Covalent Labeling Automated Data Analysis Platform for High Throughput in R (coADAPTr): A Proteome-Wide Data Analysis Platform for Covalent Labeling Experiments.

Covalent labeling methods coupled to mass spectrometry have emerged in recent years for studying the higher order structure of proteins. Quantifying the extent of modification of proteins in multiple states (i.e., ligand free vs ligand-bound) can provide information on protein interaction sites and regions of conformational change. Though there are several software platforms that are used to quantify the extent of modification, the process can still be time-consuming, particularly for proteome-wide studies. Here, we present an open-source software for quantitation called Covalent labeling Automated Data Analysis Platform for high Throughput in R (coADAPTr). coADAPTr tackles the need for more efficient data analysis in covalent labeling mass spectrometry for techniques such as hydroxyl radical protein footprinting (HRPF). Traditional methods like Excel's Power Pivot (PP) are cumbersome and time-intensive, posing challenges for large-scale analyses. coADAPTr simplifies analysis by mimicking the functions used in the previous quantitation platform using PowerPivot in Microsoft Excel but with fewer steps, offering proteome-wide insights with enhanced graphical interpretations. Several features have been added to improve the fidelity and throughput compared to those of PowerPivot. These include filters to remove any duplicate data and the use of the arithmetic mean rather than the geometric mean for quantitation of the extent of modification. Validation studies confirm coADAPTr's accuracy and efficiency while processing data up to 200 times faster than conventional methods. Its open-source design and user-friendly interface make it accessible for researchers exploring intricate biological phenomena via HRPF and other covalent labeling MS methods. coADAPTr marks a significant leap in structural proteomics, providing a versatile and efficient platform for data interpretation. Its potential to transform the field lies in its seamless handling of proteome-wide data analyses, empowering researchers with a robust tool for deciphering complex structural biology data.

CIUSuite 3: Next-Generation CCS Calibration and Automated Data Analysis Tools for Gas-Phase Protein Unfolding Data.

Ion mobility-mass spectrometry (IM-MS) has become a technology deployed across a wide range of structural biology applications despite the challenges in characterizing closely related protein structures. Collision-induced unfolding (CIU) has emerged as a valuable technique for distinguishing closely related, iso-cross-sectional protein and protein complex ions through their distinct unfolding pathways in the gas phase. With the speed and sensitivity of CIU analyses, there has been a rapid growth of CIU-based assays, especially regarding biomolecular targets that remain challenging to assess and characterize with other structural biology tools. With information-rich CIU data, many software tools have been developed to automate laborious data analysis. However, with the recent development of new IM-MS technologies, such as cyclic IM-MS, CIU continues to evolve, necessitating improved data analysis tools to keep pace with new technologies and facilitating the automation of various data processing tasks. Here, we present CIUSuite 3, a software package that contains updated algorithms that support various IM-MS platforms and supports the automation of various data analysis tasks such as peak detection, multidimensional classification, and collision cross section (CCS) calibration. CIUSuite 3 uses local maxima searches along with peak width and prominence filters to detect peaks to automate CIU data extraction. To support both the primary CIU (CIU1) and secondary CIU (CIU2) experiments enabled by cyclic IM-MS, two-dimensional data preprocessing is deployed, which allows multidimensional classification. Our data suggest that additional dimensions in classification improve the overall accuracy of class assignments. CIUSuite 3 also supports CCS calibration for both traveling wave and drift tube IM-MS, and we demonstrate the accuracy of a new single-field CCS calibration method designed for drift tube IM-MS leveraging calibrant CIU data. Overall, CIUSuite 3 is positioned to support current and next-generation IM-MS and CIU assay development deployed in an automated format.

CFD simulation study and experimental analysis of indoor air stratification in an unventilated classroom: A case study in Spain.

