ICU: Prognostic mortality factors of COVID-19 patients.

During the severe acute respiratory syndrome coronavirus 2 pandemic, hospital resources, particularly critical care units, were overburdened and this had a significant impact on both the therapies and the prognosis of these patients. This study aimed to identify factors and therapies that may improve prognosis and other factors associated with increased mortality. A secondary objective was to evaluate the impact that obesity had on these patients. An observational study was conducted on 482 patients aged 18 years or older who were diagnosed with SARS-CoV-2 pneumonia and admitted to the Intensive Care Units of 3 national hospitals registered in the CIBERESUCICOVID database between September 2020 and March 2021. After identifying the sample profile, risk factors were analyzed, the predictive model was constructed, and crude odd ratios were calculated for each factor. Additionally, logistic regression was used to build the multivariate model adjusting for potential confounders. The final model included only the variables selected using the Backward method. A sample of 335 men (69.5%) and 145 women (30.08%) aged 61.94 ±â€…12.75 years with a body mass index (BMI) of 28.05 (25.7; 31.2) was obtained. A total of 113 patients received noninvasive mechanical ventilation. The most common comorbidities were: high blood pressure (51.04%), obesity (28%), diabetes mellitus (23.44%), other metabolic diseases (21.16%), chronic heart failure (18.05%), chronic obstructive pulmonary disease (11.62%), and chronic kidney disease (10.16%). In-hospital, 3-month and 6-month post-discharge mortality in patients with BMI > 30 (n = 135) versus BMI ≤ 30 (n = 347) was significantly different (P = .06). Noninvasive mechanical ventilation failed in 42.4% of patients with BMI > 30 compared to 55% of patients with BMI ≤ 30. This study identified the factors associated with failure of mechanical ventilation. The most common comorbidities were congestive heart failure, high blood pressure, chronic kidney disease, severe liver disease, diabetes mellitus, and solid organ transplantation. In terms of ventilatory support, patients who received high-flow nasal oxygen therapy on admission had lower mortality rates. The use of renal replacement therapy was also significantly associated with higher mortality.

RNA methyltransferase SPOUT1/CENP-32 links mitotic spindle organization with the neurodevelopmental disorder SpADMiSS.

SPOUT1/CENP-32 encodes a putative SPOUT RNA methyltransferase previously identified as a mitotic chromosome associated protein. SPOUT1/CENP-32 depletion leads to centrosome detachment from the spindle poles and chromosome misalignment. Aided by gene matching platforms, we identified 24 individuals with neurodevelopmental delays from 18 families with bi-allelic variants in SPOUT1/CENP-32 detected by exome/genome sequencing. Zebrafish spout1/cenp-32 mutants showed reduction in larval head size with concomitant apoptosis likely associated with altered cell cycle progression. In vivo complementation assays in zebrafish indicated that SPOUT1/CENP-32 missense variants identified in humans are pathogenic. Crystal structure analysis of SPOUT1/CENP-32 revealed that most disease-associated missense variants mapped to the catalytic domain. Additionally, SPOUT1/CENP-32 recurrent missense variants had reduced methyltransferase activity in vitro and compromised centrosome tethering to the spindle poles in human cells. Thus, SPOUT1/CENP-32 pathogenic variants cause an autosomal recessive neurodevelopmental disorder: SpADMiSS ( SPOUT1 Associated Development delay Microcephaly Seizures Short stature) underpinned by mitotic spindle organization defects and consequent chromosome segregation errors.

Mouse and human studies support DSTYK loss of function as a low-penetrance and variable expressivity risk factor for congenital urinary tract anomalies.

PURPOSE: Previous work identified rare variants in DSTYK associated with human congenital anomalies of the kidney and urinary tract (CAKUT). Here, we present a series of mouse and human studies to clarify the association, penetrance, and expressivity of DSTYK variants. METHODS: We phenotypically characterized Dstyk knockout mice of 3 separate inbred backgrounds and re-analyzed the original family segregating the DSTYK c.654+1G>A splice-site variant (referred to as "SSV" below). DSTYK loss of function (LOF) and SSVs were annotated in individuals with CAKUT, epilepsy, or amyotrophic lateral sclerosis vs controls. A phenome-wide association study analysis was also performed using United Kingdom Biobank (UKBB) data. RESULTS: Results demonstrate ∼20% to 25% penetrance of obstructive uropathy, at least, in C57BL/6J and FVB/NJ Dstyk-/- mice. Phenotypic penetrance increased to ∼40% in C3H/HeJ mutants, with mild-to-moderate severity. Re-analysis of the original family segregating the rare SSV showed low penetrance (43.8%) and no alternative genetic causes for CAKUT. LOF DSTYK variants burden showed significant excess for CAKUT and epilepsy vs controls and an exploratory phenome-wide association study supported association with neurological disorders. CONCLUSION: These data support causality for DSTYK LOF variants and highlights the need for large-scale sequencing studies (here >200,000 cases) to accurately assess causality for genes and variants to lowly penetrant traits with common population prevalence.

