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INTRODUCTION: Since the 2018 change in the US adult heart allocation policy, more patients are bridged-to-transplant on temporary mechanical circulatory support (tMCS). Previous studies indicate that durable left ventricular assist devices (LVAD) may lead to allosensitization. The goal of this study was to assess whether tMCS implantation is associated with changes in sensitization. METHODS: We included patients evaluated for heart transplants between 2015 and 2022 who had alloantibody measured before and after MCS implantation. Allosensitization was defined as development of new alloantibodies after tMCS implant. RESULTS: A total of 41 patients received tMCS before transplant. Nine (22.0%) patients developed alloantibodies following tMCS implantation: 3 (12.0%) in the intra-aortic balloon pump group (n = 25), 2 (28.6%) in the microaxial percutaneous LVAD group (n = 7), and 4 (44.4%) in the veno-arterial extra-corporeal membrane oxygenation group (n = 9)-p = .039. Sensitized patients were younger (44.7 ± 11.6 years vs. 54.3 ± 12.5 years, p = .044), were more likely to be sensitized at baseline - 3 of 9 (33.3%) compared to 2 out of 32 (6.3%) (p = .028) and received more transfusions with red blood cells (6 (66.6%) vs. 8 (25%), p = .02) and platelets (6 (66.6%) vs. 5 (15.6%), p = .002). There was no significant difference in tMCS median duration of support (4 [3,15] days vs. 8.5 [5,14.5] days, p = .57). Importantly, out of the 11 patients who received a durable LVAD after tMCS, 5 (45.5%) became sensitized, compared to 4 out of 30 patients (13.3%) who only had tMCS-p = .028. CONCLUSIONS: Our findings suggest that patients bridged-to-transplant with tMCS, without significant blood product transfusions and a subsequent durable LVAD implant, have a low risk of allosensitization. Further studies are needed to confirm our findings and determine whether risk of sensitization varies by type of tMCS and duration of support.
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Trasplante de Corazón , Corazón Auxiliar , Isoanticuerpos , Humanos , Masculino , Femenino , Persona de Mediana Edad , Isoanticuerpos/inmunología , Isoanticuerpos/sangre , Estudios de Seguimiento , Adulto , Factores de Riesgo , Pronóstico , Estudios Retrospectivos , Insuficiencia Cardíaca/cirugía , Insuficiencia Cardíaca/terapia , Rechazo de Injerto/etiologíaRESUMEN
BACKGROUND: Inhibition of mutant KRAS challenged cancer research for decades. Recently, allele-specific inhibitors were approved for the treatment of KRAS-G12C mutant lung cancer. However, de novo and acquired resistance limit their efficacy and several combinations are in clinical development. Our study shows the potential of combining G12C inhibitors with farnesyl-transferase inhibitors. METHODS: Combinations of clinically approved farnesyl-transferase inhibitors and KRAS G12C inhibitors are tested on human lung, colorectal and pancreatic adenocarcinoma cells in vitro in 2D, 3D and subcutaneous xenograft models of lung adenocarcinoma. Treatment effects on migration, proliferation, apoptosis, farnesylation and RAS signaling were measured by histopathological analyses, videomicroscopy, cell cycle analyses, immunoblot, immunofluorescence and RAS pulldown. RESULTS: Combination of tipifarnib with sotorasib shows synergistic inhibitory effects on lung adenocarcinoma cells in vitro in 2D and 3D. Mechanistically, we present antiproliferative effect of the combination and interference with compensatory HRAS activation and RHEB and lamin farnesylation. Enhanced efficacy of sotorasib in combination with tipifarnib is recapitulated in the subcutaneous xenograft model of lung adenocarcinoma. Finally, combination of additional KRAS G1C and farnesyl-transferase inhibitors also shows synergism in lung, colorectal and pancreatic adenocarcinoma cellular models. DISCUSSION: Our findings warrant the clinical exploration of KRAS-G12C inhibitors in combination with farnesyl-transferase inhibitors.
