Analysis Of Blood Plasma Research Articles

Toxic Metal Species and 'Endogenous' Metalloproteins at the Blood-Organ Interface: Analytical and Bioinorganic Aspects.

Globally, human exposure to environmental pollutants causes an estimated 9 million deaths per year and it could also be implicated in the etiology of diseases that do not appear to have a genetic origin. Accordingly, there is a need to gain information about the biomolecular mechanisms that causally link exposure to inorganic environmental pollutants with distinct adverse health effects. Although the analysis of blood plasma and red blood cell (RBC) cytosol can provide important biochemical information about these mechanisms, the inherent complexity of these biological matrices can make this a difficult task. In this perspective, we will examine the use of metalloentities that are present in plasma and RBC cytosol as potential exposure biomarkers to assess human exposure to inorganic pollutants. Our primary objective is to explore the principal bioinorganic processes that contribute to increased or decreased metalloprotein concentrations in plasma and/or RBC cytosol. Furthermore, we will also identify metabolites which can form in the bloodstream and contain essential as well as toxic metals for use as exposure biomarkers. While the latter metal species represent useful biomarkers for short-term exposure, endogenous plasma metalloproteins represent indicators to assess the long-term exposure of an individual to inorganic pollutants. Based on these considerations, the quantification of metalloentities in blood plasma and/or RBC cytosol is identified as a feasible research avenue to better understand the adverse health effects that are associated with chronic exposure of various human populations to inorganic pollutants. Exposure to these pollutants will likely increase as a consequence of technological advances, including the fast-growing applications of metal-based engineering nanomaterials.

Alterations in lipid profile upon uterine fibroids and its recurrence

Uterine fibroids (UF) is the most common (about 70% cases) type of gynecological disease, with the recurrence rate varying from 11 to 40%. Because UF has no distinct symptomatology and is often asymptomatic, the specific and sensitive diagnosis of UF as well as the assessment for the probability of UF recurrence pose considerable challenge. The aim of this study was to characterize alterations in the lipid profile of tissues associated with the first-time diagnosed UF and recurrent uterine fibroids (RUF) and to explore the potential of mass spectrometry (MS) lipidomics analysis of blood plasma samples for the sensitive and specific determination of UF and RUF with low invasiveness of analysis. MS analysis of lipid levels in the myometrium tissues, fibroids tissues and blood plasma samples was carried out on 66 patients, including 35 patients with first-time diagnosed UF and 31 patients with RUF. The control group consisted of 15 patients who underwent surgical treatment for the intrauterine septum. Fibroids and myometrium tissue samples were analyzed using direct MS approach. Blood plasma samples were analyzed using high performance liquid chromatography hyphened with mass spectrometry (HPLC/MS). MS data were processed by discriminant analysis with projection into latent structures (OPLS-DA). Significant differences were found between the first-time UF, RUF and control group in the levels of lipids involved in the metabolism of glycerophospholipids, sphingolipids, lipids with an ether bond, triglycerides and fatty acids. Significant differences between the control group and the groups with UF and RUF were found in the blood plasma levels of cholesterol esters, triacylglycerols, (lyso) phosphatidylcholines and sphingomyelins. Significant differences between the UF and RUF groups were found in the blood plasma levels of cholesterol esters, phosphotidylcholines, sphingomyelins and triacylglycerols. Diagnostic models based on the selected differential lipids using logistic regression showed sensitivity and specificity of 88% and 86% for the diagnosis of first-time UF and 95% and 79% for RUF, accordingly. This study confirms the involvement of lipids in the pathogenesis of uterine fibroids. A diagnostically significant panel of differential lipid species has been identified for the diagnosis of UF and RUF by low-invasive blood plasma analysis. The developed diagnostic models demonstrated high potential for clinical use and further research in this direction.

