Enhancing diagnostic accuracy of multiple myeloma through ML-driven analysis of hematological slides: new dataset and identification model to support hematologists.

Multiple Myeloma (MM) is a hematological malignancy characterized by the clonal proliferation of plasma cells within the bone marrow. Diagnosing MM presents considerable challenges, involving the identification of plasma cells in cytology examinations on hematological slides. At present, this is still a time-consuming manual task and has high labor costs. These challenges have adverse implications, which rely heavily on medical professionals' expertise and experience. To tackle these challenges, we present an investigation using Artificial Intelligence, specifically a Machine Learning analysis of hematological slides with a Deep Neural Network (DNN), to support specialists during the process of diagnosing MM. In this sense, the contribution of this study is twofold: in addition to the trained model to diagnose MM, we also make available to the community a fully-curated hematological slide dataset with thousands of images of plasma cells. Taken together, the setup we established here is a framework that researchers and hospitals with limited resources can promptly use. Our contributions provide practical results that have been directly applied in the public health system in Brazil. Given the open-source nature of the project, we anticipate it will be used and extended to diagnose other malignancies.

Vaccination coverage in children in the period before and during the COVID-19 pandemic in Brazil: a time series analysis and literature review

Abstract Objective: To evaluate the behavior of VCR and VCH, per municipality and per vaccines offered at the NVC, to identify priority areas for intervention. Methods: Descriptive study of a time series, using secondary data and accompanied by a narrative review of the literature evaluating VCR and VCH. Vaccines offered to children under one year and to those aged one year in the pre-pandemic period of COVID-19 (2015 to 2019) were selected and compared to those offered during the pandemic period (2020 and 2021 ). Results and discussions: The decrease in VCR and VCH is a process that precedes the COVID-19 pandemic but was intensified during this period. In 2021, the VCR was around 70% for most vaccines. This phenomenon encompasses the entire country; however, it is more intense in the states/municipalities located in the north and northeast regions, suggesting greater difficulty in accessing health services. Conclusion: Low and heterogeneous VCR requires the adoption of practices that were previously implemented, establishing partnerships with governmental and non-governmental institutions, with adequate communication, active search for non-compliance and non-adherence to the regular vaccination program, adopting intra- and extramural vaccination strategies, to reverse the current situation and reduce the risk of recurrence of diseases that have been already controlled and eliminated.

Vaccination coverage in children in the period before and during the COVID-19 pandemic in Brazil: a time series analysis and literature review.

OBJECTIVE: To evaluate the behavior of VCR and VCH, per municipality and per vaccines offered at the NVC, to identify priority areas for intervention. METHODS: Descriptive study of a time series, using secondary data and accompanied by a narrative review of the literature evaluating VCR and VCH. Vaccines offered to children under one year and to those aged one year in the pre-pandemic period of COVID-19 (2015 to 2019) were selected and compared to those offered during the pandemic period (2020 and 2021). RESULTS AND DISCUSSIONS: The decrease in VCR and VCH is a process that precedes the COVID-19 pandemic but was intensified during this period. In 2021, the VCR was around 70% for most vaccines. This phenomenon encompasses the entire country; however, it is more intense in the states/municipalities located in the north and northeast regions, suggesting greater difficulty in accessing health services. CONCLUSIONS: Low and heterogeneous VCR requires the adoption of practices that were previously implemented, establishing partnerships with governmental and non-governmental institutions, with adequate communication, active search for non-compliance and non-adherence to the regular vaccination program, adopting intra- and extramural vaccination strategies, to reverse the current situation and reduce the risk of recurrence of diseases that have been already controlled and eliminated.

Determination and human health risk assessment of mercury in fish samples.

This work reports the determination of mercury in fish samples purchased at a public market in Belem City, Brazil. The mercury quantification was performed using the DMA method, which allows limits of detection and quantification of 0.004 and 0.012 ng, respectively. Method accuracy was confirmed using a certified reference material of fish protein from (NRCC) National Research Council, Canada. The analyzed species were: Dourada (Brachyplatystoma rousseauxii), Filhote (Brachyplatystoma filamentosum), Pescada Branca (Cynoscion leiarchus), Piramutaba (Brachyplatystoma vaillanti). The mercury contents expressed as wet sample weight varied from 0.078 to 0.150 µg g-1. Afterward, the health risk assessment indices Estimated Weekly Intake (EWI), Target Hazard Quotient (THQ), and Maximum Safe Consuming Quantity (MSCQ) were applied to the analytical data, and the results obtained were exhaustively interpreted and discussed. All the indices demonstrated that the daily consumption of 25 g of these fishes does not pose a risk to the human health of the local population. However, these conclusions are preliminary and should not be used in public policy matters.