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Adenocarcinoma del Pulmón , Adenocarcinoma , Neoplasias Colorrectales , Neoplasias Pulmonares , Neoplasias Pancreáticas , Humanos , Animales , Adenocarcinoma/tratamiento farmacológico , Adenocarcinoma/genética , Neoplasias Pancreáticas/tratamiento farmacológico , Neoplasias Pancreáticas/genética , Proteínas Proto-Oncogénicas p21(ras)/genética , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/genética , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/farmacología , Transferasas , Neoplasias Colorrectales/tratamiento farmacológico , Neoplasias Colorrectales/genética , MutaciónRESUMEN
Pulmonary fibrosis develops as a consequence of failed regeneration after injury. Analyzing mechanisms of regeneration and fibrogenesis directly in human tissue has been hampered by the lack of organotypic models and analytical techniques. In this work, we coupled ex vivo cytokine and drug perturbations of human precision-cut lung slices (hPCLS) with single-cell RNA sequencing and induced a multilineage circuit of fibrogenic cell states in hPCLS. We showed that these cell states were highly similar to the in vivo cell circuit in a multicohort lung cell atlas from patients with pulmonary fibrosis. Using micro-CT-staged patient tissues, we characterized the appearance and interaction of myofibroblasts, an ectopic endothelial cell state, and basaloid epithelial cells in the thickened alveolar septum of early-stage lung fibrosis. Induction of these states in the hPCLS model provided evidence that the basaloid cell state was derived from alveolar type 2 cells, whereas the ectopic endothelial cell state emerged from capillary cell plasticity. Cell-cell communication routes in patients were largely conserved in hPCLS, and antifibrotic drug treatments showed highly cell type-specific effects. Our work provides an experimental framework for perturbational single-cell genomics directly in human lung tissue that enables analysis of tissue homeostasis, regeneration, and pathology. We further demonstrate that hPCLS offer an avenue for scalable, high-resolution drug testing to accelerate antifibrotic drug development and translation.
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Fibrosis Pulmonar , Humanos , Fibrosis Pulmonar/genética , Fibrosis Pulmonar/patología , Análisis de Expresión Génica de una Sola Célula , Pulmón/patología , Células Epiteliales Alveolares , Células Epiteliales/metabolismoRESUMEN
Maintaining stable and transient quiescence in differentiated and stem cells, respectively, requires repression of the cell cycle. The plant RETINOBLASTOMA-RELATED (RBR) has been implicated in stem cell maintenance, presumably by forming repressor complexes with E2F transcription factors. Surprisingly we find that mutations in all three canonical E2Fs do not hinder the cell cycle, but similarly to RBR silencing, result in hyperplasia. Contrary to the growth arrest that occurs when exit from proliferation to differentiation is inhibited upon RBR silencing, the e2fabc mutant develops enlarged organs with supernumerary stem and differentiated cells as quiescence is compromised. While E2F, RBR and the M-phase regulatory MYB3Rs are part of the DREAM repressor complexes, and recruited to overlapping groups of targets, they regulate distinct sets of genes. Only the loss of E2Fs but not the MYB3Rs interferes with quiescence, which might be due to the ability of E2Fs to control both G1-S and some key G2-M targets. We conclude that collectively the three canonical E2Fs in complex with RBR have central roles in establishing cellular quiescence during organ development, leading to enhanced plant growth.
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Neoplasias de la Retina , Retinoblastoma , Humanos , Retinoblastoma/genética , División Celular , Ciclo Celular/genética , Desarrollo de la PlantaRESUMEN
BACKGROUND: Adalimumab is a fully human monoclonal antibody developed against tumor necrosis factor (TNF), used for the treatment of autoimmune and chronic inflammatory diseases. Immunogenicity to this drug may lead to therapeutic failure. Various laboratory assays are used for measuring serum adalimumab and anti-drug antibodies (ADA) to adalimumab, for therapeutic monitoring and evaluation of clinical non-responsiveness. This study compared the performance of 2 clinical assays used by different reference laboratories. METHODS: In total, 120 residual clinical samples were tested at both laboratories. A sandwich ELISA for adalimumab detecting free drug and a bridging ELISA capable of detecting both free and bound ADA were performed at the Mayo Clinic. A functional cell-based reporter gene assay (RGA) was used at ARUP Laboratories for measuring bioactive serum drug concentrations, and neutralizing ADA. RESULTS: Seventy-eight samples had measurable concentrations of adalimumab by both methods and yielded a correlation coefficient r = 0.93, slope = 0.886, and intercept = 0.950. Overall agreement of 92.5% was observed between the assays, with most discrepant drug results being attributed to a higher positivity rate with ELISA (8/9). One outlier positive with RGA and negative with ELISA was confirmed by LC-MS/MS to be attributed to infliximab. Overall agreement of 79.2% was observed between the ADA assays. Differences in ADA results may be due to the bridging ELISA detecting total ADA (free, drug-bound, neutralizing, and non-neutralizing), while RGA detects free, neutralizing ADA only. CONCLUSIONS: Although the assays are fundamentally different, the results show significant concordance between the clinically validated tests performed in different laboratories.