Aspects of transition cow metabolomics—Part I: Effects of a metaphylactic butaphosphan and cyanocobalamin treatment on the metabolome in liver, blood, and urine in cows with different liver metabotypes

Dairy cows in modern production systems are at risk to develop metabolic disorders during the transition period. Reasons for individual differences in susceptibility, as well as the underlying pathomechanisms, are still only partially understood. The development of metaphylactic treatment protocols is needed. In this context, an on-farm prospective 3-fold blinded randomized study involving 80 German Holstein cows was performed throughout 1 yr. The trial involved a thorough recording of the production and clinical traits, clinical chemistry, and liver biopsies and blood and urine sampling at d 14 (mean: 12 d, range: 1-26 d) antepartum (AP), and d 7 (7, 4-13) and 28 (28, 23-34) postpartum (PP) for metabolomics analyses. Two groups received a treatment with butaphosphan and cyanocobalamin (BCC) at either the dosage recommended by the manufacturer or the double dosage (5 or 10 mL/100 kg of body weight 10% butaphosphan and 0.005% cyanocobalamin (Catosal, Bayer Animal Health), n = 20 in each group, parity: 4.2 ± 2.0 and 3.4 ± 1.3, respectively (mean ± SD)] and one group a placebo treatment (NaCl 0.9%, n = 40, parity: 4.0 ± 1.9). The animals were treated at 6 time points (7, 6, and 5 d AP, and 1, 2, and 3 d PP) via intravenous injection. Mass spectroscopy-based targeted metabolomics analysis of blood plasma and liver samples were performed using the AbsoluteIDQ p180 kit (Biocrates Life Sciences), whereas the urine samples were analyzed by nuclear magnetic resonance spectroscopy. Statistical analysis was performed using multivariate [partial least squares discriminant analysis (PLS-DA)] and univariate methods (linear mixed model). Multivariate data analysis (PLS-DA plots) of the liver metabolome revealed 3 different metabotypes (A = medium, B = minor, C = large alterations in liver metabolome profile between AP and PP status). Metabotype B animals were characterized by higher PP lipomobilization (stronger PP body condition decrease and higher blood bilirubin, fatty acids, gamma-glutamyltransferase, and triglyceride levels) and a higher occurrence of transition cow diseases, compared with the animals in metabotype C. Analysis of the feeding data showed that the period of metabotype B animals (calving in a distinct time frame) was characterized by a decreased grass silage quality. The PP liver metabolome of the metabotype C animals was characterized by higher concentrations of AA, acylcarnitines, lysoPC and sphingomyelins compared with metabotype B. For the metaphylactic treatment with BCC a dose-dependent effect was confirmed, differing between the metabotypes. In all matrices and metabotypes at various time points significant treatment effects were observed, with different profiles in clinical chemistry and as well in metabolomics data. The most clear-cut treatment effect was observed in metabotype B in the liver at 7 d PP, characterized by an increase in several acylcarnitines and phosphatidylcholines, indicating a more efficient influx and oxidation of fatty acids in mitochondria and thereby an increase in energy supply and more efficient triglyceride export in the liver. The results from the liver metabolomics analysis support the application of an indication-based metaphylactic treatment with BCC.

A resource of lipidomics and metabolomics data from individuals with undiagnosed diseases

Every year individuals experience symptoms that remain undiagnosed by healthcare providers. In the United States, these rare diseases are defined as a condition that affects fewer than 200,000 individuals. However, there are an estimated 7000 rare diseases, and there are an estimated 25–30 million Americans in total (7.6–9.2% of the population as of 2018) affected by such disorders. The NIH Common Fund Undiagnosed Diseases Network (UDN) seeks to provide diagnoses for individuals with undiagnosed disease. Mass spectrometry-based metabolomics and lipidomics analyses could advance the collective understanding of individual symptoms and advance diagnoses for individuals with heretofore undiagnosed disease. Here, we report the mass spectrometry-based metabolomics and lipidomics analyses of blood plasma, urine, and cerebrospinal fluid from 148 patients within the UDN and their families, as well as from a reference population of over 100 individuals with no known metabolic diseases. The raw and processed data are available to the research community so that they might be useful in the diagnoses of current or future patients suffering from undiagnosed disorders.

Machine learning-based LIBS spectrum analysis of human blood plasma allows ovarian cancer diagnosis.

