Detection of a Subset of Posttranscriptional Transfer RNA Modifications in Vivo with a Restriction Fragment Length Polymorphism-Based Method

Transfer RNAs (tRNAs) are among the most heavily modified RNA species. Posttranscriptional tRNA modifications (ptRMs) play fundamental roles in modulating tRNA structure and function and are being increasingly linked to human physiology and disease. Detection of ptRMs is often challenging, expensive, and laborious. Restriction fragment length polymorphism (RFLP) analyses study the patterns of DNA cleavage after restriction enzyme treatment and have been used for the qualitative detection of modified bases on mRNAs. It is known that some ptRMs induce specific and reproducible base “mutations” when tRNAs are reverse transcribed. For example, inosine, which derives from the deamination of adenosine, is detected as a guanosine when an inosine-containing tRNA is reverse transcribed, amplified via polymerase chain reaction (PCR), and sequenced. ptRM-dependent base changes on reverse transcription PCR amplicons generated as a consequence of the reverse transcription reaction might create or abolish endonuclease restriction sites. The suitability of RFLP for the detection and/or quantification of ptRMs has not been studied thus far. Here we show that different ptRMs can be detected at specific sites of different tRNA types by RFLP. For the examples studied, we show that this approach can reliably estimate the modification status of the sample, a feature that can be useful in the study of the regulatory role of tRNA modifications in gene expression

The path to Z And-type outbursts: The case of V426 Sagittae (HBHA 1704-05)

Context. The star V426 Sge (HBHA 1704-05), originally classified as an emission-line object and a semi-regular variable, brightened at the beginning of August 2018, showing signatures of a symbiotic star outburst. Aims. We aim to confirm the nature of V426 Sge as a classical symbiotic star, determine the photometric ephemeris of the light minima, and suggest the path from its 1968 symbiotic nova outburst to the following 2018 Z And-type outburst. Methods. We re-constructed an historical light curve (LC) of V426 Sge from approximately the year 1900, and used original low- (R ∼ 500-1500; 330-880 nm) and high-resolution (R ∼ 11 000-34 000; 360-760 nm) spectroscopy complemented with Swift-XRT and UVOT, optical UBVRCIC and near-infrared JHKL photometry obtained during the 2018 outburst and the following quiescence. Results. The historical LC reveals no symbiotic-like activity from ∼1900 to 1967. In 1968, V426 Sge experienced a symbiotic nova outburst that ceased around 1990. From approximately 1972, a wave-like orbitally related variation with a period of 493.4 ± 0.7 days developed in the LC. This was interrupted by a Z And-type outburst from the beginning of August 2018 to the middle of February 2019. At the maximum of the 2018 outburst, the burning white dwarf (WD) increased its temperature to ? 2 × 105 K, generated a luminosity of ∼7 × 1037 (d/3.3 kpc)2 erg s-1 and blew a wind at the rate of ∼3 × 10-6 M yr-1. Our spectral energy distribution models from the current quiescent phase reveal that the donor is a normal M4-5 III giant characterised with Teff ∼ 3400 K, RG ∼ 106 (d/3.3 kpc) R and LG ∼ 1350 (d/3.3 kpc)2 L and the accretor is a low-mass ∼0.5 M WD. Conclusions. During the transition from the symbiotic nova outburst to the quiescent phase, a pronounced sinusoidal variation along the orbit develops in the LC of most symbiotic novae. The following eventual outburst is of Z And-type, when the accretion by the WD temporarily exceeds the upper limit of the stable burning. At this point the system becomes a classical symbiotic star.Fil: Skopal, A.. Astronomical Institute Slovak Academy Of Sciences; EslovaquiaFil: Shugarov, S. Y.. Lomonosov Moscow State University; Rusia. Astronomical Institute Slovak Academy Of Sciences; EslovaquiaFil: Munari, U.. Osservatorio Astronomico Di Padova; ItaliaFil: Masetti, N.. Inaf Istituto Di Astrofisica Spaziale E Fisica Cosmica, Bologna; Italia. Universidad Andrés Bello; ChileFil: Marchesini, Ezequiel Joaquín. Università di Torino; Italia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Astrofísica La Plata. Universidad Nacional de La Plata. Facultad de Ciencias Astronómicas y Geofísicas. Instituto de Astrofísica La Plata; ArgentinaFil: Komzík, R. M.. Astronomical Institute Slovak Academy Of Sciences; EslovaquiaFil: Kundra, E.. Astronomical Institute Slovak Academy Of Sciences; EslovaquiaFil: Shagatova, N.. Astronomical Institute Slovak Academy Of Sciences; EslovaquiaFil: Tarasova, T. N.. Crimean Astrophysical Observatory Ras; RusiaFil: Buil, C.. Castanet Tolosan Observatory; FranciaFil: Boussin, C.. Observatoire de L'eridan Et de la Chevelure de Bérénice; FranciaFil: Shenavrin, V. I.. Lomonosov Moscow State University; RusiaFil: Hambsch, F. J.. Istituto Nazionale di Astrofisica; ItaliaFil: Dallaporta, S.. Istituto Nazionale di Astrofisica; ItaliaFil: Frigolé, Cecilia Andrea. Istituto Nazionale di Astrofisica; ItaliaFil: Gardey, Juan Cruz. Observatoire de la Tourbière; FranciaFil: Zubareva, A.. Institute Of Astronomy Of The Russian Academy Of Sciences; Rusia. Lomonosov Moscow State University; RusiaFil: Dubovský, P. A.. Vihorlat Observatory; EslovaquiaFil: Kroll, P.. Sonneberg Observatory; Alemani