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Laboratorios Clínicos , Espectrometría de Masas en Tándem , Humanos , Adalimumab/uso terapéutico , Cromatografía Liquida , Anticuerpos MonoclonalesRESUMEN
In silico studies raised the possibility that farnesyltransferase inhibitors (FTIs) may have antitumoral effects on KRAS mutant cancer cells. Accordingly, we have tested FTIs (tipifarnib and lonafarnib) in G12C mutant human cancer cell lines in vitro and in vivo. We have discovered that the combination of the two drugs has a synergistic antitumoral effect. Next, we have tested FTIs on G12D mutant human cancer cell lines and found that the combination has antitumoral effect in various preclinical cancer models. At last, we have also tested FTIs on G12V mutant human cancer cells and again we have detected antitumoral effects. We suggest that FTIs may have clinical relevance outside the HRAS mutant cancers.
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Neoplasias , Proteínas Proto-Oncogénicas p21(ras) , Humanos , Farnesiltransferasa/metabolismo , Proteínas Proto-Oncogénicas p21(ras)/genética , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/uso terapéutico , Neoplasias/tratamiento farmacológico , Neoplasias/genéticaRESUMEN
OBJECTIVES: The paper proposes to identify the determinants of patients' rights awareness in mothers and to examine the relationship of health literacy with awareness of those rights. METHODS: Our results are based on data from a convenience sample of 894 non-health professional ethnic Hungarian mothers from Hungary, Romania and Slovakia. Health literacy is measured with the HLS-EU-16 questionnaire. RESULTS: Analysis of variance reveals a significant association of health literacy with patient rights awareness. Our results show that health literacy is the highest among patients who filed a complaint through formal channels and/or took legal measures to restore their rights upon violation. A logistic regression model is built to identify the likelihood of having high patient rights awareness, that is, acting formally for the restoration of rights upon infringement. The model controls for covariates. When controlled for covariates, the likelihood of having high patient rights awareness increases with age, and is higher for mothers with highest education, for inhabitants of larger towns, as well as for those with adequate health literacy. CONCLUSIONS: The findings of our study have implications for health policy, as they reveal significant inequalities in patient rights culture.