Early-stage screening and diagnosis of ovarian cancer represent an urgent need in medicine. Usual ultrasound imaging and cancer antigen CA-125 test when prescribed to a suspicious population still require reconfirmations. Spectroscopic analyses of blood, at the molecular and atomic levels, provide useful supplementary tests when coupled with effective information extraction methods. Laser-induced breakdown spectroscopy (LIBS) was employed in this work to record the elemental fingerprint of human blood plasma. A machine learning data treatment process was developed combining feature selection and regression with a back-propagation neural network, resulting in classification models for cancer detection among 176 blood plasma samples collected from patients, including also ovarian cyst and normal cases. Cancer diagnosis sensitivity and specificity of respectively 71.4% and 86.5% were obtained for randomly selected validation samples.

Preclinical Safety Evaluation of Intranasally Delivered Human Mesenchymal Stem Cells in Juvenile Mice.

Simple SummaryThe concept of utilizing mesenchymal stem cells for the treatment of central nervous system disorders has progressed from preclinical studies to clinical trials. While promising, the effectiveness of cell therapy is hampered by the route used to deliver cells into the brain. In this context, intranasal cell administration has boomed over the past few years as an effective cell delivery method. However, comprehensive safety studies are required before translation to the clinic. Our study shed light on how intranasally administrated mesenchymal stem cells may be used to safely treat neurological disorders.Mesenchymal stem cell (MSC)-based therapy is a promising therapeutic approach in the management of several pathologies, including central nervous system diseases. Previously, we demonstrated the therapeutic potential of human adipose-derived MSCs for neurological sequelae of oncological radiotherapy using the intranasal route as a non-invasive delivery method. However, a comprehensive investigation of the safety of intranasal MSC treatment should be performed before clinical applications. Here, we cultured human MSCs in compliance with quality control standards and administrated repeated doses of cells into the nostrils of juvenile immunodeficient mice, mimicking the design of a subsequent clinical trial. Short- and long-term effects of cell administration were evaluated by in vivo and ex vivo studies. No serious adverse events were reported on mouse welfare, behavioral performances, and blood plasma analysis. Magnetic resonance study and histological analysis did not reveal tumor formation or other abnormalities in the examined organs of mice receiving MSCs. Biodistribution study reveals a progressive disappearance of transplanted cells that was further supported by an absent expression of human GAPDH gene in the major organs of transplanted mice. Our data indicate that the intranasal application of MSCs is a safe, simple and non-invasive strategy and encourage its use in future clinical trials.

Comparison of plasma and cerebrospinal fluid proteomes identifies gene products guiding adult neurogenesis and neural differentiation in birds

Cerebrospinal fluid (CSF) proteins regulate neurogenesis, brain homeostasis and participate in signalling during neuroinflammation. Even though birds represent valuable models for constitutive adult neurogenesis, current proteomic studies of the avian CSF are limited to chicken embryos. Here we use liquid chromatography–tandem mass spectrometry (nLC-MS/MS) to explore the proteomic composition of CSF and plasma in adult chickens (Gallus gallus) and evolutionarily derived parrots: budgerigar (Melopsittacus undulatus) and cockatiel (Nymphicus hollandicus). Because cockatiel lacks a complete genome information, we compared the cross-species protein identifications using the reference proteomes of three model avian species: chicken, budgerigar and zebra finch (Taeniopygia guttata) and found the highest identification rates when mapping against the phylogenetically closest species, the budgerigar. In total, we identified 483, 641 and 458 unique proteins consistently represented in the CSF and plasma of all chicken, budgerigar and cockatiel conspecifics, respectively. Comparative pathways analyses of CSF and blood plasma then indicated clusters of proteins involved in neurogenesis, neural development and neural differentiation overrepresented in CSF in each species. This study provides the first insight into the proteomics of adult avian CSF and plasma and brings novel evidence supporting the adult neurogenesis in birds.

