Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) main protease (Mpro) autocatalytically releases itself out of the viral polyprotein to form a fully active mature dimer in a manner that is not fully understood. Here, we introduce several tools to help elucidate differences between cis (intramolecular) and trans (intermolecular) proteolytic processing and to evaluate inhibition of precursor Mpro. We found that many mutations at the P1 position of the N-terminal autoprocessing site do not block cis autoprocessing but do inhibit trans processing. Notably, substituting the WT glutamine at the P1 position with isoleucine retains Mpro in an unprocessed precursor form that can be purified and further studied. We also developed a cell-based reporter assay suitable for compound library screening and evaluation in HEK293T cells. This assay can detect both overall Mpro inhibition and the fraction of uncleaved precursor form of Mpro through separable fluorescent signals. We observed that inhibitory compounds preferentially block mature Mpro. Bofutrelvir and a novel compound designed in-house showed the lowest selectivity between precursor and mature Mpro, indicating that inhibition of both forms may be possible. Additionally, we observed positive modulation of precursor activity at low concentrations of inhibitors. Our findings help expand understanding of the SARS-CoV-2 viral life cycle and may facilitate development of strategies to target precursor form of Mpro for inhibition or premature activation of Mpro.

• MeSH
• Antiviral Agents * pharmacology   chemistry   MeSH
• COVID-19 Drug Treatment MeSH
• HEK293 Cells MeSH
• Protease Inhibitors pharmacology   chemistry   MeSH
• Coronavirus 3C Proteases * metabolism   antagonists & inhibitors   chemistry   genetics   MeSH
• Humans MeSH
• Mutation MeSH
• Drug Discovery * methods   MeSH
• Proteolysis MeSH
• SARS-CoV-2 * enzymology   drug effects   metabolism   genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

The glycoprotein clusterin (CLU) is involved in cell proliferation and DNA damage repair and is highly expressed in tumor cells. Here, we aimed to investigate the effects of CLU dysregulation on two human astrocytic cell lines: CCF-STTG1 astrocytoma cells and SV-40 immortalized normal human astrocytes. We observed that suppression of CLU expression by RNA interference inhibited cell proliferation, triggered the DNA damage response, and resulted in cellular senescence in both cell types tested. To further investigate the underlying mechanism behind these changes, we measured reactive oxygen species, assessed mitochondrial function, and determined selected markers of the senescence-associated secretory phenotype. Our results suggest that CLU deficiency triggers oxidative stress-mediated cellular senescence associated with pronounced alterations in mitochondrial membrane potential, mitochondrial mass, and expression levels of OXPHOS complex I, II, III and IV, indicating mitochondrial dysfunction. This report shows the important role of CLU in cell cycle maintenance in astrocytes. Based on these data, targeting CLU may serve as a potential therapeutic approach valuable for treating gliomas.

• MeSH
• Astrocytes * metabolism   pathology   MeSH
• Clusterin * metabolism   genetics   MeSH
• Humans MeSH
• Membrane Potential, Mitochondrial * physiology   MeSH
• Mitochondria * metabolism   MeSH
• Cell Line, Tumor MeSH
• Oxidative Stress physiology   MeSH
• Oxidative Phosphorylation MeSH
• DNA Damage MeSH
• Cell Proliferation * MeSH
• Reactive Oxygen Species metabolism   MeSH
• Cellular Senescence * physiology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Present study was aimed to develop an efficient microbial consortium for combating Alternaria blight disease in cumin. The research involved isolating biocontrol agents against Alternaria burnsii, characterizing their biocontrol and growth promotion traits, and assessing compatibility. A pot experiment was conducted during rabi season of 2022-2023 to evaluate the bioefficacy of four biocontrol agents (1F, 16B, 31B, and 223B) individually and in consortium, focusing on disease severity, plant growth promotion, and defense responses in cumin challenged with A. burnsii. Microbial isolates 1F, 16B, 31B, and 223B significantly inhibited A. burnsii growth in dual plate assays (~ 86%), displaying promising biocontrol and plant growth promotion activities. They were identified as Trichoderma afroharzianum 1F, Aneurinibacillus aneurinilyticus 16B, Pseudomonas lalkuanensis 31B, and Bacillus licheniformis 223B, respectively. The excellent compatibility was observed among all selected biocontrol agents. Cumin plants treated with consortia of 1F + 16B + 31B + 223B showed least percent disease index (32.47%) and highest percent disease control (64.87%). Consortia of biocontrol agents significantly enhanced production of secondary metabolites (total phenol, flavonoids, antioxidant, and tannin) and activation of antioxidant-defense enzymes (POX, PPOX, CAT, SOD, PAL, and TAL) compared to individual biocontrol treatment and infected control. Moreover, consortium treatments effectively reduced electrolyte leakage over the individual biocontrol agent and infected control treatment. The four-microbe consortium significantly enhanced chlorophyll (154%), carotenoid content (88%), plant height (78.77%), dry weight (72.81%), and seed yield (104%) compared to infected control. Based on these findings, this environmentally friendly four-microbe consortium may be recommended for managing Alternaria blight in cumin.

