Inflammatory bowel diseases (IBDs) are chronic conditions characterized by periods of spontaneous intestinal inflammation and are increasing in industrialized populations. Combined with host genetics, diet and gut bacteria are thought to contribute prominently to IBDs, but mechanisms are still emerging. In mice lacking the IBD-associated cytokine, interleukin-10, we show that a fiber-deprived gut microbiota promotes the deterioration of colonic mucus, leading to lethal colitis. Inflammation starts with the expansion of natural killer cells and altered immunoglobulin-A coating of some bacteria. Lethal colitis is then driven by Th1 immune responses to increased activities of mucin-degrading bacteria that cause inflammation first in regions with thinner mucus. A fiber-free exclusive enteral nutrition diet also induces mucus erosion but inhibits inflammation by simultaneously increasing an anti-inflammatory bacterial metabolite, isobutyrate. Our findings underscore the importance of focusing on microbial functions-not taxa-contributing to IBDs and that some diet-mediated functions can oppose those that promote disease.
Colitis , Inflammatory Bowel Diseases , Microbiota , Mice , Animals , Inflammatory Bowel Diseases/microbiology , Colitis/microbiology , Inflammation , Diet , Genetic Predisposition to Disease , Bacteria
ABSTRACT Primary graft failure (PGF) is a known complication following penetrating keratoplasty (PKP). The usual approach to treat this complication is to repeat a penetrating keratoplasty. Here, we report a case of Descemet's membrane endothelial keratoplasty (DMEK) for the treatment of PGF after PKP. A patient that underwent PKP, developed PGF with persistent graft edema and very poor visual acuity despite aggressive steroid use and a proof anti-viral treatment. Three months after the initial surgery, a DMEK was performed under the PKP graft. There was progressive early corneal clearing and, by the end of the first month, the patient already had no corneal edema. Uncorrected visual acuity (UCVA) improved to 20/40 and best corrected visual acuity (BCVA) to 20/20. DMEK may be an alternative to a second PKP for the treatment of PGF. This technique is a less invasive option when compared to the standard PKP procedure.
RESUMO A falência primária do enxerto é uma complicação conhecida que pode ocorrer após o transplante penetrante de córnea. O tratamento usual dessa complicação é com um novo transplante penetrante. Apresentamos um caso em que foi usado o transplante endotelial de membrana de Descemet (DMEK - do inglês Descemet membrane endo-thelial keratoplasty) para o tratamento da falência primária após o transplante penetrante. Uma paciente submetida a transplante penetrante evoluiu com falência primária do enxerto a despeito do uso intenso de corticoide tópico e uma prova terapêutica de antivirais. Três meses após a cirurgia inicial, foi optado pela realização do transplante endotelial de membrana de Descemet sob o transplante penetrante. Houve um clareamento precoce e progressivo do enxerto com melhora importante da visão. Após um mês, a visão sem correção era de 20/40 melhorando para 20/20 com refração. O transplante endotelial de membrana de Descemet pode ser uma alternativa a um novo transplante penetrante como tratamento da falência primária.
This study investigated the mechanical and tribological properties of 3D-printed Poly (lactic acid) (PLA) composites reinforced with different concentrations of carbon fibers (SCF) and graphene nanoparticles (GNP) (0.5 to 5 wt.% of each filler). The samples were produced using FFF (fused filament fabrication) 3D printing. The results showed a good dispersion of the fillers in the composites. SCF and GNP promoted the crystallization of the PLA filaments. The hardness, elastic modulus, and specific wear resistance grew with the increase in the filler concentration. A hardness improvement of about 30% was observed for the composite with 5 wt.% of SCF + 5 wt.% GNP (PSG-5) compared to PLA. The same trend was observed for the elastic modulus with an increase of 220%. All the composites presented lower coefficients of friction (0.49 to 0.6) than PLA (0.71). The composite PSG-5 sample showed the lowest value of specific wear rate (4.04 × 10-4 mm3/N.m), corresponding to about a five times reduction compared to PLA. Therefore, it was concluded that the addition of GNP and SCF to PLA made it possible to obtain composites with better mechanical and tribological behavior.
