Blood Plasma Samples Research Articles

Inner-filter effect of nitrogen-doped carbon quantum dots-MnO2 nanotubes for smartphone-integrated dual-mode sensing of glutathione and captopril.

Nitrogen-doped carbon quantum dots (N-CQDs) exhibit unique fluorescence properties and are considered one of the best candidates for the development of fluorescence-based sensors for the detection of many analytes. In this work, a smartphone-assisted fluorescent sensor has been developed using N-CQDs and MnO2 nanotubes (MnO2 NTs) for the detection of glutathione (GSH) and captopril (CAP). N-CQDs were facilely synthesized via the solvothermal method, where o-phenylenediamine (o-PD) and urea were used as nitrogen precursors. Likewise, MnO2 NTs were synthesized using the hydrothermal method. Relying on the excellent fluorescence quenching ability of MnO2 NTs, a nanocomposite of N-CQDs and MnO2 NTs is prepared, wherein the fluorescence intensity of N-CQDs was effectively quenched in the presence of MnO2 NTs via the inner-filter effect (IFE). The addition of thiolated compounds (GSH and CAP) helped in the recovery of the fluorescence of N-CQDs by triggering the redox reaction and decomposing the MnO2 NTs. An investigation of fluorescence along with smartphone-based studies by evaluating the gray measurement using Image J software showed a great response towards GSH and CAP providing LODs of 4.70 μM and 5.22 μM (fluorometrically) and 5.76 μM and 2.81 μM (smartphone-based), respectively. The practical applicability of the sensing system has been verified using human blood plasma samples.

An electrografted monolayer of polyaniline as a tuneable platform for a glucose biosensor.

Polyaniline (PANI), a nanostructured conducting polymer, has shown significant potential in optical and bioelectrochemical devices. However, its performance and stability on various substrates are hindered by weak adhesion to the surface. In this study, a strongly adherent polyaniline conducting polymer layer with a thickness of five nanometers was electrografted onto an initiating monolayer on gold and tin-doped indium oxide substrates. These electrografted monolayers consist of vertically oriented fully oxidized-protonated (pernigraniline salt) and deprotonated (pernigraniline base) forms of polyaniline. The monolayer exhibits pH-dependent colour changes and it is suitable for enzyme compatibility. In light of these findings, we have developed and characterized an electrochemical glucose biosensor based on the monolayer of polyaniline on a gold electrode. The biosensor utilizes glucose oxidase as the biorecognition element for the selective detection of glucose concentrations in real blood plasma samples.

Luminescent and time-resolved determination of gemifloxacin mesylate in pharmaceutical formulations and spiked blood plasma samples using a lanthanide complex as a probe.

A novel luminescence-based analytical methodology was established employing a europium(III) complex with 3-allyl-2-hydroxybenzohydrazide (HAZ) as the coordinating ligand for the quantification of gemifloxacin mesylate (GMF) in pharmaceutical preparations and human plasma samples spiked with the compound. The stoichiometry of the europium complex with HAZ was determined via the Job plot and exhibited a metal-to-ligand ratio of 1 : 2. The analytical procedure relies on a rapid and significant enhancement of luminescence by the Eu(AZ)2 complex when it interacts with gemifloxacin mesylate, which allowed for the rapid detection of 96 samples within approximately 2 minutes. The thermodynamic parameters of the complexation of GMF with Eu(AZ)2 were evaluated and showed that the complexation of GMF was spontaneous with a negative ΔG. The binding constant K was 4.27 × 105 L mol-1 and DFT calculations supported GMF binding and the formation of Eu(AZ)2-GMF without further ligand exchange. The calibration graph for the luminescence quantitation of GMF was linear over a wide concentration range of 0.11-16 μg mL-1 (2.26 × 10-7 to 3.30 × 10-5 mol L-1), with a limit of quantification (LOQ) of 110 ng mL-1 (230 nmol L-1) and a detection limit (LOD) of 40 ng mL-1 (82 nmol L-1). The proposed method showed good accuracy with an average recovery of 99% with relative standard deviations of less than 5% in spiking experiments, even in complex pharmaceutical dosage forms such as tablets and in human blood plasma. Herein, the ability of the suppression of the luminescence background by using the long lag times of the lanthanide probe in a time-resolved detection scheme provided reliable and precise results, which suggests its potential for use in further real or patient samples.

Variability of haptoglobin beta-chain proteoforms.