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Alfabetización en Salud , Femenino , Humanos , Hungría , Encuestas y Cuestionarios , Europa Oriental , Derechos del PacienteAsunto(s)
Antígenos de Plaqueta Humana , Trombocitopenia , Humanos , Transfusión de Plaquetas , Isoanticuerpos , Antígenos HLA , PlaquetasRESUMEN
Összefoglaló. Bevezetés: A SpyGlass-kolangioszkópia újonnan kifejlesztett endoszkópos technika, mely az epeutak közvetlen vizualizációját teszi lehetové. A kolangioszkóp egy 10,8 Fr átméroju, a duodenoszkóp munkacsatornáján keresztül az epeútba vezetheto, a különbözo endoszkópos tartozékok számára saját munkacsatornával bíró endoszkóp. Fo indikációs területe a bizonytalan dignitású epeúti szukületek diagnosztikája, valamint a konvencionális endoszkópos technikával nem megoldható epeúti kövesség terápiája. Célkituzés: Célunk a SpyGlass berendezés hasznosságának és hatásosságának megítélése. Módszer: A Jahn Ferenc Dél-pesti Kórház Gasztroenterológia Osztályán 2018. január 1. és 2020. december 31. között a Spyglass DS I, míg 2021 januárjában a SpyGlass DS II rendszert használtuk. 14 diagnosztikus és 15 terápiás beavatkozást végeztünk. A diagnosztikus beavatkozások beválogatási kritériuma azon bizonytalan dignitású epeúti szukületeket fogalta magában, melyek esetén a végso diagnózis korábban elvégzett endoszkópos retrográd kolangiopankreatográfiával vagy endoszkópos, ultrahangvezérelt szövettani mintavétellel nem volt megállapítható. A terápiás beavatkozás indikációja a konvencionális endoszkópos technikával nem megoldható epeúti kövesség volt. Eredmények: A makroszkópos megítélés és a végso diagnózis egyezésének tekintetében a kolangioszkópia pontossága 85% volt. A szövettani diagnózis pontossága kolangioszkópvezérelt biopsziák esetén 62,5%. A makroszkópos diagnózis szenzitivitása 100%, specificitása 71% volt, a szövettani minták szenzitivitása 60%, specificitása 100% volt. Komplett clearence-t 4 esetben értünk el, ez összesen 57,14% sikerességi rátának felel meg. Következtetés: A SpyGlass-vizsgálat lehetové teszi a bizonytalan eredetu epeúti szukületek pontos értékelését, valamint megkönnyíti a szövettani mintavételezést. A diagnosztikus specificitás és szenzitivitás tekintetében a nemzetközi irodalmi adatok eléréséhez további fejlodés és az esetszámok növelése szükséges. A SpyGlass-vezérelt elektrohidraulikus lithotripsia a konvencionális endoszkópos technikával nem megoldható nehéz epeúti kövek kezelési alternatívája. Orv Hetil. 2022; 163(4): 150-156 Summary. INTRODUCTION: SpyGlass cholangioscopy is a recently developed endoscopic technique to the direct visualization of the biliary tract. The SpyGlass cholangioscop is a 10,8 Fr diameter endoscop which can be guided to the biliary tract through the work channel of the doudenoscope and has its own work channel for the different endoscopic accessories. The main indications of the examination are the diagnosis of the uncertain dignity biliary stenosis and the therapy of the biliary stones which failed conventional therapy. OBJECTIVE: Our aim was to assess the utility and efficacy of the SpyGlass system. METHOD: In Jahn Ferenc South Pest Hospital Gastroenterology Department, we used the SpyGlass™ DS I system between 2018 and 2020 and from 2021 the SpyGlass™ DS II. 14 diagnostic and 15 therapeutic Spyglass procedures have been performed. Inclusion criterion of diagnostic procedures was indeterminate bile duct stenosis where the final diagnosis could not be confirmed by endoscopic retrograde cholangiopancreatography or endoscopic ultrasound-guided biopsy. Inclusion criteria of the therapeutic examinations were difficult bile duct stones which failed conventional therapy. RESULTS: Concerning the correspondence of the macroscopic image and the final diagnosis, the accuracy of the cholangioscope was 85%. The accuracy of the histological diagnosis in the case of cholangioscopy-guided biopsies was 62.5%. The sensitivity of the macroscopic diagnosis was 100%, specificity was 71%, while the sensitivity of histologic samples was 60% and the specificity was 100%. Complete clearence was performed four times in the case of therapeutic procedures, which refers to 57.14% success rate. CONCLUSION: The use of SpyGlass enhances the precise evaluation of indeterminate bile duct lesions and tissue acquisition is easier to perform. However, to reach the international standards of diagnostic sensitivity and specificity, further improvement and examinations are necessary. Spyglass-guided electrohydraulic lithotripsy is an alternative for difficult stones which failed conventional therapy. Orv Hetil. 2022; 163(4): 150-156.