• MeSH
• Alternaria * growth & development   physiology   MeSH
• Biological Control Agents MeSH
• Cuminum * microbiology   immunology   growth & development   MeSH
• Microbial Consortia * MeSH
• Plant Diseases * microbiology   prevention & control   immunology   MeSH
• Disease Resistance MeSH
• Publication type
• Journal Article MeSH

Inhibitory Janusových kináz (JAK) patří k novějším terapeutickým modalitám v léčbě zánětlivých revmatických onemocnění. Našly si své místo v terapii u nemocných s refrakterním onemocněním, u kterých došlo k selhání nejen konvenčních chorobu modifikujících léků, ale i biologik. Nitrobuněčná signalizace prostřednictvím JAK je aktivována navázáním celé řady cytokinů na odpovídající buněčné receptory a dochází k aktivaci transkripčních proteinů (signal transducer and activator of transcription protein, STAT) a v konečné fázi dochází k transkripci genů zapojených do imunitní odpovědi. Filgotinib je inhibitor JAK s pětinásobně vyšší selektivitu pro JAK1 než pro JAK2, JAK3 a TYK2. V současné době je schválen v České republice pro terapii revmatoidní artritidy a ulcerózní kolitidy.

Janus kinase (JAK) inhibition is a newer therapeutic approach for treating autoimmune inflammatory rheumatic diseases. It has found its place in the therapy of patients with refractory disease, in whom not only conventional disease-modifying drugs have failed, but also biologics. JAKs activate intracellular signaling by binding to cellular receptors and activating signal transducers and activator of transcription (STAT) proteins, which ultimately lead to the transcription of genes involved in the immune response. Filgotinib is a JAK inhibitor with fivefold higher selectivity for JAK1 than for JAK2, JAK3, and TYK2. It is currently approved in the Czech Republic for the treatment of rheumatoid arthritis and ulcerative colitis.

Dermatomyositis (DM) is a rare and debilitating, systemic, autoimmune disease. While heterogenous in presentation and severity, DM is primarily characterised by a spectrum of skin and muscle disease, which may include proximal muscle weakness and recalcitrant cutaneous eruptions. DM may also be associated with joint pain and stiffness, inflammatory arthritis, dysphagia, fatigue, and calcinosis. The current standard of care for DM includes glucocorticoids, immunosuppressants, and intravenous immunoglobulin (IVIg). Unfortunately, these medications are not uniformly effective and can lead to adverse events, particularly with chronic use, necessitating discontinuation of therapy. Therefore, a substantial unmet need exists for more tailored and efficacious therapies that target DM pathogenesis. Brepocitinib is an oral, once-daily, novel, and specific TYK2/JAK1 inhibitor. Brepocitinib's potent inhibition of TYK2 and JAK1 reduces the signalling of pro-inflammatory cytokines, including IFN-α/β, IL-12, IL-23, and IFNγ, that have been implicated in the pathogenesis of DM. Other JAK inhibitors have been used off-label in both case series and open-label clinical trials in patients with DM; and brepocitinib has demonstrated efficacy in phase 2 clinical trials of several other autoimmune diseases, including plaque psoriasis, psoriatic arthritis, Crohn's disease, hidradenitis suppurativa, and ulcerative colitis. Therefore, there is a strong scientific and clinical rationale for the utility and potential effectiveness of brepocitinib in the treatment of DM patients. Currently, the safety, tolerability, and efficacy of brepocitinib is being evaluated in the largest (n=225) double-blind placebo-controlled phase 3 trial in DM patients to date (VALOR - NCT0543726).