Complex network model indicates a positive effect of inspiratory muscles pre-activation on performance parameters in a judo match.

This study investigated the effects of inspiratory muscle pre-activation (IMPA) on the interactions among the technical-tactical, physical, physiological, and psychophysiological parameters in a simulated judo match, based on the centrality metrics by complex network model. Ten male athletes performed 4 experimental sessions. Firstly, anthropometric measurements, maximal inspiratory pressure (MIP) and global strenght of the inspiratory muscles were determined. In the following days, all athletes performed four-minute video-recorded judo matches, under three conditions: without IMPA (CON), after IMPA at 15% (IMPA15), and at 40% (IMPA40) of MIP using an exerciser device. Blood lactate, heart rate and rating of perceived exertion were monitored, and the technical-tactical parameters during the match were related to offensive actions and the time-motion. Based on the complex network, graphs were constructed for each scenario (CON, IMPA15, and IMPA40) to investigate the Degree and Pagerank centrality metrics. IMPA40 increased the connectivity of the physical and technical-tactical parameters in complex network and highlighted the combat frequency and average combat time in top-five ranked nodes. IMPA15 also favoured the interactions among the psychophysiological, physical, and physiological parameters. Our results suggest the positive effects of the IMPA, indicating this strategy to prepare the organism (IMPA15) and to improve performance (IMPA40) in judo match.

Prediction of Death After Noncardiac Surgery: Potential Advantage of Using High-Sensitivity Troponin T as a Continuous Variable.

Background Increased high-sensitivity cardiac troponin T (hs-cTnT) above the upper reference limit (URL) after noncardiac surgery identifies patients at risk for mortality. Prior studies have not analyzed hs-cTnT as a continuous variable or probed age- and sex-specific URLs. This study compared the prediction of 30-day mortality using continuous postoperative hs-cTnT levels to the use of the overall URL and age- and sex-specific URLs. Methods and Results Patients (876) >40 years of age who underwent noncardiac surgery were included. Hs-cTnT was measured on postoperative day 1. Cox proportional hazards models were used to compare associations between 30-day mortality and using hs-cTnT as a continuous variable, or above the overall or age- and sex-specific URLs. Comparisons were performed by the area under the receiver operating characteristic curve analysis. Mortality was 4.2%. For each 1 ng/L increase in postoperative hs-cTnT, there was a 0.3% increase in mortality (P<0.001). Patients with postoperative hs-cTnT >14 ng/L were 37% of the cohort, while those above age- and sex-specific URLs were 25.3%. Both manifested higher mortality (hazard ratio [HR], 3.19; 95% CI, 1.20-8.49; P=0.020) and (HR, 2.76; P=0.009) than those with normal levels. The area under receiver operating characteristic curve was 0.89 using hs-cTnT as a continuous variable, 0.87 for age- and sex-specific URLs, and 0.86 for the overall URL. Conclusions Hs-cTnT as a continuous variable was independently associated with 30-day mortality and had the highest accuracy. Hs-cTnT elevations using overall and/or age- and sex-specific URLs were also associated with higher mortality.

Prevalence of human papillomavirus (HPV) in Brazil: A systematic review and meta-analysis.