Diet analysis, blood plasma biochemical indicators and body compositions of first year university students in the context of adaptation to the new educational environment

Introduction. The article examines the problem of adaptation to the new educational environment and pedagogical support for first-year university students. The purpose of the study is to evaluate the body composition, nutrition and biochemical parameters of blood plasma of first-year students who are adapting to living in a university dormitory. Materials and Methods. The methods of empirical research of physical development (length, weight, body mass index), body component composition (total fat content, muscle component), nutrition structure (macronutrient composition and caloric content of the daily diet), biochemical analysis of blood plasma (plasma content of triglycerides, cholesterol, low-and high-density lipoproteins, glucose), as well as statistical methods of data comparison were used. Results. The body length of males (175.4 cm), in contrast to females (162 cm) aged between 18 and 19 years is not a definitive indicator and can increase during 2 or 3 years. The percentage of overweight and obesity among first-year students was 16.1%; total fat content exceeding the limit values was found in 35.5% of females and 6.7% of males, and insufficient fat content was found only in 10% of males. The muscle component, both in girls and boys, corresponded to the indicators of the norm. The actual nutrition of first-year students living in a university dormitory was characterized by a lack of calories, fats, including polyunsaturated fatty acids, and insufficient consumption of carbohydrates, including dietary fibers. This deficiency was more pronounced among females, who also had a deficit in the consumption of proteins, especially of animal origin. The biochemical parameters of the blood plasma of all students did not exceed the normal limits, except for the values of high-density lipoproteins. The percentage of students with low values of high-density lipoproteins was 8.3%. The content of CCS in the blood plasma was inversely proportional to the caloric content of the diet and the amount of fat consumed. The predisposition to disorders of lipid metabolism in first-year students living in a university dormitory was due to insufficient replenishment of the body's energy expenditure and an unbalanced diet. Conclusions. Early adulthood is characterized by the formation of a definitive level and the predominance of assimilation processes, so the issues of healthy nutrition, especially in the conditions of a high rhythm of students’ life, insufficient replenishment of energy consumption and unbalanced consumption of nutrients, are relevant and require attention from the group leaders for first-year students. Pedagogical support of first-year students should contain methods and techniques aimed at promoting healthy nutrition and financial literacy.

Early Detection of Pancreatic Cancer in Type 2 Diabetes Mellitus Patients Based on 1H NMR Metabolomics.

The association of pancreatic cancer with type 2 diabetes mellitus was investigated by 1H NMR metabolomic analysis of blood plasma. Concentration data of 58 metabolites enabled discrimination of pancreatic cancer (PC) patients from healthy controls (HC) and long-term type 2 diabetes mellitus (T2DM) patients. A panel of eight metabolites was proposed and successfully tested for group discrimination. Furthermore, a prediction model for the identification of at-risk individuals for future development of pancreatic cancer was built and tested on recent-onset diabetes mellitus (RODM) patients. Six of 59 RODM samples were assessed as PC with an accuracy of more than 80%. The health condition of these individuals was re-examined, and in four cases, a correlation to the prediction was found. The current health condition can be retrospectively attributed to misdiagnosed pancreatogenic diabetes or to early-stage pancreatic cancer.

Assessment of Population Immunity to SARS-CoV-2 Virus in the Rostov Region

By August 2020, more than 850000 cases of new coronavirus infection (COVID-19) caused by SARSCoV-2 were confirmed in the Russian Federation, with the Rostov Region as one of the ten most affected regions in Russia. The spread of the disease is largely determined by the state of population immunity in a certain area. Our research focuses on specific humoral immune response and estimates the level of herd immunity to SARS-CoV-2 virus among the population of the Rostov Region.Materials and methods. The study involved 3,048 people; the volunteers participating in the study were divided into seven age groups. The content of antibodies to SARS-CoV-2 was determined applying ELISA using a kit for the analysis of human serum or blood plasma for the presence of specific IgG to the nucleocapsid of the SARS-CoV-2 virus, manufactured by the State Scientific Center of Applied Microbiology and Biotechnology (Obolensk) in accordance with the instructions for use.Results and discussion. The assessment of seroprevalence to SARS-CoV-2 in the Rostov Region showed that the proportion of people positive for IgG to the new coronavirus was 16.5 %, the range of seropositive individuals in the general population was between 13.9 % and 19.1 % (p<0.05). There were no significant gender differences in the degree of seroprevalence with a positive result registered in 16.6 % of women and 16.5 % of men. A high level of humoral immunity to SARS-CoV-2 was established in individuals aged 1–17 against the background of low incidence rates, which may indicate the dominance of asymptomatic forms of the disease in this age group. The highest level of seropositivity was found in preschool children (33.6 %), students (29.3 %), employees (17.3 %), and education professionals (15.3 %).

