Tacrine, the first approved drug against Alzheimer's disease (AD), was withdrawn from clinical use due to serious adverse effects. The main concern was the human hepatotoxicity, stemming probably from the liver biotransformation and clinically manifested as hepatocellular necrosis, and lobular hepatitis. Concerning the biotransformation, 7-OH-tacrine metabolite is generally suspected of being a precursor of toxic quinone methide, which binds to intracellular -SH proteins and/or depletes intracellular glutathione, and by that probably causes the hepatotoxicity. However, to study these toxic effects, proper animal model is needed to monitor the interspecies differences of metabolism. To fully describe in vivo ADMET parameters of tacrine, five experimental pigs (Sus scrofa f. domestica), as the most physiologically human-like in vivo model showing similar tacrine biotransformation, were used. We studied tacrine and its metabolites ADMET characteristics after both acute i.g. single dose and chronic 42 days p.o daily dose administration of 200 mg of tacrine. Tacrine and its two major metabolites show Tmax in plasma of 360 min, so the absorption is much slower than in human (Tmax = 120 min) and are primarily distributed to the gastro-intestinal tract and CNS. Furthermore, due to the lower activity of CYP1A2 in pigs, tacrine is biotransformed much less efficiently than in humans. This study showed that tacrine accumulates only in adipose tissue, and organ histology and plasma biochemistry assessment revealed no signs of hepatotoxicity even after chronic tacrine administration. Pigs are therefore an unsuitable human-like animal model for evaluating tacrine toxicity.

• MeSH
• Biotransformation MeSH
• Cholinesterase Inhibitors toxicity   pharmacokinetics   MeSH
• Liver * drug effects   metabolism   pathology   MeSH
• Chemical and Drug Induced Liver Injury * metabolism   pathology   MeSH
• Humans MeSH
• Swine MeSH
• Sus scrofa MeSH
• Tacrine * pharmacokinetics   toxicity   metabolism   analogs & derivatives   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

BACKGROUND: In the ARIEL4 trial of rucaparib versus standard-of-care chemotherapy in patients with relapsed BRCA-mutated ovarian carcinoma, the primary endpoint was met, showing improved investigator-assessed progression-free survival with rucaparib. Here, we present the final overall survival analysis of the trial and other post-progression outcomes. METHODS: This open-label, randomised, controlled phase 3 trial was done at 64 hospitals and cancer centres in 12 countries, including Brazil, Canada, Czech Republic, Hungary, Israel, Italy, Poland, Russia, Spain, Ukraine, the UK, and the USA. Eligible patients were women aged 18 or older with BRCA1 or BRCA2-mutated ovarian carcinoma and had received at least two previous chemotherapy regimens. Patients had to have evaluable disease as per Response Evaluation Criteria in Solid Tumors (RECIST; version 1.1) criteria and an Eastern Cooperative Oncology Group performance status of 0 or 1. Patients were randomly assigned (2:1) using an interactive response technology and block randomisation (block size of six) and stratified by progression-free interval after the most recent platinum-containing therapy to receive oral rucaparib (600 mg twice daily administered in 28-day cycles) or chemotherapy on the basis of platinum-sensitivity status. In the chemotherapy group, patients with platinum-resistant disease (progression-free interval ≥1 to <6 months) or partially platinum-sensitive disease (progression-free interval ≥6 to <12 months) received weekly paclitaxel (starting dose 60-80 mg/m2 on days 1, 8, and 15). Patients with fully platinum-sensitive disease (progression-free interval ≥12 months) received the investigator's choice of platinum-based chemotherapy (single-agent cisplatin or carboplatin, or platinum-doublet chemotherapy), in 21-day or 28-day cycles. The primary endpoint (previously reported) was investigator-assessed progression-free survival, assessed in the efficacy population (all randomly assigned patients with deleterious BRCA1 or BRCA2 mutations without reversion mutations) and in the intention-to-treat population (all randomly assigned patients). Overall survival was a prespecified secondary endpoint and was analysed in the intention-to-treat population. Safety was assessed in all patients who received at least one dose of assigned study treatment. The cutoff date was April 10, 2022. This study is registered with ClinicalTrials.gov, NCT02855944; enrolment is complete and the study is closed. FINDINGS: Between March 1, 2017, and Sept 24, 2020, 349 eligible patients were randomly assigned to receive rucaparib (n=233) or chemotherapy (n=116). 332 (95%) of 349 patients were white and 17 (5%) patients were other or of unknown race. In the chemotherapy group, 80 (69%) of 116 patients crossed over to receive rucaparib. Median follow-up was 41·2 months (IQR 37·8-44·6). At data cutoff for this final analysis (April 10, 2022), 244 (70%) of 349 patients had died: 167 (72%) of 233 in the rucaparib group and 77 (66%) of 116 in the rucaparib group. Median overall survival was 19·4 months (95% CI 15·2-23·6) in the rucaparib group versus 25·4 months (21·4-27·6) in the chemotherapy group (hazard ratio 1·3 [95% CI 1·0-1·7], p=0·047). No new safety signals were observed, including during crossover to rucaparib. The most common grade 3-4 adverse events across treatment groups included anaemia or decreased haemoglobin (reported in 59 [25%] of 232 patients in the rucaparib group and seven [6%] of 113 in the chemotherapy group), and neutropenia or decreased neutrophil count (in 26 [11%] of 232 in the rucaparib group and 16 [14%] of 113 patients in the chemotherapy group). Serious adverse events were reported in 66 (28%) of 232 patients in the rucaparib group and 14 (12%) of 113 patients in the chemotherapy group. Ten treatment-related deaths were reported in the rucaparib group, two of which were linked to judged to be related to rucaparib (cardiac disorder and myelodysplastic syndrome), and one death related to treatment was reported in the chemotherapy group, with no specific cause linked to the treatment. INTERPRETATION: These data highlight the need for a better understanding of the most appropriate treatment for patients who have progressed on a poly(adenosine diphosphate-ribose) polymerase (PARP) inhibitor, and the optimal sequencing of chemotherapy and PARP inhibitors in advanced ovarian cancer. FUNDING: Clovis Oncology.