Existing knowledge on changes of the haptoglobin (Hp) molecule suggests that it may exist in multiple proteoforms, which obviously exhibit different functions. Using two-dimensional electrophoresis (2DE) in combination with mass spectrometry and immunodetection, we have analyzed blood plasma samples from both healthy donors and patients with primary grade IV glioblastoma (GBM), and obtained a detailed composite 2DE distribution map of β-chain proteoforms, as well as the full-length form of Hp (zonulin). Although the total level of plasma Hp exceeded normal values in cancer patients (especially patients with GBM), the presence of particuar proteoforms, detected by their position on the 2DE map, was very individual. Variability was found in both zonulin and the Hp β-chain. The presence of an alkaline form of zonulin in plasma can be considered a conditional, but insufficient, GBM biomarker. In other words, we found that at the level of minor proteoforms of Hp, even in normal conditions, there was a high individual variability. On the one hand, this raises questions about the reasons for such variability, if it is present not only in Hp, but also in other proteins. On the other hand, this may explain the discrepancy between the number of experimentally detected proteoforms and the theoretically possible ones not only in Hp, but also in other proteins.

Heat inactivation does not alter host plasma cell-free DNA characteristics in infectious disease research

BackgroundCell-free DNA (cfDNA) is a promising analyte for non-invasive liquid biopsy, carrying abundant signatures for disease diagnosis and monitoring. In infectious disease researches, blood plasma samples are routinely heat-inactivated before proceeding with downstream analyses. However, the effects of heat inactivation on cfDNA fragmentomic analysis remain largely unclear, potentially introducing biases or altering the characteristics of cfDNA. MethodsWe performed a comprehensive investigation of cfDNA concentrations and fragmentomics in 21 plasma samples from 7 healthy individuals, by comparing the sample group without the heat inactivation to those exposed to once or twice heat-inactivation at 56 °C for 30 min and following freeze–thaw. ResultsPlasma samples with once and twice heat inactivation displayed no significant deviations in primary characteristics, including cfDNA concentrations, size profiles, end motif features, and genome-wide distributions, compared to samples without heat treatment. ConclusionsHeat-inactivated cfDNA can be utilized for liquid biopsy in infectious disease researches, without substantial impact on cfDNA concentrations and fragmentomic properties. This study provides essential insights into the effects of heat inactivation on cfDNA properties and will contribute to the development of reliable non-invasive biomarkers for infectious disease.

The influence of increased protein intake and L-carnitine supply on the outcomes of term newborns in intensive therapy

We studied the possibilities of increased protein and L-carnitine supplementation in full-term newborns with perinatal diseases to improve the results of intensive care. Aim: to assess the impact of short-term increased protein consumption and L-carnitine supplementation by full-term newborns on the main indicators of treatment effectiveness in the intensive care unit. Materials and methods. To check the impact of the proposed method of feeding newborns, the life indicators of 59 full-term children, who were randomly divided into two groups, were studied. The first group (n = 30) received standard nutrition with mother’s milk or formula, the second group (n = 29) received a food product fortified with a protein supplement and a subsidy of L-carnitine during hospital stay. Results. Infants of both groups had statistically equivalent anthropometric characteristics at the beginning of the study. The proportions of infants of both sexes were within the 50 % percentile. Starting from the third week, caloric intake for the standard diet group (SG) and the enrichment group (FG), respectively, was 111.83 ± 7.19 kcal/kg/day, against 113.44 ± 5.61 kcal/kg/day, p = 0.0667. Per 1 g of protein, children in the FG group had amount of non-protein calories as 28.68 (27.50; 29.80) kcal/kg/day, compared to the indicator in the SG group of 42.37 (41.60; 38.00) kcal/kg/day, with U = 28.0000; p = 0.0001. Carnitine supplementation in the FG group led to an increase in the level of free carnitine in the last blood plasma samples, before the children were discharged home. In general, the proposed nutrition strategy was accompanied by better indicators of the physical development of children, and a statistically probable faster recovery of body weight in the fortification group. Thus, the generalized indicator of body weight of children of the SG group was 3966.90 ± 439.08 g, compared to the FG group of 3554.62 ± 452.28 g, with p = 0.0033. In general, children who consumed more protein were transferred from the intensive care unit earlier – 12.00 (11.00; 16.00) days, against 10.00 (8.00; 12.00) days, with U = 235,00; p = 0.0024; previously discharged from the hospital – 26.50 (22.00; 31.00) days, against 21.00 (19.00; 27.00) days, with U = 267.00, p = 0.0109. This strategy proved to be safe, as confirmed by normal levels of phenylalanine and urea at all stages of the examination of children. Conclusions. Short-term increased protein consumption against the background of carnitine supplementation improves the results of physical development of newborns and indicators of hospitalization and intensive care unit stay. Larger studies and catamnestic observation of children are needed.

