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.

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.

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.

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).

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.

ISG15 as a prognostic biomarker in solitary fibrous tumour.

BACKGROUND: Solitary fibrous tumour (SFT) is a rare mesenchymal malignancy that lacks robust prognostic and predictive biomarkers. Interferon-stimulated gene 15 (ISG15) is a ubiquitin-like modifier, associated with tumour progression, and with poor survival of SFT patients, as previous published by our group. Here, we describe the role of ISG15 in the biology of this rare tumour. METHODS: ISG15 expression was assessed by immunohistochemistry in tissue microarrays from SFT patients and tested for correlation with progression-free survival and overall survival (OS). The effects of ISG15 knockdown or induction were investigated for cancer stem cell (CSC) characteristics and for drug sensitivity in unique in vitro models of SFT. RESULTS: The prognostic value of ISG15 for OS was validated at protein level in malignant SFT patients, prospectively treated with pazopanib and enrolled in GEIS-32 trial. In SFT in vitro models, ISG15 knockdown lead to a decrease in the expression of CSC-related genes, including SOX2, NANOG, ALDH1A1, ABCB1 and ABCC1. Likewise, ISG15 downregulation decreased the clonogenic/ tumoursphere-forming ability of SFT cells, while enhancing apoptotic cell death after doxorubicin, pazopanib or trabectedin treatment in 3D cell cultures. The regulation of CSC-related genes by ISG15 was confirmed after inducing its expression with interferon-ß1; ISG15 induction upregulated 1.28- to 451-fold the expression of CSC-associated genes. CONCLUSIONS: ISG15 is a prognostic factor in malignant SFT, regulating the expression of CSC-related genes and CSCs maintenance. Our results suggest that ISG15 could be a novel therapeutic target in SFT, which could improve the efficacy of the currently available treatments.

Human Biomonitoring of Polychlorinated Biphenyls (PCBs) in the Breast Milk of Colombian Mothers.

Polychlorinated biphenyls (PCBs) are persistent in the environment, bioaccumulate and biomagnify throughout the food chain, and may have adverse effects on human health and wildlife. PCB indicator (PCB 28, PCB 52, PCB 101, PCB 118, PCB 138, PCB 153, and PCB 180) were monitored in human milk using 68 samples from healthy and primiparous mothers from seven cities in Colombia, and the estimated daily intake (EDI) of infants was calculated. The PCB indicator with the highest concentration was PCB 153 with a value of 7.30 ng g-1 lipids. The maximum EDI was calculated as 0.257 µg kg-1 bw-1 day-1. In general, the PCB levels found in the 68 samples were low and did not represent a risk to breastfed infants. Additionally, these results could strengthen Colombia's efforts to increase the practice of breastfeeding. Finally, the results establish a general overview of population exposure and can be a scientific tool to improve environmental health policies in the country.

Dietary carbohydrate-to-protein ratio influences growth performance, hepatic health and dynamic of gut microbiota in atlantic salmon (Salmo salar).

Atlantic salmon (Salmo salar) fed a carbohydrate-rich diet exhibit suboptimal growth performance, along with other metabolic disturbances. It is well known that gut microbes play a pivotal role in influencing metabolism of the host, and these microbes can be modified by the diet. The main goal of the present study was to determine the effect of feeding graded levels of digestible carbohydrates to Atlantic salmon on the distal intestine digesta microbiota at 3 sampling times (i.e., weeks 4, 8 and 12), during a 12-week trial. A low carbohydrate-to-high protein diet (LC/HP, 0% wheat starch), a medium carbohydrate-to-medium protein diet (MC/MP, 15% wheat starch) or a high carbohydrate-to-low protein diet (HC/LP, 30% wheat starch) was fed to triplicate fish tanks (27 to 28 fish per tank). We performed an in-depth characterization of the distal intestine digesta microbiota. Further, growth parameters, liver histology and the expression of genes involved in hepatic neolipogenesis in fish were measured. Fish fed a HC/LP diet showed greater hepatosomatic and viscerosomatic indexes (P = 0.026 and P = 0.018, respectively), lower final weight (P = 0.005), weight gain (P = 0.003), feed efficiency (P = 0.033) and growth rate (P = 0.003) compared with fish fed the LC/HP diet. Further, feeding salmon a high digestible carbohydrate diet caused greater lipid vacuolization, steatosis index (P = 0.007) and expression of fatty acid synthase (fas) and delta-6 fatty acyl desaturase (d6fad) (P = 0.001 and P = 0.001, respectively) in the liver compared with fish fed the LC/HP diet. Although, the major impact of feeding a carbohydrate-rich diet to Atlantic salmon in beta diversity of distal intestine digesta microbiota was observed at week 4 (HC/LP vs MC/MP and HC/LP vs LC/HP; P = 0.007 and P = 0.008, respectively) and week 8 (HC/LP vs MC/MP; P = 0.04), no differences between experimental groups were detected after 12 weeks of feeding. Finally, at the end of the trial, there was a negative correlation between lactic acid bacteria (LAB) members, including Leuconostoc and Lactobacillus, with hepatic steatosis level, the hepatosomatic and viscerosomatic indexes as well as the expression of fas and d6fad. Weissella showed negative correlation with hepatic steatosis level and the hepatosomatic index. Finally, further research to explore the potential use of LAB as probiotics to improve liver health in carnivorous fish fed fatty liver-induced diet is warranted.