OBJECTIVES: This study aimed to estimate the prevalence of genital, anal and oral HPV infection in Brazil through systematic review and meta-analysis. METHODS: We searched EMBASE, LILACS, MEDLINE, Web of Science and SciELO from inception to December 2018. Original research articles that assessed the prevalence of genital (i.e., cervical, penile), anal and oral HPV infection in Brazil were selected in pairs by independent authors. No sex, age, HPV vaccination, language or date restrictions were applied. HPV prevalence was estimated and stratified according to risk factors population and by geographic area throughout the country. The study prevalence was pooled using a random effects model. Analysis was performed using R (version 3.5.2), packages meta version 4.9-4 and metaphor 2.0-0. This review is registered on PROSPERO under protocol number CRD42016032751. RESULTS: We identified 3,351 references. After the screening process, 139 of them were eligible for this systematic review (57,513 total participants). Prevalence of cervical HPV was 25.41% (95% CI 22.71-28.32). Additionally, prevalence was 36.21% (95% CI 23.40, 51.33) in the penile region, 25.68% (95%CI 14.64, 41.04) in the anal region, and 11.89% (95%CI 6.26, 21.43) in the oral region. Subgroup analysis showed prevalence in each anatomic site was higher in high-risk populations. CONCLUSION: The prevalence of HPV is high in the Brazilian population and varies by population risk and anatomic body site, with lower rates in the oral cavity compared to that in the cervical, penile and anal region. Studies on HPV have primarily been developed to evaluate infection and cancer in the cervical region. There is a profound lack of HPV data in many geographic regions of Brazil and for different anatomic sites.

Trends and spatial distribution of MMR vaccine coverage in Brazil during 2007-2017.

We analyzed the time trends and spatial distribution of MMR vaccine coverage in Brazil during 2007-2017. In early 2018, a measles outbreak started in the North region of Brazil, reaching 11 of the 27 federal units by January 24, 2019. In this period, 10,302 cases were confirmed. Although the reintroduction of measles in Brazil is likely due to migration from Venezuela, the spread of the virus was made possible by the low levels of MMR coverage, as a result of significant decreases during the study period. Areas with high concentration of municipalities with low coverage are more susceptible to the spread of the virus, especially in the North and Northeast regions. Increasing vaccination coverage is essential to block the ongoing outbreak in Brazil. Vaccination strategies might target priority areas, especially those with a marked decrease in coverage. Moreover, it is essential to extend actions to travelers, migrants and refugees.

Structure-guided identification of antimicrobial peptides in the spathe transcriptome of the non-model plant, arum lily (Zantedeschia aethiopica).

Antimicrobial peptides (AMPs) are small molecules present in all living beings. Despite their huge sequence variability, AMPs present great structural conservation, mainly in cysteine-stabilized families. Moreover, in non-model plants, it is possible to detect cysteine-stabilized AMPs (cs-AMPs) with different sequences not covered by conventional searches. Here, we described a threading application for cs-AMP identification in the non-model arum lily (Zantedeschia aethiopica) plant, exploring the spathe transcriptome. By using the predicted proteins from the Z. aethiopica transcriptome as our primary source of sequences, we have filtered by using structural alignments of 12 putative cs-AMP sequences. The two unreported sequences were submitted to PCR validation, and ZaLTP7 gene was confirmed. By using the structure alignments, we classified ZaLTP7 as an LTP type 2-like. The successful threading application for cs-AMP identification is an important advance in transcriptomic and proteomic data mining. Besides, the same approach could be applied to the use of NGS public data to discover molecules to combat multidrug-resistant bacteria.

Microwave-Assisted Digestion Procedures Using Diluted Oxidant Mixture for Elemental Analysis of Crustaceans by MIP OES.

The aim of this study was to evaluate three digestion procedures using a diluted oxidant mixture of nitric acid, hydrogen peroxide, and water for determination of Cu, Fe, K, Mg, Mn, and Zn in shrimp and crab samples from Northern Brazil by microwave-induced plasma optical emission spectrometry. The efficiency of the digestion procedure was evaluated by comparing the residual carbon content, residual acidity, and elemental concentrations obtained in the samples. Digestion procedure 2 (4 mL HNO3 + 4 mL H2O2 + 4 mL H2O) was chosen due the adequate residual carbon content (5.85%) and low residual acidity (2.94 mol L-1). The recoveries obtained using the certified reference material (DORM-4) varied from 90 to 105%. High contents of K and Mg were found in the studied crustaceans. Also, it can give an excellent contribution to the recommended daily intake of Cu and Zn, indicating that these foods can be considered a good source of these minerals for the human diet.

Comparison of high-sensitivity cardiac troponin I and T for the prediction of cardiac complications after non-cardiac surgery.