Malignant and benign thyroid nodule differentiation through the analysis of blood plasma with terahertz spectroscopy.

The liquid and lyophilized blood plasma of patients with benign or malignant thyroid nodules and healthy individuals were studied by terahertz (THz) time-domain spectroscopy and machine learning. The blood plasma samples from malignant nodule patients were shown to have higher absorption. The glucose concentration and miRNA-146b level were correlated with the sample's absorption at 1 THz. A two-stage ensemble algorithm was proposed for the THz spectra analysis. The first stage was based on the Support Vector Machine with a linear kernel to separate healthy and thyroid nodule participants. The second stage included additional data preprocessing by Ornstein-Uhlenbeck kernel Principal Component Analysis to separate benign and malignant thyroid nodule participants. Thus, the distinction of malignant and benign thyroid nodule patients through their lyophilized blood plasma analysis by terahertz time-domain spectroscopy and machine learning was demonstrated.

Enhancing Nonfouling and Sensitivity of Surface-Enhanced Raman Scattering Substrates for Potent Drug Analysis in Blood Plasma via Fabrication of a Flexible Plasmonic Patch.

Surface-enhanced Raman scattering (SERS) is an ultrasensitive analytical technique, which is capable of providing high specificity; thus, it can be used for toxicological drug assay (detection and quantification). However, SERS-based drug analysis directly in human biofluids requires mitigation of fouling and nonspecificity effects that commonly appeared from unwanted adsorption of endogenous biomolecules present in biofluids (e.g., blood plasma and serum) onto the SERS substrate. Here, we report a bottom-up fabrication strategy to prepare ultrasensitive SERS substrates, first, by functionalizing chemically synthesized gold triangular nanoprisms (Au TNPs) with poly(ethylene glycol)-thiolate in the solid state to avoid protein fouling and second, by generating flexible plasmonic patches to enhance SERS sensitivity via the formation of high-intensity electromagnetic hot spots. Poly(ethylene glycol)-thiolate-functionalized Au TNPs in the form of flexible plasmonic patches show a twofold-improved signal-to-noise ratio in comparison to triethylamine (TEA)-passivated Au TNPs. Furthermore, the plasmonic patch displays a SERS enhancement factor of 4.5 ×107. Utilizing the Langmuir adsorption model, we determine the adsorption constant of drugs for two different surface ligands and observe that the drug molecules display stronger affinity for poly(ethylene glycol) ligands than TEA. Our density functional theory calculations unequivocally support the interaction between drug molecules and poly(ethylene glycol) moieties. Furthermore, the universality of the plasmonic patch for SERS-based drug detection is demonstrated for cocaine, JWH-018, and opioids (fentanyl, despropionyl fentanyl, and heroin) and binary mixture (trace amount of fentanyl in heroin) analyses. We demonstrate the applicability of flexible plasmonic patches for the selective assay of fentanyl at picogram/milliliter concentration levels from drug-of-abuse patients' blood plasma. The fentanyl concentration calculated in the patients' blood plasma from SERS analysis is in excellent agreement with the values determined using the paper spray ionization mass spectrometry technique. We believe that the flexible plasmonic patch fabrication strategy would be widely applicable to any plasmonic nanostructure for SERS-based chemical sensing for clinical toxicology and therapeutic drug monitoring.

Pharmacokinetics of primary metabolites 5-hydroxythalidomide and 5'-hydroxythalidomide formed after oral administration of thalidomide in the rabbit, a thalidomide-sensitive species.

