Translational regulation plays a pivotal role during pre-implantation development. However, the mechanisms by which messenger RNAs (mRNAs) are selectively regulated over time, along with their dynamic utilization and fate during this period, remain largely unknown. Here, we performed fraction-resolved polysome profiling and characterized translational dynamics across oocytes and early embryo development. This approach allowed us to examine the changes in translation during pre-implantation development in high resolution and uncover previously unrecognized modes of translational selectivity. We observed a stage-specific delay in translation, characterized by the postponed recruitment of stored mRNAs-either unbound or associated with light ribosomal fractions-into actively translating polysomes (heavy fraction). Comparative analysis of translatome with proteomics, RNA N6-methyladenosine modifications, and mRNA features further revealed both coordinated and distinct regulatory mechanisms during pre-implantation development. Furthermore, we identified a eukaryotic initiation factor 1A domain containing 3, Eif1ad3, which is exclusively translated at the two-cell stage and is essential for embryonic development by regulating ribosome biogenesis and protein synthesis. Collectively, our study provides a valuable resource of spatiotemporal translational regulation in mammalian pre-implantation development and highlights a previously uncharacterized translation initiation factor critical for early embryos.

• MeSH
• adenosin analogy a deriváty   metabolismus   MeSH
• blastocysta metabolismus   MeSH
• embryonální vývoj * genetika   MeSH
• eukaryotický iniciační faktor 1 metabolismus   genetika   MeSH
• messenger RNA * metabolismus   genetika   MeSH
• myši MeSH
• oocyty metabolismus   růst a vývoj   MeSH
• polyribozomy metabolismus   genetika   MeSH
• proteosyntéza * MeSH
• ribozomy metabolismus   MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adenosin MeSH
• eukaryotický iniciační faktor 1 MeSH
• messenger RNA * MeSH
• N-methyladenosine MeSH Prohlížeč

The YTHDF protein family plays a critical role in cancer development by recognizing and regulating the stability of N6-methyladenosine (m6A)-modified RNA. Here, we reveal an autophagy-dependent mechanism controlling YTHDF protein levels. Using contact inhibition as a cellular model system, we show YTHDF proteins to be rapidly degraded, coinciding with increased autophagy and decreased mTOR activity. Upon pharmacological mTOR inhibition, YTHDF2 is also downregulated via lysosomal degradation. YTHDF2 selectively interacts with the autophagy modifier GABARAP L2 through LC3-interacting region (LIR) motifs in its unstructured N- and C-terminal regions. Autophagic YTHDF2 downregulation results in the co-degradation of its bound m6A-modified RNA clients. While YTHDF depletion induces cell death in contact-inhibition-deficient HCT116 cancer cells, contact-inhibited MRC5 and RPE1 cells remain unaffected. Our findings uncover a regulatory pathway that governs YTHDF protein stability with significant implications for cancer biology and cell fate determination and suggest the existence of an autophagy-mediated degradation pathway for m6A-modified RNA.

• Klíčová slova
• CP: Cell biology, RNA binding, YTHDF, cancer, cellular homeostasis, contact inhibition, lysosomal degradation, m6A, selective autophagy,
• MeSH
• adaptorové proteiny signální transdukční metabolismus   MeSH
• adenosin * analogy a deriváty   metabolismus   MeSH
• autofagie * MeSH
• HCT116 buňky MeSH
• lidé MeSH
• lyzozomy metabolismus   MeSH
• proteiny asociované s mikrotubuly MeSH
• proteiny regulující apoptózu metabolismus   MeSH
• proteiny vázající RNA * metabolismus   genetika   MeSH
• RNA * metabolismus   MeSH
• TOR serin-threoninkinasy metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• adenosin * MeSH
• GABARAP protein, human MeSH Prohlížeč
• MTOR protein, human MeSH Prohlížeč
• N-methyladenosine MeSH Prohlížeč
• proteiny asociované s mikrotubuly MeSH
• proteiny regulující apoptózu MeSH
• proteiny vázající RNA * MeSH
• RNA * MeSH
• TOR serin-threoninkinasy MeSH
• YTHDF2 protein, human MeSH Prohlížeč