BACKGROUND: We aimed to directly compare preoperative high-sensitivity cardiac troponin (hs-cTn) I and T concentration for the prediction of major cardiac complications after non-cardiac surgery. METHODS: We measured hs-cTnI and hs-cTnT preoperatively in a blinded fashion in 1022 patients undergoing non-cardiac surgery. The primary endpoint was a composite of major cardiac complications including cardiac death, cardiac arrest, myocardial infarction, clinically relevant arrhythmias, and acute heart failure within 30 days. We hypothesized that the type of surgery may impact on the predictive accuracy of hs-cTnI/T and stratified all analyses according to the type of surgery. RESULTS: Major cardiac complications occurred in 108 (11%) patients, 58/243 (24%) patients undergoing vascular surgery and 50/779 (6%, P < .001) patients undergoing non-vascular surgery. Using regulatory-approved 99th percentile cut-off concentrations, preoperative hs-cTnI elevations were less than one-fifth as common as preoperative hs-cTnT elevations (P < .001). Among patients undergoing vascular surgery, preoperative hs-cTnI concentrations, but not hs-cTnT, was an independent predictor of cardiac complications (adjusted odds ratio (aOR) 1.5, 95% confidence interval (95% CI) 1.0-2.1). The area under the receiver-operating characteristics curve (AUC) was 0.67 (95% CI, 0.59-0.75) for hs-cTnI versus 0.59 (95% CI 0.51-0.67, P = .012) for hs-cTnT. In contrast, among patients undergoing non-vascular surgery both preoperative hs-cTnI and hs-cTnT were independent predictors of the primary endpoint (aOR 1.6, 95% CI 1.3-2.0, and aOR 3.0, 95% CI 2.0-4.6, respectively) and showed higher predictive accuracy (AUC 0.77, 95% CI, 0.71-0.83, and 0.79, 95% CI 0.73-0.85, P = ns). CONCLUSIONS: Preoperative hs-cTnI and hs-cTnT concentrations predict major cardiac complications after non-vascular surgery, while, in patients undergoing vascular surgery, hs-cTnI may have better accuracy.

In silico optimization of a guava antimicrobial peptide enables combinatorial exploration for peptide design.

Plants are extensively used in traditional medicine, and several plant antimicrobial peptides have been described as potential alternatives to conventional antibiotics. However, after more than four decades of research no plant antimicrobial peptide is currently used for treating bacterial infections, due to their length, post-translational modifications or  high dose requirement for a therapeutic effect . Here we report the design of antimicrobial peptides derived from a guava glycine-rich peptide using a genetic algorithm. This approach yields guavanin peptides, arginine-rich α-helical peptides that possess an unusual hydrophobic counterpart mainly composed of tyrosine residues. Guavanin 2 is characterized as a prototype peptide in terms of structure and activity. Nuclear magnetic resonance analysis indicates that the peptide adopts an α-helical structure in hydrophobic environments. Guavanin 2 is bactericidal at low concentrations, causing membrane disruption and triggering hyperpolarization. This computational approach for the exploration of natural products could be used to design effective peptide antibiotics.

Building a safety culture in global health: lessons from Guatemala.

Programmes to modify the safety culture have led to lasting improvements in patient safety and quality of care in high-income settings around the world, although their use in low-income and middle-income countries (LMICs) has been limited. This analysis explores (1) how to measure the safety culture using a health culture survey in an LMIC and (2) how to use survey data to develop targeted safety initiatives using a paediatric nephrology unit in Guatemala as a field test case. We used the Safety, Communication, Operational Reliability, and Engagement survey to assess staff views towards 13 health climate and engagement domains. Domains with low scores included personal burnout, local leadership, teamwork and work-life balance. We held a series of debriefings to implement interventions targeted towards areas of need as defined by the survey. Programmes included the use of morning briefings, expansion of staff break resources and use of teamwork tools. Implementation challenges included the need for education of leadership, limited resources and hierarchical work relationships. This report can serve as an operational guide for providers in LMICs for use of a health culture survey to promote a strong safety culture and to guide their quality improvement and safety programmes.

In silico analyses of deleterious missense SNPs of human apolipoprotein E3.