Monoallelic variation in DHX9, the gene encoding the DExH-box helicase DHX9, underlies neurodevelopment disorders and Charcot-Marie-Tooth disease.

DExD/H-box RNA helicases (DDX/DHX) are encoded by a large paralogous gene family; in a subset of these human helicase genes, pathogenic variation causes neurodevelopmental disorder (NDD) traits and cancer. DHX9 encodes a BRCA1-interacting nuclear helicase regulating transcription, R-loops, and homologous recombination and exhibits the highest mutational constraint of all DDX/DHX paralogs but remains unassociated with disease traits in OMIM. Using exome sequencing and family-based rare-variant analyses, we identified 20 individuals with de novo, ultra-rare, heterozygous missense or loss-of-function (LoF) DHX9 variant alleles. Phenotypes ranged from NDDs to the distal symmetric polyneuropathy axonal Charcot-Marie-Tooth disease (CMT2). Quantitative Human Phenotype Ontology (HPO) analysis demonstrated genotype-phenotype correlations with LoF variants causing mild NDD phenotypes and nuclear localization signal (NLS) missense variants causing severe NDD. We investigated DHX9 variant-associated cellular phenotypes in human cell lines. Whereas wild-type DHX9 was restricted to the nucleus, NLS missense variants abnormally accumulated in the cytoplasm. Fibroblasts from an individual with an NLS variant also showed abnormal cytoplasmic DHX9 accumulation. CMT2-associated missense variants caused aberrant nucleolar DHX9 accumulation, a phenomenon previously associated with cellular stress. Two NDD-associated variants, p.Gly411Glu and p.Arg761Gln, altered DHX9 ATPase activity. The severe NDD-associated variant p.Arg141Gln did not affect DHX9 localization but instead increased R-loop levels and double-stranded DNA breaks. Dhx9-/- mice exhibited hypoactivity in novel environments, tremor, and sensorineural hearing loss. All together, these results establish DHX9 as a critical regulator of mammalian neurodevelopment and neuronal homeostasis.

Heterozygous rare variants in NR2F2 cause a recognizable multiple congenital anomaly syndrome with developmental delays.

Nuclear receptor subfamily 2 group F member 2 (NR2F2 or COUP-TF2) encodes a transcription factor which is expressed at high levels during mammalian development. Rare heterozygous Mendelian variants in NR2F2 were initially identified in individuals with congenital heart disease (CHD), then subsequently in cohorts of congenital diaphragmatic hernia (CDH) and 46,XX ovotesticular disorders/differences of sexual development (DSD); however, the phenotypic spectrum associated with pathogenic variants in NR2F2 remains poorly characterized. Currently, less than 40 individuals with heterozygous pathogenic variants in NR2F2 have been reported. Here, we review the clinical and molecular details of 17 previously unreported individuals with rare heterozygous NR2F2 variants, the majority of which were de novo. Clinical features were variable, including intrauterine growth restriction (IUGR), CHD, CDH, genital anomalies, DSD, developmental delays, hypotonia, feeding difficulties, failure to thrive, congenital and acquired microcephaly, dysmorphic facial features, renal failure, hearing loss, strabismus, asplenia, and vascular malformations, thus expanding the phenotypic spectrum associated with NR2F2 variants. The variants seen were predicted loss of function, including a nonsense variant inherited from a mildly affected mosaic mother, missense and a large deletion including the NR2F2 gene. Our study presents evidence for rare, heterozygous NR2F2 variants causing a highly variable syndrome of congenital anomalies, commonly associated with heart defects, developmental delays/intellectual disability, dysmorphic features, feeding difficulties, hypotonia, and genital anomalies. Based on the new and previous cases, we provide clinical recommendations for evaluating individuals diagnosed with an NR2F2-associated disorder.

Two cases of MT-ND5-related mitochondrial disorder misdiagnosed as seronegative neuromyelitis optica spectrum disorder.

Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune disease primarily affecting the optic nerves and spinal cord, which is usually associated with anti-aquaporin-4 antibodies. Here, we present two individuals who were negative for anti-aquaporin-4 antibodies and were initially diagnosed with seronegative NMOSD. Each patient's clinical course and radiographic features raised suspicion for an alternative disease process. Both individuals were found to have pathogenic variants of MT-ND5, encoding subunit 5 of mitochondrial complex I, ultimately leading to a revised diagnosis of a primary mitochondrial disorder. These cases illustrate the importance of biochemical and genetic testing in atypical cases of NMOSD.