Oligomerization, trans-reduction, and instability of mutant NOTCH3 in inherited vascular dementia.

Cerebral small vessel disease (SVD) is a prevalent disease of aging and a major contributor to stroke and dementia. The most commonly inherited SVD, CADASIL, is caused by dominantly acting cysteine-altering mutations in NOTCH3. These mutations change the number of cysteines from an even to an odd number, but the impact of these alterations on NOTCH3 protein structure remain unclear. Here, we prepared wildtype and four mutant recombinant NOTCH3 protein fragments to analyze the impact of CADASIL mutations on oligomerization, thiol status, and protein stability. Using gel electrophoresis, tandem MS/MS, and collision-induced unfolding, we find that NOTCH3 mutant proteins feature increased amounts of inappropriate disulfide bridges, reduced cysteines, and structural instability. Presence of a second protein factor, an N-terminal fragment of NOTCH3 (NTF), is capable of further altering disulfide statuses of both wildtype and mutant proteins, leading to increased numbers of reduced cysteines and further destabilization of NOTCH3 structure. In sum, these studies identify specific cysteine residues alterations and quaternary structure induced by CADASIL mutations in NOTCH3; further, we validate that reductive factors alter the structure and stability of this small vessel disease protein.

The establishment of public health policies and the burden of non-alcoholic fatty liver disease in the Americas.

Non-alcoholic fatty liver disease (NAFLD) affects 20-25% of the general population and is associated with morbidity, increased mortality, and elevated health-care costs. Most NAFLD risk factors are modifiable and, therefore, potentially amenable to being reduced by public health policies. To date, there is no information about NAFLD-related public health policies in the Americas. In this study, we analysed data from 17 American countries and found that none have established national public health policies to decrease NAFLD-related burden. There is notable heterogeneity in the existence of public health policies to prevent NAFLD-related conditions. The most common public health policies were related to diabetes (15 [88%] countries), hypertension (14 [82%] countries), cardiovascular diseases (14 [82%] countries), obesity (nine [53%] countries), and dyslipidaemia (six [35%] of countries). Only seven (41%) countries had a registry of the burden of NAFLD, and efforts to raise awareness in the Americas were scarce. The implementation of public health policies are urgently needed in the Americas to decrease the burden of NAFLD.

Iduronate-2-sulfatase interactome: validation by yeast two-hybrid assay.

Mucopolysaccharidosis type II (MPS II), also known as Hunter syndrome, is a rare X-linked recessive disease caused by a deficiency of the lysosomal enzyme iduronate-2-sulfatase (IDS), which activates intracellular accumulation of nonmetabolized glycosaminoglycans such as heparan sulfate and dermatan sulfate. This accumulation causes severe damage to several tissues, principally the central nervous system. Previously, we identified 187 IDS-protein interactions in the mouse brain. To validate a subset of these interactions, we selected and cloned the coding regions of 10 candidate genes to perform a targeted yeast two-hybrid assay. The results allowed the identification of the physical interaction of IDS with LSAMP and SYT1. Although the physiological relevance of these complexes is unknown, recent advances allow us to point out that these interactions could be involved in vesicular trafficking of IDS through the interaction with SYT1, as well as to the ability to form a transcytosis module between the cellular components of the blood-brain-barrier (BBB) through its interaction with LSAMP. These results may shed light on the role of IDS on cellular homeostasis and may also contribute to the understanding of MPS II physiopathology and the development of novel therapeutic strategies to transport recombinant IDS through the brain endothelial cells toward the brain parenchyma.

Mass Spectrometry Methods for Measuring Protein Stability.

Mass spectrometry is a central technology in the life sciences, providing our most comprehensive account of the molecular inventory of the cell. In parallel with developments in mass spectrometry technologies targeting such assessments of cellular composition, mass spectrometry tools have emerged as versatile probes of biomolecular stability. In this review, we cover recent advancements in this branch of mass spectrometry that target proteins, a centrally important class of macromolecules that accounts for most biochemical functions and drug targets. Our efforts cover tools such as hydrogen-deuterium exchange, chemical cross-linking, ion mobility, collision induced unfolding, and other techniques capable of stability assessments on a proteomic scale. In addition, we focus on a range of application areas where mass spectrometry-driven protein stability measurements have made notable impacts, including studies of membrane proteins, heat shock proteins, amyloidogenic proteins, and biotherapeutics. We conclude by briefly discussing the future of this vibrant and fast-moving area of research.