BACKGROUND: To validate the clinical utility of a previously identified circulating tumor DNA methylation marker (meth-ctDNA) panel for disease detection and survival outcomes, meth-ctDNA markers were compared to PSA levels and PSMA PET/CT findings in men with different stages of prostate cancer (PCa). METHODS: 122 PCa patients who underwent [68Ga]Ga-PSMA-11 PET/CT and plasma sampling (03/2019-08/2021) were analyzed. cfDNA was extracted, and a panel of 8 individual meth-ctDNA markers was queried. PET scans were qualitatively and quantitatively assessed. PSA and meth-ctDNA markers were compared to PET findings, and their relative prognostic value was evaluated. RESULTS: PSA discriminated best between negative and tumor-indicative PET scans in all (AUC 0.77) and hormone-sensitive (hsPC) patients (0.737). In castration-resistant PCa (CRPC), the meth-ctDNA marker KLF8 performed best (AUC 0.824). CHST11 differentiated best between non- and metastatic scans (AUC 0.705) overall, KLF8 best in hsPC and CRPC (AUC 0.662, 0.85). Several meth-ctDNA markers correlated low to moderate with the tumor volume in all (5/8) and CRPC patients (6/8), while PSA levels correlated moderately to strongly with the tumor volume in all groups (all p < 0.001). CRPC overall survival was independently associated with LDAH and PSA (p = 0.0168, p < 0.001). CONCLUSION: The studied meth-ctDNA markers are promising for the minimally-invasive detection and prognostication of CRPC but do not allow for clinical characterization of hsPC. Prospective studies are warranted for their use in therapy response and outcome prediction in CRPC and potential incremental value for PCa monitoring in PSA-low settings.
- MeSH
- Circulating Tumor DNA genetics blood MeSH
- Edetic Acid analogs & derivatives MeSH
- Gallium Isotopes * MeSH
- Middle Aged MeSH
- Humans MeSH
- DNA Methylation * genetics MeSH
- Biomarkers, Tumor * genetics blood MeSH
- Prostatic Neoplasms, Castration-Resistant genetics blood diagnostic imaging MeSH
- Prostatic Neoplasms * genetics blood diagnostic imaging MeSH
- Positron Emission Tomography Computed Tomography * methods MeSH
- Prognosis MeSH
- Prostate-Specific Antigen * blood genetics MeSH
- Cross-Sectional Studies MeSH
- Gallium Radioisotopes * MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged, 80 and over MeSH
- Aged MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
BACKGROUND: Antimicrobial lock therapy is recommended for preventing and treating catheter-related bloodstream infections, but different solutions have uncertain efficacy. METHODS: Two locks, 1.35% taurolidine and 4% ethylenediaminetetraacetic acid (EDTA), were tested on Staphylococcus epidermidis, Staphylococcus aureus, methicillin-resistant S. aureus, Pseudomonas aeruginosa, multidrug-resistant P. aeruginosa, vancomycin-resistant Enterococcus faecium, Klebsiella oxytoca (carbapenemase producing), K. pneumoniae (extended-spectrum β-lactamase producing), Candida albicans, and Candida glabrata. Broviac catheter segments were incubated with these organisms and then exposed to various lock solutions. Colony-forming units (CFUs) were counted after 2, 4, and 24 h of incubation. RESULTS: Taurolidine showed a significant decrease in CFUs after 2 h in S. aureus, S. epidermidis, methicillin-resistant S. aureus, vancomycin-resistant E. faecium, P. aeruginosa (both sensitive and multidrug-resistant strains), K. oxytoca, C. albicans, and C. glabrata. After 4 h, significant reductions were noted in S. aureus, S. epidermidis, methicillin-resistant S. aureus, P. aeruginosa, multidrug-resistant P. aeruginosa, K. pneumoniae, K. oxytoca, and C. albicans. Taurolidine was also effective after 24 h, especially against methicillin-resistant S. aureus and multidrug-resistant P. aeruginosa. Four percent EDTA acid showed a significant reduction in CFUs after 2 h in S. aureus, vancomycin-resistant E. faecium, P. aeruginosa, K. oxytoca, C. albicans, and C. glabrata. After 4 h, reductions occurred in P. aeruginosa, multidrug-resistant P. aeruginosa, K. oxytoca, and C. albicans and after 24 h in methicillin-resistant S. aureus, P. aeruginosa, and K. oxytoca. CONCLUSION: Taurolidine is more effective than 4% EDTA acid in eradicating Gram-positive and Gram-negative microorganisms and fungi.