Effect of viral storm in patients admitted to intensive care units with severe COVID-19 in Spain: a multicentre, prospective, cohort study.

BACKGROUND: The contribution of the virus to the pathogenesis of severe COVID-19 is still unclear. We aimed to evaluate associations between viral RNA load in plasma and host response, complications, and deaths in critically ill patients with COVID-19. METHODS: We did a prospective cohort study across 23 hospitals in Spain. We included patients aged 18 years or older with laboratory-confirmed SARS-CoV-2 infection who were admitted to an intensive care unit between March 16, 2020, and Feb 27, 2021. RNA of the SARS-CoV-2 nucleocapsid region 1 (N1) was quantified in plasma samples collected from patients in the first 48 h following admission, using digital PCR. Patients were grouped on the basis of N1 quantity: VIR-N1-Zero (<1 N1 copies per mL), VIR-N1-Low (1-2747 N1 copies per mL), and VIR-N1-Storm (>2747 N1 copies per mL). The primary outcome was all-cause death within 90 days after admission. We evaluated odds ratios (ORs) for the primary outcome between groups using a logistic regression analysis. FINDINGS: 1068 patients met the inclusion criteria, of whom 117 had insufficient plasma samples and 115 had key information missing. 836 patients were included in the analysis, of whom 403 (48%) were in the VIR-N1-Low group, 283 (34%) were in the VIR-N1-Storm group, and 150 (18%) were in the VIR-N1-Zero group. Overall, patients in the VIR-N1-Storm group had the most severe disease: 266 (94%) of 283 patients received invasive mechanical ventilation (IMV), 116 (41%) developed acute kidney injury, 180 (65%) had secondary infections, and 148 (52%) died within 90 days. Patients in the VIR-N1-Zero group had the least severe disease: 81 (54%) of 150 received IMV, 34 (23%) developed acute kidney injury, 47 (32%) had secondary infections, and 26 (17%) died within 90 days (OR for death 0·30, 95% CI 0·16-0·55; p<0·0001, compared with the VIR-N1-Storm group). 106 (26%) of 403 patients in the VIR-N1-Low group died within 90 days (OR for death 0·39, 95% CI 0·26-0·57; p<0·0001, compared with the VIR-N1-Storm group). INTERPRETATION: The presence of a so-called viral storm is associated with increased all-cause death in patients admitted to the intensive care unit with severe COVID-19. Preventing this viral storm could help to reduce poor outcomes. Viral storm could be an enrichment marker for treatment with antivirals or purification devices to remove viral components from the blood. FUNDING: Instituto de Salud Carlos III, Canadian Institutes of Health Research, Li Ka-Shing Foundation, Research Nova Scotia, and European Society of Clinical Microbiology and Infectious Diseases. TRANSLATION: For the Spanish translation of the abstract see Supplementary Materials section.

Combined heart and liver transplantation in a patient supported by left ventricular assist device (LVAD) with propionic acidemia.

Propionic acidemia (PA) is a rare inherited metabolic disease due to inborn errors of metabolism. PA results in the accumulation of abnormal organic acid metabolites in multiple systems, mainly the central nervous system and the heart. Cardiac complications include dilated cardiomyopathy (DCM) and carry a 40-50% increased mortality risk. Liver transplantation (LT) is required in PA patients when medical treatment fails and may prevent or slow down the cardiomyopathy progression. However, severe heart disease may be a serious contraindication to LT. We present a complicated case of a PA patient, supported with a Left Ventricular Assist Device, who underwent a heart and Liver transplant. PA patients are at increased risk for metabolic acidosis during surgery, with increased anion gap and hyperammonemia. A strict multi-disciplinary approach is needed to prevent and treat metabolic decompensation. The patient had a successful heart and liver transplant after a strict treatment protocol in the pre, intra, and post-operative periods. His case highlights the complexity of PA patients and the increased risk for metabolic decompensation during surgery and provides an insight into how to manage such complicated patients.

Seudoartrosis después de una osteotomía en Chevron: incidencia, tratamiento, seguimiento y evolución / Nonunion after Chevron osteotomy. Incidence, treatment, follow-up, and outcomes