PURPOSE: Radiomics is a promising method for advancing imaging assessment in rectal cancer. This review aims to describe the emerging role of radiomics in the imaging assessment of rectal cancer, including various applications of radiomics based on CT, MRI, or PET/CT. METHODS: We conducted a literature review to highlight the progress of radiomic research to date and the challenges that need to be addressed before radiomics can be implemented clinically. RESULTS: The results suggest that radiomics has the potential to provide valuable information for clinical decision-making in rectal cancer. However, there are still challenges in terms of standardization of imaging protocols, feature extraction, and validation of radiomic models. Despite these challenges, radiomics holds great promise for personalized medicine in rectal cancer, with the potential to improve diagnosis, prognosis, and treatment planning. Further research is needed to validate the clinical utility of radiomics and to establish its role in routine clinical practice. CONCLUSION: Overall, radiomics has emerged as a powerful tool for improving the imaging assessment of rectal cancer, and its potential benefits should not be underestimated.
Positron Emission Tomography Computed Tomography , Rectal Neoplasms , Humans , Radiomics , Rectal Neoplasms/diagnostic imaging , Rectal Neoplasms/therapy , Prognosis , Magnetic Resonance Imaging/methods
ABSTRACT Purpose: The aim of this study was to validate the Portuguese version of Catquest-9SF through its application in a native Brazilian population with cataracts and to determine the correlation of the questionnaire scores with preoperative visual acuity. Methods: A prospective study was conducted to validate the Catquest-9SF questionnaire, which was translated and back-translated, generating a final version in Portuguese. A total of 120 Brazilian patients awaiting cataract surgery were recruited to answer the questionnaire and to document their preoperative data and visual acuity. The Rasch analysis was used to assess the instrument's psychometric properties. Results: The Portuguese version of Catquest-9SF demonstrated an acceptable adjustment of the items (item fit statistics ranging from 0.7 to 1.3), unidimensionality (principal component analysis), and good organization in the item response categories (thresholds of the categories: -2.79, 0.57, and 2.22, respectively). The questionnaire contains items with stable relationships if considered at the same level of visual impairment in the comparison between the two groups (absence of differential item functioning). The separation of people (person separation index, 3.07) was adequate. The visual acuity in the logarithm of the minimum angle of resolution (logMAR) in the best eye with the best optical correction showed a statistically significant correlation with the Catquest-9SF logit score (r=0.282 and p=0.004). Conclusions: The Portuguese version of Catquest-9SF presents evidence of validity and reliability, in addition to being linguistically and culturally understandable for Portuguese-speaking patients born in Brazil. The questionnaire is easy to understand and quick to apply, as it could adequately estimate the subjective visual functioning in patients with cataracts. We found a significant correlation between visual acuity and the questionnaire score.
RESUMO Objetivos: Validar a versão em português do Catquest-9SF através de sua aplicação em uma população nativa do Brasil com catarata e determinar a correlação da pontuação obtida no questionário com a acuidade visual pré-operatória. Métodos: Realizou-se um estudo prospectivo para validação de questionário. O Catquest-9SF foi traduzido e retro traduzido gerando uma versão final em português. Um total de 120 pacientes brasileiros que aguardavam realização de cirurgia de catarata foram recrutados para responder ao questionário e para documentação de dados pré-operatórios e acuidade visual. Análise Rasch foi utilizada para acessar as propriedades psicométricas do instrumento. Resultados: A versão em português do Catquest-9SF demonstrou ajuste aceitável dos itens (item fit statistics variando entre 0,7 e 1,3), unidimensionalidade (Principal Component Analisis) e boa organização nas categorias de resposta dos itens (limiares das categorias: -2,79; 0,57; 2,22). O questionário contém itens com relação estável, se considerado um mesmo nível de deficiência visual, na comparação entre dois grupos (ausência de differential item functioning). Existe adequada separação de pessoas (Person Separation Index 3,07). A acuidade visual em LogMAR no melhor olho com melhor correção óptica mostrou correlação estatisticamente significativa com a pontuação em logit do Catquest-9SF (r=0,282 e p=0,004). Conclusões: A versão em português do Catquest-9SF apresenta evidência de validade e confiabilidade, além de ser linguística e culturalmente compreensível para pacientes de língua portuguesa naturais do Brasil. Trata-se de questionário de fácil entendimento e rápida aplicação, sendo capaz de estimar de maneira adequada o funcionamento visual subjetivo em pacientes com catarata. Existe correlação significativa com a acuidade visual e a pontuação obtida no questionário.
