RESUMEN
Abnormal assembly of tau, α-synuclein, TDP-43 and amyloid-ß proteins into amyloid filaments defines most human neurodegenerative diseases. Genetics provides a direct link between filament formation and the causes of disease. Developments in cryo-electron microscopy (cryo-EM) have made it possible to determine the atomic structures of amyloids from postmortem human brains. Here we review the structures of brain-derived amyloid filaments that have been determined so far and discuss their impact on research into neurodegeneration. Whereas a given protein can adopt many different filament structures, specific amyloid folds define distinct diseases. Amyloid structures thus provide a description of neuropathology at the atomic level and a basis for studying disease. Future research should focus on model systems that replicate the structures observed in disease to better understand the molecular mechanisms of disease and develop improved diagnostics and therapies.
Asunto(s)
Amiloide , Microscopía por Crioelectrón , Enfermedades Neurodegenerativas , Patología Molecular , Pliegue de Proteína , Humanos , alfa-Sinucleína , Amiloide/química , Amiloide/clasificación , Amiloide/ultraestructura , Péptidos beta-Amiloides , Enfermedades Neurodegenerativas/metabolismo , Enfermedades Neurodegenerativas/patologíaRESUMEN
The ISA Nomenclature Committee met electronically before and directly after the XVII ISA International Symposium on Amyloidosis, which, unfortunately, had to be virtual in September 2020 due to the ongoing COVID-19 pandemic instead of a planned meeting in Tarragona in March. In addition to confirmation of basic nomenclature, several additional concepts were discussed, which are used in scientific amyloid literature. Among such concepts are cytotoxic oligomers, protofibrils, primary and secondary nucleation, seeding and cross-seeding, amyloid signature proteins, and amyloid plaques. Recommendations for their use are given. Definitions of amyloid and amyloidosis are confirmed. Possible novel human amyloid fibril proteins, appearing as 'classical' in vivo amyloid, were discussed. It was decided to include fibulin-like extracellular matrix protein 1 (amyloid protein: AEFEMP1), which appears as localised amyloid in portal veins. There are several possible amyloid proteins under investigation, and these are included in a new Table.
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Amiloide/clasificación , Proteínas Amiloidogénicas/clasificación , Amiloidosis/clasificación , Terminología como Asunto , Amiloide/genética , Amiloide/metabolismo , Proteínas Amiloidogénicas/genética , Proteínas Amiloidogénicas/metabolismo , Amiloidosis/diagnóstico , Amiloidosis/genética , Amiloidosis/patología , COVID-19 , Congresos como Asunto , Infecciones por Coronavirus , Educación a Distancia/organización & administración , Expresión Génica , Humanos , Pandemias , Neumonía ViralRESUMEN
OBJECTIVE: To map the occurrence of amyloid types in a large clinical cohort using mass spectrometry-based shotgun proteomics, an unbiased method that unambiguously identifies all amyloid types in a single assay. METHODS: A mass spectrometry-based shotgun proteomics assay was implemented in a central reference laboratory. We documented our experience of typing 16,175 amyloidosis specimens over an 11-year period from January 1, 2008, to December 31, 2018. RESULTS: We identified 21 established amyloid types, including AL (n=9542; 59.0%), ATTR (n=4600; 28.4%), ALECT2 (n=511; 3.2%), AA (n=463; 2.9%), AH (n=367; 2.3%), AIns (n=182; 1.2%), KRT5-14 (n=94; <1%), AFib (n=71; <1%), AApoAIV (n=57; <1%), AApoA1 (n=56; <1%), AANF (n=47; <1%), Aß2M (n=38; <1%), ASem1 (n=34; <1%), AGel (n=29; <1%), TGFB1 (n=29; <1%), ALys (n=15; <1%), AIAPP (n=13; <1%), AApoCII (n=11; <1%), APro (n=8; <1%), AEnf (n=6; <1%), and ACal (n=2; <1%). We developed the first comprehensive organ-by-type map showing the relative frequency of 21 amyloid types in 31 different organs, and the first type-by-organ map showing organ tropism of 18 rare types. Using a modified bioinformatics pipeline, we detected amino acid substitutions in cases of hereditary amyloidosis with 100% specificity. CONCLUSION: Amyloid typing by proteomics, which effectively recognizes all amyloid types in a single assay, optimally supports the diagnosis and treatment of amyloidosis patients in routine clinical practice.