• MeSH
• Progression-Free Survival MeSH
• Adult MeSH
• Indoles * therapeutic use   administration & dosage   adverse effects   MeSH
• Middle Aged MeSH
• Humans MeSH
• Neoplasm Recurrence, Local * drug therapy   genetics   pathology   MeSH
• Mutation * MeSH
• Ovarian Neoplasms * drug therapy   genetics   pathology   mortality   MeSH
• Paclitaxel administration & dosage   adverse effects   MeSH
• Poly(ADP-ribose) Polymerase Inhibitors therapeutic use   adverse effects   administration & dosage   MeSH
• BRCA1 Protein * genetics   MeSH
• BRCA2 Protein * genetics   MeSH
• Antineoplastic Combined Chemotherapy Protocols * therapeutic use   MeSH
• Aged MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Clinical Trial, Phase III MeSH
• Multicenter Study MeSH
• Randomized Controlled Trial MeSH
• Comparative Study MeSH

Targeting ubiquitin E3 ligases is therapeutically attractive; however, the absence of an active-site pocket impedes computational approaches for identifying inhibitors. In a large, unbiased biochemical screen, we discover inhibitors that bind a cryptic cavity distant from the catalytic cysteine of the homologous to E6-associated protein C terminus domain (HECT) E3 ligase, SMAD ubiquitin regulatory factor 1 (SMURF1). Structural and biochemical analyses and engineered escape mutants revealed that these inhibitors restrict an essential catalytic motion by extending an α helix over a conserved glycine hinge. SMURF1 levels are increased in pulmonary arterial hypertension (PAH), a disease caused by mutation of bone morphogenetic protein receptor-2 (BMPR2). We demonstrated that SMURF1 inhibition prevented BMPR2 ubiquitylation, normalized bone morphogenetic protein (BMP) signaling, restored pulmonary vascular cell homeostasis, and reversed pathology in established experimental PAH. Leveraging this deep mechanistic understanding, we undertook an in silico machine-learning-based screen to identify inhibitors of the prototypic HECT E6AP and confirmed glycine-hinge-dependent allosteric activity in vitro. Inhibiting HECTs and other glycine-hinge proteins opens a new druggable space.

• MeSH
• Allosteric Regulation drug effects   MeSH
• Humans MeSH
• Mice MeSH
• Pulmonary Arterial Hypertension drug therapy   MeSH
• Bone Morphogenetic Protein Receptors, Type II MeSH
• Signal Transduction drug effects   MeSH
• Ubiquitination drug effects   MeSH
• Ubiquitin-Protein Ligases * antagonists & inhibitors   metabolism   chemistry   genetics   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Stresová kardiomyopatie je klinický syndrom, při kterém dochází k rozvoji myokardiální dysfunkce v reakci na stres. Častým spouštěčem bývá neurologické onemocnění, nejčastěji netraumatické subarachnoidální krvácení a ischemická CMP. Rozvoj stresové kardiomyopatie může být u těchto pacientů příčinou arteriální hypotenze, arytmií nebo akutního srdečního selhání. I přes reverzibilitu onemocnění je zvláště její sekundární forma nebezpečná pro riziko rozvoje závažných komplikací. Kauzální léčba se zaměřuje na eliminaci příčiny, další terapie je symptomatická, vedená echokardiografickým nálezem. Cílem tohoto přehledového článku je shrnutí dosavadních znalostí ohledně stresové kardiomyopatie se zaměřením na recentní postupy v diagnostice a terapii a zdůraznění jejich odlišností u pacientů s neurologickým onemocněním.