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Biopsia Guiada por Imagen , Laparoscopía , Humanos , HungríaRESUMEN
Signaling networks represent the molecular mechanisms controlling a cell's response to various internal or external stimuli. Most currently available signaling databases contain only a part of the complex network of intertwining pathways, leaving out key interactions or processes. Hence, we have developed SignaLink3 (http://signalink.org/), a value-added knowledge-base that provides manually curated data on signaling pathways and integrated data from several types of databases (interaction, regulation, localisation, disease, etc.) for humans, and three major animal model organisms. SignaLink3 contains over 400 000 newly added human protein-protein interactions resulting in a total of 700 000 interactions for Homo sapiens, making it one of the largest integrated signaling network resources. Next to H. sapiens, SignaLink3 is the only current signaling network resource to provide regulatory information for the model species Caenorhabditis elegans and Danio rerio, and the largest resource for Drosophila melanogaster. Compared to previous versions, we have integrated gene expression data as well as subcellular localization of the interactors, therefore uniquely allowing tissue-, or compartment-specific pathway interaction analysis to create more accurate models. Data is freely available for download in widely used formats, including CSV, PSI-MI TAB or SQL.
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Bases de Datos Genéticas , Redes Reguladoras de Genes/genética , Mapas de Interacción de Proteínas/genética , Transducción de Señal/genética , Animales , Caenorhabditis elegans/genética , Drosophila melanogaster/genética , Humanos , Pez Cebra/genéticaRESUMEN
Allogeneic hematopoietic stem cell transplantation (HSCT) and coronavirus disease 2019 (COVID-19) infection can both lead to severe cytokine release syndrome (sCRS) resulting in critical illness and death. In this single institution, preliminary comparative case-series study we compared clinical and laboratory co-variates as well as response to tocilizumab (TCZ)-based therapy of 15 allogeneic-HSCT- and 17 COVID-19-associated sCRS patients. Reaction to a TCZ plus posttransplant cyclophosphamide (PTCY) consolidation therapy in the allogeneic-HSCT-associated sCRS group yielded significantly inferior long-term outcome as compared to TCZ-based therapy in the COVID-19-associated group (P = 0.003). We report that a TCZ followed by consolidation therapy with a Janus kinase/signal transducer and activator of transcription (JAK/STAT) inhibitor given to 4 out of 8 critically ill COVID-19 patients resulted in their complete recovery. Non-selective JAK/STAT inhibitors influencing the action of several cytokines exhibit a broader effect than TCZ alone in calming down sCRS. Serum levels of cytokines and chemokines show similar changes in allogeneic-HSCT- and COVID-19-associated sCRS with marked elevation of interleukin-6 (IL-6), regulated upon activation normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein-1 (MCP-1) and interferon γ-induced protein 10 kDa (IP-10) levels. In addition, levels of IL-5, IL-10, IL-15 were also elevated in allogeneic-HSCT-associated sCRS. Our multi-cytokine expression data indicate that the pathophysiology of allogeneic-HSCT and COVID-19-associated sCRS are similar therefore the same clinical grading system and TCZ-based treatment approaches can be applied. TCZ with JAK/STAT inhibitor consolidation therapy might be highly effective in COVID-19 sCRS patients.
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The morphology and histology of the soft tissue around the implant are different from the periodontal tissue, but the difference in the regulation of blood flow is not known. The aim of the study was to compare the resting blood flow and the vasodilatation capacity of the gingiva between implants and teeth. Twenty-six healthy volunteers with single-tooth implants were involved. The implant-borne crown was retained on either a zirconia or titanium abutment. The vasodilatation capacity of the gingiva was assessed by a postocclusive reactive hyperemia test. Blood flow was measured by a laser speckle contrast imager at the buccal gingiva of the implant-borne crown and an analog natural tooth. No significant differences in baseline gingival blood flow were observed between the different abutments and the teeth in either region. The hyperemia after compression was significantly attenuated at the zirconia abutments in all regions during the entire investigation period (20 minutes) compared to the titanium abutments and the teeth. No differences were observed between titanium abutments and the teeth. The resting microcirculation seems to be the same at implants and teeth. However, the vascular reactivity might be disturbed at the zirconia, but not at the titanium, abutment.