Introducción: El hallux valgus es el trastorno más común del primer dedo del pie. Provoca dolor, discapacidad funcional y altera los patrones de la marcha. Las deformidades leves o moderadas se han corregido con osteotomías distales del primer metatarsiano, como la osteotomía en chevron, un procedimiento seguro, pero no exento de complicaciones. Los objetivos de este estudio fueron determinar la incidencia de seudoartrosis por dicha osteotomía y comunicar nuestro método terapéutico, el seguimiento y la evolución. materiales y métodos: Se realizó un estudio multicéntrico, retrospectivo que incluyó a pacientes operados entre 2009 y 2018. Se evaluaron 1156 osteotomías en chevron como tratamiento del hallux valgus leve o moderado en 1017 pacientes (rango etario 16-83 años; promedio 57.5) realizadas por 4 cirujanos experimentados. El criterio de inclusión fue que el paciente contara con estudios por imágenes compatibles con seudoartrosis a los 6 meses de la cirugía. Resultados: Se evaluó a 5 pacientes con diagnóstico de seudoartrosis después de una osteotomía en chevron para tratar el hallux valgus. Los puntajes promedio de la AOFAS fueron 51 antes del tratamiento del hallux valgus y 87,8 después del tratamiento de la seudoartrosis. Conclusiones: La incidencia de seudoartrosis fue del 0,4% en el posoperatorio alejado. Nuestro abordaje y el tratamiento de la seudoartrosis lograron una excelente mejoría clínica y funcional en todos los pacientes operados. Nivel de Evidencia: III

Introduction: Hallux valgus is the most common disorder of the first toe. It causes pain, functional impairment, and alters gait patterns. Mild to moderate deformities are tipically corrected with distal osteotomies of the first metatarsal, such as the chevron osteotomy, a safe procedure, but not without complications. The objectives of this study were to determine the incidence of pseudarthrosis following this osteotomy and report our therapeutic method, follow-up, and outcomes. Materials and methods: A retrospective multicenter study was carried out, which included patients operated on between 2009 and 2018. A total of 1156 chevron osteotomies were evaluated as a treatment for mild to moderate hallux valgus in 1017 patients (age range 16 -83 years; average 57.5 years) performed by 4 experienced surgeons. The inclusion criterion was that the patient had imaging studies compatible with pseudarthrosis six months after surgery. Results: We evaluated five patients who met our criterion. The average AOFAS (American Orthopedic Foot and Ankle Society) scores were 51 before hallux valgus treatment and 87.8 after pseudarthrosis treatment. Conclusion: The incidence of pseudarthrosis was 0.4% in the distant postoperative period. Our approach and treatment of pseudarthrosis achieved excellent clinical and functional improvements in all operated patients. Level of Evidence: III

A novel biallelic loss-of-function variant in DAND5 causes heterotaxy syndrome.

The majority of heterotaxy cases do not obtain a molecular diagnosis, although pathogenic variants in more than 50 genes are known to cause heterotaxy. A heterozygous missense variant in DAND5, a nodal inhibitor, which functions in early development for establishment of right-left patterning, has been implicated in heterotaxy. Recently, the first case was reported of a DAND5 biallelic loss-of-function (LoF) variant in an individual with heterotaxy. Here, we describe a second unrelated individual with heterotaxy syndrome and a homozygous frameshift variant in DAND5 (NM_152654.2:c.197del [p.Leu66ArgfsTer22]). Using an in vitro assay, we demonstrate that the DAND5 c.197del variant is unable to inhibit nodal signaling when compared with the wild-type expression construct. This work strengthens the genetic and functional evidence for biallelic LoF variants in DAND5 causing an autosomal recessive heterotaxy syndrome.

Variants in the zinc transporter TMEM163 cause a hypomyelinating leukodystrophy.

Hypomyelinating leukodystrophies comprise a subclass of genetic disorders with deficient myelination of the CNS white matter. Here we report four unrelated families with a hypomyelinating leukodystrophy phenotype harbouring variants in TMEM163 (NM_030923.5). The initial clinical presentation resembled Pelizaeus-Merzbacher disease with congenital nystagmus, hypotonia, delayed global development and neuroimaging findings suggestive of significant and diffuse hypomyelination. Genomic testing identified three distinct heterozygous missense variants in TMEM163 with two unrelated individuals sharing the same de novo variant. TMEM163 is highly expressed in the CNS particularly in newly myelinating oligodendrocytes and was recently revealed to function as a zinc efflux transporter. All the variants identified lie in highly conserved residues in the cytoplasmic domain of the protein, and functional in vitro analysis of the mutant protein demonstrated significant impairment in the ability to efflux zinc out of the cell. Expression of the mutant proteins in an oligodendroglial cell line resulted in substantially reduced mRNA expression of key myelin genes, reduced branching and increased cell death. Our findings indicate that variants in TMEM163 cause a hypomyelinating leukodystrophy and uncover a novel role for zinc homeostasis in oligodendrocyte development and myelin formation.

Comparative functional genomics identifies unique molecular features of EPSCs.