BIOCHEMICAL PROFILE OF FULL-TERM NEWBORNS WITH DIFFERENT PROTEIN SUPPLY ON THE BACKGROUND OF THE TREATMENT OF CRITICAL DISEASES OF THE PERINATAL PERIOD

Study of the effect of increased protein and L-carnitine supplementation in term neonates with critical illnesses on the biochemical profile and improvement of intensive care outcomes.
 Objective: assessment of the impact of increased L-carnitine and protein supplementation on the physical development and biochemical profile of newborns who were in a critical condition.
 Materials and methods. The life indicators of 59 full-term children, who were randomly divided into two groups, were studied. The first group (n = 30) received standard nutrition with mother's milk or formula, the second group (n = 29) received protein-fortified food and L-carnitine supplementation during the hospital stay.
 The results. Babies of both groups had statistically equivalent characteristics at the beginning of the study. Against the background of higher consumption of proteins, the number of non-protein calories per 1 g of protein in the study group was 28.68 kcal/kg/day (27.50; 29.80), against the indicator of the standard diet group of 42.37 kcal/kg/day (41 .60; 38.00), with U = 28.0000; p = 0.000001. Carnitine supplementation in the GZ group led to an increase in the level of free carnitine in the last blood plasma samples of newborns. In general, the proposed nutrition strategy was accompanied by better indicators of physical development of children. Children who consumed more protein were transferred from the intensive care unit earlier – 10.00 (8.00; 12.00) days, against 12.00 (11.00; 16.00) days, with U = 235.00; p = 0.0024; previously discharged from the hospital -21.00 (19.00; 27.00) days, against 26.50 (22.00; 31.00) days at U = 267.00, p = 0.0109. This strategy proved to be safe, as confirmed by normal levels of phenylalanine and blood biochemical indicators at all stages of the examination of children, including urea nitrogen and creatinine.
 Conclusions. Short-term increased consumption of protein against the background of carnitine supplementation improves the results of the physical development of newborns, does not lead to deviations in the biochemical profile, and is safe.

Neonatal adiposity is associated with microRNAs in adipocyte-derived extracellular vesicles in maternal and cord blood, a discovery analysis.

Maternal body size, nutrition, and hyperglycemia contribute to neonatal body size and composition. There is little information on maternal-fetal transmission of messages which influence fetal growth. We analyzed adipocyte-derived small extracellular vesicular (ADsEV) microRNAs in maternal and cord blood to explore their adipogenic potential. There were 279 mother-neonate pairs with all phenotypic data (normal glucose tolerant NGT = 148, gestational diabetes mellitus GDM = 131). Neonates with adiposity were those in the highest tertile (T3) of sex-specific sum of skinfolds and those without adiposity (lean) in the lowest tertile T1 of NGT pregnancies. We studied ADsEV miRNAs in 76 and 51 neonates with and without adiposity respectively and their mothers based on power calculations (68 NGT and 59 GDM pregnancies). ADsEV miRNAs from maternal and cord blood plasma samples were profiled on Agilent 8*60 K microarray. Differential expression (DE) of ADsEV miRNAs in adipose vs. lean groups was studied before and after adjustment for maternal GDM, adiposity, and vitamin B12-folate status. Multiple miRNAs were common in maternal and cord blood and positively correlated. We identified 24 maternal and 5 cord blood miRNAs differentially expressed (discovery p ≤ 0.1) in the adipose group in unadjusted, and 19 and 26, respectively, in the adjusted analyses. Even though DE miRNAs were different in maternal and cord blood, they targeted similar adipogenic pathways (e.g., the forkhead box O (FOXO) family of transcription factors, mitogen‑activated protein kinase (MAPK) pathway, transforming growth factor beta (TGF-β) pathway). Maternal GDM and adiposity were associated with many DE ADsEV miRNAs. Our results suggest that the ADsEV miRNAs in mothers are potential regulators of fetal adiposity. The expression and functionality of miRNAs appear to be influenced by maternal adiposity, hyperglycemia, and micronutrient status during pregnancy.