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Amiloide/clasificación , Amiloidosis/diagnóstico , Espectrometría de Masas , Proteómica/instrumentación , Secuencia de Aminoácidos , Amiloide/metabolismo , Amiloidosis/metabolismo , Femenino , Humanos , Masculino , Estudios Retrospectivos , Distribución por SexoRESUMEN
CONTEXT.: Amyloidosis is an uncommon but important entity. A protein-based classification of amyloidosis defines the underlying disease process, directing clinical management and providing prognostic information. However, in routine surgical pathology there often is no attempt to classify amyloid other than staining to determine light chain-associated amyloidosis. Systemic and localized amyloidosis vary with respect to frequency of organ involvement by different amyloid types, and most amyloid proteins have commercial antibodies available for identification. OBJECTIVE.: To provide a guide for the likelihood of amyloid type by organ system. DATA SOURCES.: Literature review based on PubMed searches containing the word amyloid, specifically addressing the prevalence and significance of amyloid proteins in each organ system other than the brain, and the authors' practice experience. CONCLUSIONS.: In patients with amyloidosis, determination of the responsible protein is critical for appropriate patient care. In large subspecialty practices and reference laboratories with experience in using and analyzing relevant immunohistochemistry, most amyloid proteins can be identified with an organ-specific algorithm. Referring to an organ-based algorithm may be helpful in providing clinicians with a more specific differential diagnosis regarding amyloid type to help guide clinical evaluation and treatment. When the protein cannot be characterized, mass spectrometry can be performed to definitively classify the amyloid type.
Asunto(s)
Amiloide/biosíntesis , Amiloidosis/metabolismo , Tracto Gastrointestinal/metabolismo , Inmunohistoquímica/métodos , Patología Quirúrgica/métodos , Amiloide/clasificación , Amiloidosis/diagnóstico , Huesos/metabolismo , Diagnóstico Diferencial , Humanos , Riñón/metabolismo , Hígado/metabolismo , Pulmón/metabolismo , Miocardio/metabolismo , Especificidad de Órganos , Sistema Nervioso Periférico/metabolismoRESUMEN
Amyloid is a diverse group of unrelated peptides or proteins that have positive functionality or are associated with various pathologies. Despite vast differences, all amyloids share several features that together uniquely define the group. 1) All amyloids possess a characteristic cross-ß pattern with X-ray diffraction typical of ß-sheet secondary protein structures. 2) All amyloids are birefringent and dichroic under polarizing microscopy after staining with Congo red, which indicates a crystalline-like (ordered) structure. 3) All amyloids cause a spectral shift in the peak wavelength of Congo red with conventional light microscopy due to perturbation of π electrons of the dye. 4) All amyloids show heightened intensity of fluorescence with Congo red, which suggests an unusual degree of packing of the dye onto the substrate. The ß portion of amyloid molecules, the only logical substrate for specific Congo red staining under histochemical conditions, consists of a stack of ß-sheets laminated by hydrophilic and hydrophobic interactions between adjacent pairs. Only the first and last ß-sheets are accessible to dyes. Each sheet is composed of numerous identical peptides running across the width of the sheet and arranged in parallel with side chains in register over the length of the fibril. Two sets of grooves are bordered by side chains. X grooves run perpendicular to the long axis of the fibril; these grooves are short (the width of the sheet) and number in the hundreds or thousands. Y grooves are parallel with the long axis. Each groove runs the entire length of the fibril, but there are very few of them. While Congo red is capable of ionic bonding with proteins via two sulfonic acid groups, physical constraints on the staining solution preclude ionic interactions. Hydrogen bonding between dye amine groups and peptide carbonyls is the most likely primary bonding mechanism, because all ß-sheets possess backbone carbonyls. Various amino acid residues may form secondary bonds to the dye via any of three van der Waals forces. It is possible that Congo red binds within the Y grooves, but that would not produce the characteristic staining features that are the diagnostic hallmarks of amyloid. Binding in the X grooves would produce a tightly packed series of dye molecules over the entire length of the fibril. This would account for the signature staining of amyloid by Congo red: dichroic birefringence, enhanced intensity of fluorescence and a shift in visible absorption wavelength.