- MeSH
- Anti-Infective Agents * pharmacology MeSH
- Candida albicans drug effects MeSH
- Edetic Acid * pharmacology MeSH
- Catheter-Related Infections * prevention & control microbiology MeSH
- Humans MeSH
- Methicillin-Resistant Staphylococcus aureus drug effects MeSH
- Microbial Sensitivity Tests MeSH
- Colony Count, Microbial MeSH
- Pseudomonas aeruginosa drug effects MeSH
- Staphylococcus aureus drug effects MeSH
- Staphylococcus epidermidis drug effects MeSH
- Taurine * analogs & derivatives pharmacology MeSH
- Thiadiazines * pharmacology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
CXCL12 and CXCR4 proteins and mRNAs were monitored in the dorsal root ganglia (DRGs) of lumbar (L4-L5) and cervical (C7-C8) spinal segments of naïve rats, rats subjected to sham operation, and those undergoing unilateral complete sciatic nerve transection (CSNT) on post-operation day 7 (POD7). Immunohistochemical, Western blot, and RT-PCR analyses revealed bilaterally increased levels of CXCR4 protein and mRNA in both lumbar and cervical DRG neurons after CSNT. Similarly, CXCL12 protein levels increased, and CXCL12 mRNA was upregulated primarily in lumbar DRGs ipsilateral to the nerve lesion. Intrathecal application of the CXCR4 inhibitor AMD3100 following CSNT reduced CXCL12 and CXCR4 protein levels in cervical DRG neurons, as well as the length of afferent axons regenerated distal to the ulnar nerve crush. Furthermore, treatment with the CXCR4 inhibitor decreased levels of activated Signal Transducer and Activator of Transcription 3 (STAT3), a critical transforming factor in the neuronal regeneration program. Administration of IL-6 increased CXCR4 levels, whereas the JAK2-dependent STAT3 phosphorylation inhibitor (AG490) conversely decreased CXCR4 levels. This indicates a link between the CXCL12/CXCR4 signaling axis and IL-6-induced activation of STAT3 in the sciatic nerve injury-induced pro-regenerative state of cervical DRG neurons. The role of CXCR4 signaling in the axon-promoting state of DRG neurons was confirmed through in vitro cultivation of primary sensory neurons in a medium supplemented with CXCL12, with or without AMD3100. The potential involvement of conditioned cervical DRG neurons in the induction of neuropathic pain is discussed.
- MeSH
- Benzylamines MeSH
- Chemokine CXCL12 * metabolism MeSH
- Cyclams pharmacology MeSH
- Heterocyclic Compounds pharmacology MeSH
- Interleukin-6 metabolism MeSH
- Rats MeSH
- Sciatic Neuropathy metabolism MeSH
- Sensory Receptor Cells * metabolism MeSH
- Sciatic Nerve * injuries metabolism MeSH
- Rats, Sprague-Dawley MeSH
- Receptors, CXCR4 * metabolism MeSH
- Nerve Regeneration * MeSH
- Signal Transduction * MeSH
- Ganglia, Spinal * metabolism MeSH
- STAT3 Transcription Factor * metabolism MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
Aminophylline, a bronchodilator mainly used to treat severe asthma attacks, may induce arrhythmias. Unfortunately, the underlying mechanism is not well understood. We have recently described a significant, on average inhibitory effect of aminophylline on inward rectifier potassium current IK1, known to substantially contribute to arrhythmogenesis, in rat ventricular myocytes at room temperature. This study was aimed to examine whether a similar effect may be observed under clinically relevant conditions. Experiments were performed using the whole cell patch clamp technique at 37°C on enzymatically isolated healthy porcine and failing human ventricular myocytes. The effect of clinically relevant concentrations of aminophylline (10-100 μM) on IK1 did not significantly differ in healthy porcine and failing human ventricular myocytes. IK1 was reversibly inhibited by ∼20 and 30 % in the presence of 30 and 100 μM aminophylline, respectively, at -110 mV; an analogical effect was observed at -50 mV. To separate the impact of IK1 changes on AP configuration, potentially interfering ionic currents were blocked (L-type calcium and delayed rectifier potassium currents). A significant prolongation of AP duration was observed in the presence of 100 μM aminophylline in porcine cardiomyocytes which well agreed with the effect of a specific IK1 inhibitor Ba2+ (10 μM) and with the result of simulations using a porcine ventricular cell model. We conclude that the observed effect of aminophylline on healthy porcine and failing human IK1 might be involved in its proarrhythmic action. To fully understand the underlying mechanism, potential aminophylline impact on other ionic currents should be explored.