To study the bioequivalence of the drugs Doxitron chewable tablets and Ronaxan when used in dogs

A study was carried out to investigate the bioequivalence of the drugs "Doxitron chewable tablets" and "Ronaxan" when administered to dogs. Two groups of animals were formed with 4 dogs in each group. A cross-sectional study design was used for the experiment. Biological material (blood) was sampled before drug administration, and 15, 30, 45 min, 1, 1.5, 2, 3, 4, 6, 8, 8, 10, 24, 30, 48, 72 h after drug administration. In the course of the study the following parameters were controlled: concentrations of the active substance of the preparations in the blood plasma of dogs. The article describes in detail the scheme of the experiment, reagents and standard samples, the process of sample preparation of blood plasma samples and HPLC-MS/MS analysis. The bioequivalence of the preparations "Doxitron chewable tablets" and "Ronaxan" is evaluated. The time to reach the maximum plasma concentration (Tmax) averaged 3.0 h for the investigational drug "Doxitron chewable tablets" and 3.0 h for the reference drug "Ronaxan". The maximum plasma concentration (Cmax) was 4007.4±964.4 ng/ml for the study drug Doxitron Chewable Tablets and 3794.9±1121.5 ng/ml for the reference drug Ronaxan. The 90% confidence intervals of CmaxT/CmaxR and AUC0-tT/AUC0-tR ratios of doxycycline were within the range of 80-125%. The preparations "Doxitron chewable tablets" and "Ronaxan" are bioequivalent.

Bioresource collection of blood components from pregnant women to identify fetal genetic features, as well as to search for genetic markers of gestational complications

Aim. To create collection of plasma and leukocyte blood samples from pregnant women to improve methods of non-invasive prenatal testing, as well as to search for deoxyribonucleic acid markers and study the molecular mechanisms of gestational complications.Material and methods. The collection creation process uses original standard operating procedures. The biomaterial for storage was plasma and leukocytes from the blood of pregnant women.Results. As of July 2023, the collection contains samples of blood plasma and buffy coat suspension from 5814 pregnant women. For 5692 women, whole- genome sequencing of extracellular deoxyribonucleic acid from blood plasma was performed on an Ion GeneStudio S5 system (Thermo Fisher Scientific Inc, USA). Data on cytogenetic examination of fetal material after invasive prenatal diagnosis were collected from 522 women. In 241 cases, samples were obtained from pregnant women with fetuses with a normal karyotype and in 282 cases from pregnant women with fetuses with chromosomal abnormalities. After pregnancy ended with childbirth, data on the course and outcomes of pregnancy were collected from 590 women. In 212 cases, pregnancy was complicated by great obstetrical syndromes. Based on the collection, a previously developed bioinformatic algorithm for non-invasive prenatal analysis was tested on a DNBSEQ-G50 sequencer (MGI, China). The obtained specificity corresponded to the expected one and was >99,9%. The sensitivity of the method is 100%.Conclusion. The created collection, as well as the clinical and genomic data associated with it, is a valuable resource for basic and applied research.

Comprehensive spectroscopic, metabolomic, and proteomic liquid biopsy in the diagnostics of hepatocellular carcinoma

Liquid biopsy is a very topical issue in clinical diagnostics research nowadays. In this study, we explored and compared various analytical approaches to blood plasma analysis. Finally, we proposed a comprehensive procedure, which, thanks to the utilization of multiple analytical techniques, allowed the targeting of various biomolecules in blood plasma reflecting diverse biological processes underlying disease development. The potential of such an approach, combining proteomics, metabolomics, and vibrational spectroscopy along with preceding blood plasma fractionation, was demonstrated on blood plasma samples of patients suffering from hepatocellular carcinoma in cirrhotic terrain (n = 20) and control subjects with liver cirrhosis (n = 20) as well as healthy subjects (n = 20). Most of the applied methods allowed the classification of the samples with an accuracy exceeding 80.0 % and therefore have the potential to be used as a stand-alone method in clinical diagnostics. Moreover, a final panel of 48 variables obtained by a combination of the utilized analytical methods enabled the discrimination of the hepatocellular carcinoma samples from cirrhosis with 94.3 % cross-validated accuracy. Thus, this study, although limited by the cohort size, clearly demonstrated the benefit of the multimethod approach in clinical diagnosis.