Extended pluripotent or expanded potential stem cells (EPSCs) possess superior developmental potential to embryonic stem cells (ESCs). However, the molecular underpinning of EPSC maintenance in vitro is not well defined. We comparatively studied transcriptome, chromatin accessibility, active histone modification marks, and relative proteomes of ESCs and the two well-established EPSC lines to probe the molecular foundation underlying EPSC developmental potential. Despite some overlapping transcriptomic and chromatin accessibility features, we defined sets of molecular signatures that distinguish EPSCs from ESCs in transcriptional and translational regulation as well as metabolic control. Interestingly, EPSCs show similar reliance on pluripotency factors Oct4, Sox2, and Nanog for self-renewal as ESCs. Our study provides a rich resource for dissecting the regulatory network that governs the developmental potency of EPSCs and exploring alternative strategies to capture totipotent stem cells in culture.

Differential Immune Checkpoint and Ig-like V-Type Receptor Profiles in COVID-19: Associations with Severity and Treatment.

Identifying patients' immune system status has become critical to managing SARS-CoV-2 infection and avoiding the appearance of secondary infections during a hospital stay. Despite the high volume of research, robust severity and outcome markers are still lacking in COVID-19. We recruited 87 COVID-19 patients and analyzed, by unbiased automated software, 356 parameters at baseline emergency department admission including: high depth immune phenotyping and immune checkpoint expression by spectral flow cytometry, cytokines and other soluble molecules in plasma as well as routine clinical variables. We identified 69 baseline alterations in the expression of immune checkpoints, Ig-like V type receptors and other immune population markers associated with severity (O2 requirement). Thirty-four changes in these markers/populations were associated with secondary infection appearance. In addition, through a longitudinal sample collection, we described the changes which take place in the immune system of COVID-19 patients during secondary infections and in response to corticosteroid treatment. Our study provides information about immune checkpoint molecules and other less-studied receptors with Ig-like V-type domains such as CD108, CD226, HVEM (CD270), B7H3 (CD276), B7H5 (VISTA) and GITR (CD357), defining these as novel interesting molecules in severe and corticosteroids-treated acute infections.

Accelerated artificial aging and color stability in resin-based cements.

The aim of this study was to determine color change after accelerated artificial ageing (AAA) of different composite cements that are used with veneers. Five cylindrical test specimens, 15 mm in diameter and 2 mm thick, were made from a single layer of each of the following: RelyX Veneer 3M ESPE (RX), Paracore White Coltene (PC), Solocem White Opaque Coltene (SO), Resin Duo Cement Densell (DC), Panavia V5 Paste Kuraray Noritake (PA) and Panavia F2.0 Kuraray Noritake (PF) (30 specimens altogether). The specimens were light cured following manufacturers' instructions using a Coltolux LED (Coltene) unit. Initial color was determined using an Easyshade - Vita Zahnfabrik Spectrophotometer. Then, the specimens were subjected to AAA for two weeks (336 hours) with cycles of 4 hours of UV light at 60 °C and 4 hours of vapor condensation at 50 °C, successively, after which color was recorded again. Color change was determined for each specimen according to the differenceinshadeon the Vita scale before and after AAA. Results were analyzed using Kruskal Wallis test. Mean and standard deviation for each group were: RX 8.40 (1.52); PC 8.60 (3.13); SO 6.40 (3.51); DC 10.00 (0.00); PA 7.60 (3.29); PF 2.00 (0.00). The Kruskal Wallis test showed significant difference for material (p<0.05), and comparison of means showed difference between Panavia F2.0 and the other materials. A table providing equivalence between the Vita Classic and CIELAB scales was used to transfer the recorded colors to the CIELAB scale, and the color difference ΔE was calculated for each group, where ΔL, Δa and Δb are the differences in the L, a and b values before and after the AAA. The mean and standard deviation were analyzed statistically by the ANOVA test and Tukey's test. Mean and standard deviation for each group were: RX 14.94 (2.02); PC 14.51 (4.02); SO 12.08 (4.53); DC 16.31 (0.00); PA 10.9 (3.38); PF 7.24 (0.00). The ANOVA test showed significantdifferenceformaterial (p<0.05). Tukey's test showed two groups (PF-DC, RX, PA). Under the experimental conditions of this study, it can be concluded that accelerated ageing significantly affects the color stability of the resin based cements tested.