Inflammatory bowel disease (IBD) is a chronic condition characterized by periods of spontaneous intestinal inflammation and is increasing in industrialized populations. Combined with host genetic predisposition, diet and gut bacteria are thought to be prominent features contributing to IBD, but little is known about the precise mechanisms involved. Here, we show that low dietary fiber promotes bacterial erosion of protective colonic mucus, leading to lethal colitis in mice lacking the IBD-associated cytokine, interleukin-10. Diet-induced inflammation is driven by mucin-degrading bacteria-mediated Th1 immune responses and is preceded by expansion of natural killer T cells and reduced immunoglobulin A coating of some bacteria. Surprisingly, an exclusive enteral nutrition diet, also lacking dietary fiber, reduced disease by increasing bacterial production of isobutyrate, which is dependent on the presence of a specific bacterial species, Eubacterium rectale. Our results illuminate a mechanistic framework using gnotobiotic mice to unravel the complex web of diet, host and microbial factors that influence IBD.
The aim of this study was to fabricate different self-lubricating poly (lactic acid)-based bio-composites reinforced with mono- and multi-fillers of carbon fibers, graphene nanoparticles, and a soft Sn-based brazing alloy (Sn89-Zn8-Bi3) using a two-step process consisting of mechanical alloying followed by casting. The results showed that the incorporation of the different fillers on the PLA surface by mechanical alloying was quite homogenous. The volume ratio between the PLA and the fillers was 1:0.02, respectively. The PLA sample reinforced with short carbon fibers and graphene nanoparticles presented the highest hardness (84.5 Shore D, corresponding to a 10% increase compared to PLA) and the lowest specific wear rate (1.5 × 10-4 mm3/N·m, one order of magnitude lower than PLA). With regard to the coefficient of friction, the lowest value was obtained for the sample reinforced with graphene (0.43, corresponding to a decrease of 12% compared to PLA).
PURPOSE: The aim of this study was to validate the Portuguese version of Catquest-9SF through its application in a native Brazilian population with cataracts and to determine the correlation of the questionnaire scores with preoperative visual acuity. METHODS: A prospective study was conducted to validate the Catquest-9SF questionnaire, which was translated and back-translated, generating a final version in Portuguese. A total of 120 Brazilian patients awaiting cataract surgery were recruited to answer the questionnaire and to document their preoperative data and visual acuity. The Rasch analysis was used to assess the instrument's psychometric properties. RESULTS: The Portuguese version of Catquest-9SF demonstrated an acceptable adjustment of the items (item fit statistics ranging from 0.7 to 1.3), unidimensionality (principal component analysis), and good organization in the item response categories (thresholds of the categories: -2.79, 0.57, and 2.22, respectively). The questionnaire contains items with stable relationships if considered at the same level of visual impairment in the comparison between the two groups (absence of differential item functioning). The separation of people (person separation index, 3.07) was adequate. The visual acuity in the logarithm of the minimum angle of resolution (logMAR) in the best eye with the best optical correction showed a statistically significant correlation with the Catquest-9SF logit score (r=0.282 and p=0.004). CONCLUSIONS: The Portuguese version of Catquest-9SF presents evidence of validity and reliability, in addition to being linguistically and culturally understandable for Portuguese-speaking patients born in Brazil. The questionnaire is easy to understand and quick to apply, as it could adequately estimate the subjective visual functioning in patients with cataracts. We found a significant correlation between visual acuity and the questionnaire score.