Coronaviruses can spread rapidly to new host species and cause severe respiratory and enteric diseases in vertebrates, including humans. To date, seven coronaviruses have been identified in humans, with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) being the most notorious due to its substantial social and economic impact. Although anti-SARS-CoV-2 vaccines are available, infections remain widespread, highlighting the ongoing need for antiviral treatments. Here, we report the synthesis and evaluation of the activity of uridine glycoconjugates, designed as glycosyltransferase donor-type inhibitors incorporating a 1,2,3-triazole moiety. These compounds were tested against two model coronaviruses: murine hepatitis virus strain A59 (MHV) and human coronavirus strain NL63 (HCoV-NL63). Four of the synthesized compounds demonstrated strong antiviral activity against both viruses, and their efficacy was further confirmed against SARS-CoV-2. Our results suggest that these compounds interfere with the coronavirus infectivity and replication process. Thus, these novel compounds may prove to be effective broad-spectrum antiviral inhibitors.

• MeSH
• antivirové látky * farmakologie   chemie   chemická syntéza   MeSH
• Cercopithecus aethiops MeSH
• farmakoterapie COVID-19 MeSH
• glykokonjugáty * farmakologie   chemie   chemická syntéza   MeSH
• lidé MeSH
• myši MeSH
• replikace viru účinky léků   MeSH
• SARS-CoV-2 účinky léků   MeSH
• triazoly * chemie   farmakologie   MeSH
• uridin * farmakologie   chemie   analogy a deriváty   MeSH
• Vero buňky MeSH
• virus myší hepatitidy účinky léků   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antivirové látky * MeSH
• glykokonjugáty * MeSH
• triazoly * MeSH
• uridin * MeSH

Higher-risk myelodysplastic syndrome (HR-MDS) with RARA gene overexpression is a subset of patients (pts) with an actionable target for tamibarotene, an oral and a selective retinoic acid receptor-α (RAR-α) agonist. Tamibarotene with azacitidine (AZA) showed complete remission (CR) rates in myeloid leukemia. SELECT-MDS-1 was a phase 3 study comparing the activity of tamibarotene + AZA to placebo + AZA in these pts with newly diagnosed HR-MDS with RARA overexpression. Eligible pts had confirmed RARA overexpression, untreated MDS with higher-risk features by revised International Prognostic Scoring System (IPSS-R), and marrow blast count >5%. Pts were randomized 2:1 to receive tamibarotene + AZA or placebo + AZA, respectively. A total of 246 participants were randomized with 164 and 82 in the tamibarotene + AZA and placebo + AZA groups, respectively. Baseline characteristics included: 69.9% male; median age 75 years (range, 38-93); primary MDS, 89.8%; MDS-excess blasts-1, 48% and MDS-excess blasts-2, 52%; and IPSS-R risk category intermediate (25.5%), high (35.7%), and very high (38.9%). The study did not meet the primary end point of CR, with a P value of .2084 for the treatment effect in the tamibarotene + AZA group. The CR rates were 23.81% and 18.75% in the tamibarotene + AZA and placebo + AZA groups, respectively. The use of tamibarotene-based therapy to target RAR-α as a novel approach in pts with HR-MDS with RARA gene overexpression is not a paradigm, which can augment response rates beyond AZA monotherapy. Further explorations of alternative approaches, including those with a biomarker, to alter the natural history of this disease are warranted. This trial was registered at www.clinicaltrials.gov as #NCT04797780.