Stress cardiomyopathy stands for a clinical syndrome characterized by the onset of myocardial dysfunction caused by stressful event. A common trigger is neurological disease, most commonly non-traumatic subarachnoid hemorrhage and ischemic stroke. The development of stress cardiomyopathy may cause arterial hypotension, arrhythmias, or acute heart failure in these patients. Despite the reversibility of the disease, its secondary form is particularly dangerous because of the risk of developing serious complications. Causal treatment focuses on eliminating the cause; further therapy is symptomatic, guided by echocardiographic findings. The aim of this review article is to summarize the current knowledge regarding stress cardiomyopathy focusing on up- -to-date diagnostics and treatment and highlight their differences in patients with neurological disease.

• MeSH
• Acute Coronary Syndrome etiology   MeSH
• Stroke complications   MeSH
• Humans MeSH
• Risk Factors MeSH
• Subarachnoid Hemorrhage complications   MeSH
• Takotsubo Cardiomyopathy * diagnosis   etiology   complications   therapy   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH

Hypokalcemií indukovaná kardiomyopatie je vzácné, avšak závažné onemocnění, náročné z hlediska diagnostiky i léčby. těžká hypokalcemie na podkladě hypoparatyreózy může vyvolat vzácnou, nicméně potenciálně reverzibilní, formu dilatační kardiomyopatie. tato kazuistika popisuje případ 31leté ženy se srdečním selháním vyvolaným hypoparatyreózou po tyreoidektomii. Přes klasickou léčbu srdečního selhání přetrvával u pacientky její závažný stav až do úpravy hypokalcemie. léčba zahrnovala suplementaci kalcia a terapii vedenou podle příslušných doporučených postupů; následně došlo k významnému zlepšení. Proto je pro dosažení zlepšení – pokud byla klasická léčba srdečního selhání neúčinná – naprosto nezbytné monitorovat hodnoty kalcia v séru, zvláště u pacientů s tyreoidektomií v anamnéze. Při léčbě srdečního selhání je třeba být ostražitý a mít neustále na paměti možnou přítomnost hypokalcemie jako reverzibilní příčinu uvedené komplikace.

Hypocalcemia-associated cardiomyopathy, a rare but serious condition, presents challenges in diagnosis and management. Severe hypocalcemia resulting from hypoparathyroidism can precipitate rare yet reversible cases of dilated cardiomyopathy. This case report reported a 31-year-old woman with heart failure precipitated by hypoparathyroidism following thyroidectomy. Despite conventional heart failure treatments, her condition persisted until correction of hypocalcemia. Treatment involved calcium supplementation and guideline-directed heart failure therapy. Significant improvement was observed then. Hence, monitoring serum calcium levels is imperative when conventional heart failure therapies fail to yield improvement, particularly in patients with a history of thyroidectomy. Sustaining clinical awareness and vigilance concerning hypocalcemia is crucial in managing heart failure, considering its possibility as a reversible cause.

• MeSH
• Adult MeSH
• Hypocalcemia * etiology   MeSH
• Hypoparathyroidism complications   MeSH
• Cardiomyopathies * diagnosis   etiology   complications   MeSH
• Humans MeSH
• Thyroxine administration & dosage   MeSH
• Thyroidectomy adverse effects   MeSH
• Calcium administration & dosage   MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Female MeSH
• Publication type
• Case Reports MeSH

Syphilis is a multistage sexually transmitted disease caused by Treponema pallidum ssp. pallidum (TPA). This study analyzed clinical samples collected from patients with a diagnosed syphilis infection from 2004-2022, isolated in the Czech Republic. Mucocutaneous swab samples (n = 543) from 543 patients were analyzed, and from these samples, 80.11 % (n = 435) were PCR positive, and 19.89 % (n = 108) were PCR negative for TPA DNA. Swabs were more often positive when collected from syphilis patients in the primary and secondary stages, compared to the latent or unknown stage. There was no significant difference in PCR positivity between the primary and secondary stages (p = 0.099). In IgM-positive patients, a statistically significant association with PCR-positivity was found in samples from seropositive (p = 0.033) and serodiscrepant (RPR negative) patients (p = 0.0006). When assessing our laboratory-defined cases of syphilis, the RPR, IgM, and PCR tests were similarly effective (within the range of 80.1-86.1 %). However, parallel testing with these methods was even more effective, i.e., RPR + PCR was 96.1 % effective and RPR + IgM + PCR was 97.8 % effective. A combination of RPR + PCR, or a combination of all three tests (RPR, IgM, and PCR) can therefore be used to reliably detect active syphilis cases, including reinfections. Our findings show that the reverse algorithm for detecting syphilis could be substantially improved by adding IgM and PCR testing.