Cataract , Humans , Reproducibility of Results , Prospective Studies , Portugal , Cataract/diagnosis , Cataract/epidemiology , Surveys and Questionnaires , Psychometrics , Quality of Life
Sulforaphane is a bioactive metabolite with anti-inflammatory activity and is derived from the glucosinolate glucoraphanin, which is highly abundant in broccoli sprouts. However, due to its inherent instability its use as a therapeutic against inflammatory diseases has been limited. There are few studies to investigate a whole food approach to increase sulforaphane levels with therapeutic effect and reduce inflammation. In the current study, using a mouse model of inflammatory bowel disease, we investigated the ability of steamed broccoli sprouts to ameliorate colitis and the role of the gut microbiota in mediating any effects. We observed that despite inactivation of the plant myrosinase enzyme responsible for the generation of sulforaphane via steaming, measurable levels of sulforaphane were detectable in the colon tissue and feces of mice after ingestion of steamed broccoli sprouts. In addition, this preparation of broccoli sprouts was also capable of reducing chemically-induced colitis. This protective effect was dependent on the presence of an intact microbiota, highlighting an important role for the gut microbiota in the metabolism of cruciferous vegetables to generate bioactive metabolites and promote their anti-inflammatory effects.
Brassica , Colitis , Gastrointestinal Microbiome , Isothiocyanates/pharmacology , Diet , Brassica/metabolism , Colitis/chemically induced , Colitis/prevention & control , Glucosinolates
AISI 420 martensitic stainless steel is widely used in the mould industry due to its high tensile strength, hardness, and corrosion properties. Another requirement concerning any material used for this type of application is high thermal conductivity to minimise the time between consecutive injection cycles. The surfaces of some parts of the mould may be textured and reinforced with a material with higher thermal conductivity to achieve this aim. The results of a detailed study on the texturing of annealed 420 stainless steel using a Nd:YVO4 fibre laser are presented in this work. The influence of the laser's processing parameters (laser power, scanning speed, number of passes, and line spacing) on the dimensions of the track, microstructure, and hardness of the modified surfaces was studied. Based on the continuity and dimensions of the machined grooves, several promising textures could be produced with laser power values from 5 to 30 W, scanning speeds of 500 to 2000 mm/s, 8 passes or more, and line spacings of 40 and 50 µm. High laser powers were responsible for the dissolution of chromium carbides in the laser tracks, the incorporation of chromium in austenite, and the consequent hardening of the microstructure.
Primary graft failure (PGF) is a known complication following penetrating keratoplasty (PKP). The usual approach to treat this complication is to repeat a penetrating keratoplasty. Here, we report a case of Descemet's membrane endothelial keratoplasty (DMEK) for the treatment of PGF after PKP. A patient that underwent PKP, developed PGF with persistent graft edema and very poor visual acuity despite aggressive steroid use and a proof anti-viral treatment. Three months after the initial surgery, a DMEK was performed under the PKP graft. There was progressive early corneal clearing and, by the end of the first month, the patient already had no corneal edema. Uncorrected visual acuity (UCVA) improved to 20/40 and best corrected visual acuity (BCVA) to 20/20. DMEK may be an alternative to a second PKP for the treatment of PGF. This technique is a less invasive option when compared to the standard PKP procedure.