El objetivo de este trabajo fue determinar el cambio de coloración de distintos medios de fijación a base de resinas con indicación para carillas estéticas luego de someterse al envejecimiento artificial acelerado (AAA). Se confeccionaron 30 probetas cilíndricas de 15 mm de diámetro y 2 mm de espesor con RelyX Veneer 3M ESPE (RX), Paracore White Coltene (PC), Solocem White Opaque Coltene (SO), Resin Duo Cement Densell (DC), Panavia V5 Paste Kuraray Noritake (PA) y Panavia F2.0 Kuraray Noritake (PF) de una sola capa de material. Se polimerizaron según las indicaciones del fabricante con unidad de curado Coltolux LED (Coltene). Cada grupo quedó conformado con 5 probetas de cada material. Se tomó el color con el espectrofotómetro Easyshade de Vita Zahnfabrik. A continuación, se sometieron a AAA por dos semanas (336 horas) con ciclos de 4 horas de radiaciones ultravioletas a 60°C Y 4 horas de condensación de vapor de agua a 50°C sucesivamente. Unavezterminado esteproceso se volvió a registrar el color. El cambio de color se evaluó dentro de la escala de color, ordenada en función del valor. El dato registrado fue la diferencia en la posición inicial y final en esta escala. Los resultados obtenidos para cada grupo fueron analizados por medio de la prueba de Kruskal Wallis. Los valores de media y desvío estándar de cada grupo fueron: RX 8,40 (1,52); PC 8,60 (3,13); SO 6,40 (3,51); DC 10,00 (0,00); PA 7,60 (3,29); PF 2,00 (0,00). El análisis con la prueba de Kruskal Wallis mostró diferencia significativa para el factor material (p<0.05) y la comparación de medias mostró diferencia entre PF y el resto de los materiales. Por otra parte, con una tabla de equivalencia entre los colores en escala de Vita Classic y CIE Lab, se hicieron el pasaje de los valores registrados a la escala de CIE Lab y se calculó la diferencia de color ΔE de cada grupo antes y después del AAA. La media y el desvío estándarfueronanalizadosestadísticamente por el test de ANOVA y prueba de Tukey. Los valores de media y desvío estándar de cada grupo fueron: RX 14,94 (2,02); PC 14,51 (4,02); SO 12,08 (4,53); DC 16,31 (0,00); PA 10,9 (3,38); PF 7,24 (0,00). En la evaluación de los resultados con ANOVA se encontró diferencia significativa entre los materiales evaluados (p<0,05). La comparación de medias con prueba de Tukey mostró la presencia de dos grupos (PF-DC, RX, PA). En las condiciones experimentales de este trabajo puede concluirse que el envejecimiento acelerado afecta significativamente la estabilidad de color de los cementos utilizados en este trabajo.

A homozygous splice variant in ATP5PO, disrupts mitochondrial complex V function and causes Leigh syndrome in two unrelated families.

Mitochondrial complex V plays an important role in oxidative phosphorylation by catalyzing the generation of ATP. Most complex V subunits are nuclear encoded and not yet associated with recognized Mendelian disorders. Using exome sequencing, we identified a rare homozygous splice variant (c.87+3A>G) in ATP5PO, the complex V subunit which encodes the oligomycin sensitivity conferring protein, in three individuals from two unrelated families, with clinical suspicion of a mitochondrial disorder. These individuals had a similar, severe infantile and often lethal multi-systemic disorder that included hypotonia, developmental delay, hypertrophic cardiomyopathy, progressive epileptic encephalopathy, progressive cerebral atrophy, and white matter abnormalities on brain MRI consistent with Leigh syndrome. cDNA studies showed a predominant shortened transcript with skipping of exon 2 and low levels of the normal full-length transcript. Fibroblasts from the affected individuals demonstrated decreased ATP5PO protein, defective assembly of complex V with markedly reduced amounts of peripheral stalk proteins, and complex V hydrolytic activity. Further, expression of human ATP5PO cDNA without exon 2 (hATP5PO-∆ex2) in yeast cells deleted for yATP5 (ATP5PO homolog) was unable to rescue growth on media which requires oxidative phosphorylation when compared to the wild type construct (hATP5PO-WT), indicating that exon 2 deletion leads to a non-functional protein. Collectively, our findings support the pathogenicity of the ATP5PO c.87+3A>G variant, which significantly reduces but does not eliminate complex V activity. These data along with the recent report of an affected individual with ATP5PO variants, add to the evidence that rare biallelic variants in ATP5PO result in defective complex V assembly, function and are associated with Leigh syndrome.

Accelerated artificial aging and color stability in resin-based cements / Envejecimiento artificial acelerado y estabilidad de color de medios de fijación a base de resinas