• MeSH
• azacytidin * aplikace a dávkování   terapeutické užití   škodlivé účinky   MeSH
• benzoáty * aplikace a dávkování   terapeutické užití   škodlivé účinky   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• myelodysplastické syndromy * farmakoterapie   diagnóza   mortalita   MeSH
• protokoly protinádorové kombinované chemoterapie * terapeutické užití   škodlivé účinky   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• tetrahydronaftaleny * aplikace a dávkování   terapeutické užití   škodlivé účinky   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze III MeSH
• multicentrická studie MeSH
• randomizované kontrolované studie MeSH
• Názvy látek
• azacytidin * MeSH
• benzoáty * MeSH
• tamibarotene MeSH Prohlížeč
• tetrahydronaftaleny * MeSH

Cytidine deaminase (CDA) and deoxycytidine monophosphate deaminase (DCTD) play crucial roles in pyrimidine metabolism, affecting DNA synthesis, cell cycle progression, and the efficacy of numerous nucleoside analog-based chemotherapeutics. Given their significance, accurate and sensitive measurement of their enzymatic activity is paramount for both fundamental biochemical research and clinical applications. This review provides a comprehensive overview of the methodologies used to assess CDA and DCTD activity, both established and emerging. We systematically categorize and discuss various approaches, including spectrophotometric, fluorimetric, liquid chromatography-based (Ultraviolet-Visible, fluorescence, and mass spectrometry), radiometric, and cell-based assays. For each method, we present its underlying principles, advantages, and limitations. Furthermore, we draw comparisons across the techniques to highlight their suitability for specific research questions.

• Klíčová slova
• LC-MS, cell-based approaches, cytidine deaminase, dCMP deaminase, modified nucleosides, spectrophotometry,
• MeSH
• chromatografie kapalinová MeSH
• cytidin metabolismus   MeSH
• cytidindeaminasa * metabolismus   MeSH
• enzymatické testy * metody   MeSH
• hmotnostní spektrometrie MeSH
• lidé MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• cytidin MeSH
• cytidindeaminasa * MeSH

Nucleos(t)ides are essential for DNA/RNA synthesis, energy metabolism, and signaling, yet their roles in placental development remain poorly understood. The placenta undergoes dynamic metabolic adaptations throughout gestation to support fetal growth. This study investigates gene expression shifts in nucleos(t)ide metabolism, transport, and adenosine signaling during placental development and in the pathological condition of spontaneous preterm birth (PTB). We analyzed gene expression in first-trimester (n = 10) and term (n = 10), and PTB (n = 10) human placentas, and in cytotrophoblast and syncytiotrophoblast stage in primary human trophoblasts (n = 3) and BeWo (n = 5) cells. For developmental context, rat placentas were examined at gestation days (GD) GD12, GD15, and GD20 (n = 5 per group) that correspond to early second trimester in the human placenta. We found that genes involved in nucleos(t)ide metabolism and adenosine signaling were dominantly upregulated from early gestation to term in the human placenta. PTB placentas revealed further elevation compared to the term placenta. Differentiation from cytotrophoblast to syncytiotrophoblast was accompanied by only minor changes. Pearson's correlation analysis revealed strong gene-metabolite and gene-gene associations, highlighting an integrated metabolic network regulating placental function. Gene expression also differed among the tested GDs in the rat placenta. These findings demonstrate dynamic changes of nucleos(t)ide metabolism during healthy placental development and enhanced expression in PTB placentas, suggesting increasing needs for nucleos(t)ides during placental growth and metabolic shifts in the PTB placenta. Our data also indicate that nucleos(t)ide metabolism is preserved in both proliferative and differentiated states.

• Klíčová slova
• Adenosine receptors, Cytotrophoblast and syncytiotrophoblast, Nucleoside transporters, Nucleotide and nucleoside metabolism in placenta, Placental gene expression, Preterm birth,
• MeSH
• adenosin * metabolismus   MeSH
• krysa rodu Rattus MeSH
• lidé MeSH
• placenta metabolismus   MeSH
• předčasný porod * metabolismus   genetika   MeSH
• signální transdukce MeSH
• těhotenství MeSH
• trofoblasty * metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adenosin * MeSH