ApoE3 is the major chylomicron apolipoprotein, binding in a specific liver peripheral cell receptor, allowing transport and normal catabolism of triglyceride-rich lipoprotein constituents. Point mutations in ApoE3 have been associated with Alzheimer's disease, type III hyperlipoproteinemia, atherosclerosis, telomere shortening and impaired cognitive function. Here, we evaluate the impact of missense SNPs in APOE retrieved from dbSNP through 16 computational prediction tools, and further evaluate the structural impact of convergent deleterious changes using 100 ns molecular dynamics simulations. We have found structural changes in four analyzed variants (Pro102Arg, Arg132Ser, Arg176Cys and Trp294Cys), two of them (Pro102Arg and Arg176Cys) being previously associated with human diseases. In all cases, except for Trp294Cys, there was a loss in the number of hydrogen bonds between CT and NT domains that could result in their detachment. In conclusion, data presented here could increase the knowledge of ApoE3 activity and be a starting point for the study of the impact of variations on APOE gene.

Antimicrobial activity predictors benchmarking analysis using shuffled and designed synthetic peptides.

The antimicrobial activity prediction tools aim to help the novel antimicrobial peptides (AMP) sequences discovery, utilizing machine learning methods. Such approaches have gained increasing importance in the generation of novel synthetic peptides by means of rational design techniques. This study focused on predictive ability of such approaches to determine the antimicrobial sequence activities, which were previously characterized at the protein level by in vitro studies. Using four web servers and one standalone software, we evaluated 78 sequences generated by the so-called linguistic model, being 40 designed and 38 shuffled sequences, with ∼60 and ∼25% of identity to AMPs, respectively. The ab initio molecular modelling of such sequences indicated that the structure does not affect the predictions, as both sets present similar structures. Overall, the systems failed on predicting shuffled versions of designed peptides, as they are identical in AMPs composition, which implies in accuracies below 30%. The prediction accuracy is negatively affected by the low specificity of all systems here evaluated, as they, on the other hand, reached 100% of sensitivity. Our results suggest that complementary approaches with high specificity, not necessarily high accuracy, should be developed to be used together with the current systems, overcoming their limitations.

Theoretical structural characterization of lymphoguanylin: A potential candidate for the development of drugs to treat gastrointestinal disorders.

Guanylin peptides (GPs) are small cysteine-rich peptide hormones involved in salt absorption, regulation of fluids and electrolyte homeostasis. This family presents four members: guanylin (GN), uroguanylin (UGN), lymphoguanylin (LGN) and renoguanylin (RGN). GPs have been used as templates for the development of drugs for the treatment of gastrointestinal disorders. Currently, LGN is the only GP with only one disulfide bridge, making it a remarkable member of this family and a potential drug template; however, there is no structural information about this peptide. In fact, LGN is predicted to be highly disordered and flexible, making it difficult to obtain structural information using in vitro methods. Therefore, this study applied a series of 1µs molecular dynamics simulations in order to understand the structural behavior of LGN, comparing it to the C115Y variant of GN, which shows the same Cys to Tyr modification. LGN showed to be more flexible than GN C115Y. While the negatively charged N-terminal, despite its repellent behavior, seems to be involved mainly in pH-dependent activity, the hydrophobic core showed to be the determinant factor in LGN's flexibility, which could be essential in its activity. These findings may be determinant in the development of new medicines to help in the treatment of gastrointestinal disorders. Moreover, our investigation of LGN structure clarified some issues in the structure-activity relationship of this peptide, providing new knowledge of guanylin peptides and clarifying the differences between GN C115Y and LGN.

Classificação de risco de transmissão de doenças imunopreveníveis a partir de indicadores de coberturas vacinais nos municípios brasileiros / Clasificación de riesgo de transmisión de enfermedades prevenibles a partir de indicadores de cobertura de vacunación en municipios / Classification of transmission risk of vaccine-preventable diseases based on vaccination indicators in Brazilian municipalities

OBJETIVO: descrever a classificação de risco de doenças imunopreveníveis nos municípios brasileiros. MÉTODOS: estudo epidemiológico descritivo com dados do Sistema de Informações do Programa Nacional de Imunizações (SI-PNI) para 2014; os indicadores de coberturas vacinais foram utilizados para classificar o risco de transmissão de doenças imunopreveníveis nos municípios. RESULTADOS: dos 5.570 municípios brasileiros, 12,0% foram classificados como de risco muito baixo, 29,6% de risco baixo, 2,2% de risco médio, 54,3% de risco alto e 1,8% de risco muito alto. CONCLUSÃO: a vigilância das coberturas vacinais permitiu identificar a maioria dos municípios em situação de alto risco e a minoria das crianças vivendo em municípios com cobertura adequada; a vigilância das coberturas utilizando indicadores pactuados no Sistema Único de Saúde (SUS) oferece nova ferramenta para identificação de áreas prioritárias, onde as ações poderão ter maiores chances de acerto pelos gestores e melhorar a qualidade e o sucesso do PNI.