ABSTRACT The aim of this study was to determine color change after accelerated artificial ageing (AAA) of different composite cements that are used with veneers. Five cylindrical test specimens, 15 mm in diameter and 2 mm thick, were made from a single layer of each of the following: RelyX Veneer 3M ESPE (RX), Paracore White Coltene (PC), Solocem White Opaque Coltene (SO), Resin Duo Cement Densell (DC), Panavia V5 Paste Kuraray Noritake (PA) and Panavia F2.0 Kuraray Noritake (PF) (30 specimens altogether). The specimens were light cured following manufacturers' instructions using a Coltolux LED (Coltene) unit. Initial color was determined using an Easyshade - Vita Zahnfabrik Spectrophotometer. Then, the specimens were subjected to AAA for two weeks (336 hours) with cycles of 4 hours of UV light at 60 °C and 4 hours of vapor condensation at 50 °C, successively, after which color was recorded again. Color change was determined for each specimen according to the difference in shade on the Vita scale before and after AAA. Results were analyzed using Kruskal Wallis test. Mean and standard deviation for each group were: RX 8.40 (1.52); PC 8.60 (3.13); SO 6.40 (3.51); DC 10.00 (0.00); PA 7.60 (3.29); PF 2.00 (0.00). The Kruskal Wallis test showed significant difference for material (p<0.05), and comparison of means showed difference between Panavia F2.0 and the other materials. A table providing equivalence between the Vita Classic and CIELAB scales was used to transfer the recorded colors to the CIELAB scale, and the color difference ΔE was calculated for each group, where ΔL, Δa and Δb are the differences in the L, a and b values before and after the AAA. The mean and standard deviation were analyzed statistically by the ANOVA test and Tukey's test. Mean and standard deviation for each group were: RX 14.94 (2.02); PC 14.51 (4.02); SO 12.08 (4.53); DC 16.31 (0.00); PA 10.9 (3.38); PF 7.24 (0.00). The ANOVA test showed significant difference for material (p<0.05). Tukey's test showed two groups (PF-DC, RX, PA). Under the experimental conditions of this study, it can be concluded that accelerated ageing significantly affects the color stability of the resin based cements tested.

RESUMEN El objetivo de este trabajo fue determinar el cambio de coloración de distintos medios de fijación a base de resinas con indicación para carillas estéticas luego de someterse al envejecimiento artificial acelerado (AAA). Se confeccionaron 30 probetas cilíndricas de 15 mm de diámetro y 2 mm de espesor con RelyX Veneer 3M ESPE (RX), Paracore White Coltene (PC), Solocem White Opaque Coltene (SO), Resin Duo Cement Densell (DC), Panavia V5 Paste Kuraray Noritake (PA) y Panavia F2.0 Kuraray Noritake (PF) de una sola capa de material. Se polimerizaron según las indicaciones del fabricante con unidad de curado Coltolux LED (Coltene). Cada grupo quedó conformado con 5 probetas de cada material. Se tomó el color con el espectrofotómetro Easyshade de Vita Zahnfabrik. A continuación, se sometieron a AAA por dos semanas (336 horas) con ciclos de 4 horas de radiaciones ultravioletas a 60°C Y 4 horas de condensación de vapor de agua a 50°C sucesivamente. Una vez terminado este proceso se volvió a registrar el color. El cambio de color se evaluó dentro de la escala de color, ordenada en función del valor. El dato registrado fue la diferencia en la posición inicial y final en esta escala. Los resultados obtenidos para cada grupo fueron analizados por medio de la prueba de Kruskal Wallis. Los valores de media y desvío estándar de cada grupo fueron: RX 8,40 (1,52); PC 8,60 (3,13); SO 6,40 (3,51); DC 10,00 (0,00); PA 7,60 (3,29); PF 2,00 (0,00). El análisis con la prueba de Kruskal Wallis mostró diferencia significativa para el factor material (p<0.05) y la comparación de medias mostró diferencia entre PF y el resto de los materiales. Por otra parte, con una tabla de equivalencia entre los colores en escala de Vita Classic y CIE Lab, se hicieron el pasaje de los valores registrados a la escala de CIE Lab y se calculó la diferencia de color ΔE de cada grupo antes y después del AAA. La media y el desvío estándar fueron analizados estadísticamente por el test de ANOVA y prueba de Tukey. Los valores de media y desvío estándar de cada grupo fueron: RX 14,94 (2,02); PC 14,51 (4,02); SO 12,08 (4,53); DC 16,31 (0,00); PA 10,9 (3,38); PF 7,24 (0,00). En la evaluación de los resultados con ANOVA se encontró diferencia significativa entre los materiales evaluados (p<0,05). La comparación de medias con prueba de Tukey mostró la presencia de dos grupos (PF-DC, RX, PA). En las condiciones experimentales de este trabajo puede concluirse que el envejecimiento acelerado afecta significativamente la estabilidad de color de los cementos utilizados en este trabajo.

Intertwined leukocyte balances in tumours and peripheral blood as robust predictors of right and left colorectal cancer survival.