INTRODUCTION: It is well known that platelets from diabetic patients can be resistant to clinically used antiplatelet drugs. METHODS: To assess the phenomenon in more detail, 50 adult patients suffering from type 1 diabetes mellitus (T1D) were recruited and their responses to 7 platelet aggregation inducers, as well as to 3 clinically used antiplatelet drugs (acetylsalicylic acid /ASA/, ticagrelor and vorapaxar) and one experimental compound, 4-methylcatechol, were assessed ex vivo. A control group of 50 generally healthy age-matched controls was also included for comparison. RESULTS: T1D patients exhibited a lower aggregation reaction to 3 inducers but were conversely more resistant to the effect of ASA and vorapaxar than controls. Ticagrelor tended to be less active in T1D as well. On the other hand, 4-methylcatechol was equally or even more potent in T1D than in healthy controls. Plasma glucose levels above 7 mM were associated with lower platelet aggregation responses to four aggregation inducers. In contrast, the effect of 4-methylcatechol, unlike that of ASA, did not appear to be strongly influenced by glycemia. Further subanalyses, excluding hypertensive patients and significantly more frequently administered drugs, did not substantially modify the results. CONCLUSION: Conclusively, 4-methylcatechol seems to be a prototypical antiplatelet compound with a strong effect even in diabetic patients.

• Klíčová slova
• 4-methylcatechol, Aggregation, Diabetes mellitus, Platelets,
• MeSH
• agregace trombocytů * účinky léků   MeSH
• Aspirin * terapeutické užití   farmakologie   škodlivé účinky   MeSH
• diabetes mellitus 1. typu * krev   diagnóza   farmakoterapie   MeSH
• dospělí MeSH
• inhibitory agregace trombocytů * terapeutické užití   škodlivé účinky   farmakologie   MeSH
• katecholy * terapeutické užití   farmakologie   MeSH
• krevní glukóza metabolismus   účinky léků   MeSH
• laktony * terapeutické užití   farmakologie   MeSH
• léková rezistence MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• pyridiny * terapeutické užití   farmakologie   MeSH
• studie případů a kontrol MeSH
• ticagrelor terapeutické užití   MeSH
• trombocyty * účinky léků   metabolismus   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• Aspirin * MeSH
• inhibitory agregace trombocytů * MeSH
• katecholy * MeSH
• krevní glukóza MeSH
• laktony * MeSH
• pyridiny * MeSH
• ticagrelor MeSH
• vorapaxar MeSH Prohlížeč

Patients with acute myeloid leukemia (AML) ineligible for intensive chemotherapy (IC) have limited treatment options. The phase 3 ENHANCE-3 study aimed to determine whether magrolimab (magrolimab arm) was superior to placebo (control arm) when either was combined with venetoclax and azacitidine. Adults with previously untreated AML who were ineligible for IC were randomized to receive magrolimab (1 mg/kg on days 1 and 4, 15 mg/kg on day 8, 30 mg/kg on days 11 and 15, then weekly for 5 weeks, and then every 2 weeks) or placebo, venetoclax (100 mg on day 1, 200 mg on day 2, and 400 mg daily thereafter), and azacitidine (75 mg/m2 days 1-7) in 28-day cycles. The primary end point was overall survival (OS); key secondary end points included complete remission (CR) rate and safety. After randomization of 378 patients, the trial was stopped at a prespecified interim analysis owing to futility. At final analysis, with median follow-up of 7.6 months (magrolimab arm) vs 7.4 months (control arm), median OS was 10.7 vs 14.1 months (hazard ratio, 1.178; 95% confidence interval, 0.848-1.637). The CR rate within 6 cycles was 41.3% vs 46.0%. Addition of magrolimab to venetoclax and azacitidine resulted in more fatal adverse events (19.0% vs 11.4%), primarily driven by grade 5 infections (11.1% vs 6.5%) and respiratory events (2.6% vs 0%). There were similar incidences of any-grade infections, febrile neutropenia, and neutropenia between arms. These results highlight the difficulty in improving outcomes for patients with AML who were ineligible for IC. This trial was registered at www.clinicaltrials.gov as #NCT05079230.