- MeSH
- Action Potentials drug effects MeSH
- Aminophylline * pharmacology MeSH
- Potassium Channels, Inwardly Rectifying * metabolism MeSH
- Myocytes, Cardiac * drug effects metabolism MeSH
- Humans MeSH
- Patch-Clamp Techniques MeSH
- Swine MeSH
- Heart Ventricles drug effects metabolism MeSH
- Heart Failure metabolism drug therapy MeSH
- Dose-Response Relationship, Drug MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
The combination of aminophylline and salbutamol is frequently used in clinical practice in the treatment of obstructive lung diseases. While the side effects (including arrhythmias) of the individual bronchodilator drugs were well described previously, the side effects of combined treatment are almost unknown. We aimed to study the arrhythmogenic potential of combined aminophylline and salbutamol treatment in vitro. For this purpose, we used the established atomic force microscopy (AFM) model coupled with cardiac organoids derived from human pluripotent stem cells (hPSC-CMs). We focused on the chronotropic, inotropic, and arrhythmogenic effects of salbutamol alone and aminophylline and salbutamol combined treatment. We used a method based on heart rate/beat rate variability (HRV/BRV) analysis to detect arrhythmic events in the hPSC-CM based AFM recordings. Salbutamol and aminophylline had a synergistic chronotropic and inotropic effect compared to the effects of monotherapy. Our main finding was that salbutamol reduced the arrhythmogenic effect of aminophylline, most likely mediated by endothelial nitric oxide synthase activated by beta-2 adrenergic receptors. These findings were replicated and confirmed using hPSC-CM derived from two cell lines (CCTL4 and CCTL12). Data suggest that salbutamol as an add-on therapy may not only deliver a bronchodilator effect but also increase the cardiovascular safety of aminophylline, as salbutamol reduces its arrhythmogenic potential.
- MeSH
- Albuterol * pharmacology MeSH
- Aminophylline * pharmacology MeSH
- Bronchodilator Agents pharmacology MeSH
- Cell Line MeSH
- Myocytes, Cardiac drug effects metabolism MeSH
- Humans MeSH
- Microscopy, Atomic Force MeSH
- Pluripotent Stem Cells drug effects cytology MeSH
- Arrhythmias, Cardiac * drug therapy MeSH
- Heart Rate drug effects MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
Neonatal hypoxic-ischemic (HI) brain insult is a major cause of neonatal mortality and morbidity. To assess the underlying pathological mechanisms, we mapped the spatiotemporal changes in polyamine, amino acid, and neurotransmitter levels, following HI insult (by the Rice-Vannucci method) in the brains of seven-day-old rat pups. Matrix-assisted laser desorption/ionization mass spectrometry imaging of chemically modified small-molecule metabolites by 4-(anthracen-9-yl)-2-fluoro-1-methylpyridin-1-ium iodide revealed critical HI-related metabolomic changes of 22 metabolites in 14 rat brain subregions, much earlier than light microscopy detected signs of neuronal damage. For the first time, we demonstrated excessive polyamine oxidation and accumulation of 3-aminopropanal in HI neonatal brains, which was later accompanied by neuronal apoptosis enhanced by increases in glycine and norepinephrine in critically affected brain regions. Specifically, putrescine, cadaverine, and 3-aminopropanal increased significantly as early as 12 h postinsult, mainly in motor and somatosensory cortex, hippocampus, and midbrain, followed by an increase in norepinephrine 24 h postinsult, which was predominant in the caudate putamen, the region most vulnerable to HI. The decrease of γ-aminobutyric acid (GABA) and the continuous dysregulation of the GABAergic system together with low taurine levels up to 36 h sustained progressive neurodegenerative cellular processes. The molecular alterations presented here at the subregional rat brain level provided unprecedented insight into early metabolomic changes in HI-insulted neonatal brains, which may further aid in the identification of novel therapeutic targets for the treatment of neonatal HI encephalopathy.