In the present work, we developed and evaluated the performance of a new flow cytometry-based single platform, referred to as "FC-Duplex IgG1 (HTLV-1/2)", for universal and differential serodiagnosis of HTLV-1/2 infection. The proposed technology employs a system for detection of IgG1 antibodies in a single competitive immunofluorescence platform by flow cytometry using fluorescently labeled MT-2/MoT cell line mix coupled to a highly sensitive development system (Biotin/Streptavidin/Phycoerythrin). The stability of fluorescent labeling and the antigenicity of MT-2 and MoT cell lines were confirmed upon storage at -20°C for 2, 6, and 12 months. The anti-HTLV-1/2 IgG1 reactivity, expressed as percentage of positive fluorescent cells (PPFC), was evaluated for each target antigen along the titration curve of test serum samples (1:32 to 1:4,096). Upon selection of target cell line and serum dilutions with higher segregation score between groups, the performance of "FIX" and "FIX & PERM" protocols was evaluated. The "FIX" protocol presented excellent performance indices (Se = 92%/Sp = 94%/AUC = 0.96; Se = 96%/Sp = 100%/AUC = 0.99) for the universal (HTLV-1/2 vs. NI) and differential (HTLV-1 vs. HTLV-2) diagnosis of HTLV-1 infection, respectively. Optimization of the "FIX" protocol using the principle of synchronous and asynchronous pairwise analysis further improved the performance of "FC-Duplex IgG1 (HTLV-1/2)", using the "FIX" protocol for differential diagnosis of HTLV-1 and HTLV-2 infections (Se = 100%/Sp = 100%/AUC = 1.00). In conclusion, the "FC-Duplex IgG1 (HTLV-1/2)" method represents an innovation in the biotechnology segment with the potential to compose a serological kit for differential diagnosis of HTLV-1/2 infection for reference laboratories and blood centers.
HTLV-I Infections , Human T-lymphotropic virus 1 , Flow Cytometry/methods , HTLV-I Infections/diagnosis , Humans , Immunoglobulin G , Serologic Tests
Cytomegalovirus (CMV) is a member of the Herpesviridae family, including viruses that are well-known agents of keratitis, anterior uveitis, scleritis and retinitis. CMV is usually associated with ocular diseases in immunosuppressed individuals, with a notable exception of hypertensive anterior uveitis with distinctive clinical features in immunocompetent patients. This syndrome was characterized in the last two decades in Europe and Southeast Asia, and then documented in the rest of world. Definitive diagnosis in these cases is usually made by Polymerase Chain Reaction (PCR) of the anterior chamber fluid. We report three immunocompetent Brazilian adults with history of multiple glaucomatocyclitic crises and presenting with chronic hypertensive anterior uveitis invariably with mild anterior chamber inflammation and characteristic scarce nummular keratic precipitates. CMV DNA was successfully amplified and detected in the aqueous humor of all patients. Corneal endothelial counts were significantly reduced in the involved eyes, with one patient developing bullous keratopathy. All patients were then treated with topical ganciclovir gel and corticosteroids, with subsequent control of the intraocular inflammation. CMV may represent an overlooked / underestimated etiology of hypertensive anterior uveitis that may progressively lead to endothelial dysfunction, culminating in bullous keratopathy. Management of patients is challenging, with the potential use of topical antivirals to decrease the number of relapses, and corticosteroids to control anterior uveitis / endotheliitis and to protect the corneal endothelium.