Health problems and respiratory diseases are associated with poor indoor air ventilation. We investigated the air quality inside a classroom-laboratory where no ventilation is provided. The case of study, consisting of an internal enclosure, is located at the Escuela Técnica Superior de Edificación (ETSEM) of Madrid (Spain). The high height favours air stratification which is analysed in terms of temperature and CO2 spatial distribution. Temperature, air humidity, atmospheric pressure and CO2 concentration measurements were taken in time at three different height locations. A CFD numerical model was established to analyse air quality. Flow circulation is derived by solving full 3D Navier - Stokes governing equations, coupled with the thermal problem. The diffusion problem of the CO2 produced by the inner occupants is then derived from the kinematics solution. Three scenarios were taken into account: occupants seated (1), standing (2), half seated, half standing (3). Results clearly show the air stratification as a result of density gradient, which is in turn determined by temperature difference between the occupants and the surrounding air. Temperature prediction maximum relative error is contained to 3.5 %. As expected, CO2 concentration increases over time, reaching maximum values depending on the configuration considered and height location.

Are Internal Fragments Observable in Electron Based Top-Down Mass Spectrometry?

Protein tandem mass spectrometry (MS/MS) often generates sequence-informative fragments from backbone bond cleavages near the termini. This lack of fragmentation in the protein interior is particularly apparent in native top-down mass spectrometry (MS). Improved sequence coverage, critical for reliable annotation of posttranslational modifications and sequence variants, may be obtained from internal fragments generated by multiple backbone cleavage events. However, internal fragment assignments can be error prone due to isomeric/isobaric fragments from different parts of a protein sequence. Also, internal fragment generation propensity depends on the chosen MS/MS activation strategy. Here, we examine internal fragment formation in electron capture dissociation (ECD) and electron transfer dissociation (ETD) following native and denaturing MS, as well as LC/MS of several proteins. Experiments were undertaken on multiple instruments, including quadrupole time-of-flight, Orbitrap, and high-field Fourier-transform ion cyclotron resonance (FT-ICR) across four laboratories. ECD was performed at both ultrahigh vacuum and at similar pressure to ETD conditions. Two complementary software packages were used for data analysis. When feasible, ETD-higher energy collision dissociation MS3 was performed to validate/refute potential internal fragment assignments, including differentiating MS3 fragmentation behavior of radical versus even-electron primary fragments. We show that, under typical operating conditions, internal fragments cannot be confidently assigned in ECD or ETD. On the other hand, such fragments, along with some b-type terminal fragments (not typically observed in ECD/ETD spectra) appear at atypical ECD operating conditions, suggesting they originate from a separate ion-electron activation process. Furthermore, atypical fragment ion types, e.g., x ions, are observed at such conditions as well as upon EThcD, presumably due to vibrational activation of radical z-type ions.

Personalized brain circuit scores identify clinically distinct biotypes in depression and anxiety.

There is an urgent need to derive quantitative measures based on coherent neurobiological dysfunctions or 'biotypes' to enable stratification of patients with depression and anxiety. We used task-free and task-evoked data from a standardized functional magnetic resonance imaging protocol conducted across multiple studies in patients with depression and anxiety when treatment free (n = 801) and after randomization to pharmacotherapy or behavioral therapy (n = 250). From these patients, we derived personalized and interpretable scores of brain circuit dysfunction grounded in a theoretical taxonomy. Participants were subdivided into six biotypes defined by distinct profiles of intrinsic task-free functional connectivity within the default mode, salience and frontoparietal attention circuits, and of activation and connectivity within frontal and subcortical regions elicited by emotional and cognitive tasks. The six biotypes showed consistency with our theoretical taxonomy and were distinguished by symptoms, behavioral performance on general and emotional cognitive computerized tests, and response to pharmacotherapy as well as behavioral therapy. Our results provide a new, theory-driven, clinically validated and interpretable quantitative method to parse the biological heterogeneity of depression and anxiety. Thus, they represent a promising approach to advance precision clinical care in psychiatry.

HMGA1 regulates trabectedin sensitivity in advanced soft-tissue sarcoma (STS): A Spanish Group for Research on Sarcomas (GEIS) study.

