Case Study of Diesters of o-Phthalic Acid in Surface Waters with Background Levels of Pollution.

Lake Baikal was studied as a model for elucidating the general pattern of o-phthalic acid diester (PAE) distributions in surface waters with background pollution levels. The influence of factors including congeners, concentrations, sampling points, seasons, years, and potential sources was considered and the environmental risk for various hydrobionts was established. Priority PAEs in Baikal waters are represented by dimethyl phthalate (DMP), diethyl phthalates (DEP), di-n-butyl phthalate (DnBP) and di-(2-ethylhexyl)phthalate (DEHP). Statistically valuable average concentrations and ranges for DMP, DEP, DnBP, and DEHP were 0.02 (0.01-0.02), 0.07 (0.06-0.09), 0.55 (0.47-0.66), and 0.30 (0.26-0.34) µg/L, respectively. The main factors determining PAE concentrations were the year and season of sampling, whereas sampling points were not among the factors influencing PAE levels. The distribution of PAEs in the water body was characterized by (i) an even distribution of minor hydrophilic DMP and DEP congeners in the whole water body, (ii) a maximum concentration of hydrophobic DnBP and DEHP congeners in the upper and near-bottom layers of the water column, and (iii) a low concentration of hydrophobic congeners in the near-shore area. The main PAE source was found to be the atmospheric transfer of polluted air masses, while the supply of PAEs from coastal sources to the pelagic zone was low. The contribution of biogenic sources to the background level of PAEs in the surface waters of Lake Baikal was established. The ecological risk of the background concentration level of PAEs for Lake Baikal biota was estimated. It was found that (i) DMP and DEP congeners do not represent a risk, or represent a very low risk, (ii) the concentration levels of dominant DnBP and DEHP congeners represent a low risk for crustaceans and fishes but (iii) a rather high risk for algae at a DEHP concentration of 0.30 µg/L.

Effects of COVID-19 Infection in Healthy Subjects on Cardiac Function and Biomarkers of Oxygen Transport, Blood Coagulation and Inflammation.

BACKGROUND: The manifestations, severity, and mortality of COVID-19 are considered to be associated with the changes in various hematological parameters and in immunity. Associations of immunoglobulin G antibodies against severe acute respiratory syndrome-linked coronavirus (IgG-SARS)-positive status with cardiac function and hematological and biochemical parameters in apparently health subjects are poorly understood. METHODS: The present cross-sectional study included 307 healthy volunteers (24-69 years of age; 44.8 ± 8.6 years; 80.4% men) and was initiated in 2019 before the COVID-19 pandemic. COVID-19 episodes were confirmed by detection of IgG-SARS against SARS-CoV-2 S1 RBD to reveal 70 IgG-SARS-positive and 237 negative participants. Numerous ultrasound characteristics were assessed by echocardiography, and 15 hematological and biochemical parameters were assayed in the blood. Descriptive and comparative analysis was based on the IgG-SARS status of the participants. RESULTS: The left ventricular mass index, mitral ratio of peak early to late diastolic filling velocity or flow velocity across the mitral valve, and deceleration time of early mitral inflow were decreased (p < 0.05) in IgG-SARS-positive participants versus those in IgG-SARS-negative participants according to multivariate logistic regression analysis. Erythrocyte sedimentation rate and platelet count were slightly increased, and blood hemoglobin was decreased in IgG-SARS-positive participants compared with those in IgG-SARS-negative participants. CONCLUSIONS: LV filling, inflammation, blood coagulation, and hemoglobin appear to be influenced by COVID-19 infection in healthy participants. Our observations contribute to the definition of vulnerabilities in the apparently healthy subjects with long COVID-19. These vulnerabilities may be more severe in patients with certain chronic diseases.

Assessment of stable carbon isotope 13С/12С ratio in phthalates from surface waters using HPLC-HRMS-TOF approach.

A method for estimating the ratio of stable carbon isotopes 13С/12С in the composition of phthalates from surface water at a trace concentration level is proposed. It is based on the concentration of hydrophobic components of water using an analytical reversed phase HPLC column followed by their gradient separation and detection of eluted phthalates using a high-resolution time-of-flight mass spectrometer (ESI-HRMS-TOF) in the form of molecular ions. The ratio of stable carbon isotopes 13С/12C in phthalates is calculated as a ratio of integrals under the monoisotopic [M+1+H]+ and [M+H]+ peaks. The Δ13C value is calculated relatively to the 13C/12C ratio in commercial DnBP and DEHP phthalates used as standards. The minimal concentration of DnBP and DEHP in water required for a reliable determination of Δ13C value is estimated by the level of ca. 0.2 µg L-1. The technique has been verified during the monitoring of priority phthalates in the waters of Lake Baikal.