Androgen receptor condensates as drug targets

Transcription factors are among the most attractive therapeutic targets, but are considered largely undruggable. Here we provide evidence that small molecule-mediated partitioning of the androgen receptor, an oncogenic transcription factor, into phase-separated condensates has therapeutic effect in prostate cancer models. We show that the phase separation capacity of the androgen receptor is driven by aromatic residues and short unstable helices in its intrinsically disordered activation domain. Based on this knowledge, we developed tool compounds that covalently attach aromatic moieties to cysteines in the receptors’ activation domain. The compounds enhanced partitioning of the receptor into condensates, facilitated degradation of the receptor, inhibited androgen receptor-dependent transcriptional programs, and had antitumorigenic effect in models of prostate cancer and castration-resistant prostate cancer in vitro and in vivo. These results establish a generalizable framework to target the phase- separation capacity of intrinsically disordered regions in oncogenic transcription factors and other disease-associated proteins with therapeutic intent

Multi-isotopic and morphometric evidence for the migration of farmers leading up to the Inka conquest of the southern Andes

We present isotopic and morphometric evidence suggesting the migration of farmers in the southern Andes in the period AD 1270–1420, leading up to the Inka conquest occurring ~ AD 1400. This is based on the interdisciplinary study of human remains from archaeological cemeteries in the Andean Uspallata Valley (Argentina), located in the southern frontier of the Inka Empire. The studied samples span AD 800–1500, encompassing the highly dynamic Late Intermediate Period and culminating with the imperial expansion. Our research combines a macro-regional study of human paleomobility and migration based on a new strontium isoscape across the Andes that allows identifying locals and migrants, a geometric morphometric analysis of cranio-facial morphology suggesting separate ancestral lineages, and a paleodietary reconstruction based on stable isotopes showing that the migrants had diets exceptionally high in C4 plants and largely based on maize agriculture. Significantly, this migration influx occurred during a period of regional demographic increase and would have been part of a widespread period of change in settlement patterns and population movements that preceded the Inka expansion. These processes increased local social diversity and may have been subsequently utilized by the Inka to channel interaction with the local societies.Fil: Barberena, Ramiro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; Argentina. Universidad Nacional de Cuyo. Facultad de Filosofía y Letras; ArgentinaFil: Menéndez, Lumila. Universitat Bonn; AlemaniaFil: le Roux, Petrus J.. University of Cape Town; SudáfricaFil: Marsh, Erik Johnson. Universidad Nacional de Cuyo. Facultad de Filosofía y Letras; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; ArgentinaFil: Tessone, Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geocronología y Geología Isotópica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geocronología y Geología Isotópica; ArgentinaFil: Novellino, Paula Silvana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mendoza. Museo de Ciencias Naturales y Antropológicas J. Cornelio Moyano; ArgentinaFil: Lucero, Gustavo. Universidad Católica de Temuco; ChileFil: Luyt, Julie. University of Cape Town; SudáfricaFil: Sealy, Judith. University of Cape Town; SudáfricaFil: Cardillo, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Saavedra 15. Instituto Multidisciplinario de Historia y Ciencias Humanas; ArgentinaFil: Gasco, Alejandra Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; Argentina. Universidad Nacional de Cuyo. Facultad de Filosofía y Letras; ArgentinaFil: Llano, Carina Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; Argentina. Universidad Nacional de Cuyo. Facultad de Ciencias Aplicadas a la Industria; ArgentinaFil: Frigolé, Cecilia Andrea. Universidad Nacional de Cuyo. Facultad de Filosofía y Letras; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; ArgentinaFil: Guevara, Daniela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mendoza. Museo de Ciencias Naturales y Antropológicas J. Cornelio Moyano; ArgentinaFil: Da Peña, Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Mendoza. Museo de Ciencias Naturales y Antropológicas J. Cornelio Moyano; ArgentinaFil: Winocur, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Estudios Andinos "Don Pablo Groeber". Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Estudios Andinos "Don Pablo Groeber"; ArgentinaFil: Benítez, Anahí. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Estudios Andinos "Don Pablo Groeber". Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Estudios Andinos "Don Pablo Groeber"; ArgentinaFil: Cornejo, Luis. Universidad Alberto Hurtado; ChileFil: Falabella, Fernanda. Universidad de Chile; ChileFil: Méndez, César. Centro de Investigación en Ecosistemas de la Patagonia; ChileFil: Nuevo Delaunay, Amalia. Centro de Investigación en Ecosistemas de la Patagonia; ChileFil: Sanhueza, Lorena. Universidad de Chile; ChileFil: Santana Sagredo, Francisca. Pontificia Universidad Católica de Chile; ChileFil: Troncoso, Andrés. Universidad de Chile; ChileFil: Zárate, Sol. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; ArgentinaFil: Duran, Victor Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; Argentina. Universidad Nacional de Cuyo. Facultad de Filosofía y Letras; ArgentinaFil: Cortegoso, Valeria. Universidad Nacional de Cuyo. Facultad de Filosofía y Letras; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; Argentin