The teratogenicity of the chemotherapeutic drug thalidomide is species-specific and affects humans, non-human primates, and rabbits. The primary oxidation of thalidomide in previously investigated rodents predominantly resulted in the formation of deactivated 5'-hydroxythalidomide. In the current study, similar in vivo biotransformations to 5-hydroxythalidomide and 5'-hydroxythalidomide were confirmed by the analysis of blood plasma from male rabbits, a thalidomide-sensitive species, after oral administration of thalidomide (2.0 mg/kg). Similar levels of thalidomide in seminal plasma and in blood plasma were detected using liquid chromatography-tandem mass spectrometry at 4 hr and 7 hr after oral doses in male rabbits. Seminal plasma concentrations of 5-hydroxythalidomide and 5'-hydroxythalidomide were also seen in male rabbits in a roughly similar time-dependent manner to those in the blood plasma after oral doses of thalidomide (2.0 mg/kg). Furthermore, the values generated by a simplified physiologically based pharmacokinetic rabbit model were in agreement with the measured in vivo blood plasma data under metabolic ratios of 0.01 for the hepatic intrinsic clearance of thalidomide to both unconjugated 5-hydroxythalidomide and 5'-hydroxythalidomide. These results suggest that metabolic activation of thalidomide may be dependent on rabbit liver enzymes just it was for cytochrome P450 enzymes in humanized-liver mice; in contrast, rodent livers predominantly mediate biotransformation of thalidomide to 5'-hydroxythalidomide. A developmental toxicity test system with experimental animals that involves intravaginal exposures to the chemotherapeutic drug thalidomide via semen should be considered in the future.

Improving broiler chickens' health by using lecithin and lysophosphatidylcholine emulsifiers: a comparative analysis of physiological indicators.

Lipids play a vital function in a bird's body and to improve the lipids utilization and their absorption in a bird's digestive system, emulsifiers are suggested. This study evaluated and compared the effects of lecithin (LEC) and lysophosphatidylcholine (LPC) emulsifiers on broiler chicken's productivity traits and physiological indicators such as blood plasma parameters, intestine traits, short-chain fatty acids (SCFA) profile in caecum's content, caecum's villus height and crypt depth and their ratio. 900 Ross 308 broiler chickens were assigned to 3 groups with 6 replicate pens and fed with a standard compound diet (SCD) and an SCD supplemented with 0.05% LEC and 0.05% LPC. Body weight (BW), average daily gain (ADG), daily feed intake (DFI), feed conversion ratio (FCR), and dry matter (DM) content of the litter were recorded. At the end of the trial, 10 birds from each group were randomly selected and euthanized. Blood samples were collected, and blood plasma analysis was performed. Intestinal samples were collected post-mortem and intestinal traits, SCFA profiles, and intestinal histomorphometric were measured. The inclusion of lysophosphatidylcholine significantly increased the broilers' BW and ADG at their fifth week of age (P<0.05). Lecithin increased the low-density lipoprotein cholesterol (LDL/C) concentration in blood plasma (P<0.05). Butyric and isovaleric acid concentrations significantly increased by LPC and reduced by LEC (P<0.05). Lecithin and LPC caecum's villus heights were significantly increased (P<0.05), and caecum's crypt depth was also increased in LEC compared to SCD (P<0.05). As an emulsifier, lysophosphatidylcholine can improve the broilers' weight, but LEC showed better effects on their physiological indicators by improving intestinal mucosal absorption areas in caecum.

ВЛИЯНИЕ ЭЛЕКТРОМАГНИТНОГО ИЗЛУЧЕНИЯ НА ПРОНИЦАЕМОСТЬ ЭРИТРОЦИТАРНЫХ МЕМБРАН

The purpose of the survey is to experimentally study the influence of electromagnetic radiation from the decimeter range on the permeability of erythrocyte membranes. The object of the study was the blood of 80 white rats, 60 of them for three months were under the influence of electromagnetic radiation of the decimeter range. 20 animals of the control group were not exposed to electromagnetic fields. The intensity of free radical oxidation was evaluated using chemiluminescent analysis of red blood cells and blood plasma. The permeability of erythrocyte membranes and the content of reduced glutathione in blood erythrocytes were determined spectrophotometrically. A prolonged stay of animals under the influence of an electromagnetic field leads to pronounced changes in chemiluminescence indices in erythrocytes: a decrease in the maximum intensity of chemiluminescence (Imax), light sum (S), and light sum after the maximum value of chemiluminescence (Simax). Despite the increase in the content of glutathione and the chemiluminescent analysis index tg2 reflecting the antioxidant potential, there is an increase in the permeability of erythrocyte membranes under the influence of electromagnetic radiation