• MeSH
• Dermatomyositis * drug therapy   diagnosis   immunology   MeSH
• Protein Kinase Inhibitors * therapeutic use   adverse effects   MeSH
• Janus Kinase 1 * antagonists & inhibitors   MeSH
• TYK2 Kinase * antagonists & inhibitors   MeSH
• Humans MeSH
• Signal Transduction drug effects   MeSH
• Treatment Outcome MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

Drug resistance is a growing problem for many pathogens, including mycobacteria. Small heterocyclic molecules are among the leading scaffolds for developing potential antimycobacterial agents. Therefore, based on the molecular hybridization approach, we have prepared an extensive series of N-substituted 5-(3,5-dinitrophenyl)-1,3,4-oxadiazol-2-amine derivatives. We also investigated their isosteres and acyclic synthetic precursors. The compounds were evaluated for their in vitro activity against Mycobacterium tuberculosis (Mtb) H37Rv, a panel of multidrug- and extensively drug-resistant Mtb isolates and two nontuberculous mycobacterial strains (NTM; M. avium and M. kansasii). The ability to inhibit mycobacterial growth was quantified using minimum inhibitory concentration (MIC) values. Many compounds achieved MIC values ≤ 0.03 μM for NTM and Mtb, regardless of their resistance profile. The highest activity was associated with oxadiazole and thiadiazole scaffolds with benzylamino or C5-C9 alkylamino substitution. The experimentally confirmed mechanism of action of these compounds consists of disruption of mycobacterial cell wall biosynthesis via inhibition of decaprenylphosphoryl-β-D-ribose 2'-epimerase (DprE1). In vitro toxicity evaluation was performed in a hepatocyte model (HepG2), while in vivo toxicity was evaluated using Danio rerio embryos. These findings identify a promising new chemotype with potent, broad-spectrum and selective antimycobacterial activity, including efficacy against resistant strains, and support its further development as a potential therapeutic candidate.

• MeSH
• Antitubercular Agents * pharmacology   chemical synthesis   chemistry   toxicity   MeSH
• Zebrafish MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Mycobacterium tuberculosis drug effects   MeSH
• Oxadiazoles * pharmacology   chemical synthesis   chemistry   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Topoisomerase II alpha and beta (TOP2A and TOP2B) isoenzymes perform essential and non-redundant cellular functions. Anthracyclines induce their potent anti-cancer effects primarily via TOP2A, but at the same time they induce a dose limiting cardiotoxicity through TOP2B. Here we describe the development of the obex class of TOP2 inhibitors that bind to a previously unidentified druggable pocket in the TOP2 ATPase domain to act as allosteric catalytic inhibitors by locking the ATPase domain conformation with the capability of isoform-selective inhibition. Through rational drug design we have developed topobexin, which interacts with residues that differ between TOP2A and TOP2B to provide inhibition that is both selective for TOP2B and superior to dexrazoxane. Topobexin is a potent protectant against chronic anthracycline cardiotoxicity in an animal model. This demonstration of TOP2 isoform-specific inhibition underscores the broader potential to improve drug specificity and minimize adverse effects in various medical treatments.

• MeSH
• Anthracyclines * adverse effects   pharmacology   MeSH
• DNA Topoisomerases, Type II * metabolism   chemistry   MeSH
• Topoisomerase II Inhibitors * pharmacology   chemistry   MeSH
• Cardiotonic Agents * pharmacology   chemistry   MeSH
• Cardiotoxicity * prevention & control   MeSH
• Humans MeSH
• Mice MeSH
• Poly-ADP-Ribose Binding Proteins antagonists & inhibitors   metabolism   chemistry   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

INTRODUCTION: The immunosuppressive roles of galectin-3 (Gal-3) in carcinogenesis make this lectin an attractive target for pharmacological inhibition in immunotherapy. Although current clinical immunotherapies appear promising in the treatment of solid tumors, their efficacy is significantly weakened by the hostile immunosuppressive tumor microenvironment (TME). Gal-3, a prominent TME modulator, efficiently subverts the elimination of cancer, either directly by inducing apoptosis of immune cells or indirectly by binding essential effector molecules, such as interferon-gamma (IFNγ). METHODS: N-(2-Hydroxypropyl)methacrylamide (HPMA)-based glycopolymers bearing poly-N-acetyllactosamine-derived tetrasaccharide ligands of Gal-3 were designed, synthesized, and characterized using high-performance liquid chromatography, dynamic light scattering, UV-Vis spectrophotometry, gel permeation chromatography, nuclear magnetic resonance, high-resolution mass spectrometry and CCK-8 assay for evaluation of glycopolymer non-toxicity. Pro-immunogenic effects of purified glycopolymers were tested by apoptotic assay using flow cytometry, competitive ELISA, and in vitro cell-free INFγ-based assay. RESULTS: All tested glycopolymers completely inhibited Gal-3-induced apoptosis of monocytes/macrophages, of which the M1 subtype is responsible for eliminating cancer cells during immunotherapy. Moreover, the glycopolymers suppressed Gal-3-induced capture of glycosylated IFNγ by competitive inhibition to Gal-3 carbohydrate recognition domain (CRD), which enables further inherent biological activities of this effector, such as differentiation of monocytes into M1 macrophages and repolarization of M2-macrophages to the M1 state. CONCLUSION: The prepared glycopolymers are promising inhibitors of Gal-3 and may serve as important supportive anti-cancer nanosystems enabling the infiltration of proinflammatory macrophages and the reprogramming of unwanted M2 macrophages into the M1 subtype.