Asunto(s)
Amiloide/química , Amiloide/clasificación , Péptidos/química , Amiloide/genética , Péptidos beta-Amiloides/química , Péptidos beta-Amiloides/genética , Rojo Congo , Enlace de Hidrógeno , Péptidos/genéticaRESUMEN
Amyloids are unbranched protein fibrils with a characteristic spatial structure. Although the amyloids were first described as protein deposits that are associated with the diseases, today it is becoming clear that these protein fibrils play multiple biological roles that are essential for different organisms, from archaea and bacteria to humans. The appearance of amyloid, first of all, causes changes in the intracellular quantity of the corresponding soluble protein(s), and at the same time the aggregate can include other proteins due to different molecular mechanisms. The co-aggregation may have different consequences even though usually this process leads to the depletion of a functional protein that may be associated with different diseases. The protein co-aggregation that is related to functional amyloids may mediate important biological processes and change of protein functions. In this review, we survey the known examples of the amyloid-related co-aggregation of proteins, discuss their pathogenic and functional roles, and analyze methods of their studies from bacteria and yeast to mammals. Such analysis allow for us to propose the following co-aggregation classes: (i) titration: deposition of soluble proteins on the amyloids formed by their functional partners, with such interactions mediated by a specific binding site; (ii) sequestration: interaction of amyloids with certain proteins lacking a specific binding site; (iii) axial co-aggregation of different proteins within the same amyloid fibril; and, (iv) lateral co-aggregation of amyloid fibrils, each formed by different proteins.
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Amiloide/metabolismo , Enfermedades Neurodegenerativas/metabolismo , Amiloide/química , Amiloide/clasificación , Animales , Sitios de Unión , Humanos , Proteínas Priónicas/química , Proteínas Priónicas/clasificación , Proteínas Priónicas/metabolismoRESUMEN
The nomenclature committee of the International Society of Amyloidosis (ISA) meets every second year to discuss and formulate recommendations. The conclusions from the discussion at the XVI International Symposium on Amyloidosis in Kumamoto, Japan, 25-29 March 2018 and afterwards are summarized in this Nomenclature Article. From having recommended the use of the designation "amyloid fibril" for in vivo material only, ISA's nomenclature committee now accepts its use more broadly following the international scientific literature. However, it is important always to stress the origin of the ß-fibrils in order to avoid misunderstanding. Given the more broad use of the word "amyloid" several classes of amyloid fibrils may be distinguished. For the medical in vivo situation, and to be included in the amyloid nomenclature list, "amyloid" still means mainly extracellular tissue deposits of protein fibrils, recognized by specific properties, such as green-yellow birefringence after staining with Congo red. It should also be underlined that in vivo amyloid fibrils, in addition to the main protein contain associated compounds, particularly serum amyloid P-component (SAP) and proteoglycans, mainly heparan sulfate proteoglycan. With this definition there are presently 36 human amyloid proteins of which 14 appear only associated with systemic amyloidosis and 19 as localized forms. Three proteins can occur both as localized and systemic amyloidosis. Strictly intracellular aggregates are not included in this list.
Asunto(s)
Amiloide/clasificación , Amiloidosis/clasificación , Terminología como Asunto , Humanos , Agencias Internacionales , Sociedades CientíficasRESUMEN
The molecular architecture of amyloids formed in vivo can be interrogated using luminescent conjugated oligothiophenes (LCOs), a unique class of amyloid dyes. When bound to amyloid, LCOs yield fluorescence emission spectra that reflect the 3D structure of the protein aggregates. Given that synthetic amyloid-ß peptide (Aß) has been shown to adopt distinct structural conformations with different biological activities, we asked whether Aß can assume structurally and functionally distinct conformations within the brain. To this end, we analyzed the LCO-stained cores of ß-amyloid plaques in postmortem tissue sections from frontal, temporal, and occipital neocortices in 40 cases of familial Alzheimer's disease (AD) or sporadic (idiopathic) AD (sAD). The spectral attributes of LCO-bound plaques varied markedly in the brain, but the mean spectral properties of the amyloid cores were generally similar in all three cortical regions of individual patients. Remarkably, the LCO amyloid spectra differed significantly among some of the familial and sAD subtypes, and between typical patients with sAD and those with posterior cortical atrophy AD. Neither the amount of Aß nor its protease resistance correlated with LCO spectral properties. LCO spectral amyloid phenotypes could be partially conveyed to Aß plaques induced by experimental transmission in a mouse model. These findings indicate that polymorphic Aß-amyloid deposits within the brain cluster as clouds of conformational variants in different AD cases. Heterogeneity in the molecular architecture of pathogenic Aß among individuals and in etiologically distinct subtypes of AD justifies further studies to assess putative links between Aß conformation and clinical phenotype.