Cytomegalovirus Infections , Eye Infections, Viral , Uveitis, Anterior , Adult , Antiviral Agents/therapeutic use , Brazil , Cytomegalovirus/genetics , Cytomegalovirus Infections/diagnosis , Cytomegalovirus Infections/drug therapy , DNA, Viral/genetics , Eye Infections, Viral/diagnosis , Eye Infections, Viral/drug therapy , Humans , Uveitis, Anterior/diagnosis , Uveitis, Anterior/drug therapy
Diets high in cholesterol alter intestinal immunity. Here, we examined how the cholesterol metabolite 25-hydroxycholesterol (25-HC) impacts the intestinal B cell response. Mice lacking cholesterol 25-hydroxylase (CH25H), the enzyme generating 25-HC, had higher frequencies of immunoglobulin A (IgA)-secreting antigen-specific B cells upon immunization or infection. 25-HC did not affect class-switch recombination but rather restrained plasma cell (PC) differentiation. 25-HC was produced by follicular dendritic cells and increased in response to dietary cholesterol. Mechanistically, 25-HC restricted activation of the sterol-sensing transcription factor SREBP2, thereby regulating B cell cholesterol biosynthesis. Ectopic expression of SREBP2 in germinal center B cells induced rapid PC differentiation, whereas SREBP2 deficiency reduced PC output in vitro and in vivo. High-cholesterol diet impaired, whereas Ch25h deficiency enhanced, the IgA response against Salmonella and the resulting protection from systemic bacterial dissemination. Thus, a 25-HC-SREBP2 axis shapes the humoral response at the intestinal barrier, providing insight into the effect of high dietary cholesterol in intestinal immunity.
Cell Differentiation/immunology , Hydroxycholesterols/metabolism , Immunoglobulin A/immunology , Plasma Cells/immunology , Sterol Regulatory Element Binding Protein 2/metabolism , Animals , Cholesterol, Dietary/immunology , Cholesterol, Dietary/metabolism , Hydroxycholesterols/immunology , Immunoglobulin A/metabolism , Intestinal Mucosa/immunology , Intestinal Mucosa/metabolism , Mice , Peyer's Patches/immunology , Peyer's Patches/metabolism , Plasma Cells/metabolism
The impact of HIV co-infection on the plasma immunological biomarker profile of HTLV-1 infected patients was evaluated. The plasma levels of leukotrienes and chemokines/cytokines were quantified by ELISA and Cytometric Bead Array. A total of 138 volunteers were enrolled and divided into two subgroups ("HTLV-1(+)HIV(-)" and "HTLV-1(+)(HIV(+)"), which were categorized according to the HTLV-1-associated neurological disease (AS, pHAM and HAM). Reference controls were BD and HIV mono-infected patients. HAM(+) exhibited higher CD4+ T-cell counts as compared to HIV+ mono-infected patients and lower HTLV-1 proviral load as compared to mono-infected HAM(-) patients. AS(+) exhibited higher levels of CysLT, CXCL8/IL-8 and lower levels of CCL5/RANTES as compared to AS(-). Increased levels of IL-6 and TNF with reduced levels of CXCL10/IP10 and CCL5/RANTES were observed in co-infected pHAM(+) as compared to mono-infected pHAM(-). HAM(+) patients revealed an increase in CXCL8/IL-8, CCL2/MCP-1, CXCL-10/IP-10, TNF and a decrease in IL-2 as compared to HAM(-) subgroup.
Coinfection , HIV Infections/immunology , HIV Infections/metabolism , HTLV-I Infections/immunology , HTLV-I Infections/metabolism , Host-Pathogen Interactions/immunology , Human T-lymphotropic virus 1/immunology , Adult , Biomarkers , CD4 Lymphocyte Count , Cross-Sectional Studies , Cytokines/blood , Cytokines/metabolism , Disease Susceptibility , Female , HIV Infections/virology , HTLV-I Infections/virology , Host-Pathogen Interactions/genetics , Humans , Leukotrienes/metabolism , Male , Middle Aged , Viral Load
INTRODUCTION: Descemet membrane endothelial keratoplasty (DMEK) is the current gold standard surgical procedure to treat Fuchs endothelial keratoplasty. Cataract removal and intraocular lens (IOL) implantation can be safely combined with this corneal procedure. This paper presents a case series in which a toric intraocular lens (IOL) was used in combined DMEK and cataract surgery to correct corneal astigmatism. METHODS: Four eyes that had a toric IOL implanted in combined cataract and DMEK surgery were included. RESULTS: There was a reduction in manifest refractive cylinder and improvement in uncorrected and best-corrected vision in all cases. DISCUSSION: Surgeons may consider the use of toric IOLs in selected cases of combined DMEK and cataract surgery for Fuchs endothelial dystrophy.