- MeSH
- Rats MeSH
- Metabolomics MeSH
- Brain * metabolism MeSH
- Hypoxia-Ischemia, Brain * metabolism pathology MeSH
- Neurons metabolism MeSH
- Neurotransmitter Agents * metabolism MeSH
- Animals, Newborn * MeSH
- Polyamines * metabolism MeSH
- Rats, Sprague-Dawley MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
BACKGROUND: Circulating-tumor DNA (ctDNA) and prostate-specific membrane antigen (PSMA) ligand positron-emission tomography (PET) enable minimal-invasive prostate cancer (PCa) detection and survival prognostication. The present study aims to compare their tumor discovery abilities and prognostic values. METHODS: One hundred thirty men with confirmed PCa (70.5 ± 8.0 years) who underwent [68Ga]Ga-PSMA-11 PET/CT (184.8 ± 19.7 MBq) imaging and plasma sample collection (March 2019-August 2021) were included. Plasma-extracted cell-free DNA was subjected to whole-genome-based ctDNA analysis. PSMA-positive tumor lesions were delineated and their quantitative parameters extracted. ctDNA and PSMA PET/CT discovery rates were compared, and the prognostic value for overall survival (OS) was evaluated. RESULTS: PSMA PET discovery rates according to castration status and PSA ranges did differ significantly (P = 0.013, P < 0.001), while ctDNA discovery rates did not (P = 0.311, P = 0.123). ctDNA discovery rates differed between localized and metastatic disease (P = 0.013). Correlations between ctDNA concentrations and PSMA-positive tumor volume (PSMA-TV) were significant in all (r = 0.42, P < 0.001) and castration-resistant (r = 0.65, P < 0.001), however not in hormone-sensitive patients (r = 0.15, P = 0.249). PSMA-TV and ctDNA levels were associated with survival outcomes in the Logrank (P < 0.0001, P < 0.0001) and multivariate Cox regression analysis (P = 0.0023, P < 0.0001). CONCLUSION: These findings suggest that PSMA PET imaging outperforms ctDNA analysis in detecting prostate cancer across the whole spectrum of disease, while both modalities are independently highly prognostic for survival outcomes.
- MeSH
- Circulating Tumor DNA * blood genetics MeSH
- Edetic Acid * analogs & derivatives MeSH
- Gallium Isotopes * MeSH
- Middle Aged MeSH
- Humans MeSH
- Prostatic Neoplasms * diagnostic imaging genetics blood MeSH
- Oligopeptides MeSH
- Positron Emission Tomography Computed Tomography * MeSH
- Prognosis MeSH
- Cross-Sectional Studies MeSH
- Gallium Radioisotopes * MeSH
- Aged MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
Multidrug resistant (MDR) bacteria are recognized to be one of the most important problems in public health. The outer membrane permeability is a critical intrinsic mechanism of bacterial resistance. In addition, bacteria produce a small number of dormant persister cells causing multidrug tolerance that reduces antimicrobial efficacy. This study aimed to evaluate the inhibitory effects of the combination of aromatic isothiocyanates (ITCs) with membrane-active agents on bacterial persisters and MDR Gram-negative bacteria. Our study demonstrated that membrane-active agents, particularly ethylenediaminetetraacetic acid (EDTA) synergistically enhanced the inhibitory activity of aromatic benzyl ITC and phenethyl ITC against most Gram-negative bacteria strains with fractional inhibitory concentration index values ranging from 0.18 to 0.5 and 0.16 to 0.5, respectively, and contributed to an 8- to 64-fold minimal inhibitory concentration reduction compared with those of aromatic ITCs alone. The EDTA-aromatic ITCs combination effectively reduced the survival rates of tested bacteria and significantly eradicated bacterial persisters (p = 0.033 and 0.037, respectively). The growth kinetics analysis also supported the enhanced inhibitory effect of EDTA-aromatic ITCs combination against tested bacteria. Our results suggested an alternate treatment strategy against Gram-negative bacteria, promoting the entry of aromatic ITCs into bacterial cytoplasm to facilitate bacterial clearance and thus preventing the development of bacterial resistance.
- MeSH
- Anti-Bacterial Agents * pharmacology MeSH
- Edetic Acid * pharmacology MeSH
- Gram-Negative Bacteria * drug effects MeSH
- Isothiocyanates * pharmacology MeSH
- Microbial Sensitivity Tests * MeSH
- Microbial Viability drug effects MeSH
- Drug Resistance, Multiple, Bacterial * drug effects MeSH
- Cell Membrane Permeability drug effects MeSH
- Drug Synergism MeSH
- Bacterial Outer Membrane drug effects MeSH
- Publication type
- Journal Article MeSH
Functional imaging with prostate-specific membrane antigen (PSMA) ligands has emerged as the standard imaging method for prostate cancer (PCA). In parallel, the analysis of blood-derived, cell-free DNA (cfDNA) has been shown to be a promising quantitative biomarker of PCA aggressiveness and patient outcome. This study aimed to evaluate the relationship and prognostic value of cfDNA concentrations and the PSMA-positive tumor volume (PSMA-TV) in men with PCA undergoing [68Ga]Ga-PSMA-11 PET/CT imaging. Methods: We recruited 148 men with histologically proven PCA (mean age, 70.7 ± 7.7 y) who underwent [68Ga]Ga-PSMA-11 PET/CT (184.9 ± 18.9 MBq) and blood sampling between March 2019 and August 2021. Among these, 74 (50.0%) had hormone-sensitive PCA and 74 (50.0%) had castration-resistant PCA (CRPC). All patients provided written informed consent before blood sample collection and imaging. The cfDNA was extracted and quantified, and PSMA-expressing tumor lesions were delineated to extract the PSMA-TVs. The Spearman coefficient assessed correlations between PSMA-TV and cfDNA concentrations and cfDNA's relation with clinical parameters. The Kruskal-Wallis test examined the mean cfDNA concentration differences based on PSMA-TV quartiles for significantly correlated patient groups. Log-rank and multivariate Cox regression analyses evaluated the prognostic significance of high and low cfDNA and PSMA-TV levels for overall survival. Results: Weak positive correlations were found between cfDNA concentration and PSMA-TV in the overall group (r = 0.16, P = 0.049) and the CRPC group (r = 0.31, P = 0.007) but not in hormone-sensitive PCA patients (r = -0.024, P = 0.837). In the CRPC cohort, cfDNA concentrations significantly differed between PSMA-TV quartiles 4 and 1 (P = 0.002) and between quartiles 4 and 2 (P = 0.016). Survival outcomes were associated with PSMA-TV (P < 0.0001, P = 0.004) but not cfDNA (P = 0.174, P = 0.12), as per the log-rank and Cox regression analysis. Conclusion: These findings suggest that cfDNA might serve as a biomarker of advanced, aggressive CRPC but does not reliably reflect total tumor burden or prognosis. In comparison, [68Ga]Ga-PSMA-11 PET/CT provides a highly granular and prognostic assessment of tumor burden across the spectrum of PCA disease progression.
- MeSH
- Biomarkers MeSH
- Edetic Acid MeSH
- Hormones MeSH
- Gallium Isotopes MeSH
- Middle Aged MeSH
- Humans MeSH
- Prostatic Neoplasms, Castration-Resistant * diagnostic imaging MeSH
- Prostatic Neoplasms * diagnostic imaging pathology MeSH
- Positron Emission Tomography Computed Tomography methods MeSH
- Prognosis MeSH
- Prospective Studies MeSH
- Gallium Radioisotopes MeSH
- Retrospective Studies MeSH
- Aged MeSH
- Tumor Burden MeSH
- Cell-Free Nucleic Acids * MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Publication type
- Journal Article MeSH
Impairments of N-methyl-D-aspartate receptor (NMDAR) activity have been implicated in several neuropsychiatric disorders, with pharmacological inhibition of NMDAR-mediated currents and associated neurobehavioral changes considered as a model of schizophrenia. We analyzed the effects of brief and long-term exposure of rat cortical cultures to the most prevalent endogenous modulators of NMDAR (kynurenic acid, pregnenolone sulfate, spermidine, and zinc) on neuronal viability, stimulation-induced release of glutamate, and dendritic morphology with synaptic density. Both, glutamate release and neuronal viability studies revealed no difference between the test and control groups. No differences were also observed in the number of dendritic branching and length, or density of synaptic connections and neuronal soma size. Comparison of the extent of dendritic projections and branching patterns, however, revealed enhanced distal arborization with the expansion of the dendritic area under prolonged treatment of cultures with physiological concentrations of NMDAR modulators, with differences reaching significance in spermidine and pregnenolone sulfate tests. Measurements of the density of glutamatergic synapses showed consistency across all neuronal groups, except those treated with pregnenolone sulfate, which showed a reduction of PSD-95-positive elements. Overall, our data suggest that constitutive glutamatergic activity mediated by NMDAR controls the dendritic field expansion and can influence the integrative properties of cortical neurons.
- MeSH
- Glutamates MeSH
- Rats MeSH
- Neurons metabolism MeSH
- Receptors, N-Methyl-D-Aspartate * metabolism MeSH
- Signal Transduction MeSH
- Spermidine * MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