Indicators of the blood clotting system in patients with acantholytic pemphigus

Research of the level of some indicators of the blood coagulation system in patients with acantholytic pemphigus is extremely important for the prevention of complications. Due to laboratory studies of patients’ blood plasma and statistical analysis of the obtained results, the relationship between the severity of dermatosis and changes in the hemostasis system was considered (level of prothrombin index, fibrinogen, partial thromboplastin time, soluble fibrin-monomer complexes, factor XII). Educed reliable dependence that correlates with weight of dermatosis and development of thrombotic complications.

Metabolomic profiling of blood plasma of patients with lung cancer and malignant tumors with metastasis in the lungs showed similar features and promising statistical discrimination against controls.

Targeting metabolomic pathways is a promising strategy for cancer treatment. Alterations in the metabolomic state have also an epigenetic impact, making the metabolomic studies even more interesting. We explored metabolomic changes in the blood plasma of patients with primary and secondary lung cancer and tried to explore their origin. We also applied a discrimination algorithm to the data. In the study, blood samples from 132 patients with primary lung cancer, 47 with secondary lung cancer, and 77 subjectively healthy subjects without any cancer history were used. The samples were measured by NMR spectroscopy. PCA and PLS-DA analyses did not distinguish between patients with primary and secondary lung tumors. Accordingly, no significantly changed levels of plasmatic metabolites were found between these groups. When comparing with healthy controls, significantly increased glucose, citrate, acetate, 3-hydroxybutyrate, and creatinine balanced with decreased pyruvate, lactate, alanine, tyrosine, and tryptophan were found as a common feature of both groups. Metabolomic analysis of blood plasma showed considerable proximity of patients with primary and secondary lung cancer. The changes observed can be partially explained as cancer-derived and also as changes showing ischemic nature. Random Forrest discrimination based on the relative concentration of metabolites in blood plasma performed very promising with AUC of 0.95 against controls; however noticeable parts of differencing metabolites are overlapping with those observed after ischemic injury in other studies.

Multi-residue analysis of pesticides in blood plasma using hollow fiber solvent bar microextraction and gas chromatography with a flame ionization detector

The challenges faced on pesticide extraction from biological samples are finding a method that allows a multi-residue extraction, pre-concentration, clean-up, and isolation of analytes in just one step. In this sense, the hollow fiber – liquid phase microextraction method (HF- LPME) in the "solvent bar" mode was used to optimize and validate a method for pesticide multi-residue analysis in blood plasma at trace levels, through gas chromatography coupled with a flame ionization detector (GC-FID). Hollow fiber solvent bar microextraction HF-SBME was carried out with octanol immobilized into the pores of hydrophobic polypropylene fiber and disposed within a matrix of blood plasma, spiked with a mixture of pesticides (monocrotophos, lindane, aldrin, methyl parathion, endosulfan, dieldrin, DDD, DDT, and endrin). The optimization parameters evaluated were: extraction temperature and time, stirring speed, and salt concentration. A principal component analysis was performed to visualize the analytes' behaviour based on their explained variance, and then, a Box-Behnken analysis was generated to identify the optimum parameters. According to the PCA, all pesticides showed similar responses to the extraction method and the response of dieldrin exhibit the lowest variance. Moreover, the stationary points selected from the Box-Behnken analysis were 25.5 °C for the extraction temperature, 870 rpm for stirring speed, 16 min for extraction time, and 8.3 % w/v of salt concentration. Moreover, the validation results proved that HF-SBME is an alternative technique for pesticide multi-residue extraction in blood plasma. The analytes were able to concentrate, reaching 46 fold enrichment. The solvent type, sample and solvent volume were narrowed down without changing the method's precision or accuracy. The relative standard deviation was under 10 %, and the recovery was between 55 % and 105 % for the different analytes excepting lindane, which had lower recovery (27 %). The detection limits were 0.02 until 0.13 μg mL−1 for most of the pesticides used. Finally, HF-SBME is a good alternative for pesticide multi-residue extraction in complex matrices like plasma.