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Implantes Dentales de Diente Único , Implantes Dentales , Coronas , Pilares Dentales , Humanos , Membrana Mucosa , Titanio , CirconioRESUMEN
BACKGROUND: Due to the strong association between ankylosing spondylitis and Human Leukocyte Antigen (HLA)-B27, accurate identification of HLA-B27 is important in the diagnosis of patients with suspected spondyloarthritides. For this study, we compared a high-resolution HLA-B typing method to the clinical flow cytometry and allele-specific PCR melting assays to determine clinical benefits of high-resolution testing. METHODS: Residual clinical samples submitted for HLA-B27 testing by flow cytometry were tested by single-locus HLA-B genotyping using next-generation sequencing (NGS), and PCR with melting curve analysis, currently used as a reflex test for indeterminate flow cytometry results. RESULTS: Fifty out of the 51 samples (98%) positive by flow cytometry confirmed as HLA-B27 positive by PCR melting assay and by NGS. The sample that did not confirm was genotyped as HLA-B*07:02. All the samples negative by flow cytometry were confirmed as HLA-B27 negative by both PCR melting assay and NGS. For the group that was indeterminate by flow cytometry, 84.5% (n = 49) typed as positive for HLA-B27, while 15.5% (n = 9) were negative for HLA-B27 but positive for HLA-B*07:02. NGS was the only method able to distinguish between pathogenic and nonpathogenic HLA-B27 variants, in contrast to the flow cytometry or the PCR melting assays. CONCLUSIONS: Single-locus NGS is superior to flow cytometry and PCR melting assay for the unambiguous identification of HLA-B27 variants, and uniquely able to distinguish between pathogenic and nonpathogenic B27 alleles. Due to its high accuracy, it may be a feasible superior alternative to flow cytometry and traditional molecular methods for clinical HLA-B27 testing.
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Antígeno HLA-B27 , Secuenciación de Nucleótidos de Alto Rendimiento , Alelos , Citometría de Flujo , Genotipo , Antígeno HLA-B27/genética , Humanos , Reacción en Cadena de la PolimerasaRESUMEN
The RAS/RAF and PI3K/Akt pathways play a key regulatory role in cancer and are often hit by oncogenic mutations. Despite molecular targeting, the long-term success of monotherapy is often hampered by de novo or acquired resistance. In the case of concurrent mutations in both pathways, horizontal combination could be a reasonable approach. In our study, we investigated the MEK inhibitor selumetinib and PI3K/mTOR dual inhibitor BEZ235 alone and in combination in BRAF-only mutant and BRAF + PI3K/PTEN double mutant cancer cells using short- and long-term 2D viability assays, spheroid assays, and immunoblots. In the 2D assays, selumetinib was more effective on BRAF-only mutant lines when compared to BRAF + PI3K/PTEN double mutants. Furthermore, combination therapy had an additive effect in most of the lines while synergism was observed in two of the double mutants. Importantly, in the SW1417 BRAF + PI3K double mutant cells, synergism was also confirmed in the spheroid and in the in vivo model. Mechanistically, p-Akt level decreased only in the SW1417 cell line after combination treatment. In conclusion, the presence of concurrent mutations alone did not predict a stronger response to combination treatment. Therefore, additional investigations are warranted to identify predictive factors that can select patients who can benefit from the horizontal combinational inhibition of these two pathways.