Assessment of Blood Microcirculation Changes after COVID-19 Using Wearable Laser Doppler Flowmetry.

The present work is focused on the study of changes in microcirculation parameters in patients who have undergone COVID-19 by means of wearable laser Doppler flowmetry (LDF) devices. The microcirculatory system is known to play a key role in the pathogenesis of COVID-19, and its disorders manifest themselves long after the patient has recovered. In the present work, microcirculatory changes were studied in dynamics on one patient for 10 days before his disease and 26 days after his recovery, and data from the group of patients undergoing rehabilitation after COVID-19 were compared with the data from a control group. A system consisting of several wearable laser Doppler flowmetry analysers was used for the studies. The patients were found to have reduced cutaneous perfusion and changes in the amplitude-frequency pattern of the LDF signal. The obtained data confirm that microcirculatory bed dysfunction is present in patients for a long period after the recovery from COVID-19.

Detection of Anti-SARS-CoV-2-S1 RBD-Specific Antibodies Prior to and during the Pandemic in 2011-2021 and COVID-19 Observational Study in 2019-2021.

BACKGROUND: Longitudinal surveys to monitor the seroprevalence are required to support efforts for assessment of the levels of endemic stability in certain countries. We investigated seroprevalence of anti-SARS-CoV-2-S1 receptor-binding domain (RBD)-specific antibodies in the serum samples in 2011-2021, including a cohort study of 2019-2021, to evaluate the vaccination and anti-IgG-SARS-CoV-2-S1 RBD-positive statuses to assess the resistance and severity of COVID-19. MATERIALS AND METHODS: Anti-SARS-CoV-2-S1 RBD-specific antibodies were assayed in the serum samples (N = 565) randomly selected from various cohorts previously recruited from 2011 to 2021 from the city of Moscow and Moscow Region. Among them there were the participants (N = 310) recruited in 2019-2021 with an endpoint of 30 October 2021 when these participants were interviewed over phone with relevant questionnaire. RESULTS: Obtained data indicated a percentage of 3-6% of SARS-CoV-2-S1 RBD-specific antibodies detected in participants recruited in 2011-2019. The percentage of SARS-CoV-2-S1 RBD-specific antibodies was increased to 16.5% in 2020 and to 46% in 2021. The vaccination rate of 238 respondents of this cohort was 58% from August 2020 to October 2021. In total, 12% of respondents were hospitalized. The morbidity rate in the subgroup of anti-SARS-CoV-2-S1 RBD-positive respondents was 5.4-fold higher than that in the subgroup of vaccinated respondents. CONCLUSIONS: A small percentage of SARS-CoV-2-S1 RBD-specific antibodies detected in 2011-2019 indicated possible spreading of coronaviruses during the pre-pandemic period. Collective immunity in Moscow and the Moscow region was able to reach 69% from August 2020 to October 2021 if this rate is added to the rate of not vaccinated SARS-CoV-2-S1 RBD-positive subjects.

Early structural and functional changes in Baikal Sculpin gills exposed to suspended soot microparticles in experiment.

The toxic influence of soot microparticles on terrestrial organisms has been well studied, although there is scarce data on how microparticles could affect hydrobionts. We performed a first-ever study of the short-term (5 days) impact of furnace soot (0.005 g/L) on the structural and functional features of gill cells in the Baikal Sculpin species Paracottus knerii, Dybowski, 1874. The soot samples used in the experiment were composed of small (10-100 nm) particles and larger (up to 20 µm) aggregates. The dominant fractions of the polycyclic aromatic hydrocarbons of these microparticles were phenanthrene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzofluoranthenes, benzopyrenes, indeno[1,2,3-c,d]pyrenes, and benzo[ghi]perylene. Trace element analysis of the soot detected the presence of C, S, Si, Al, Ca, K, Mg, P, and Fe. The gill condition was assessed with electron scanning, transmission, and laser confocal microscopy. Soot induces degenerative changes in the macrostructure and surface of secondary lamellae and increases mucus production in fish gills. A decrease in mitochondrial activity, an increase in reactive oxygen species production, and an increase in the frequency of programmed cell death in gill epithelium were observed under the influence of soot. In chloride cells, an induction of macroautophagy was detected. In general, the changes in fish gills after the short-term influence of soot microparticles indicate the stress of respiratory and osmotic regulation systems in fish. The data obtained are important for forming a coherent picture of the impact of soot on hydrobionts and for developing bioindication methods for evaluating the risks of their influence on aquatic ecosystems.

Modelling the seismic potential of the Indo-Burman megathrust.