HMGA1 is a structural epigenetic chromatin factor that has been associated with tumor progression and drug resistance. Here, we reported the prognostic/predictive value of HMGA1 for trabectedin in advanced soft-tissue sarcoma (STS) and the effect of inhibiting HMGA1 or the mTOR downstream pathway in trabectedin activity. The prognostic/predictive value of HMGA1 expression was assessed in a cohort of 301 STS patients at mRNA (n = 133) and protein level (n = 272), by HTG EdgeSeq transcriptomics and immunohistochemistry, respectively. The effect of HMGA1 silencing on trabectedin activity and gene expression profiling was measured in leiomyosarcoma cells. The effect of combining mTOR inhibitors with trabectedin was assessed on cell viability in vitro studies, whereas in vivo studies tested the activity of this combination. HMGA1 mRNA and protein expression were significantly associated with worse progression-free survival of trabectedin and worse overall survival in STS. HMGA1 silencing sensitized leiomyosarcoma cells for trabectedin treatment, reducing the spheroid area and increasing cell death. The downregulation of HGMA1 significantly decreased the enrichment of some specific gene sets, including the PI3K/AKT/mTOR pathway. The inhibition of mTOR, sensitized leiomyosarcoma cultures for trabectedin treatment, increasing cell death. In in vivo studies, the combination of rapamycin with trabectedin downregulated HMGA1 expression and stabilized tumor growth of 3-methylcholantrene-induced sarcoma-like models. HMGA1 is an adverse prognostic factor for trabectedin treatment in advanced STS. HMGA1 silencing increases trabectedin efficacy, in part by modulating the mTOR signaling pathway. Trabectedin plus mTOR inhibitors are active in preclinical models of sarcoma, downregulating HMGA1 expression levels and stabilizing tumor growth.

A close-up view of the Hunter syndrome.

The Lysosomal Storage disease known as Mucopolysaccharidosis type II, is caused by mutations affecting the iduronate-2-sulfatase required for heparan and dermatan sulfate catabolism. The central nervous system (CNS) is mostly and severely affected by the accumulation of both substrates. The complexity of the CNS damage observed in MPS II patients has been limitedly explored. The use of mass spectrometry (MS)-based proteomics tools to identify protein profiles may yield valuable information about the pathological mechanisms of Hunter syndrome. In this further study, we provide a new comparative proteomic analysis of MPS II models by using a pipeline consisting of the identification of native protein complexes positioned selectively by using a specific antibody, coupled with mass spectrometry analysis, allowing us to identify changes involving in a significant number of new biological functions, including a specific brain antioxidant response, a down-regulated autophagic, the suppression of sulfur catabolic process, a prominent liver immune response and the stimulation of phagocytosis among others.

Unusual Trisomy X Phenotype Associated with a Concurrent Heterozygous 16p11.2 Deletion: Importance of an Integral Approach for Proper Diagnosis.

Trisomy X is the most frequent sex chromosome anomaly in women, but it is often underdiagnosed postnatally because most patients do not show any clinical manifestation. It is estimated that only 10% of patients with trisomy X are diagnosed by clinical findings. Thus, it has been proposed that the clinical spectrum is not yet fully delimited, and additional uncommon or atypical clinical manifestations could be related to this entity. The present report describes a female carrying trisomy X but presenting atypical manifestations, including severe intellectual disability, short stature, thymus hypoplasia, and congenital hypothyroidism (CH). These clinical findings were initially attributed to trisomy X. However, chromosome microarray analysis (CMA) subsequently revealed that the patient also bears a heterozygous 304-kb deletion at 16p11.2. This pathogenic copy-number variant (CNV) encompasses 13 genes, including TUFM. Some authors recommend that when a phenotype differs from that described for an identified microdeletion, the presence of pathogenic variants in the non-deleted allele should be considered to assess for an autosomal recessive disorder; thus, we used a panel of 697 genes to rule out a pathogenic variant in the non-deleted TUFM allele. We discuss the possible phenotypic modifications that might be related to an additional CNV in individuals with sex chromosome aneuploidy (SCA), as seen in our patient. The presence of karyotype-demonstrated trisomy X and CMA-identified 16p11.2 deletion highlights the importance of always correlating a patient's clinical phenotype with the results of genetic studies. When the phenotype includes unusual manifestations and/or exhibits discrepancies with that described in the literature, as exemplified by our patient, a more extensive analysis should be undertaken to enable a correct diagnosis that will support proper management, genetic counseling, and medical follow-up.

Dimensions of reading: a study of the beliefs of language and literature preservice teachers.