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Quinasas Quinasa Quinasa PAM/metabolismo , Melanoma/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas B-raf/genética , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo , Animales , Antineoplásicos/farmacología , Bencimidazoles/farmacología , Línea Celular Tumoral , Resistencia a Antineoplásicos , Femenino , Humanos , Imidazoles/farmacología , Quinasas Quinasa Quinasa PAM/antagonistas & inhibidores , Quinasas Quinasa Quinasa PAM/genética , Melanoma/metabolismo , Ratones , Ratones Desnudos , Ratones SCID , Mutación , Fosfohidrolasa PTEN/genética , Fosfohidrolasa PTEN/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Inhibidores de Proteínas Quinasas/farmacología , Quinolinas/farmacología , Esferoides Celulares/efectos de los fármacos , Serina-Treonina Quinasas TOR/antagonistas & inhibidores , Serina-Treonina Quinasas TOR/genéticaRESUMEN
BACKGROUND: Immunoassays based on label-free technologies (label-free immunoassay [LFIA]) offer an innovative approach to clinical diagnostics and demonstrate great promise for therapeutic drug monitoring (TDM) of monoclonal antibody (mAb) drugs. An LFIA measures immunocomplex formation in real time and allows for quantification on initial binding rate, which facilitates fast measurement within a few minutes. METHODS: Based on thin-film interferometry (TFI) technology, open-access LFIAs were developed for the quantification of the mAb drugs adalimumab (ADL) and infliximab (IFX) and for the detection of the antidrug antibodies (ADAs) to the mAb drugs (ADL-ADAs and IFX-ADAs). RESULTS: The LFIAs for active mAb drugs (ADL and IFX) and for ADAs (ADL-ADAs and IFX-ADAs) were validated. The analytical measurement range (AMR) for both ADL and IFX was from 2 to 100 µg/mL. The AMR for ADL-ADAs was from 5 to 100 µg/mL and for IFX-ADAs was 10 to 100 µg/mL. In the comparison of LFIAs and reporter gene assays, the correlation coefficient was 0.972 for the quantification of ADL and 0.940 for the quantification of IFX. The concordance rate was 90% for the detection of ADL-ADAs and 76% for the detection of IFX-ADAs. CONCLUSIONS: The LFIAs for active mAb drugs and ADAs were appropriate for the TDM of ADL and IFX. The TFI technology has unique advantages compared with other technologies used for the measurement of mAb drugs. Label-free technologies, especially those allowing for open-access LFIAs, have great potential for clinical diagnostics.
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Adalimumab/sangre , Monitoreo de Drogas/métodos , Inmunoensayo/métodos , Infliximab/sangre , Adalimumab/inmunología , Biosimilares Farmacéuticos/sangre , Humanos , Infliximab/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Multinucleate cells can be produced in Dictyostelium by electric pulse-induced fusion. In these cells, unilateral cleavage furrows are formed at spaces between areas that are controlled by aster microtubules. A peculiarity of unilateral cleavage furrows is their propensity to join laterally with other furrows into rings to form constrictions. This means cytokinesis is biphasic in multinucleate cells, the final abscission of daughter cells being independent of the initial direction of furrow progression. Myosin-II and the actin filament cross-linking protein cortexillin accumulate in unilateral furrows, as they do in the normal cleavage furrows of mononucleate cells. In a myosin-II-null background, multinucleate or mononucleate cells were produced by cultivation either in suspension or on an adhesive substrate. Myosin-II is not essential for cytokinesis either in mononucleate or in multinucleate cells but stabilizes and confines the position of the cleavage furrows. In fused wild-type cells, unilateral furrows ingress with an average velocity of 1.7 µm × min-1, with no appreciable decrease of velocity in the course of ingression. In multinucleate myosin-II-null cells, some of the furrows stop growing, thus leaving space for the extensive broadening of the few remaining furrows.
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Citocinesis/fisiología , Dictyostelium/citología , Dictyostelium/fisiología , División Celular/genética , División Celular/fisiología , Fusión Celular/métodos , Membrana Celular/fisiología , Citocinesis/genética , Dictyostelium/genética , Técnicas de Inactivación de Genes , Genes Protozoarios , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/fisiología , Miosina Tipo II/deficiencia , Miosina Tipo II/genética , Miosina Tipo II/fisiología , Proteínas Protozoarias/genética , Proteínas Protozoarias/fisiología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismoRESUMEN
During the past few months, a pandemic originating from China named new coronavirus disease (COVID-19) has shown how vulnerable the world is. To date, no medication supported by randomized clinical trials has been approved for the treatment of COVID-19. At the time of writing of this paper, severe acute respiratory syndrome caused by coronavirus-2 (SARS-CoV-2) has been responsible - according to modest estimations - for around 4 million of infections and 300 000 deaths. Unveiling details of patomechanism, in fatal cases the role of immune dysregulation, namely cytokine release syndrome (CRS) has been discovered. Based on the current knowledge, interleukin-6 (IL6) plays a pivotal role in COVID-19 associated CRS. Case reports and result of small case series suggest efficacy of an IL6 inhibitor monoclonal antibody (tocilizumab) in treating CRS. Authors describe a case and review recent knowledge on the treatment of COVID-19. To our knowledge, the first case of severe COVID-19-associated cytokine storm syndrome - treated succesfully with IL6 monoclocal antibody at a Hungarian department of infectology - is presented here. Orv Hetil. 2020; 161(26): 1070-1077.