Mycophenolate mofetil versus cyclophosphamide for remission induction in ANCA-associated vasculitis: a randomised, non-inferiority trial

OBJECTIVES: Cyclophosphamide induction regimens are effective for antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), but are associated with infections, malignancies and infertility. Mycophenolate mofetil (MMF) has shown high remission rates in small studies of AAV. METHODS: We conducted a randomised controlled trial to investigate whether MMF was non-inferior to cyclophosphamide for remission induction in AAV. 140 newly diagnosed patients were randomly assigned to MMF or pulsed cyclophosphamide. All patients received the same oral glucocorticoid regimen and were switched to azathioprine following remission. The primary endpoint was remission by 6 months requiring compliance with the tapering glucocorticoid regimen. Patients with an eGFR <15 mL/min were excluded from the study. RESULTS: At baseline, ANCA subtype, disease activity and organ involvement were similar between groups. Non-inferiority was demonstrated for the primary remission endpoint, which occurred in 47 patients (67%) in the MMF group and 43 patients (61%) in the cyclophosphamide group (risk difference 5.7%, 90% CI -7.5% to 19%). Following remission, more relapses occurred in the MMF group (23 patients, 33%) compared with the cyclophosphamide group (13 patients, 19%) (incidence rate ratio 1.97, 95% CI 0.96 to 4.23, p=0.049). In MPO-ANCA patients, relapses occurred in 12% of the cyclophosphamide group and 15% of the MMF group. In PR3-ANCA patients, relapses occurred in 24% of the cyclophosphamide group and 48% of the MMF group. Serious infections were similar between groups (26% MMF group, 17% cyclophosphamide group) (OR 1.67, 95% CI 0.68 to 4.19, p=0.3). CONCLUSION: MMF was non-inferior to cyclophosphamide for remission induction in AAV, but resulted in higher relapse rate. TRIAL REGISTRATION NUMBER: NCT00414128

Genome-wide association study of eosinophilic granulomatosis with polyangiitis reveals genomic loci stratified by ANCA status

Eosinophilic granulomatosis with polyangiitis (EGPA) is a rare inflammatory disease of unknown cause. 30% of patients have anti-neutrophil cytoplasmic antibodies (ANCA) specific for myeloperoxidase (MPO). Here, we describe a genome-wide association study in 676 EGPA cases and 6,809 controls, that identifies 4 EGPA-associated loci through conventional case-control analysis, and 4 additional associations through a conditional false discovery rate approach. Many variants are also associated with asthma and six are associated with eosinophil count in the general population. Through Mendelian randomisation, we show that a primary tendency to eosinophilia contributes to EGPA susceptibility. Stratification by ANCA reveals that EGPA comprises two genetically and clinically distinct syndromes. MPO+ ANCA EGPA is an eosinophilic autoimmune disease sharing certain clinical features and an HLA-DQ association with MPO+ ANCA-associated vasculitis, while ANCA-negative EGPA may instead have a mucosal/barrier dysfunction origin. Four candidate genes are targets of therapies in development, supporting their exploration in EGPA

Mepolizumab for Eosinophilic Granulomatosis With Polyangiitis: A European Multicenter Observational Study.