BACKGROUND: Colorectal cancer (CRC) accounts for 9.4% of overall cancer deaths, ranking second after lung cancer. Despite the large number of factors tested to predict their outcome, most patients with similar variables show big differences in survival. Moreover, right-sided CRC (RCRC) and left-sided CRC (LCRC) patients exhibit large differences in outcome after surgical intervention as assessed by preoperative blood leukocyte status. We hypothesised that stronger indexes than circulating (blood) leukocyte ratios to predict RCRC and LCRC patient outcomes will result from combining both circulating and infiltrated (tumour/peritumour fixed tissues) concentrations of leukocytes. AIM: To seek variables involving leukocyte balances in peripheral blood and tumour tissues and to predict the outcome of CRC patients. METHODS: Sixty-five patients diagnosed with colon adenocarcinoma by the Digestive Surgery Service of the La Paz University Hospital (Madrid, Spain) were enrolled in this study: 43 with RCRC and 22 with LCRC. Patients were followed-up from January 2017 to March 2021 to record overall survival (OS) and recurrence-free survival (RFS) after surgical interventions. Leukocyte concentrations in peripheral blood were determined by routine laboratory protocols. Paraffin-fixed samples of tumour and peritumoural tissues were assessed for leukocyte concentrations by immunohistochemical detection of CD4, CD8, and CD14 marker expression. Ratios of leukocyte concentration in blood and tissues were calculated and evaluated for their predictor values for OS and RFS with Spearman correlations and Cox univariate and multivariate proportional hazards regression, followed by the calculation of the receiver-operating characteristic and area under the curve (AUC) and the determination of Youden's optimal cutoff values for those variables that significantly correlated with either RCRC or LCRC patient outcomes. RCRC patients from the cohort were randomly assigned to modelling and validation sets, and clinician-friendly nomograms were developed to predict OS and RFS from the respective significant indexes. The accuracy of the model was evaluated using calibration and validation plots. RESULTS: The relationship of leukocyte ratios in blood and peritumour resulted in six robust predictors of worse OS in RCRC: CD8+ lymphocyte content in peritumour (CD8pt, AUC = 0.585, cutoff < 8.250, P = 0.0077); total lymphocyte content in peritumour (CD4CD8pt, AUC = 0.550, cutoff < 10.160, P = 0.0188); lymphocyte-to-monocyte ratio in peritumour (LMRpt, AUC = 0.807, cutoff < 3.185, P = 0.0028); CD8+ LMR in peritumour (CD8MRpt, AUC = 0.757, cutoff < 1.650, P = 0.0007); the ratio of blood LMR to LMR in peritumour (LMRb/LMRpt, AUC = 0.672, cutoff > 0.985, P = 0.0244); and the ratio of blood LMR to CD8+ LMR in peritumour (LMRb/CD8MRpt, AUC = 0.601, cutoff > 1.485, P = 0.0101). In addition, three robust predictors of worse RFS in RCRC were found: LMRpt (AUC = 0.737, cutoff < 3.185, P = 0.0046); LMRb/LMRpt (AUC = 0.678, cutoff > 0.985, P = 0.0155) and LMRb/CD8MRpt (AUC = 0.615, cutoff > 1.485, P = 0.0141). Furthermore, the ratio of blood LMR to CD4+ LMR in peritumour (LMRb/CD4MRpt, AUC = 0.786, cutoff > 10.570, P = 0.0416) was found to robustly predict poorer OS in LCRC patients. The nomograms showed moderate accuracy in predicting OS and RFS in RCRC patients, with concordance index of 0.600 and 0.605, respectively. CONCLUSION: Easily obtainable variables at preoperative consultation, defining the status of leukocyte balances between peripheral blood and peritumoural tissues, are robust predictors for OS and RFS of both RCRC and LCRC patients.

Cellular and humoral functional responses after BNT162b2 mRNA vaccination differ longitudinally between naive and subjects recovered from COVID-19.

We have analyzed BNT162b2 vaccine-induced immune responses in naive subjects and individuals recovered from coronavirus disease 2019 (COVID-19), both soon after (14 days) and later after (almost 8 months) vaccination. Plasma spike (S)-specific immunoglobulins peak after one vaccine shot in individuals recovered from COVID-19, while a second dose is needed in naive subjects, although the latter group shows reduced levels all along the analyzed period. Despite how the neutralization capacity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mirrors this behavior early after vaccination, both groups show comparable neutralizing antibodies and S-specific B cell levels late post-vaccination. When studying cellular responses, naive individuals exhibit higher SARS-CoV-2-specific cytokine production, CD4+ T cell activation, and proliferation than do individuals recovered from COVID-19, with patent inverse correlations between humoral and cellular variables early post-vaccination. However, almost 8 months post-vaccination, SARS-CoV-2-specific responses are comparable between both groups. Our data indicate that a previous history of COVID-19 differentially determines the functional T and B cell-mediated responses to BNT162b2 vaccination over time.