This study aims to analyze the beliefs that future Language and Literature teachers hold regarding reading. This work is part of a broader research endeavor focused on the reading habits and practices of teachers in training and their role as prospective mediators since the way in which they perceive reading significantly impacts the mediation processes they undertake in their teaching practices to cultivate readers. To achieve these objectives, a multiple case study is conducted, involving interviews with 1st-year students (n = 15), 3rd-year students (n = 15), and 5th-year students (n = 15) enrolled in Language Pedagogy programs across three universities affiliated with the Chilean Council of Rectors. For data analysis, a content analysis approach is employed, supported by NVivo 12. The findings reveal that beliefs about reading primarily fall into two dimensions: academic and personal, with the former exhibiting clearer definition and characterization. This can be attributed to the influence of the disciplines integrated into their education, namely literature and linguistics. In conclusion, it is imperative to address the social dimension of reading during the initial teacher education program, as this aspect is not emphasized by preservice teachers, despite its pivotal role in shaping their identity as reading mediators within the context of their teaching practice.

Efficient Analysis of Proteome-Wide FPOP Data by FragPipe.

Monitoring protein structure before and after environmental alterations (e.g., different cell states) can give insights into the role and function of proteins. Fast photochemical oxidation of proteins (FPOP) coupled with mass spectrometry (MS) allows for monitoring of structural rearrangements by exposing proteins to OH radicals that oxidize solvent-accessible residues, indicating protein regions undergoing movement. Some of the benefits of FPOP include high throughput and a lack of scrambling due to label irreversibility. However, the challenges of processing FPOP data have thus far limited its proteome-scale uses. Here, we present a computational workflow for fast and sensitive analysis of FPOP data sets. Our workflow, implemented as part of the FragPipe computational platform, combines the speed of the MSFragger search with a unique hybrid search method to restrict the large search space of FPOP modifications. Together, these features enable more than 10-fold faster FPOP searches that identify 150% more modified peptide spectra than previous methods. We hope this new workflow will increase the accessibility of FPOP to enable more protein structure and function relationships to be explored.

Life Cycle Assessment of Raw and Fe-Modified Biochars: Contributing to Circular Economy.

Biochar is a carbonaceous material, which can be decorated with metals, that has been garnering attention to be used in the treatment of water due to its contribution to waste management and circular economy. This study presents the life cycle assessment (LCA) regarding the generation of Pinus patula raw biochar and its modification with iron (Fe-modified biochar). SimaPro 9.3.0.3 software was used to simulate the environmental impacts of both carbonaceous materials. The potential environmental effects obtained from the production of Pinus patula raw biochar were mainly ascribed to the source of energy utilized during this process. The potential impacts demonstrated that the generation of gases and polycyclic aromatic hydrocarbons are the main concern. In the case of Fe-modified biochar, the potential environmental effects differed only in the stage of the biomass modification with the metal. These effects are associated with the extraction of Fe and the generation of wastewater. These findings provide an insight into the environmental effects linked to the production of raw and Fe-modified biochar. However, further LCA research should be performed concerning other materials and compounds than can be generated during the biomass thermochemical conversion.

MELD 3.0 adequately predicts mortality and renal replacement therapy requirements in patients with alcohol-associated hepatitis.