PURPOSE: To evaluate the safety and efficacy of bilateral implantation of a supplementary small-aperture device to treat irregular corneal astigmatism. SETTING: Private practice. DESIGN: Retrospective consecutive case series. METHODS: Patients with bilateral irregular corneal astigmatism secondary to multiple causes and consented for implantation of the XtraFocus intraocular pinhole (IOPH) were enrolled. The mean follow-up was 27 months (range 5 to 66 months). Patients were assessed in their scheduled follow-up visits and monocular and binocular uncorrected and corrected distance and near visual acuities were recorded. Assessment of darkening vision complaints was also performed after implantation in the first eye and repeated after second-eye surgery. RESULTS: Thirty-two eyes of 16 patients were analyzed. The mean monocular and binocular uncorrected distance visual acuities improved from logMAR 1.091 ± 0.208 and 1.078 ± 0.259 preoperatively to 0.342 ± 0.091 (P < .001) and 0.342 ± 0.147 (P = .001) 1 year postoperatively. Three patients were excluded because of darkening vision complaints after surgery in the first eye. No major complications were noted after implantation of the IOPH. CONCLUSIONS: Bilateral implantation of the XtraFocus IOPH is a safe technique in a selected group of patients. There was improvement in visual acuity sustained over the analyzed period. Postoperative darkening vision complaints vary between individuals and can limit the application of this approach in certain patients.
Astigmatism , Lenses, Intraocular , Phacoemulsification , Astigmatism/surgery , Humans , Lens Implantation, Intraocular , Prosthesis Design , Retrospective Studies , Visual Acuity
One of the main goals of the Spanish and Portuguese-Speaking Working Group of the International Society for Forensic Genetics (GHEP-ISFG) is to promote and contribute to the development and dissemination of scientific knowledge in the field of forensic genetics. The GHEP-ISFG supports several Working Commissions which develop different scientific activities. One of them, the Working Commission on "Massively Parallel Sequencing (MPS): Forensic Applications", organized its first collaborative exercise on forensic applications of MPS technology in 2019. The aim of this exercise was to assess the concordance between the MPS results and those obtained with conventional technologies (capillary electrophoresis and Sanger sequencing), as well as to compare the results obtained within the different MPS platforms and/or the different kits/panels and analysis software packages (commercial and open-access) available on the market. The seven participating laboratories analyzed some samples of the annual GHEP-ISFG proficiency test (EIADN No. 27 (2019)), using Ion Torrent™ or MiSeq FGx® platforms. Six of them sent autosomal STR sequence data, five laboratories performed MPS analysis of individual identification SNPs, four laboratories reported MPS data of Y-chromosomal STRs, and X-chromosomal STRs, three laboratories performed MPS analysis of ancestry informative SNPs and phenotype informative SNPs, two labs performed MPS analysis of the mitochondrial DNA control region, and only one lab produced MPS data of lineage informative SNPs. Autosomal STR sequencing results were highly concordant to the consensus obtained by capillary electrophoresis in the EIADN No. 27 (2019) exercise. Furthermore, in general, a high level of concordance was observed between the results of the participating laboratories, regardless of the platform used. The main discordances were due to errors during the analysis process or from sequence data obtained with low depth of coverage. In this paper we highlight some issues that still arise, such as standardization of the nomenclature for STRs analyzed by sequencing with MPS, the universal uptake of a nomenclature framework by the analysis software, and well established validation and accreditation of the new MPS platforms for use in routine forensic case-work.
High-Throughput Nucleotide Sequencing/standards , Laboratories/standards , Chromosomes, Human, X , Chromosomes, Human, Y , DNA, Mitochondrial/genetics , Forensic Genetics/standards , Humans , Microsatellite Repeats , Polymorphism, Single Nucleotide , Sequence Analysis, DNA , Societies, Scientific