Asunto(s)
Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/química , Amiloide/química , Placa Amiloide/metabolismo , Agregado de Proteínas , Enfermedad de Alzheimer/clasificación , Enfermedad de Alzheimer/genética , Enfermedad de Alzheimer/patología , Amiloide/clasificación , Amiloide/ultraestructura , Péptidos beta-Amiloides/metabolismo , Precursor de Proteína beta-Amiloide/genética , Precursor de Proteína beta-Amiloide/metabolismo , Animales , Modelos Animales de Enfermedad , Femenino , Colorantes Fluorescentes/química , Lóbulo Frontal/química , Lóbulo Frontal/metabolismo , Lóbulo Frontal/patología , Expresión Génica , Humanos , Masculino , Ratones , Lóbulo Occipital/química , Lóbulo Occipital/metabolismo , Lóbulo Occipital/patología , Péptido Hidrolasas/química , Placa Amiloide/clasificación , Placa Amiloide/genética , Placa Amiloide/patología , Presenilina-1/genética , Presenilina-1/metabolismo , Unión Proteica , Conformación Proteica , Proteolisis , Espectrometría de Fluorescencia , Lóbulo Temporal/química , Lóbulo Temporal/metabolismo , Lóbulo Temporal/patología , Tiofenos/químicaRESUMEN
Amyloidosis is the result of various, differently approachable diseases. It is vital to subtype the amyloid deposits in order to establish and finally treat the underlying disease properly. Besides the classical staining with Congo red, further procedures like immunohistochemical staining are needed for classification. Here, we present a more accurate approach using Congo red/immunohistochemical double staining easily applicable in routine diagnostics. Modifications of the Congo red staining technique and the immunohistochemical procedures were needed in order to combine both staining procedures on one slide. The evaluation was done using conventional light and fluorescence microscopy. By shortening the staining time for Congo red to 10 s and by modification regarding endogenous peroxidase blockage, accurate results could be obtained for evaluating the Congo red/immunohistochemistry double staining using a fluorescence microscope. Sections of 2 µm instead of 4 µm thickness were superior for evaluation, since they increased staining specificity. The combination of Congo red and immunohistochemistry as in situ double staining on one slide is a feasible approach in the diagnosis of amyloidosis. It allows focusing on the fluorescent Congo red-positive areas when evaluating immunohistochemistry, thus avoiding signing out false-positive results. Additionally, it increases the signal-to-noise ratio of the immunohistochemically stained sections on conventional microscopy.
Asunto(s)
Amiloide/análisis , Amiloidosis/diagnóstico , Amiloide/clasificación , Amiloide/metabolismo , Amiloidosis/metabolismo , Médula Ósea/patología , Colorantes , Rojo Congo , Mucosa Gástrica/patología , Humanos , Inmunohistoquímica , Microscopía Fluorescente , Mucosa Bucal/patología , Miocardio/patología , Glándulas Salivales/patología , Sensibilidad y Especificidad , Coloración y EtiquetadoRESUMEN
Polypeptide aggregation into amyloid is linked with several debilitating human diseases. Despite the inherent risk of aggregation-induced cytotoxicity, bacteria control the export of amyloid-prone subunits and assemble adhesive amyloid fibres during biofilm formation. An Escherichia protein, CsgC potently inhibits amyloid formation of curli amyloid proteins. Here we unlock its mechanism of action, and show that CsgC strongly inhibits primary nucleation via electrostatically-guided molecular encounters, which expands the conformational distribution of disordered curli subunits. This delays the formation of higher order intermediates and maintains amyloidogenic subunits in a secretion-competent form. New structural insight also reveal that CsgC is part of diverse family of bacterial amyloid inhibitors. Curli assembly is therefore not only arrested in the periplasm, but the preservation of conformational flexibility also enables efficient secretion to the cell surface. Understanding how bacteria safely handle amyloidogenic polypeptides contribute towards efforts to control aggregation in disease-causing amyloids and amyloid-based biotechnological applications.