OBJECTIVE: Mepolizumab proved to be an efficacious treatment for eosinophilic granulomatosis with polyangiitis (EGPA) at a dose of 300 mg every 4 weeks in the randomized, controlled MIRRA trial. In a few recently reported studies, successful real-life experiences with the approved dose for treating severe eosinophilic asthma (100 mg every 4 weeks) were observed. We undertook this study to assess the effectiveness and safety of mepolizumab 100 mg every 4 weeks and 300 mg every 4 weeks in a large European EGPA cohort. METHODS: We included all patients with EGPA treated with mepolizumab at the recruiting centers in 2015-2020. Treatment response was evaluated from 3 months to 24 months after initiation of mepolizumab. Complete response to treatment was defined as no disease activity (Birmingham Vasculitis Activity Score [BVAS] = 0) and a prednisolone or prednisone dose (or equivalent) of ≤4 mg/day. Respiratory outcomes included asthma and ear, nose, and throat (ENT) exacerbations. RESULTS: Two hundred three patients, of whom 191 received a stable dose of mepolizumab (158 received 100 mg every 4 weeks and 33 received 300 mg every 4 weeks) were included. Twenty-five patients (12.3%) had a complete response to treatment at 3 months. Complete response rates increased to 30.4% and 35.7% at 12 months and 24 months, respectively, and rates were comparable between mepolizumab 100 mg every 4 weeks and 300 mg every 4 weeks. Mepolizumab led to a significant reduction in BVAS score, prednisone dose, and eosinophil counts from 3 months to 24 months, with no significant differences observed between 100 mg every 4 weeks and 300 mg every 4 weeks. Eighty-two patients (40.4%) experienced asthma exacerbations (57 of 158 [36%] who received 100 mg every 4 weeks; 17 of 33 [52%] who received 300 mg every 4 weeks), and 31 patients (15.3%) experienced ENT exacerbations. Forty-four patients (21.7%) experienced adverse events (AEs), most of which were nonserious AEs (38 of 44). CONCLUSION: Mepolizumab at both 100 mg every 4 weeks and 300 mg every 4 weeks is effective for the treatment of EGPA. The 2 doses should be compared in the setting of a controlled trial

Genome-wide association study of eosinophilic granulomatosis with polyangiitis reveals genomic loci stratified by ANCA status

Abstract: Eosinophilic granulomatosis with polyangiitis (EGPA) is a rare inflammatory disease of unknown cause. 30% of patients have anti-neutrophil cytoplasmic antibodies (ANCA) specific for myeloperoxidase (MPO). Here, we describe a genome-wide association study in 676 EGPA cases and 6809 controls, that identifies 4 EGPA-associated loci through conventional case-control analysis, and 4 additional associations through a conditional false discovery rate approach. Many variants are also associated with asthma and six are associated with eosinophil count in the general population. Through Mendelian randomisation, we show that a primary tendency to eosinophilia contributes to EGPA susceptibility. Stratification by ANCA reveals that EGPA comprises two genetically and clinically distinct syndromes. MPO+ ANCA EGPA is an eosinophilic autoimmune disease sharing certain clinical features and an HLA-DQ association with MPO+ ANCA-associated vasculitis, while ANCA-negative EGPA may instead have a mucosal/barrier dysfunction origin. Four candidate genes are targets of therapies in development, supporting their exploration in EGPA

A Large-Scale Genetic Analysis Reveals a Strong Contribution of the HLA Class II Region to Giant Cell Arteritis Susceptibility

We conducted a large-scale genetic analysis on giant cell arteritis (GCA), a polygenic immune-mediated vasculitis. A case-control cohort, comprising 1,651 case subjects with GCA and 15,306 unrelated control subjects from six different countries of European ancestry, was genotyped by the Immunochip array. We also imputed HLA data with a previously validated imputation method to perform a more comprehensive analysis of this genomic region. The strongest association signals were observed in the HLA region, with rs477515 representing the highest peak (p = 4.05 × 10−40, OR = 1.73). A multivariate model including class II amino acids of HLA-DRβ1 and HLA-DQα1 and one class I amino acid of HLA-B explained most of the HLA association with GCA, consistent with previously reported associations of classical HLA alleles like HLA-DRB1∗04. An omnibus test on polymorphic amino acid positions highlighted DRβ1 13 (p = 4.08 × 10−43) and HLA-DQα1 47 (p = 4.02 × 10−46), 56, and 76 (both p = 1.84 × 10−45) as relevant positions for disease susceptibility. Outside the HLA region, the most significant loci included PTPN22 (rs2476601, p = 1.73 × 10−6, OR = 1.38), LRRC32 (rs10160518, p = 4.39 × 10−6, OR = 1.20), and REL (rs115674477, p = 1.10 × 10−5, OR = 1.63). Our study provides evidence of a strong contribution of HLA class I and II molecules to susceptibility to GCA. In the non-HLA region, we confirmed a key role for the functional PTPN22 rs2476601 variant and proposed other putative risk loci for GCA involved in Th1, Th17, and Treg cell function