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Anticuerpos Monoclonales Humanizados/uso terapéutico , Infecciones por Coronavirus/tratamiento farmacológico , Síndrome de Liberación de Citoquinas/tratamiento farmacológico , Neumonía Viral/tratamiento farmacológico , COVID-19 , Infecciones por Coronavirus/complicaciones , Infecciones por Coronavirus/epidemiología , Síndrome de Liberación de Citoquinas/virología , Humanos , Hungría/epidemiología , Pandemias , Neumonía Viral/complicaciones , Neumonía Viral/epidemiología , Índice de Severidad de la Enfermedad , Resultado del TratamientoRESUMEN
HLA associations have been linked to many diseases and are important for risk assessment of drug hypersensitivity reactions. The increasing number of HLA alleles discovered generated a list of ambiguities that cannot be resolved with the current clinical assays, which commonly include sequence-specific oligonucleotide probe (SSOP) genotyping, and real-time PCR with melting curve analysis. HLA typing by next-generation sequencing (NGS) has recently been adopted by clinical laboratories for transplantation testing, as it provides unambiguous and cost-effective HLA typing. The goal of this study was to evaluate the feasibility of using NGS-based HLA-B and DQ genotyping for clinical HLA disease association testing, and provide direct comparison with the currently used clinical tests, including SSOP genotyping, and real-time PCR with melting curve analysis. While the real-time PCR method is easy and inexpensive to perform, ambiguities are rapidly increasing as more and more HLA alleles are discovered. SSOP genotyping identifies the alleles present but limitations include ambiguities and underreporting less common alleles. Our data show that HLA typing by NGS is superior to the existing clinical methods for identifying HLA alleles associated with disease or drug hypersensitivity, and offers a viable approach for high volume clinical diagnostic laboratories.
Asunto(s)
Enfermedad Celíaca/inmunología , Hipersensibilidad a las Drogas/inmunología , Antígenos HLA-B/genética , Antígenos HLA-DQ/genética , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Prueba de Histocompatibilidad/métodos , Narcolepsia/inmunología , Pruebas de Farmacogenómica/métodos , Espondilitis Anquilosante/inmunología , Alelos , Enfermedad Celíaca/genética , Hipersensibilidad a las Drogas/genética , Estudios de Factibilidad , Genotipo , Técnicas de Genotipaje/métodos , Humanos , Narcolepsia/genética , Análisis de Secuencia de ADN/métodos , Espondilitis Anquilosante/genéticaRESUMEN
INTRODUCTION: At present, neither specific curative treatment nor vaccines for novel coronavirus 2019 (COVID-19) are available. There is an urgent need to look for alternative strategies for COVID-19 treatment especially in the case of severe and/or critically ill patients with cytokine release syndrome (CRS). AIM: Convalescent plasma proved to increase survival rates in other severe viral infections. Therefore, convalescent plasma could be a promising treatment option for severe COVID-19 patients. METHOD: In our article, we present the first two critically ill Hungarian patients with COVID-19 infection treated with convalescent fresh frozen plasma. RESULTS: At the time of plasma therapy both patients were on mechanical ventilation and received antiviral agents and a full scale of supportive care. Each patient received 3 × 200 mL of convalescent plasma of recently recovered donors with sufficient novel anti-coronavirus IgG titers. Subsequent to convalescent plasma infusion, oxygenization improved and inflammatory markers decreased in both individuals. As compared to pretransfusion, lymphocyte counts increased and interleukin-6 level lessened. Both patients were weaned from mechanical ventilation within 2 weeks of treatment. No severe adverse effects were observed. CONCLUSIONS: Our experience indicates that convalescent plasma therapy is well tolerated and could potentially improve clinical outcomes. Optimal dose and timing as well as precise assessment of clinical benefit of convalescent plasma therapy will need further investigation in larger, well-controlled trials. This is the first report of the successful use of convalescent plasma in the treatment of critically ill patients with COVID-19 infection in Hungary. Orv Hetil. 2020; 161(27): 1111-1121.