Background & Aims: Model for End-Stage Liver Disease (MELD) score better predicts mortality in alcohol-associated hepatitis (AH) but could underestimate severity in women and malnourished patients. Using a global cohort, we assessed the ability of the MELD 3.0 score to predict short-term mortality in AH. Methods: This was a retrospective cohort study of patients admitted to hospital with AH from 2009 to 2019. The main outcome was all-cause 30-day mortality. We compared the AUC using DeLong's method and also performed a time-dependent AUC with competing risks analysis. Results: A total of 2,124 patients were included from 28 centres from 10 countries on three continents (median age 47.2 ± 11.2 years, 29.9% women, 71.3% with underlying cirrhosis). The median MELD 3.0 score at admission was 25 (20-33), with an estimated survival of 73.7% at 30 days. The MELD 3.0 score had a better performance in predicting 30-day mortality (AUC:0.761, 95%CI:0.732-0.791) compared with MELD sodium (MELD-Na; AUC: 0.744, 95% CI: 0.713-0.775; p = 0.042) and Maddrey's discriminant function (mDF) (AUC: 0.724, 95% CI: 0.691-0.757; p = 0.013). However, MELD 3.0 did not perform better than traditional MELD (AUC: 0.753, 95% CI: 0.723-0.783; p = 0.300) and Age-Bilirubin-International Normalised Ratio-Creatinine (ABIC) (AUC:0.757, 95% CI: 0.727-0.788; p = 0.765). These results were consistent in competing-risk analysis, where MELD 3.0 (AUC: 0.757, 95% CI: 0.724-0.790) predicted better 30-day mortality compared with MELD-Na (AUC: 0.739, 95% CI: 0.708-0.770; p = 0.028) and mDF (AUC:0.717, 95% CI: 0.687-0.748; p = 0.042). The MELD 3.0 score was significantly better in predicting renal replacement therapy requirements during admission compared with the other scores (AUC: 0.844, 95% CI: 0.805-0.883). Conclusions: MELD 3.0 demonstrated better performance compared with MELD-Na and mDF in predicting 30-day and 90-day mortality, and was the best predictor of renal replacement therapy requirements during admission for AH. However, further prospective studies are needed to validate its extensive use in AH. Impact and implications: Severe AH has high short-term mortality. The establishment of treatments and liver transplantation depends on mortality prediction. We evaluated the performance of the new MELD 3.0 score to predict short-term mortality in AH in a large global cohort. MELD 3.0 performed better in predicting 30- and 90-day mortality compared with MELD-Na and mDF, but was similar to MELD and ABIC scores. MELD 3.0 was the best predictor of renal replacement therapy requirements. Thus, further prospective studies are needed to support the wide use of MELD 3.0 in AH.

Efficient Analysis of Proteome-wide FPOP Data by FragPipe.

Monitoring protein structure before and after perturbations can give insights into the role and function of proteins. Fast photochemical oxidation of proteins (FPOP) coupled with mass spectrometry (MS) allows monitoring of structural rearrangements by exposing proteins to OH radicals that oxidize solvent accessible residues, indicating protein regions undergoing movement. Some of the benefits of FPOP include high throughput and lack of scrambling due to label irreversibility. However, the challenges of processing FPOP data have thus far limited its proteome-scale uses. Here, we present a computational workflow for fast and sensitive analysis of FPOP datasets. Our workflow combines the speed of MSFragger search with a unique hybrid search method to restrict the large search space of FPOP modifications. Together, these features enable more than 10-fold faster FPOP searches that identify 50% more modified peptide spectra than previous methods. We hope this new workflow will increase the accessibility of FPOP to enable more protein structure and function relationships to be explored.

ALPL-1 is a target for chimeric antigen receptor therapy in osteosarcoma.

Osteosarcoma (OS) remains a dismal malignancy in children and young adults, with poor outcome for metastatic and recurrent disease. Immunotherapies in OS are not as promising as in some other cancer types due to intra-tumor heterogeneity and considerable off-target expression of the potentially targetable proteins. Here we show that chimeric antigen receptor (CAR) T cells could successfully target an isoform of alkaline phosphatase, ALPL-1, which is highly and specifically expressed in primary and metastatic OS. The target recognition element of the second-generation CAR construct is based on two antibodies, previously shown to react against OS. T cells transduced with these CAR constructs mediate efficient and effective cytotoxicity against ALPL-positive cells in in vitro settings and in state-of-the-art in vivo orthotopic models of primary and metastatic OS, without unexpected toxicities against hematopoietic stem cells or healthy tissues. In summary, CAR-T cells targeting ALPL-1 show efficiency and specificity in treating OS in preclinical models, paving the path for clinical translation.

Análisis de los polimorfismos ECA y ACTN3 con la agilidad en seleccionados nacionales de deportes de conjunto de Costa Rica. Estudio piloto / ACE and ACTN3 polymorphisms analysis on agility test in national players in team sports of Costa Rica. Pilot study / Análise dos polimorfismos ECA e ACTN3 com agilidade em jogadores esportivos selecionados da seleção costarriquenha. Estudo piloto