• MeSH
• DNA, Bacterial genetics   MeSH
• Adult MeSH
• Immunoglobulin M * blood   MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Polymerase Chain Reaction * methods   MeSH
• Antibodies, Bacterial blood   MeSH
• Sensitivity and Specificity MeSH
• Syphilis Serodiagnosis methods   MeSH
• Syphilis * diagnosis   microbiology   MeSH
• Treponema pallidum * genetics   isolation & purification   immunology   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Czech Republic MeSH

Angelman Syndrome (AS) is a neurodevelopmental disorder caused by the loss of function of ubiquitin-protein ligase E3A (UBE3A), resulting in marked changes in synaptic plasticity. In AS mice, a dysregulation of Ca2+/calmodulin-dependent protein kinase II alpha (CaMKIIα) was previously described. This has been convincingly validated through genetic rescue of prominent phenotypes in mouse cross-breeding experiments. Selective ligands that specifically stabilize the CaMKIIα central association (hub) domain and affect different conformational states in vitro are now available. Two of these ligands, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) and (E)-2-(5-hydroxy-2-phenyl-5,7,8,9-tetrahydro-6H-benzo[7]annulen-6-ylidene)acetic acid (Ph-HTBA), confer neuroprotection after ischemic stroke in mice where CaMKIIα is known to be dysregulated. Here, we sought to investigate whether pharmacological modulation with these prototypical CaMKIIα hub ligands presents a viable approach to alleviate AS symptoms. We performed an in vivo functional evaluation of AS mice treated for a total of 14 days with either HOCPCA or Ph-HTBA (7 days pre-treatment and 7 days of behavioural assessment). Both compounds were well-tolerated but unable to revert robust phenotypes of motor performance, anxiety, repetitive behaviour or seizures in AS mice. Biochemical experiments subsequently assessed CaMKIIα autophosphorylation in AS mouse brain tissue. Taken together our results indicate that pharmacological modulation of CaMKIIα via the selective hub ligands used here is not a viable treatment strategy in AS.

• MeSH
• Angelman Syndrome * drug therapy   genetics   MeSH
• Behavior, Animal drug effects   MeSH
• Phenotype * MeSH
• Ligands MeSH
• Disease Models, Animal * MeSH
• Brain drug effects   metabolism   MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Neuroprotective Agents pharmacology   MeSH
• Calcium-Calmodulin-Dependent Protein Kinase Type 2 * metabolism   MeSH
• Ubiquitin-Protein Ligases metabolism   genetics   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Despite lower virulence, the omicron variant of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes coronavirus disease 2019 (COVID-19) still poses a relevant threat for immunocompromised patients. A retrospective multicentric study was conducted to evaluate the efficacy of pre-exposure prophylaxis with tixagevimab/cilgavimab (Evusheld) with a 6-month follow-up for preventing severe COVID-19 in adult patients with hematology malignancy. Among the 606 patients in the cohort, 96 (16%) contracted COVID-19 with a median of 98.5 days after Evusheld administration. A total of 75% of patients had asymptomatic or mild severity of COVID-19, while just 25% of patients with SARS-CoV-2 positivity had to be hospitalized. Two patients (2%) died directly, and one patient (1%) in association with COVID-19. Eight patients (1.3%) of every cohort experienced adverse events related to Evusheld, mostly grade 1 and of reversible character. It was found that complete vaccination status or positive seroconversion was not associated with lower risk of COVID-19 infection. Previous treatment with an anti-CD20 monoclonal antibody was associated with higher rates of COVID-19, while previous treatment with anti-CD38 monoclonal antibody was not, as was the case for recipients of hematopoietic stem cell transplantation or CAR-T cell therapy. Presence of other comorbidities was not associated with more severe COVID-19. The results support the growing evidence for Evusheld's efficacy against severe COVID-19 in patients with hematology malignancies.