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Amiloide/química , Proteínas de Escherichia coli/química , Chaperonas Moleculares/química , Electricidad Estática , Transporte Activo de Núcleo Celular , Amiloide/clasificación , Amiloide/genética , Amiloide/metabolismo , Proteínas de Escherichia coli/metabolismo , Cinética , Chaperonas Moleculares/metabolismo , Concentración Osmolar , Unión Proteica , Conformación Proteica , Pliegue de ProteínaRESUMEN
Laser microdissection followed by mass spectrometry has been successfully used for amyloid typing. However, sample contamination can interfere with proteomic analysis, and overnight digestion limits the analytical throughput. Moreover, current quantitative analysis methods are based on the spectrum count, which ignores differences in protein length and may lead to misdiagnoses. Here, we developed a microwave-assisted filter-aided sample preparation (maFASP) method that can efficiently remove contaminants with a 10-kDa cutoff ultrafiltration unit and can accelerate the digestion process with the assistance of a microwave. Additionally, two parameters (P- and D-scores) based on the exponentially modified protein abundance index were developed to define the existence of amyloid deposits and those causative proteins with the greatest abundance. Using our protocol, twenty cases of systemic amyloidosis that were well-typed according to clinical diagnostic standards (training group) and another twenty-four cases without subtype diagnoses (validation group) were analyzed. Using this approach, sample preparation could be completed within four hours. We successfully subtyped 100% of the cases in the training group, and the diagnostic success rate in the validation group was 91.7%. This maFASP-aided proteomic protocol represents an efficient approach for amyloid diagnosis and subtyping, particularly for serum-contaminated samples.
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Amiloide/clasificación , Amiloidosis/diagnóstico , Métodos Analíticos de la Preparación de la Muestra , Cromatografía Liquida/métodos , Filtración/instrumentación , Microondas , Espectrometría de Masas en Tándem/métodos , Biomarcadores/metabolismo , Estudios de Casos y Controles , Hemoglobinas/metabolismo , Humanos , Amiloidosis de Cadenas Ligeras de las Inmunoglobulinas , Reproducibilidad de los Resultados , Albúmina Sérica/metabolismoAsunto(s)
Amiloide , Amiloidosis , Ácidos Carboxílicos/farmacología , Prealbúmina/metabolismo , Pirrolidinas/farmacología , Proteína Amiloide A Sérica/metabolismo , Amiloide/clasificación , Amiloide/metabolismo , Amiloidosis/tratamiento farmacológico , Amiloidosis/etiología , Amiloidosis/metabolismo , Amiloidosis/patología , Manejo de la Enfermedad , Terapia Genética , Humanos , Terapia Molecular Dirigida/métodos , Placa Amiloide/metabolismo , Placa Amiloide/patología , Placa Amiloide/ultraestructura , Estabilidad Proteica/efectos de los fármacos , Secuestrantes/farmacología , Componente Amiloide P Sérico/metabolismoRESUMEN
BACKGROUND: Amyloids are proteins capable of forming aberrant intramolecular contact sites, characteristic of beta zipper configuration. Amyloids can underlie serious health conditions, e.g. Alzheimer's or Parkinson's diseases. It has been proposed that short segments of amino acids can be responsible for protein amyloidogenicity, but no more than two hundred such hexapeptides have been experimentally found. The authors of the computational tool Pafig published in BMC Bioinformatics a method for extending the amyloid hexapeptide dataset that could be used for training and testing models. They assumed that all hexapeptides belonging to an amyloid protein can be regarded as amylopositive, while those from proteins never reported as amyloid are always amylonegative. Here we show why the above described method of extending datasets is wrong and discuss the reasons why the incorrect data could lead to falsely correct classification. RESULTS: The amyloid classification of hexapeptides by Pafig was confronted with the classification results from different state of the art computational methods and the outputs of all methods were studied by clustering analysis. The clustering methods show that Pafig is an outlier with regard to other approaches. Our study of the statistical patterns of its training and testing datasets showed a strong bias towards STVIIE hexapeptide in their positive part. Different statistical patterns of seemingly amylo-positive and -negative hexapeptides allow for a repeatable classification, which is not related to amyloid propensity of the hexapetides. CONCLUSIONS: Our study on recognition of amyloid hexapeptides showed that occurrence of incidental patterns in wrongly selected datasets can produce falsely correct results of classification. The assumption that all hexapeptides belonging to amyloid protein can be regarded as amylopositive and those from proteins never reported as amyloid are always amylonegative is not supported by any other computational method. This is in line with experimental observations that amyloid propensity of a full protein can result from only one amyloidogenic fragment in this protein, while the occurrence of amyliodogenic part that is well hidden inside the protein may never lead to fibril formation. This leads to the conclusion that Pafig does not provide correct classification with regard to amyloidogenicity.
Asunto(s)
Amiloide/química , Bases de Datos de Proteínas , Oligopéptidos/química , Enfermedad de Alzheimer/metabolismo , Amiloide/biosíntesis , Amiloide/clasificación , Biología Computacional/métodos , Bases de Datos de Proteínas/tendencias , Reacciones Falso Positivas , Humanos , Simulación de Dinámica Molecular , Oligopéptidos/biosíntesis , Oligopéptidos/clasificación , Enfermedad de Parkinson/metabolismo , Fragmentos de Péptidos/biosíntesis , Fragmentos de Péptidos/química , Fragmentos de Péptidos/clasificaciónRESUMEN
Functional bacterial amyloids (FuBA) are important components in many environmental biofilms where they provide structural integrity to the biofilm, mediate bacterial aggregation and may function as virulence factor by binding specifically to host cell molecules. A novel FuBA system, the Fap system, was previously characterized in the genus Pseudomonas, however, very little is known about the phylogenetic diversity of bacteria with the genetic capacity to apply this system. Studies of genomes and public metagenomes from a diverse range of habitats showed that the Fap system is restricted to only three classes in the phylum Proteobacteria, the Beta-, Gamma- and Deltaproteobacteria. The structural organization of the fap genes into a single fapABCDEF operon is well conserved with minor variations such as a frequent deletion of fapA. A high degree of variation was seen within the primary structure of the major Fap fibril monomers, FapC, whereas the minor monomers, FapB, showed less sequence variation. Comparison of phylogenetic trees based on Fap proteins and the 16S rRNA gene of the corresponding bacteria showed remarkably similar overall topology. This indicates, that horizontal gene transfer is an infrequent event in the evolution of the Fap system.
Asunto(s)
Amiloide/genética , Proteínas Bacterianas/genética , Evolución Molecular , Pseudomonas/genética , Secuencia de Aminoácidos , Amiloide/clasificación , Amiloide/metabolismo , Proteínas Bacterianas/metabolismo , Biopelículas/crecimiento & desarrollo , Variación Genética , Genoma Bacteriano/genética , Cadenas de Markov , Metagenoma/genética , Datos de Secuencia Molecular , Operón , Filogenia , Proteobacteria/clasificación , Proteobacteria/genética , Proteobacteria/metabolismo , Pseudomonas/clasificación , Pseudomonas/fisiología , Homología de Secuencia de Aminoácido , Especificidad de la EspecieRESUMEN
BACKGROUND: Cardiac amyloidosis is a fatal disease whose prognosis and treatment rely on identification of the amyloid type. In our aging population transthyretin amyloidosis (ATTRwt) is common and must be differentiated from other amyloid types. We report the clinical presentation, natural history, and prognostic features of ATTRwt compared with cardiac-isolated AL amyloidosis and calculate the probability of disease diagnosis of ATTRwt from baseline factors. METHODS AND RESULTS: All patients with biopsy-proven ATTRwt (102 cases) and isolated cardiac AL (36 cases) seen from 2002 to 2011 at the UK National Amyloidosis Center were included. Median survival from the onset of symptoms was 6.07 years in the ATTRwt group and 1.7 years in the AL group. Positive troponin, a pacemaker, and increasing New York Heart Association (NYHA) class were associated with worse survival in ATTRwt patients on univariate analysis. All patients with isolated cardiac AL and 24.1% of patients with ATTRwt had evidence of a plasma cell dyscrasia. Older age and lower N-terminal pro-B-type natriuretic peptide (NT pro-BNP) were factors significantly associated with ATTRwt. Patients aged 70 years and younger with an NT pro-BNP <183 pmol/L were more likely to have ATTRwt, as were patients older than 70 years with an NT pro-BNP <1420 pmol/L. CONCLUSIONS: Factors at baseline associated with a worse outcome in ATTRwt are positive troponin T, a pacemaker, and NYHA class IV symptoms. The age of the patient at diagnosis and NT pro-BNP level can aid in distinguishing ATTRwt from AL amyloidosis.
Asunto(s)
Amiloide/clasificación , Amiloidosis/diagnóstico , Cardiomiopatías/diagnóstico , Miocardio/metabolismo , Anciano , Amiloide/genética , Amiloidosis/complicaciones , Amiloidosis/mortalidad , Biopsia , Cardiomiopatías/etiología , Cardiomiopatías/mortalidad , Progresión de la Enfermedad , Ecocardiografía , Electrocardiografía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Miocardio/patología , Péptido Natriurético Encefálico/sangre , Fragmentos de Péptidos/sangre , Prealbúmina , Pronóstico , Derivación y Consulta/estadística & datos numéricosRESUMEN
BACKGROUND: Amyloids are proteins capable of forming fibrils. Many of them underlie serious diseases, like Alzheimer disease. The number of amyloid-associated diseases is constantly increasing. Recent studies indicate that amyloidogenic properties can be associated with short segments of aminoacids, which transform the structure when exposed. A few hundreds of such peptides have been experimentally found. Experimental testing of all possible aminoacid combinations is currently not feasible. Instead, they can be predicted by computational methods. 3D profile is a physicochemical-based method that has generated the most numerous dataset - ZipperDB. However, it is computationally very demanding. Here, we show that dataset generation can be accelerated. Two methods to increase the classification efficiency of amyloidogenic candidates are presented and tested: simplified 3D profile generation and machine learning methods. RESULTS: We generated a new dataset of hexapeptides, using more economical 3D profile algorithm, which showed very good classification overlap with ZipperDB (93.5%). The new part of our dataset contains 1779 segments, with 204 classified as amyloidogenic. The dataset of 6-residue sequences with their binary classification, based on the energy of the segment, was applied for training machine learning methods. A separate set of sequences from ZipperDB was used as a test set. The most effective methods were Alternating Decision Tree and Multilayer Perceptron. Both methods obtained area under ROC curve of 0.96, accuracy 91%, true positive rate ca. 78%, and true negative rate 95%. A few other machine learning methods also achieved a good performance. The computational time was reduced from 18-20 CPU-hours (full 3D profile) to 0.5 CPU-hours (simplified 3D profile) to seconds (machine learning). CONCLUSIONS: We showed that the simplified profile generation method does not introduce an error with regard to the original method, while increasing the computational efficiency. Our new dataset proved representative enough to use simple statistical methods for testing the amylogenicity based only on six letter sequences. Statistical machine learning methods such as Alternating Decision Tree and Multilayer Perceptron can replace the energy based classifier, with advantage of very significantly reduced computational time and simplicity to perform the analysis. Additionally, a decision tree provides a set of very easily interpretable rules.
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Amiloide/clasificación , Inteligencia Artificial , Péptidos/clasificación , Algoritmos , Amiloide/química , Árboles de Decisión , Humanos , Redes Neurales de la Computación , Péptidos/química , Curva ROCRESUMEN
Escherichia coli and a few other members of the Enterobacteriales can produce functional amyloids known as curli. These extracellular fibrils are involved in biofilm formation and studies have shown that they may act as virulence factors during infections. It is not known whether curli fibrils are restricted to the Enterobacteriales or if they are phylogenetically widespread. The growing number of genome-sequenced bacteria spanning many phylogenetic groups allows a reliable bioinformatic investigation of the phylogenetic diversity of the curli system. Here we show that the curli system is phylogenetically much more widespread than initially assumed, spanning at least four phyla. Curli fibrils may consequently be encountered frequently in environmental as well as pathogenic biofilms, which was supported by identification of curli genes in public metagenomes from a diverse range of habitats. Identification and comparison of curli subunit (CsgA/B) homologs show that these proteins allow a high degree of freedom in their primary protein structure, although a modular structure of tightly spaced repeat regions containing conserved glutamine, asparagine and glycine residues has to be preserved. In addition, a high degree of variability within the operon structure of curli subunits between bacterial taxa suggests that the curli fibrils might have evolved to fulfill specific functions. Variations in the genetic organization of curli genes are also seen among different bacterial genera. This suggests that some genera may utilize alternative regulatory pathways for curli expression. Comparison of phylogenetic trees of Csg proteins and the 16S rRNA genes of the corresponding bacteria showed remarkably similar overall topography, suggesting that horizontal gene transfer is a minor player in the spreading of the curli system.