Resumen: Objetivo: Analizar las diferencias de los polimorfismos de los genes ECA y ACTN3 en el rendimiento de una prueba de agilidad en jugadores élite de deportes colectivos pertenecientes a selecciones nacionales de Costa Rica. Metodología: Se contó con una muestra de 33 jugadores hombres, de deportes colectivos (fútbol sala, rugby, voleibol y balonmano). Para la evaluación de la agilidad se utilizó el test de Illinois. Se realizaron dos visitas, en la primera se obtuvo muestras de células por medio de un enjuague y en la segunda se aplicó la prueba de agilidad. Se utilizó la prueba de Chi-cuadrado (X2) para conocer las diferencias entre las frecuencias de los polimorfismos de los genes ECA y ACTN3 y el tipo de deporte. Resultados: La mayor distribución de los polimorfismos del gen ECA, de jugadores de selecciones nacionales de deportes de conjunto, se encuentra en el ID (X2= 6.87, p= .334) y en ACTN3 el RX (X2= 6.33, p= .388). Además, tampoco se encontraron diferencias significativas entre el tiempo efectuado en el test de Illinois y los polimorfismos del gen ECA (F= 2.150, p= .134), de igual forma para los polimorfismos del gen ACTN3 (F= .950, p= .339). Conclusiones: Los polimorfismos de los genes ECA y ACTN3 no se relacionaron estadísticamente con el tipo de deporte colectivo. La agilidad no se ve asociada por un tipo de polimorfismo, lo que indica que, de forma independiente al gen, esta cualidad física se puede entrenar y generar buenos resultados en la población en general.

Abstract: Objective: To analyze the differences in the polymorphisms of the ACE and ACTN3 genes on agility test performance in elite players of collective sports from National teams of Costa Rica. Methods: a sample of 33 male team sports players (futsal, rugby, volleyball, and handball). All subjects were tested with the Illinois Agility Test. Two days of measurements were made; on the first day, cell samples were obtained and on the second day, the agility test was applied. The Chi-square test (x2) was used to determine the differences between the frequencies of the polymorphisms of the ACE and ACTN3 genes and the type of sport. Results: The highest distribution of polymorphisms of the ECA gene of players from national teams of collective sports was found in the ACE ID (X2 = 6.87, p = .334), and in ACTN3 the RX (X2 = 6.33, p =. 388). Furthermore, no significant relationship was found between the Illinois test performance and the polymorphisms of the ECA gene (F = 2,150, p = .134). Conclusions: The ACE and ACTN3 genes polymorphisms were not statistically related to the type of team sport. Agility is not associated with the type of polymorphism, which indicates that regardless of the gene, this physical quality can be trained and generate good results in the general population.

Resumo: Objetivo: analisar as diferenças dos polimorfismos dos genes ECA e ACTN3 na realização de um teste de agilidade em jogadores de elite de equipes esportivas pertencentes a equipes nacionais costarriquenhas. Metodologia: foi utilizada uma amostra de 33 jogadores de futebol masculino (futsal, rúgbi, vôlei e handebol). O teste de Illinois foi usado para avaliar a agilidade. Foram feitas duas visitas; na primeira foram obtidas amostras de uma célula por lavagem e na segunda foi aplicado o teste de agilidade. O teste qui-quadrado (X2) foi usado para determinar as diferenças entre as frequências dos polimorfismos dos genes ECA e ACTN3 e o tipo de esporte. Resultados: A maior distribuição de polimorfismos do gene ECA em jogadores de equipes nacionais de esportes coletivos é encontrada no ID (X2= 6,87, p= 0,334) e no ACTN3 no RX (X2= 6,33, p= 0,388). Além disso, não foram encontradas diferenças significativas entre o tempo gasto no teste de Illinois e os polimorfismos do gene ECA (F= 2,150, p= 0,134), assim como para os polimorfismos do gene ACTN3 (F= 0,950, p= 0,339). Conclusões: Os polimorfismos dos genes ECA e ACTN3 não estavam estatisticamente relacionados com o tipo de esporte coletivo. A agilidade não está associada pelo tipo de polimorfismo, indicando que, independentemente do gene, essa qualidade física pode ser treinada e gerar bons resultados na população em geral.

Personalized Interventional Surgery of the Lumbar Spine: A Perspective on Minimally Invasive and Neuroendoscopic Decompression for Spinal Stenosis.