Analysis of α- and β-amanitin in Human Plasma at Subnanogram per Milliliter Levels by Reversed Phase Ultra-High Performance Liquid Chromatography Coupled to Orbitrap Mass Spectrometry.

Amatoxins are known to be one of the main causes of serious to fatal mushroom intoxication. Thorough treatment, analytical confirmation, or exclusion of amatoxin intake is crucial in the case of any suspected mushroom poisoning. Urine is often the preferred matrix due to its higher concentrations compared to other body fluids. If urine is not available, analysis of human blood plasma is a valuable alternative for assessing the severity of intoxications. The aim of this study was to develop and validate a liquid chromatography (LC)-high resolution tandem mass spectrometry (HRMS/MS) method for confirmation and quantitation of α- and β-amanitin in human plasma at subnanogram per milliliter levels. Plasma samples of humans after suspected intake of amatoxin-containing mushrooms should be analyzed and amounts of toxins compared with already published data as well as with matched urine samples. Sample preparation consisted of protein precipitation, aqueous liquid-liquid extraction, and solid-phase extraction. Full chromatographical separation of analytes was achieved using reversed-phase chromatography. Orbitrap-based MS allowed for sufficiently sensitive identification and quantification. Validation was successfully carried out, including analytical selectivity, carry-over, matrix effects, accuracy, precision, and dilution integrity. Limits of identification were 20 pg/mL and calibration ranged from 20 pg/mL to 2000 pg/mL. The method was applied to analyze nine human plasma samples that were submitted along with urine samples tested positive for amatoxins. α-Amanitin could be identified in each plasma sample at a range from 37–2890 pg/mL, and β-amanitin was found in seven plasma samples ranging from <20–7520 pg/mL. A LC-HRMS/MS method for the quantitation of amatoxins in human blood plasma at subnanogram per milliliter levels was developed, validated, and used for the analysis of plasma samples. The method provides a valuable alternative to urine analysis, allowing thorough patient treatment but also further study the toxicokinetics of amatoxins.

Assessment of the Herd Immunity to SARS-CoV-2 among the Population of the Leningrad Region during the COVID-19 Epidemic

The first case of COVID-19 was registered in the Leningrad Region on March 13, 2020. The period of increasing intensity of the epidemic process lasted 8 weeks. One month after reaching the maximum incidence rate, a study was organized to determine seroprevalence to COVID-19 among the population of the Region.Objective of the study was to determine the level and structure of community immunity to SARS-CoV-2 virus in the population of the Leningrad Region in the period of intensive COVID-19 transmission.Materials and methods. The work was carried out within the framework of the Rospotrebnadzor project on assessment of community immunity to the SARS-CoV-2 virus in the population of the Russian Federation. The content of antibodies to SARS-CoV-2 was determined applying ELISA using a kit for the analysis of human serum or blood plasma for the presence of specific IgG to the nucleocapsid of the SARS-CoV-2 virus , manufactured by the State Scientific Center of Applied Microbiology and Biotechnology (Obolensk) in accordance with the instructions for use.Results and discussion. The study has showed that the herd immunity of the population of the Leningrad Region was 20.7 %. The maximum level has been established in children 1–6 years old (42.3 %) and people over 70 years old (29.0 %). The highest level of seropositivity, except for children and older people, was found among the unemployed (25.1 %). The lowest level of seroprevalence was detected in civil servants (12.8 %) and military personnel (16.7 %). It has been shown that the risk of infection increases by 1.5 times in case of contacts with COVID-19 patients. After exposure to COVID-19 virus, antibodies are produced in 82.1 % of the cases. In individuals with a positive PCR test result obtained earlier, antibodies are detected in 82.8 % of cases. The share of asymptomatic forms among seropositive residents of the Leningrad Region was 86.9 %. The results of assessing the herd immunity to SARS-CoV-2 in the population of the Leningrad Region indicate that during the period of intensive COVID-19 transmission, an average level of seroprevalence was formed. A significant proportion of asymptomatic forms of infection characterizes the high intensity of the latently developing epidemic process. The results obtained should be taken into account when organizing preventive measures, including vaccination, and predicting morbidity rates.