• MeSH
• akutní myeloidní leukemie * farmakoterapie   mortalita   MeSH
• azacytidin * aplikace a dávkování   škodlivé účinky   MeSH
• bicyklické sloučeniny heterocyklické aplikace a dávkování   škodlivé účinky   MeSH
• dospělí MeSH
• dvojitá slepá metoda MeSH
• humanizované monoklonální protilátky * aplikace a dávkování   škodlivé účinky   MeSH
• lidé středního věku MeSH
• lidé MeSH
• protokoly protinádorové kombinované chemoterapie * terapeutické užití   škodlivé účinky   aplikace a dávkování   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• sulfonamidy aplikace a dávkování   škodlivé účinky   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky, fáze III MeSH
• multicentrická studie MeSH
• randomizované kontrolované studie MeSH
• Názvy látek
• azacytidin * MeSH
• bicyklické sloučeniny heterocyklické MeSH
• humanizované monoklonální protilátky * MeSH
• sulfonamidy MeSH
• venetoclax MeSH Prohlížeč

The RNA-binding protein La is found in most eukaryotes, and despite being essential in many organisms, its function is not completely clear. Trypanosoma brucei, the causative agent of human African trypanosomiasis, encodes a 'classical' La protein (TbLa) composed of a La-motif, two RNA recognition motifs (RRM1 and RRM2α), a C-terminal short basic motif (SBM), and a nuclear localization signal (NLS). In T. brucei, like in most eukaryotes, position 34 of tRNATyr, -Asp, -Asn and -His is modified with queuosine (Q34). The steady-state levels of queuosine-modified tRNA in the insect form (procyclic) of T. brucei can fluctuate dynamically depending on growth conditions, but the mechanism(s) controlling Q34 levels are not well understood. A well-established function of La is in precursor-tRNA 3'-end metabolism, but in this work, we demonstrate that La also controls Q34-tRNA levels. Individual domain deletions showed that while deletion of La motif or RRM1 causes dysregulation of Q34-tRNA levels, no other domain plays a similar role. We also show that La is important for the normal balance of several additional tRNA modifications. These findings are discussed in the context of substrate competition between La and modification enzymes, also highlighting subcellular localization as a key determinant of tRNA function.

• MeSH
• nukleosid Q metabolismus   analogy a deriváty   MeSH
• posttranskripční úpravy RNA * MeSH
• proteinové domény MeSH
• proteiny vázající RNA * metabolismus   chemie   genetika   MeSH
• protozoální proteiny * metabolismus   chemie   genetika   MeSH
• RNA transferová * metabolismus   genetika   MeSH
• Trypanosoma brucei brucei * genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• nukleosid Q MeSH
• proteiny vázající RNA * MeSH
• protozoální proteiny * MeSH
• RNA transferová * MeSH

Cytosine (DNA) methylation plays important roles in silencing transposable elements, plant development, genomic imprinting, stress responses, and maintenance of genome stability. To better understand the functions of this epigenetic modification, several tools have been developed to manipulate DNA methylation levels. These tools include mutants of DNA methylation writers and readers, targeted manipulation of locus-specific methylation, and the use of chemical inhibitors. Here, we summarize the effects of commonly used cytidine analog chemical inhibitors, represented by zebularine, 5-azacytidine, and their related compounds, on plants. These analogs are incorporated into chromosomal DNA, where they block the activity of the replicative CG DNA methyltransferase 1 (MET1). This leads to manifold alterations in the plant epigenome, modified developmental programs, or suppression of hybridization barriers. We also highlight the DNA-damaging effects of cytidine analogs, particularly the formation of stable DNA-protein crosslinks between DNA and MET1. This phenomenon sheds new light on specific phenotypes observed upon treatment with cytidine analogs. In conclusion, cytidine analogs are a vital tool for plant genome research and have the potential to open new promising avenues for applications in plant biotechnology and breeding.

• Klíčová slova
• 5-Methylcytosine, DNA damage, DNA methylation, DNA–protein crosslinks, cytidine analogs, epigenetics, plants, repetitive DNA, transcriptional gene silencing, transposons,
• MeSH
• cytidin * analogy a deriváty   farmakologie   metabolismus   MeSH
• epigeneze genetická * účinky léků   MeSH
• genom rostlinný MeSH
• metylace DNA účinky léků   MeSH
• rostliny * genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• cytidin * MeSH
• pyrimidin-2-one beta-ribofuranoside MeSH Prohlížeč