Pain generator-based lumbar spinal decompression surgery is the backbone of modern spine care. In contrast to traditional image-based medical necessity criteria for spinal surgery, assessing the severity of neural element encroachment, instability, and deformity, staged management of common painful degenerative lumbar spine conditions is likely to be more durable and cost-effective. Targeting validated pain generators can be accomplished with simplified decompression procedures associated with lower perioperative complications and long-term revision rates. In this perspective article, the authors summarize the current concepts of successful management of spinal stenosis patients with modern transforaminal endoscopic and translaminar minimally invasive spinal surgery techniques. They represent the consensus statements of 14 international surgeon societies, who have worked in collaborative teams in an open peer-review model based on a systematic review of the existing literature and grading the strength of its clinical evidence. The authors found that personalized clinical care protocols for lumbar spinal stenosis rooted in validated pain generators can successfully treat most patients with sciatica-type back and leg pain including those who fail to meet traditional image-based medical necessity criteria for surgery since nearly half of the surgically treated pain generators are not shown on the preoperative MRI scan. Common pain generators in the lumbar spine include (a) an inflamed disc, (b) an inflamed nerve, (c) a hypervascular scar, (d) a hypertrophied superior articular process (SAP) and ligamentum flavum, (e) a tender capsule, (f) an impacting facet margin, (g) a superior foraminal facet osteophyte and cyst, (h) a superior foraminal ligament impingement, (i) a hidden shoulder osteophyte. The position of the key opinion authors of the perspective article is that further clinical research will continue to validate pain generator-based treatment protocols for lumbar spinal stenosis. The endoscopic technology platform enables spine surgeons to directly visualize pain generators, forming the basis for more simplified targeted surgical pain management therapies. Limitations of this care model are dictated by appropriate patient selection and mastering the learning curve of modern MIS procedures. Decompensated deformity and instability will likely continue to be treated with open corrective surgery. Vertically integrated outpatient spine care programs are the most suitable setting for executing such pain generator-focused programs.

Spatial variation of cardiovascular mortality in Cali, Colombia, between 2010 and 2017.

BACKGROUND: Cardiovascular disease (CVD) is the leading cause of mortality worldwide and in Colombia. The analysis of CVD mortality has been mainly relied on individual factors and rates, but occurrence is also related to contextual conditions. Understanding the distribution of CVD in a region will contribute to implement more focused-preventive and care interventions. METHODS: Using the national mortality register established by the Department of National Statistics, standardized rates and spatial distribution of CVD mortality were estimated for Cali, Colombia, between 2010-2017. Global and local spatial aggregation was assessed using the Geary's C test and for each district standardized mortality ratios smoothed by the Bayesian empirical were estimated. RESULTS: Over the period, CVD was the main cause of mortality with 28,804 deaths accounting for 23,8% of total deaths. The global CVD mortality rate varied from 235.9 to 257.4 per 100.000 habitants, with an average increase of 9.1% in the percentage change. The main cause of mortality were hypertensive diseases following by ischemic heart diseases and stroke. The standardized mortality ratios smoothed by the Bayesian empirical method showed that the districts 7, 13, 14, 15 and 16 located at the eastern area with the lowest socio-economic strata and the district 22 at the south of the city with the highest socio-economic strata had the high risks of CVD mortality. All these areas were at the boundary of the city. The the lowest risk was observed in districts 1 and 2 at the northwest area with the upper socio-economic strata. Over the study period, a spatial autocorrelation was found in the districts 1,9 10, 11, 12, 13, 14, 15, 19, and 21 by using the Geary's C test. The highest significant spatial association was found in the districts 1 and 21. CONCLUSION: Of 22 districts in Cali, the highest risk of CVD mortality was found in three at the lowest and one in the upper socio-economic strata between 2013 and 2017. Over the period a global spatial aggregation was identified due mainly to districts peripherical located suggesting that there could be contextual conditions influencing the risk. Therefore, there is a need for considering local conditions to prevent CVD mortality.

Flexural Capacity of Concrete Beams with Basalt Fiber-Reinforced Polymer Bars and Stirrups.