• MeSH
• Acrylamides chemistry   pharmacology   MeSH
• Apoptosis drug effects   MeSH
• Galectin 3 * antagonists & inhibitors   MeSH
• Galectins MeSH
• Interferon-gamma * metabolism   MeSH
• Blood Proteins MeSH
• Humans MeSH
• Macrophages drug effects   MeSH
• Monocytes * drug effects   MeSH
• Tumor Microenvironment drug effects   MeSH
• Polymers * chemistry   pharmacology   MeSH
• Antineoplastic Agents * pharmacology   chemistry   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

Lipoprotein (a) [Lp(a)] has been recognized as an independent, inherited, causal risk factor for atherosclerotic cardiovascular disease (ASCVD) and aortic valve stenosis, thus representing a major target of residual CV risk. Currently, no drug has been officially approved for lowering Lp(a) levels, and in clinical practice, Lp(a) is mainly used to (re)define CV risk, particularly in individuals at borderline CV risk and people with a family history of premature coronary heart disease, according to various guidelines. Specific Lp(a)-targeted antisense oligonucleotides (ASOs) and small interfering RNA (siRNA) agents have been developed to produce substantial Lp(a) reductions via the inhibition of apo(a) synthesis in the liver. These drugs are conjugated to N-acetylgalactosamine (GalNAc) to ensure their binding to asialoglycoproteins, which are specifically expressed on the surface of the hepatocytes. Such drugs include pelacarsen (an injectable ASO) and olpasiran, zerlasiran, and lepodisiran (injectable siRNA agents). Muvalaplin represents another therapeutic option to lower Lp(a) levels, since it is an oral selective small molecule inhibitor of Lp(a) formation, thus potentially exerting certain advantages in terms of its clinical use. The present narrative review summarizes the available clinical data on the efficacy and safety of these investigational Lp(a)-lowering therapies, as reported in phase 1 and 2 trials. The effects of these drugs on other [aside from Lp(a)] lipid parameters are also discussed. The phase 3 CV trial outcomes are ongoing for some of these agents (i.e., pelacarsen, olpasiran, and lepodisiran) and are briefly mentioned. Overall, there is an urgent need for evidence-based guidelines on Lp(a) reduction in daily clinical practice, following the results of the phase 3 CV trials, as well as for establishing the ideal Lp(a) quantification method (i.e., using an apo(a) isoform-independent assay with appropriate calibrators, reporting the Lp(a) level in molar units).

• Publication type
• Journal Article MeSH
• Review MeSH

Miřátský, P, Hank, M, Gryc, T, Brožka, M, Cabell, L, Escamilla, R, Zahálka, F, and Malý, T. Fitness differences among professional firefighters utilizing various conditioning regimens throughout the year. J Strength Cond Res 39(7): e898-e908, 2025-The aim of this study was to compare selected parameters of physical fitness of Czech frontline professional firefighters ( n = 92), who performed various physical activities according to their preferences within their regular physical preparation for 1 year. These firefighters were assigned to 1 of 3 groups based on their training preferences and sport focus: General Sports Activities (GSA; n = 30), Fire Sport (FS; n = 36), and Toughest Firefighter Alive (TFA; n = 26). All subjects completed (in period September) a battery of laboratory fitness tests including body composition, lower limb muscular strength and power, and cardiorespiratory fitness. Firefighters in the FS and TFA groups were found to have significantly higher values ( p < 0.01) for the following variables: fat-free mass, isokinetic strength, muscle power, maximum aerobic capacity, minute ventilation, and heart rate at the anaerobic threshold than their counterparts in the GSA group. There were no significant differences in body mass ( p = 0.21) and body mass index (BMI) ( p = 0.48) between the firefighter groups. Professional frontline firefighters' level of physical fitness is improved by the incorporation of specific physical training activities, duties, and disciplines. This can have a direct impact on frontline firefighters' capacity to fight fires safely and effectively.

• MeSH
• Anaerobic Threshold physiology   MeSH
• Adult MeSH
• Firefighters * MeSH
• Cardiorespiratory Fitness physiology   MeSH
• Physical Conditioning, Human * methods   physiology   MeSH
• Humans MeSH
• Body Composition physiology   MeSH
• Heart Rate physiology   MeSH
• Muscle Strength physiology   MeSH
• Physical Fitness * physiology   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH