Plasma Components Research Articles

The relationship between disorders of the plasma and platelet components of hemostasis with the severity and timing of coronavirus infection in patients with chronic coronary syndromes

The relationship between disorders of the plasma and platelet components of hemostasis with the severity and timing of coronavirus infection in patients with chronic coronary syndromes

Relationship between semen quality and seminal plasma components: Alpha-glucosidase, fructose and zinc in Morrocan infertile men

Relationship between semen quality and seminal plasma components: Alpha-glucosidase, fructose and zinc in Morrocan infertile men

X-Rays from a Central “Exhaust Vent” of the Galactic Center Chimney

Using deep archival observations from the Chandra X-ray Observatory, we present an analysis of linear X-ray-emitting features located within the southern portion of the Galactic center chimney and oriented orthogonal to the Galactic plane, centered at coordinates l = 0.°08, b = −1.°42. The surface brightness and hardness ratio patterns are suggestive of a cylindrical morphology, which may have been produced by a plasma outflow channel extending from the Galactic center. Our fits of the feature’s spectra favor a complex two-component model consisting of thermal and recombining plasma components, possibly a sign of shock compression or heating of the interstellar medium by outflowing material. Assuming a recombining plasma scenario, we further estimate the cooling timescale of this plasma to be on the order of a few hundred to thousands of years, leading us to speculate that a sequence of accretion events onto the Galactic black hole may be a plausible quasi-continuous energy source to sustain the observed morphology.

Dust magnetoacoustic waves in an inhomogeneous cylindrical four-component dusty plasma in the presence of polarization force

Abstract Dust magnetoacoustic waves have been examined in an inhomogeneous, bounded, cylindrical dusty plasma containing oppositely polarized dust particles. Considering polarization force, dust dynamics in r − θ plane is studied in the presence of inhomogeneous external magnetic field along z axis. At equilibrium, the dusty plasma components are supposed to follow Gaussian density distribution. Using reductive perturbation method (RPM), a variable coefficient cylindrical Kadomtsev–Petviashvili (VCCKP) equation has been derived. For weak azimuthal perturbation, an analytical solution, obtained by Zhang (“Exact solutions of a kdv equation with variable coefficients via exp-function method,” Nonlinear Dynam., vol. 52, nos. 1–2, pp. 11–17, 2008) using Exp-function method, is chosen. Phase velocity of dust magnetoacoustic wave is found to be modified by the density inhomogeneities, polarization force, dust charge state ratio and ion-to-electron temperature ratio. Spatio-temporal evolution of the dust number densities has been noticed. Existence of the compressive electromagnetic solitary waves is observed numerically for the chosen dusty plasma parameter range. The impacts of the inhomogeneity, polarization force, dust charge state ratio and ion-to-electron temperature ratio on the relative amplitude of the dust magnetoacoustic wave are also discussed.

Integrated pharmacokinetic-pharmacodynamic modeling and metabolomic research on polyphenol-rich fraction of Thymus quinquecostatus Celak. Alleviating cerebral ischemia-reperfusion injury

Ethnopharmacological relevanceThymus quinquecostatus Celak., a member of thymus genus in Lamiaceae family, has been used as a folk medicine for relieving exterior syndrome and alleviating pain in China. The polyphenol-rich fraction (PRF) derived from Thymus quinquecostatus Celak. had been validated that it can protect cerebral ischemia-reperfusion injury (CIRI) by activating Keap1/Nrf2/HO-1 signaling pathway.Aim of this study: To explore effective components and their pharmacokinetic and pharmacodynamic characteristics as well as possible mechanisms of PRF in treating CIRI. Materials and methodsNormal treated group (NTG) and tMCAO model treated group (MTG) rats were administrated PRF intragastrically. The prototype components and metabolites of PRF in plasma and brain were analyzed by the UPLC-Q-Exactive Orbitrap MSn method. Subsequently, the pharmacokinetics properties of indicative components were performed based on HPLC-QQQ-MS/MS. SOD and LDH activities were determined to study the pharmacodynamic (PD) properties of PRF. The PK-PD relationship of PRF was constructed. In addition, the effect of PRF on endogenous metabolites in plasma and brain was investigated using metabolomic method. ResultsSalvianic acid A, caffeic acid, rosmarinic acid, scutellarin, and apigenin-7-O-glucuronide were selected as indicative components based on metabolic analysis. The non-compartmental parameters were calculated for indicative components in plasma and brain of NTG and MTG rats. Furthermore, single-component and multi-component PK-PD modeling involved Emax, Imax PD models for effect indexes were fitted as well as ANN models were established, which indicated that these components can work together to regulate SOD and LDH activities in plasma and SOD activity in brain tissue to improve CIRI. Additionally, PRF may ameliorate CIRI by regulating the disorder of endogenous metabolites in lipid metabolism, amino acid metabolism, and purine metabolism pathways in vivo, among which lipid metabolism and purine metabolism are closely related to oxidative stress. ConclusionThe PK-PD properties of effect substances and mechanisms of PRF anti-CIRI were further elaborated. The findings provide a convincing foundation for the application of T. quinquecostatus Celak. in the maintenance of human health disorders.

Photocoagulation or sham laser in addition to conventional anti-VEGF therapy in macular edema associated with TelCaps due to diabetic macular edema or retinal vein occlusion (TalaDME): a study protocol for a multicentric, French, two-group, non-commercial, active-control, observer-masked, non-inferiority, randomized controlled clinical trial

BackgroundMacular edema (ME) results from hyperpermeability of retinal vessels, leading to chronic extravasation of plasma components into the retina and hence potentially severe visual acuity loss. Current standard of care consists in using intravitreal injections (IVI), which results in a significant medical and economic burden. During diabetic retinopathy (DR) or retinal vein occlusion (RVO), it has recently been shown that focal vascular anomalies (capillary macro-aneurysms, also termed TelCaps) for telangiectatic capillaries may play a central role in the onset, early recurrence, and/or persistence of ME. Since targeted photocoagulation of TelCaps may improve vision, identification, and photocoagulation of TelCaps, it may represent a way to improve management of ME.ObjectiveThe Targeted Laser in (Diabetic) Macular Edema (TalaDME) study aims to evaluate whether ICG-guided targeted laser (IGTL), in association with standard of care by IVI, allows reducing the number of injections during the first year of treatment compared with IVI only, while remaining non-inferior for visual acuity.MethodsTalaDME is a French, multicentric, two-arms, randomized, sham laser-controlled, double-masked trial evaluating the effect of photocoagulation of TelCaps combined to IVI in patients with ME associated with TelCaps. Patients with vision loss related to center involved ME secondary to RVO or DR and presenting TelCaps are eligible. Two hundred and seventy eyes of 270 patients are randomized in a 1:1 ratio to standard care, i.e., IVI of anti-VEGF solely (control group) or combined with IGTL therapy (experimental group). Stratification is done on the cause of ME (i.e., RVO versus diabetes). Anti-VEGF IVI are administered to both groups monthly for 3 months (loading dose) and then with a pro re nata regimen with a monthly follow-up for 12 months. The primary endpoint will be the number of IVI and the change in visual acuity from baseline to 12 months. Secondary endpoints will be the changes in central macular thickness, impact on quality of life, cost of treatment, and incremental cost-utility ratio in each groups.Key safetyRare but severe AE linked to the use of IVI and laser, and previously described, are expected. In the sham group, rescue laser photocoagulation may be administered by the unmasked investigator if deemed necessary at month 3.DiscussionThe best management of ME associated with TelCaps is debated, and there have been no randomized study designed to answer this question. Given the fact that TelCaps may affect 30 to 60% of patients with chronic ME due to DR or RVO, a large number of patients could benefit from a specific management of TelCaps. TalaDME aims to establish the clinical and medico-economic benefits of additional targeted laser. The results of TalaDME may raise new recommendations for managing ME and impact healthcare costs.Trial registrationEudraCT: 2018-A00800-55/ NCT03751501. Registration date: Nov. 23, 2018.

Performance assessment of spectroscopic measurements of hydrogen and beryllium influxes and ionization fronts in ITER divertor plasmas using the divertor impurity monitor

Characteristics of Hα and Be II emissions in ITER divertor plasmas have been evaluated, and the applicability of a spectroscopic evaluation method of influxes and ionization front positions by using the divertor impurity monitor (DIM) has been systematically verified for the first time based on plasma profile datasets with parameter scans. In addition, practical values of the ionization/photon factor (S/XB) for DIM to evaluate influxes have been determined. In attached divertor plasmas and the onset of plasma detachment, H influx integrated over the entire area of the divertor can be evaluated within ±30 % accuracy using Hα intensities measured by the DIM and S/XB value of 32, which is higher than the S/XB in most divertor plasmas in previous devices owing to high electron density. The ionization front position can also be evaluated to within a few mm by using peak channel position. However, when electron temperature further decreases from the detachment onset, the ionizing plasma component (caused by electron impact excitation) in the total Hα emission decreases and the spatial profiles of emission and ionization diverge, making spectroscopic evaluation difficult. In all conditions, Hα intensities could be measured with a more than sufficient number of photons at the required time resolution using the current DIM design. For Be influx integrated over the entire divertor area, it has been found that Be I is not suitable for the DIM since its intensity is similar to the background continuous spectra. Replacing Be I with Be II solves this problem. Even in detached divertor plasmas, it is possible to evaluate Be1+ ionization flux within ±30 % accuracy using Be II intensities and S/XB, the value of which depends on the DIM’s fields of view. However, the evaluated Be1+ ionization flux is lower than Be influx by more than 80 %, meaning that the evaluation of Be influx using Be II would be an underestimate. But, Be II can be measured with a more than sufficient number of photons.

Charge-Based Isolation of Extracellular Vesicles from Human Plasma.

Extracellular vesicles (EVs) have garnered significant attention due to their potential applications in disease diagnostics and management. However, the process of isolating EVs, primarily from blood samples, is still suboptimal. This is mainly attributed to the abundant nature of soluble proteins and lipoproteins, which are often separated together with EVs in the end products of conventional isolation methods. As such, we devise a single-step charge-based EV isolation method by utilizing positively charged beads to selectively remove negatively charged major impurities from human plasma via electrostatic interaction. By carefully controlling the buffer pH, we successfully collected EVs from undesired plasma components with superior purity and yield compared to conventional EV collection methods. Moreover, the developed process is rapid, taking only about 20 min for overall EV isolation. The charge-based isolation can ultimately benefit the EV-based liquid biopsy field for the early diagnosis of various diseases.

Enhanced absorption of prenylated cinnamic acid derivatives from Brazilian green propolis by turmeric in humans and rats

AbstractPrenylated cinnamic acid derivatives are the bioactive components of Brazilian green propolis (BGP). The effect of other botanical components on the pharmacokinetic profiles of these derivatives remains relatively unexplored. In the present study, we investigated the influence of several herbal extracts (turmeric, ginkgo leaf, coffee fruit, soybean, and gotu kola) on the plasma concentrations of cinnamic acid derivatives after BGP consumption. When the herbal extracts were co‐administered with BGP in the clinical study, the area under the curve (AUC) values of artepillin C and drupanin, the major BGP components in plasma, were significantly increased by 1.7‐ and 1.5‐fold, respectively, compared to those after BGP administration alone. Among the herbal extracts administered to rats, turmeric extract increased the AUC. Furthermore, a bidirectional transport assay suggested that artepillin C and drupanin are substrates of breast cancer resistance protein (BCRP), a drug elimination transporter. These results suggest that curcumin‐containing turmeric extract may increase the plasma concentrations of artepillin C and drupanin via BCRP. Our findings enabled us to estimate the food–herb and herb–herb interactions in vivo in foods and herbal medicines containing cinnamic acid derivatives and prenylated compounds.

Surface Heating Calculation of Composite Material Under the Influence of an Electron Beam on the Surface

Introduction. Modern plasma magnetic confinement systems use tungsten as a material in contact with plasma. Under the influence of high-density plasma irradiation, tungsten undergoes cracking, intense erosion, and macro-particle emission. High-temperature ceramics are considered a promising material for protective coating of plasma components, as they are resistant to thermal loads. One possible solution could be a boron carbide coating, which has a high melting temperature.Materials and Methods. The impact of an electron beam on samples of rolled tungsten and boron carbide and tungsten composite was studied in experiments on the BETA setup. The heat from the beam propagates into the samples, with the maximum temperature reached at the center and decreasing towards the edges. The modeling area represents a cross-section of the samples, optimal for a task with a cylindrical coordinate system. The numerical implementation is based on the correction scheme and the marching method.Results. A new model of heating the boron carbide and tungsten composite sample under the influence of surface heating by an electron beam is presented. The model is based on solving the heat conduction equation in an axially symmetric setup with constant values of specific heat capacity, density, and thermal conductivity of metals.Discussion and Conclusions. An analysis of the model of heating the composite material under the influence of surface heating by an electron beam at constant values of density, thermal conductivity, and specific heat capacity has been conducted. The modeling results are in demand for analyzing experimental results and planning experiments at the Beam of Electrons for Materials Test Applications (BETA) facility, created at the Budker Institute of Nuclear Physics SB RAS.

Hif-1α expression targets the TMA/Fmo3/TMAO axis to participate in gallbladder cholesterol stone formation in individuals living in plateau regions

The incidence of gallbladder cholesterol stones (GCS) increases rapidly among people living in high-altitude hypoxic environments compared to those in normoxic areas. Upregulation of hepatic hypoxia inducible factor 1α (Hif-1α) plays a key role in the formation of GCS. High plasma trimethylamine-N-oxide (TMAO) levels are positively correlated with the occurrence of GCS. We hypothesized that HIF-1α may upregulate TMAO levels by promoting the transcription of flavin-containing monooxygenase 3 (Fmo3), which eventually leads to GCS formation. Our study shows that in women, high plasma total cholesterol and apolipoprotein B were positively correlated with cholecystolithiasis and hypoxia. Hif-1α binds to the Fmo3 promoter and promotes Fmo3 expression. Hypoxia and lithogenic diet induce the expression of Hif-1α, Fmo3, TMAO and cholesterol tube transporters in the livers of mice, disturb the proportion of bile and plasma components, and induce the formation of GCS. In cell experiments, silencing Hif-1α downregulates the expression of Fmo3, TMAO and cholesterol tube transporters. In a mouse model of hypoxic cholecystolithiasis, silencing Hif-1α downregulates the expression of related genes, restores the proportion of bile and plasma lipid components, and reduces the formation of GCS. Our study shows that Hif-1α binds to the promoter region of Fmo3 and promotes Fmo3 transcription. Thus, it mediates the transcriptional activation of the TMA/Fmo3/TMAO pathway, upregulates the expression of ATP-binding cassettes (Abc) g5 and g8, and participates in the regulation of the occurrence of GCS in the plateau region.

Optical identification and follow-up observations of SRGA J213151.5+491400

Context. The paper is comprised of optical identification and multiwavelength studies of a new X-ray source discovered by the Spectrum Roentgen-Gamma (SRG) observatory during the ART-XC survey and its follow-up optical and X-ray observations. Aims. We aim to identify SRGA J213151.5+491400 in the optical wavelengths. We determine spectra and light curves in the optical high and low states to find periodicities in the light curves and resolve emission lines in the system using optical ground-based data. We intend to study the spectral and temporal X-ray characteristics of the new source using the SRG surveys in the high and low states and NICER data in the low state. Methods. We present optical data from telescopes in Türkiye (RTT-150 and T100 at the TÜBİTAK National Observatory) and in Russia (6-m and 1-m at SAO RAS), together with the X-ray data obtained with ART-XC and eROSITA telescopes aboard SRG and the NICER observatory. Using the optical data, we performed astrometry, photometry, spectroscopy, and power spectral analysis of the optical time series. We present optical Doppler tomography along with X-ray data analysis producing light curves and spectra. Results. We detected SRGA J213151.5+491400 in a high state in 2020 (17.9 mag) that decreased by about 3 mag into a low state (21 mag) in 2021. We find only one significant period using optical photometric time series analysis, which reveals the white dwarf spin (orbital) period to be 0.059710(1) days (85.982 min). The long slit spectroscopy in the high state yields a power-law continuum increasing towards the blue with a prominent He II line along with the Balmer line emissions with no cyclotron humps, which is consistent with a magnetic cataclysmic variable (MCV) nature. Doppler Tomography confirms the polar nature revealing ballistic stream accretion along with magnetic stream during the high state. These characteristics show that the new source is a polar-type MCV source. ART-XC detections yield an X-ray flux of (4.0−7.0) × 10−12 erg s−1 cm−2 in the high state. eROSITA detects a dominating hot plasma component (kTmax > 21 keV in the high state) declining to (4.0−6.0) × 10−13 erg s−1 cm−2 in 2021 (low state). The NICER data obtained in the low state reveal a two-pole accretor showing a soft X-ray component at (6−7)σ significance with a blackbody temperature of 15−18 eV. A soft X-ray component has never been detected for a polar in the low state before.

Impact of Mass‐Loading Effect on the Competition in the Energy Conversion Rate During Magnetic Reconnection

AbstractHeavy particles are extensively detected in the space environment, especially in the solar wind and interplanetary magnetosphere. Various physical processes can be affected by the physical dynamics of heavy particles. One of the processes is magnetic reconnection, which converts the energy from the magnetic field to the particles. In the present study, we investigate the impact of heavy particles with increasing mass (i.e., mass‐loading effect) on energy conversion rate during magnetic reconnection. The declined reconnection rate and energy conversion rates by this effect are captured as reported previously. After considering three major regions in reconnection, it reveals that the mass‐loading effect decreases more energy conversion rate of positive species at the separatrix, affects the heavier species less in the inner electron diffusion region (EDR), and propels the formation of a non‐zero electric field at reconnection front (RF). Our results provide a more comprehensive understanding of the magnetic reconnection with complicated plasma components.

Revisiting the abundance pattern and charge-exchange emission in the centre of M82

The interstellar medium (ISM) in starburst galaxies contains many chemical elements that are synthesised by core-collapse supernova explosions. By measuring the abundances of these metals, we can study the chemical enrichment within the galaxies and the transportation of metals into the circumgalactic environment through powerful outflows. We performed a spectral analysis of the X-ray emissions from the core of M82 using the Reflection Grating Spectrometer (RGS) on board XMM-Newton to accurately estimate the metal abundances in the ISM. We analysed over 300\,ks of RGS data observed with 14 position angles, covering a cross-dispersion width of 80\,arcsec. We employed multi-temperature thermal plasma components in collisional ionisation equilibrium (CIE) to reproduce the observed spectra, each of which exhibited a different spatial broadening. The O VII band CCD image shows a broader distribution that those for the O VIII and Fe-L bands. The O VIII line profiles have a prominent double-peaked structure that corresponds to the north- and southward outflows. The O VII triplet feature exhibits marginal peaks. A single CIE component that is convolved with the O VII band image approximately reproduces the spectral shape. A CIE model combined with a charge-exchange emission model also successfully reproduces the O VII line profiles. However, the ratio of these two components varies significantly with the observed position angles, which is physically implausible. Spectral fitting of the broadband spectra suggests a multi-temperature phase in the ISM that is approximated by three components at 0.1, 0.4, and 0.7\,keV. Notably, the 0.1\,keV component exhibits a broader distribution than the 0.4 and 0.7\,keV plasmas. The derived abundance pattern shows super-solar N/O, solar Ne/O and Mg/O, and half-solar Fe/O ratios. These results indicate the chemical enrichment by core-collapse supernovae in starburst galaxies.

Characterization of lipid peroxidation processes and antioxidant defense in parturients with COVID-19

Aim. The aim of the study was to investigate the functional state of the «lipid peroxidation - antioxidant defense» system in the peripheral blood of parturients with mild and severe COVID-19.Materials and methods. This casecontrol study examined 95 parturients with confirmed COVID-19 diagnosis in the third trimester, divided into two groups based on the severity of the disease. Group 1a included parturients with severe COVID-19 (community-acquired pneumonia, n=48), while group 1b consisted of patients with mild COVID-19 (acute respiratory viral infection, n=47). The control group was composed of 45 parturients without a prior or current diagnosis of COVID-19. The average age of women in the study groups was 23.9±0.50 years, and 24.4±0.60 years in the control group (p>0.05). Peripheral venous blood served as the material for the study. Spectrophotometric methods were used to examine the concentrations of primary (diene conjugates - DC) and final (active thiobarbituric acid products - TBARS) products of lipoperoxidation. The content of 8-isoprostane, total antioxidant capacity of plasma components, and phospholipase A2 were investigated using enzymelinked immunosorbent assay (ELISA).Results. Study showed that parturients with severe COVID-19 had increased levels of phospholipase A2 by 2.1 times (p<0.001), oxidative stress markers: diene conjugate concentrations by 1.94 times (p><0.001), TBARS by 2.2 times (p><0.001), and 8-isoprostane by more than 2 times (p><0.001), along with a simultaneous decrease in the total antioxidant capacity of blood plasma components by 1.7 times (p><0.001) compared to the control group. In the group of parturients with mild COVID-19, the studied indicators changed less significantly. Conclusion. The study established the pathogenetic role of SARS-CoV-2 in disrupting the functional system of «lipid peroxidation - antioxidant defense» and the development of oxidative stress in parturients who had COVID-19 in the third trimester. A dependency of oxidative stress marker concentrations in peripheral blood on the severity of COVID-19 was identified. It was shown that COVID-19 in the third trimester of pregnancy is associated with high levels of phospholipase A2, an imbalance in the prooxidant-antioxidant system, characterized by a reduction in the level of total antioxidant capacity and an increase in oxidative modification products of biomolecules, expressed by the growth of DC, TBARS, and 8-isoprostane levels, leading to oxidative stress correlated with the severity of lung damage. Key words: COVID-19, parturients, lipid peroxidation, antioxidant defense, oxidative stress> ˂ 0.001), oxidative stress markers: diene conjugate concentrations by 1.94 times (p ˂ 0.001), TBARS by 2.2 times (p ˂ 0.001), and 8-isoprostane by more than 2 times (p ˂ 0.001), along with a simultaneous decrease in the total antioxidant capacity of blood plasma components by 1.7 times (p ˂ 0.001) compared to the control group. In the group of parturients with mild COVID-19, the studied indicators changed less significantly.Conclusion. The study established the pathogenetic role of SARS-CoV-2 in disrupting the functional system of «lipid peroxidation - antioxidant defense» and the development of oxidative stress in parturients who had COVID-19 in the third trimester. A dependency of oxidative stress marker concentrations in peripheral blood on the severity of COVID-19 was identified. It was shown that COVID-19 in the third trimester of pregnancy is associated with high levels of phospholipase A2, an imbalance in the prooxidant-antioxidant system, characterized by a reduction in the level of total antioxidant capacity and an increase in oxidative modification products of biomolecules, expressed by the growth of DC, TBARS, and 8-isoprostane levels, leading to oxidative stress correlated with the severity of lung damage.

A Phase I, Open-Label, Mass Balance Study of [14C]-Iberdomide in Healthy Subjects.

Iberdomide is a novel potent cereblon modulator (CELMoD®) agent, which is currently under clinical development for hematological malignancies. A human mass balance study was conducted to characterize the biotransformation and excretion pathways of iberdomide. After a single dose of radiolabelled [14C]-iberdomide (1 mg) in six healthy subjects. Blood, urine, and fecal samples were collected for pharmacokinetics, mass balance, and clinical laboratory assessments. Results showed that a single oral dose of 1 mg iberdomide was generally well tolerated in healthy subjects. The recovery of [14C]-iberdomide-derived radioactivity in humans was 45.9% in urine and 42.6% in feces. Based on exposure (area under the concentration-time curve [AUC0-24]), iberdomide and M12 (metabolites) accounted for approximately 59% and 14% of circulating total radioactivity (TRA) exposure, respectively. Of the 88.5% TRA excreted, approximately 27% was excreted as unchanged iberdomide and 62% as metabolites, with similar amounts of excreted metabolites in the urine (16%) and feces (11%). Biotransformation of iberdomide in humans included multiple oxidations of the morpholino moiety as well as glutarimide ring hydrolysis of parent and oxidized metabolites and a combination of these pathways. Iberdomide was the predominant component in human plasma, with metabolite M12 being the most prominent circulating metabolite. In excreta, similar iberdomide-derived radioactivity was found in urine and feces. NCT03294603.

Abstract 6562: Comprehensive liquid biopsy: Multi-analyte evaluation of cellular and plasma components of blood in prostate cancer from a single blood collection tube

Abstract Background Liquid biopsy samples offer a non-invasive method for investigating drug targets and biomarkers in solid tumors. In this study, we describe a comprehensive liquid biopsy analysis of whole blood to identify circulating tumor cells (CTCs), large extracellular vesicles and CTC fragments, and CTC-white blood cell hybrids; quantify protein biomarker expression on CTCs; perform targeted mutation profiling on CTCs and cell-free DNA (cfDNA) from plasma; and analyze protein expression on white blood cells (WBCs) from a single 7.5 mL blood sample. This array of tests was applied to blood samples from patients with Stage IV prostate cancer to demonstrate clinical feasibility. Methods A novel dual biomarker CTC assay was developed to measure protein expression of PSMA and ARv7 on CTCs from stage IV prostate cancer patients. Nucleated cells from clinical samples were processed to microscope slides using AccuCyte®, an unbiased, density-based separation method that also allows for plasma collection for downstream analysis. Slides were fixed and stained with antibodies to cytokeratin, EpCAM, and CD45 to detect CTCs and exclude WBCs, and with PSMA and ARv7 to evaluate protein biomarker expression on identified CTCs. The slides were imaged with the CyteFinder® automated immunofluorescence scanning microscope that identifies CTCs for confirmation by trained reviewers and quantitatively measures protein mean fluorescence intensity (MFI) on any cells detected on the slides (CTCs, CTC-WBC hybrids, large extracellular vesicles and CTC fragments and WBCs). Individual CTCs and control pools of WBCs from clinical samples with at least 4 CTCs were picked using CytePicker®, a needle-based system used to isolate single cells. cfDNA was isolated from plasma removed during the AccuCyte® process using a QIAamp MinElute ccfDNA kit. Picked CTCs, WBC pools, and isolated ctDNA were sequenced using the OncoZoom Cancer HotSpot panel from Paragon Genomics. Results Nine progressing Stage IV prostate cancer patient blood samples were stained with a novel PSMA/ARv7 dual biomarker assay. CTCs were identified in all clinical samples (median 4, range 1 - 384). PSMA positivity in clinical samples ranged from 0% to 75%, and ARv7 positivity ranged from 33% to 100%, and. Additional analysis using QuPath was performed to determine PSMA/ARv7 protein expression on WBCs and CTC fragments. ctDNA analysis from 5 individual samples was compared to sequencing results from individual CTCs from the same samples. Conclusions Using a single 7.5 mL blood liquid biopsy, we can measure protein expression on enumerated CTCs and quantify these same protein biomarkers on WBCs and CTC fragments/oncosomes, determine the genetic makeup of the tumor through the sequencing of cfDNA, and monitor genetic mutations in intact CTCs in circulation. Citation Format: Jon Ladd, Erin Bayer, Brock Bartels, Arista Tischner, Rachel Ponting, Eric Kaldjian, Arturo Ramirez. Comprehensive liquid biopsy: Multi-analyte evaluation of cellular and plasma components of blood in prostate cancer from a single blood collection tube [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6562.

Pharmacokinetics, mass balance, and metabolism of [14C]envonalkib (TQ-B3139), a novel ALK tyrosine kinase inhibitor, in healthy Chinese subjects.

Envonalkib (TQ-B3139) is a novel, potent anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitor used to treat ALK-positive non-small cell lung cancer. This phase I mass balance study investigated the pharmacokinetics, metabolism, and excretion of 14C-radiolabeled envonalkib in healthy Chinese male subjects. A single oral dose of 600mg (150µCi) [14C]envonalkib was administered to healthy male subjects under fasted state. Samples of blood, urine and feces were collected for quantitative determination of total radioactivity and unchanged envonalkib, and the metabolites identification. After dosing, the median Tmax of radioactivity was 4h and the mean t1/2 was 65.2h in plasma. The exposure of total radioactivity was much higher than that of unchanged envonalkib in plasma. The mean total recovery of the radiolabeled dose was 93.93% over 504h post-dose, with 15.23% in urine and 78.71% in feces. Envonalkib underwent extensive metabolism and a total of 15 metabolites were identified in plasma, urine, and feces. Unchanged envonalkib and its major metabolite M315 were the main components in plasma, accounting for 20.37% and 33.33% of total plasma radioactivity. In urine, O-dealkylation metabolite M315 was the major component accounted for 7.98% of dose. In feces, 16.01% of dose was excreted as cysteine conjugate M434-1. Envonalkib was well tolerated and there were no serious adverse events observed in the study. Envonalkib was extensively metabolized prior to excretion and eliminated primarily as metabolites via feces.

Quantitative analysis of mRNA-lipid nanoparticle stability in human plasma and serum by size-exclusion chromatography coupled with dual-angle light scattering

Understanding the stability of mRNA loaded lipid nanoparticles (mRNA-LNPs) is imperative for their clinical development. Herein, we propose the use of size-exclusion chromatography coupled with dual-angle light scattering (SEC-MALS) as a new approach to assessing mRNA-LNP stability in pure human serum and plasma. By applying a dual-column configuration to attenuate interference from plasma components, SEC-MALS was able to elucidate the degradation kinetics and physical property changes of mRNA-LNPs, which have not been observed accurately by conventional dynamic light scattering techniques. Interestingly, both serum and plasma had significantly different impacts on the molecular weight and radius of gyration of mRNA-LNPs, suggesting the involvement of clotting factors in desorption of lipids from mRNA-LNPs. We also discovered that a trace impurity (~1 %) in ALC-0315, identified as its O-tert-butyloxycarbonyl-protected form, greatly diminished mRNA-LNP stability in serum. These results demonstrated the potential utility of SEC-MALS for optimization and quality control of LNP formulations.

Factors to take into account when interpreting 25-hydroxy-vitamin D serum levels

ABSTRACT Background Assessing vitamin D status, typically evaluated using serum or plasma 25-hydroxy vitamin D [25(OH)D] concentration, is complex because of various influencing factors. Methods Seasonality significantly affects intra-individual variability in 25(OH)D levels. This variation can be addressed by employing cosinor functions that are tailored to the geographical location of the patient to correct for seasonal effects. In addition to seasonality, genetic factors, such as DBP polymorphism and body composition, particularly adiposity, play crucial roles. Dialysis patients with DBP 2–2 phenotype exhibit higher vitamin D requirements. Genotyping/phenotyping of DBP allows for better tailored vitamin D supplementation. The lipid-soluble nature of vitamin D also interacts with plasma components such as serum triglycerides, which can influence vitamin D measurements. Adiposity, which is negatively correlated with vitamin D concentration, necessitates body mass-based mathematical adjustments for accurate vitamin D assessment in subjects with extreme BMI values. Conclusions Accordingly, vitamin D replacement therapy must be personalized, taking into account factors such as body size and seasonal variations, to effectively reach the target serum 25(OH)D concentrations.