The flexural properties of six 120 × 300 × 4500 mm concrete beams reinforced with bars made from basalt fiber-reinforced polymer (BFRP) basalt fibers and concrete stirrups were investigated. The beams contained different concrete compositions (with or without basalt fibers). Steel and BFRP bars were used as longitudinal and shear reinforcement. As expected, all the beams failed by the crushing of the concrete in the top compression fibers because of using BFRP bars. Beams with BFRP bars should be designed to fail by concrete crushing because it is safer than a brittle failure of the bars. The beams with composite reinforcement were characterized by the greatest number of cracks with the largest crack width. The use of basalt fibers resulted in slightly reduced cracks in beams. The most significant deflections were recorded for the beams with BFRC composite reinforcement, the smallest for FRC beams. Adding basalt fibers to the concrete resulted in slightly reduced deflection of FRC beams compared to RC beams and significantly reduced deflection compared to BFRC beams. Results showed that introducing basalt fibers to the concrete increased curvature ductility of these beams. A theoretical analysis of flexural capacity showed that the ACI standard design is more similar to experimental values (0.87). A more restrictive standard, as it turns out, is the fib Model Code (0.68).

Free Radical-Based Sequencing for Native Top-Down Mass Spectrometry.

Native top-down proteomics allows for both proteoform identification and high-order structure characterization for cellular protein complexes. Unfortunately, tandem MS-based fragmentation efficiencies for such targets are low due to an increase in analyte ion mass and the low ion charge states that characterize native MS data. Multiple fragmentation methods can be integrated in order to increase protein complex sequence coverage, but this typically requires use of specialized hardware and software. Free-radical-initiated peptide sequencing (FRIPS) enables access to charge-remote and electron-based fragmentation channels within the context of conventional CID experiments. Here, we optimize FRIPS labeling for native top-down sequencing experiments. Our labeling approach is able to access intact complexes with TEMPO-based FRIPS reagents without significant protein denaturation or assembly disruption. By combining CID and FRIPS datasets, we observed sequence coverage improvements as large as 50% for protein complexes ranging from 36 to 106 kDa. Fragment ion production in these experiments was increased by as much as 102%. In general, our results indicate that TEMPO-based FRIPS reagents have the potential to dramatically increase sequence coverage obtained in native top-down experiments.

Preferencias alimentarias en tiempos de pandemia: El caso de personas mayores chilenas / Food preferences in times of pandemic. The case of chilean older people

Resumen: Objetivo: El aislamiento social originado por el COVID-19 ha potenciado modificaciones en los estilos de vida, afectando el bienestar de las personas mayores. Es por esto que el objetivo del estudio es el interpretar las influencias de factores emocionales y socioeconómicos en las preferencias alimentarias de personas mayores en Gran Concepción, Chile, en tiempos de pandemia. Materiales y Métodos: Para el logro de los propósitos, se realizó una investigación que responde al paradigma cualitativo con enfoque fenomenológico interpretativo de Heidegger. Los participantes existieron seleccionados mediante un muestreo intencional en base a los criterios de selección y considerando como tamaño muestras el resultante del punto de saturación. El estudio incluyó a un total de 12 personas mayores, las que fueron entrevistadas de manera virtual; los datos resultantes, se codificaron, reagruparon y analizaron mediante técnica de análisis de contenido. Resultados: Los platillos preferidos responden a los propios de la cultura local; por otra parte, aspectos emocionales y económicos no son reconocidos en la selección de alimentos y preferencias alimentarias. La comunicación lograda por el uso de redes sociales con sus familias es identificada como positiva y no influye en sus conductas alimentarias. Conclusiones: Para este grupo de personas mayores, la vivencia de aislamiento social por COVID-19, no fue suficiente para modificar las preferencias alimentarias.

Abstract: Objective: The social isolation caused by COVID-19 has promoted changes in lifestyles, affecting the well-being of the elderly. Therefore, the objective of the study is to interpret the influence of emotional and socioeconomic factors in food preferences of older people of the Gran Concepcion, Chile, in times of pandemic. Materials and methods: For the achievement of its purposes, an investigation was carried out that responds to the qualitative paradigm with the interpretive phenomenological approach of Heidegger. Participants were selected by intentional sampling based on the selection criteria and considering the sample size resulting from the saturation point. The study included a total of 12 elderly, who were interviewed virtually; the resulting data was coded, regrouped and analyzed using a content analysis technique. Results: The favorite dishes respond to those of the local culture; on the other hand, emotional and economic aspects are not recognized in the selection of food and food preferences. The communication achieved using social networks with their families is identified as positive and does not influence their eating behaviors. Conclusions: For this group of older people, the experience of social isolation due to COVID-19 was not enough to modify food preferences.