OBJETIVO: describir la clasificación de riesgo de enfermedades prevenibles en municipios brasileños. MÉTODOS: estudio epidemiológico descriptivo con datos del Sistema de informaciones del programa nacional de inmunizaciones (PNI) de 2014; los indicadores de cobertura fueron utilizados para clasificar el riesgo de transmisión de enfermedades prevenibles en los municipios. RESULTADOS: de los 5.570 municipios brasileños, 12,0% fueron clasificados de muy bajo riesgo, 29,6% de bajo riesgo, 2,2% de riesgo medio, 54,3% de alto riesgo y 1,8% de riesgo muy alto. CONCLUSIÓN: la vigilancia de la cobertura de vacunación permitió identificar la mayoría de los municipios en situaciones de alto riesgo y la minoría de niños que viven en municipios con una cobertura adecuada; la vigilancia de la cobertura de vacunación según indicadores acordados en el sistema de salud pública ofrece una nueva herramienta para la identificación de áreas prioritarias en las que la acción pueda tener mayores posibilidades de éxito por los administradores de salud para mejorar la calidad y el éxito del programa de inmunización.

OBJECTIVE: to describe the transmission risk classification of vaccine-preventable diseases in Brazilian municipalities. METHODS: this was a descriptive epidemiologic study using 2014 data of the Brazilian National Immunization Program Information System; the vaccine coverage indicators were used to classify the transmission risk of vaccine-preventable diseases in the municipalities. RESULTS: of the 5,570 Brazilian municipalities, 12.0% were classified as very low risk, 29.6% as low risk, 2.2% as medium risk, 54.3% as high risk and 1.8% as very high risk. CONCLUSION: the vaccination coverage surveillance allowed to identify most of the municipalities in high risk situation and the minority of children living in municipalities with appropriate coverage; the vaccination coverage surveillance using indicators of the Brazilian National Health System (SUS) is a new tool for identifying priority areas where the actions can be more successful for health managers and improve the quality and the success of the immunizations program.

Classification of transmission risk of vaccine-preventable diseases based on vaccination indicators in Brazilian municipalities. / Classificação de risco de transmissão de doenças imunopreveníveis a partir de indicadores de coberturas vacinais nos municípios brasileiros.

OBJECTIVE: to describe the transmission risk classification of vaccine-preventable diseases in Brazilian municipalities. METHODS: this was a descriptive epidemiologic study using 2014 data of the Brazilian National Immunization Program Information System; the vaccine coverage indicators were used to classify the transmission risk of vaccine-preventable diseases in the municipalities. RESULTS: of the 5,570 Brazilian municipalities, 12.0% were classified as very low risk, 29.6% as low risk, 2.2% as medium risk, 54.3% as high risk and 1.8% as very high risk. CONCLUSION: the vaccination coverage surveillance allowed to identify most of the municipalities in high risk situation and the minority of children living in municipalities with appropriate coverage; the vaccination coverage surveillance using indicators of the Brazilian National Health System (SUS) is a new tool for identifying priority areas where the actions can be more successful for health managers and improve the quality and the success of the immunizations program.

Structural impact analysis of missense SNPs present in the uroguanylin gene by long-term molecular dynamics simulations.

The guanylate cyclase activator 2B, also known as uroguanylin, is part of the guanylin peptide family, which includes peptides such as guanylin and lymphoguanylin. The guanylin peptides could be related to sodium absorption inhibition and water secretion induction and their dysfunction may be related to various pathologies such as chronic renal failure, congestive heart failure and nephrotic syndrome. Besides, uroguanylin point mutations have been associated with essential hypertension. However, currently there are no studies on the impact of missense SNPs on uroguanylin structure. This study applied in silico SNP impact prediction tools to evaluate the impact of uroguanylin missense SNPs and to filter those considered as convergent deleterious, which were then further analyzed through long-term molecular dynamics simulations of 1µs of duration. The simulations suggested that all missense SNPs considered as convergent deleterious caused some kind of structural change to the uroguanylin peptide. Additionally, four of these SNPs were also shown to cause modifications in peptide flexibility, possibly resulting in functional changes.