• MeSH
• COVID-19 * MeSH
• Adult MeSH
• Hematologic Neoplasms * complications   drug therapy   epidemiology   MeSH
• Humans MeSH
• Antibodies, Monoclonal MeSH
• Pre-Exposure Prophylaxis * MeSH
• Retrospective Studies MeSH
• SARS-CoV-2 MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Publication type
• Journal Article MeSH
• Multicenter Study MeSH
• Geographicals
• Czech Republic MeSH

Linoleic acid (LA), an essential fatty acid, has emerged as a pivotal regulator in disorders associated with inflammation in recent years; however, the underlying mechanisms are still not completely understood. We utilized network pharmacology and experimental methodologies to elucidate the mechanisms underlying the anti-inflammatory effects of LA. Our network pharmacology analysis revealed that LA shares common targets with sepsis. These targets are enriched in various pathways comprising C-type signaling pathway, PI3K-Akt signaling pathway, toll-like receptor signaling pathway, neutrophil extracellular trap formation, AMPK signaling pathway, and autophagy-animal. These findings suggest that LA may exert regulatory effects on inflammation and autophagy during sepsis. Subsequently, we established in vivo and ex vivo models of sepsis using lipopolysaccharide (LPS) in experimental study. Treatment with LA reduced lung damage in mice with LPS-induced lung injury, and reduced tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in plasma, bronchoalveolar lavage fluid (BALF), and peritoneal lavage fluid (PLF). LA also decreased the production of TNF-α and IL-6 in RAW264.7 macrophages exposed to LPS. In LPS-induced RAW264.7 macrophages, LA induced an elevation in LC3-II while causing a reduction in p62, which was associated with downregulation of toll-like receptor 4 (TLR4). We utilized 3-methyladenine (3-MA) to inhibit the autophagic activity, which reversed the modulatory effects of LA on LC3-II and p62. 3-MA also prevented the decline in TLR4 expression along with reduction in pro-inflammatory cytokines secretion. Our findings suggest that the activation of autophagy by LA may lead to the downregulation of TLR4, thereby exerting its anti-inflammatory effects.

• MeSH
• Autophagy * drug effects   MeSH
• Linoleic Acid * pharmacology   MeSH
• Lipopolysaccharides * toxicity   MeSH
• Macrophages * drug effects   metabolism   immunology   MeSH
• Mice MeSH
• RAW 264.7 Cells MeSH
• Sepsis chemically induced   drug therapy   metabolism   immunology   MeSH
• Signal Transduction drug effects   MeSH
• Toll-Like Receptor 4 * metabolism   MeSH
• Inflammation * metabolism   drug therapy   chemically induced   pathology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Epigenetic DNA modifications are pivotal in eukaryotic gene expression, but their regulatory significance in bacteria is less understood. In Synechocystis 6803, the DNA methyltransferase M.Ssp6803II modifies the first cytosine in the GGCC motif, forming N4-methylcytosine (GGm4CC). Deletion of the sll0729 gene encoding M.Ssp6803II (∆sll0729) caused a bluish phenotype due to reduced chlorophyll levels, which was reversed by suppressor mutations. Re-sequencing of 7 suppressor clones revealed a common GGCC to GGTC mutation in the slr1790 promoter's discriminator sequence, encoding protoporphyrinogen IX oxidase, HemJ, crucial for tetrapyrrole biosynthesis. Transcriptomic and qPCR analyses indicated aberrant slr1790 expression in ∆sll0729 mutants. This aberration led to the accumulation of coproporphyrin III and protoporphyrin IX, indicative of impaired HemJ activity. To confirm the importance of DNA methylation in hemJ expression, hemJ promoter variants with varying discriminator sequences were introduced into the wild type, followed by sll0729 deletion. The sll0729 deletion segregated in strains with the GGTC discriminator motif, resulting in wild-type-like pigmentation, whereas freshly prepared ∆sll0729 mutants with the native hemJ promoter exhibited the bluish phenotype. These findings demonstrate that hemJ is tightly regulated in Synechocystis and that N4-methylcytosine is essential for proper hemJ expression. Thus, cytosine N4-methylation is a relevant epigenetic marker in Synechocystis and likely other cyanobacteria.

• MeSH
• Bacterial Proteins metabolism   genetics   MeSH
• Epigenesis, Genetic * MeSH
• DNA Methylation * MeSH
• Mutation MeSH
• Promoter Regions, Genetic * MeSH
• Gene Expression Regulation, Bacterial MeSH
• Synechocystis * genetics   metabolism   MeSH
• Tetrapyrroles * metabolism   biosynthesis   MeSH
• Publication type
• Journal Article MeSH