Electrochemical detection of myeloperoxidase (MPO) in blood plasma with surface-modified electroless nickel immersion gold (ENIG) printed circuit board (PCB) electrodes

Printed circuit board (PCB) based biosensors have often utilized hard gold electroplating, that nullifies the cost advantages of this technology as compared to screen printed electrodes. Electroless nickel immersion gold (ENIG) is a popular gold deposition process widely used in PCB manufacturing, but vulnerable to pinhole defects and large surface roughness, which compromises biosensor performance. In this work, we present a method to address these challenges through electrodeposition of methylene blue (MB) to cover surface defects and improve electroactivity of ENIG PCB electrodes. We also demonstrate a process to realize in situ synthesis of gold nanoparticles (AuNPs) using acid-functionalized multi-walled carbon nanotubes (MWCNTs) as scaffold, that are used to immobilize antibody for the target molecule (myeloperoxidase: MPO, early warning biomarker for cardiovascular diseases) through a modified cysteamine/gluteraldehyde based process. The processing steps on the electrode surface are developed in a manner that do not compromise the integrity of the electrode, resulting in repeatable and reliable performance of the sensors. Further, we demonstrate a cost-effective microfluidic packaging process to integrate a capillary pump driven microfluidic channel on the PCB electrode for seamless introduction of samples for testing. We demonstrate the ability of the sensor to distinguish clinically abnormal concentrations of MPO from normal concentrations through extensive characterization using spiked serum and blood plasma samples, with a limit of detection of 15.79 ng/mL.

Can Plasma Optical Density or Transmission be used for helping AD Diagnosis?

AbstractBackgroundCurrently, diagnosis of Alzheimer’s disease (AD) mainly relies on clinical and neuropsychological tests, which are not only costly but also time‐consuming and burdensome to the patients. The only way to confirm an AD diagnosis is the post‐mortem detection of amyloid plaques and neurofibrillary tangles. The research on biomarkers has some significant progresses in recent years but none is available to be used clinically that can meet the sensitivity and specificity requirements.MethodBlood samples were collected from participants with different diagnoses: early‐onset AD, late‐onset AD, and those without an AD diagnosis (controls). 22 participants had early‐onset AD, 72 participants had late‐onset AD, and 48 participants were in the control group. The optical density (OD), as well as transmission, were measured from blood plasma samples using a densitometer. An analysis of variance (ANOVA) was performed using the diagnosis group as the independent variable with post hoc Bonferroni correction.ResultThe optical density for the late‐onset AD group was 0.356 ± 0.009, which is significantly higher than the counterpart measure for the control group of 0.313 ± 0.012 (p = 0.015). By contrast, the optical density for the early‐onset AD group was 0.361 ± 0.017, which is not significantly different from the late‐onset group (p = 1) or the control group (p = 0.065). As expected, the late‐onset AD group had an average transmission of 0.947 ± 0.002, which is at the level compared with the early‐onset AD group of 0.947 ± 0.004 (p = 1) but significantly lower than the same measure from the control group of 0.958 ± 0.003 (p = 0.006).ConclusionThe physical properties of blood plasma: OD or transmission can help with an AD diagnosis, which is particularly useful for diagnosing late‐onset AD.

Examination of plasma YKL‐40 as a clinical biomarker for Alzheimer’s disease

AbstractBackgroundBlood biomarkers represent a potential non‐invasive and cost‐effective strategy for the clinical detection and monitoring of neurodegenerative conditions including Alzheimer’s disease (AD). YKL‐40 is a secreted glycoprotein associated with inflammatory processes, and elevated cerebrospinal fluid levels have been observed in AD and other neurodegenerative conditions. Fewer studies have examined plasma YKL‐40. We examined relationships between plasma YKL‐40 and clinical indices of AD in a large cross‐sectional sample from a clinical cohort registry.MethodThe sample included 569 participants from the Boston University Alzheimer’s Disease Research Center, including participants with normal cognition (NC; n = 235), mild cognitive impairment due to suspected AD (MCI; n = 181), and AD dementia (n = 153). Blood plasma samples were analyzed using standard ELISA methods; values were log transformed and standardized into z‐scores. Cross‐sectional analyses investigated the relationships between YKL‐40 and clinical indices of functioning including diagnostic status, dementia severity (on the Clinical Dementia Rating scale [CDR]), and neuropsychological test performance. Age, sex, education, race, and APOE ε4 carrier status were included as covariates in all analyses.ResultNo significant differences in plasma YKL‐40 were observed between participants with NC, MCI, and AD dementia after accounting for covariates. The comparison between MCI and dementia was marginally significant (p = .07), with an adjusted mean difference of Z = 0.19. For participants with dementia (CDR scores 1‐3), non‐significant trends were observed for relationships between plasma YKL‐40 values and dementia severity, including for Sum of Boxes (Pearson partial correlation: r = .14, p = .10) and Total Score (ordinal logistic regression: OR = 1.38; 95% CI: [0.98,1.93]), p = .06. Modest but significant correlations were observed between YKL‐40 and performance on 4/12 neuropsychological tests including Animal Fluency (r = ‐.10), Vegetable Fluency (r = ‐.09), and Neuropsychological Assessment Battery List Learning Trials 1‐3 (r = ‐.10) and Short Delay (r = ‐.12).ConclusionPlasma YKL‐40 biomarker levels were modestly related to neuropsychological assessments of language and memory but not significantly related to cognitive diagnosis of AD or dementia severity. This biomarker may reflect underlying inflammatory processes in the context of AD. However, the use of plasma YKL‐40 to assess the presence and severity of AD in a larger cross‐sectional sample size, and using a multiple biomarker approach, warrant further study.

ОСОБЕННОСТИ СИСТЕМНОГО ОТВЕТА НА АДЪЮВАНТНУЮ ЛУЧЕВУЮ ТЕРАПИЮ У НОСИТЕЛЕЙ ПОЛИМОРФИЗМА -308(G/A)TNF БОЛЬНЫХ РАКОМ МОЛОЧНОЙ ЖЕЛЕЗЫ

Background: Adjuvant radiation therapy (ART), an integral part of locoregional breast cance (BC) therapy, acting not only locally, but also systemically and leads to a shift in homeostasis, which is reflected in routine general clinical tests. Tumor necrosis factor (TNF) is a proinflammatory cytokine, the production of which can be influenced by the single-nucleotide substitution -308(G/A)TNF. The minor allele -308A can be included in the stable inherited haplotype AH8.1 of the HLA gene complex. At the same time, the carriage of the -308A without the AH8.1 haplotype is associated with a poor prognosis in patients with BC. The aim of the study was to evaluate the features of the systemic response to the course of ART in carriers of TNF-associated genotypes with BC. Material and methods: The sample is represented by 147 BC patients who underwent a course of ART (2 Gy in 25 fractions). Clinical and morphological characteristics and data of general clinical blood analysis were obtained from medical histories. Venous blood samples for the study were obtained at the beginning and at the end of the ART course. Alleles -308(G/A)TNF and marker alleles of haplotype AH8.1 (HLA-A×01, HLA-B×08 and HLA-DRB1×03) were determined by allele-specific PCR. sTNF concentrations were determined by the ELISA in 102 blood plasma samples. Results: TNF-associated comparison groups were identified based on genotyping: (1) 114 carriers -308GG of the TNF gene, regardless of the AH8.1 haplotype (77,6 %); (2) 23 carrier -308A(AH8.1pos) had at least one AH8.1 marker allele (15.6 %); (3) 10 carriers -308A(AH8.1neg) did not have any AH8.1 marker allele (6.8 %). In the -308A(AH8.1neg) group the average concentration of sTNF both at the beginning and at the end of ART was significantly higher and, unlike other comparison groups, did not significantly decrease at the end of the ART course. A significant decrease in absolute values was revealed during ART in a number of cases for leukocytes, platelets and lymphocytes, however within the reference values. In the group -308A(AH8.1neg) correlation analysis revealed a high strength of positive connections between sTNF and leukocytes (r=0.71; p=0.027), platelets (r=0.67; p=0.04), neutrophils (r=0.70; p=0.027) only at the end of ART, whereas at the beginning ART these correlations were weak (r≤0.3) and statistically unreliable. For other genetic groups, the revealed correlations were not strong enough. Conclusion: The revealed features of the systemic response to ART for carriers of a prognostically unfavorable genotype -308A(AH8.1neg) – a high concentration of sTNF and a positive correlation with the content of leukocytes (probably due to neutrophils) and platelets – can be considered as targets of individualized therapy.

Phase Ib/II Study (NCT02488408 / BGBC003) of Bemcentinib Monotherapy or in Combination with Cytarabine or Decitabine in Acute Myeloid Leukemia (AML) or Myelodysplastic Syndrome (MDS): FINAL Results

Background: The standard of care (SOC) in newly-diagnosed (ND) AML patients (pts) unfit for intensive chemotherapy (IC) yields median overall survival (mOS) of 14.7 months. However, beyond 1 st line (1L), pts have limited treatment options and dismal mOS of 2.3 months (4.7 months in pre-venetoclax era), highlighting the need for more effective treatments in 2 nd line (2L) and beyond. Activation of AXL, a receptor tyrosine kinase, is associated with poor prognosis, resistance to chemotherapy and decreased antitumor immune response in AML. Bemcentinib (BEM) is a first-in-class, oral, highly selective AXL inhibitor. Aims: The BGBC003 PhIb/II trial studied the safety and efficacy of BEM as monotherapy in AML (cohort B1) and MDS (cohort B4) and in combination with decitabine (cohort B3) or low-dose cytarabine (LDAC) (cohorts B2 and B5) in ND and relapsed/refractory (R/R) AML pts unfit for IC. Pharmacokinetic/pharmacodynamic (PK/PD) analysis was conducted in pts treated with BEM monotherapy in the safety run-in phase (part A). Here, we present final efficacy and safety data, as well as PK/PD analysis from part A. Methods: In part A, 36 pts received BEM monotherapy in a dose escalation manner. In part B (cohorts B1-B5), pts received 3 loading doses at daily 400 mg BEM followed by 200 mg BEM daily. Study endpoints included OS, objective response rate (ORR), clinical benefit rate (CBR) (ORR+unchanged [UC]). Plasma and peripheral blood (PB) samples were used for PK and phospho-AXL (pAXL) analyses. Results: From Oct-14 to Jul-21, 86 pts were enrolled in part B (72 efficacy evaluable), with a median age of 76 years (range 50-86), 77% being R/R at study entry. BEM monotherapy and in combination was well tolerated, with a safety profile comparable to the known safety profile of the combination agents. The ORR was 18% (B1), 0% (B3), 25% (B2+B5), and 19% (B4). The CBR was 45% (B1), 31% (B3), 31% (B2+B5) and 56% (B4). The mOS (months) was 18 (B1), 6.4 (B3), 8.0 (B2+B5) and 9.2 (B4). BEM+LDAC (B2+B5) showed promising survival benefit in ND (mOS 16.5 months, n=5) and R/R pts (mOS 7.8 months, n=27). The PK/PD analysis showed inhibition of the biological target pAXL in PB following monotherapy. BEM showed dose-dependent inhibition of pAXL and downstream targets of pAXL (pAKT, pERK and pNFkB) with EC 50 values of 89-162 ng/mL. Summary/Conclusions: BEM demonstrated on-target effect by inhibition of pAXL and downstream signalling in a concentration-dependant manner. BEM monotherapy and in combination was well tolerated across all cohorts. The overall efficacy observed is comparable with historical data in 2L AML and MDS. Although the shift in 1L SOC in AML during the study limits adequate comparisons, a survival benefit was observed both in ND and R/R AML, warranting further investigation. Keywords: Acute myeloid leukemia, Clinical trial, Receptor tyrosine kinase

Gold nanoparticles coated with PVP as a novel colorimetric sensor for sensitive and selective determination of Atenolol

In this work, gold nanoparticles coated with polyvinylpyrrolidone (PVP-AuNPs) were used as a colorimetric probe for the sensitive, selective, simple, and rapid determination of atenolol (ATN). Indeed, atenolol triggered the aggregation of PVP-AuNPs via hydrogen bonding, electrostatic interactions, and dipole-dipole forces with PVP on the surface of AuNPs, causing the colloidal solution's color to shift from red to blue. SEM, TEM, FT-IR, UV–Vis, zeta potential, and other methods were used to characterise the PVP-AuNPs that were created as well as their aggregates. An excellent linear relationship between the absorption ratio (A690/A521) and the concentration of ATN in the range of 0.07–3.0 μM with a detection limit of 0.023 μM was discovered by optimising the experimental settings. The influence of potential interfering species on the measurement of ATN was investigated, and it was discovered that the developed colorimetric sensor had satisfactory selectivity. Finally, the presented method was used to measure ATN in tablet and blood plasma samples, and the obtained recovery values showed great promise for the use of the proposed sensor in clinical applications.

Passive antibody transfer from pregnant women to their fetus are maximized after SARS-CoV-2 vaccination irrespective of prior infection

Pregnancy is associated with a higher risk of adverse symptoms and outcomes for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection for both mother and neonate. Antibodies can provide protection against SARS-CoV-2 infection and are induced in pregnant women after vaccination or infection. Passive transfer of these antibodies from mother to fetus in utero may provide protection to the neonate against infection. However, it is unclear whether the magnitude or quality and kinetics of maternally derived fetal antibodies differs in the context of maternal infection or vaccination. We aimed to determine whether antibodies transferred from maternal to fetus differed in quality or quantity between infection- or vaccination-induced humoral immune responses. We evaluated 93 paired maternal and neonatal umbilical cord blood plasma samples collected between October 2020 and February 2022 from a birth cohort of pregnant women from New Orleans, Louisiana, with histories of SARS-CoV-2 infection and/or vaccination. Plasma was profiled for the levels of spike-specific antibodies and induction of antiviral humoral immune functions, including neutralization and Fc-mediated innate immune effector functions. Responses were compared between 4 groups according to maternal infection and vaccination. We found that SARS-CoV-2 vaccination or infection during pregnancy increased the levels of antiviral antibodies compared to naive subjects. Vaccinated mothers and cord samples had the highest anti-spike antibody levels and antiviral function independent of the time of vaccination during pregnancy. These results show that the most effective passive transfer of functional antibodies against SARS-CoV-2 in utero is achieved through vaccination, highlighting the importance of vaccination in pregnant women.

Russian development for drug independence in endocrinology: comparative analysis of bioequivalence, safety and tolerability of the first domestic liraglutide

Liraglutide is one of the analogues of the incretin hormone human glucagon-like peptide-1 (GLP-1) and is currently a priority treatment for diseases such as type 2 diabetes mellitus (mono- and combination therapy), obesity and overweight in the presence of at least one concomitant disease.The aim of the work was to assess the bioequivalence and comparability of the safety and tolerability profile of the drug Enligria® (liraglutide 6 mg/ml, Promomed RUS LLC, Russia) and the drug Saxenda® (liraglutide 6 mg/ml, Novo Nordisk AS, Denmark) after a single dose in healthy volunteers.Materials and methods. This study was an open-label, randomized, crossover comparative study to evaluate pharmacokinetic parameters, safety, tolerability and immunogenicity. The study comprised 26 healthy volunteers, 26 of whom were included in the bioequivalence assessment population. The study consisted of 2 periods, in each of which the volunteers received either the test drug (liraglutide at a single dose of 0.6 mg) or the reference drug (liraglutide at a single dose of 0.6 mg) once. The washout period between each dose was 7 days. Blood plasma samples were taken to determine the concentration of liraglutide in the range from 0 to 72 hours in each study period. Liraglutide concentrations were determined using a previously validated enzyme-linked immunosorbent assay (ELISA) method. A quantitative determination of antibodies to liraglutide in the blood serum samples was carried out using a microplate photometer and ready-made ELISA kits pre-validated by the manufacturer. The conclusion about the equivalence of the compared drugs was made based on the ratio of the parameters Cmax, AUC0→t and AUC0→t of the studied drug in relation to the reference one.Results. The pharmacokinetic parameters of the drugs were comparable to each other. The resulting 90% confidence intervals for the ratio of the values of Cmax, AUC0-t and AUC0-∞ of the Russian test and reference drug were 87.18–110.46, 84.40–104.11 and 86.69–103.22% respectively, which satisfied the criteria for assessing bioequivalence. The tolerability of the drugs in the volunteers was notified as good. The incidence of adverse events was comparable for the test and reference drugs. No serious adverse events were reported throughout the study. According to the results of the immunogenicity analysis, no antibodies to russian produced liraglutide were detected in the blood serum of the volunteers, which indicated the lack of the drug immunogenicity.Conclusion. During the study, the pharmacokinetic equivalence of the test and reference drugs was confirmed. The Russian drug Enligria® (liraglutide 6 mg/ml, Promomed RUS LLC, Russia) in comparison with a foreign drug Saxenda® (liraglutide 6 mg/ml, Novo Nordisk AS, Denmark).

Clinical Chemistry and Haematology Values of a Captive Population of Humboldt Penguins (Spheniscus humboldti).

Reference intervals for clinical chemistry and haematology analytes are essential for the interpretation of blood results, but limited data are available for Humboldt penguins (Spheniscus humboldti) in zoological collections as well as in the wild. The aim of the present study was therefore to establish reference intervals for a series of clinical chemistry and haematology analytes in a single zoological population of Humboldt penguins. Furthermore, possible variations of the analytes influenced by the age and sex of the individuals were investigated. Lithium heparinized plasma and whole blood samples from 39 animals were analysed and statistically evaluated. The following clinical chemistry analytes were significantly higher in females: glutamate dehydrogenase (p = 0.0003), alanine transaminase (p = 0.0005), alkaline phosphatase (p = 0.0245), aspartate aminotransferase (p = 0.0043), creatine kinase (p = 0.0016), lactate dehydrogenase (p < 0.0001), inorganic phosphorus (p = 0.0223), and sodium (p = 0.0415). No significant differences between males and females were found for any haematology analyte. Cholesterol (p = 0.0233; -0.39), white blood cell count (p = 0.0208; -0.40), and absolute heterophiles (p = 0.0148; -0.42) had significant negative correlations with the age of the penguins. The results indicate that the factors of sex and age should be considered when interpreting the results of haematological and clinical biochemistry assays, and further studies are needed to determine whether there are more differences in other age groups or seasons.