The Indo-Burman arc is the boundary between the India and Burma plates, north of the Sumatra-Andaman subduction zone. The existence of active subduction in the Indo-Burman arc is a debatable issue because the Indian plate converges very obliquely beneath the Burma plate. Recent GPS measurements in Bangladesh, Myanmar, and northeast India indicate 13-17 mm/y of plate convergence along a shallow dipping megathrust while most of the strike-slip motion occurs on several steep faults, consistent with patterns of strain partitioning at subduction zones. A short period of instrumentally recorded seismicity and sparse historical records are insufficient to assess the possibility of great earthquakes at the Indo-Burman megathrust. Using the advantage of the Block-and-Fault Dynamics model allowing simultaneous simulation of slow tectonic motions and earthquakes, we test the hypothesis whether the India-Burma detachment is locked and able to produce great earthquakes, or it slips aseismically? We have shown that the model of locked detachment is preferred because it more adequately reproduces observed tectonic velocities. The integral characteristics of synthetic seismicity, the earthquake size distribution, and the rate of seismic activity are consistent with those derived from observations. Our results suggest that the megathrust is locked and can generate great M8+ earthquakes. The estimated average return period of great events exceeds one thousand years. Earthquakes of this size pose a great threat to NE India, Bangladesh and Myanmar, the most densely populated areas of the world.

Activation of Anthracite Combustion Using Pyrolysis Oil from Thermal Conversion of Waste Car Tires.

The ignition and combustion of anthracite modified by the addition of pyrolysis oil obtained during thermal processing of waste car tires (WCTs) had been studied. The mass fraction of WCT pyrolysis oil was varied in the range from 5 to 30 wt %. The additive was applied by the drop impregnation method. Ignition and combustion of obtained samples were carried out in a combustion chamber at temperatures of the heating medium T g = 600-800 °C. The gas-phase combustion products were analyzed using an in-line gas analyzer. The application of WCT pyrolysis oil as a combustion modifier contributed to an increase in the reactivity of anthracite, which was expressed in a decrease in the minimum ignition temperature (by 23-104 °C) and a reduction in the ignition delay time. The high-speed video recording indicated that the combustion of both initial and modified with 5 wt % pyrolysis oil anthracite samples was realized in oxidation mode. For samples with more than 10 wt % pyrolysis oil additive, the formation of a visible flame was observed near the sample surface. With an increase in the mass fraction of the additive, the rate of combustion front propagation was increased. The application of WCT pyrolysis oil as a combustion modifier also contributed to the reduction or even the almost complete elimination of unburnt carbon content in the ash residue formed after anthracite combustion.

Associations of endothelial biomarkers, nitric oxide metabolites and endothelin, with blood pressure and coronary lesions depend on cardiovascular risk and sex to mark endothelial dysfunction on the SCORE scale.

OBJECTIVES: Endothelial dysfunction contributes to the onset and progression of cardiovascular diseases. However, direct associations of vasoactive mediators with cardiovascular risk are poorly understood. METHODS: We have determined associations of circulating levels of stable metabolites of nitric oxide, nitrate and nitrite (NOx), endothelin-1, and the endothelin-1/NOx ratio with blood pressure in 177 asymptomatic subjects without signs of coronary atherosclerosis; associations with blood pressure and with presence of coronary lesions were also evaluated in 457 patients suspected to have coronary heart disease with or without coronary lesions confirmed by coronary angiography. All participants were on a low nitrate diet 24 h prior to blood sampling. RESULTS: In men, NOx levels were inversely correlated with blood pressure similar to women with low (0-4%) European Systematic Coronary Risk Estimation (SCORE). However, the correlation was not significant in women with high SCORE (5-8%). High systolic blood pressure over 140 mm Hg was negatively associated with NOx levels in asymptomatic men (p=0.05) but not in women. This association is disrupted in male and female patients with coronary atherosclerosis. In male patients, NOx (p=0.05), endothelin (p=0.01), and the endothelin/NOx ratio (p=0.04) were associated with presence of coronary lesions. CONCLUSIONS: Thus, elevated cardiovascular risk according to SCORE over 4% in asymptomatic women, but not in men, is associated with a shift in markers of endothelial dysfunction. Presence of coronary lesions in patients is associated with significant changes in circulating levels of markers of endothelial dysfunction in men but not in women.

Protatranes, effective growth biostimulants of hydrocarbon-oxidizing bacteria from Lake Baikal, Russia.

Under natural conditions, biodegradation processes proceed slowly, especially in regions with low temperature. To activate vital processes in hydrocarbon-oxidizing microorganisms at low temperatures, biologically active compounds can be employed as growth stimulants. A low-temperature (10 °C) study has shown that tris-(2 hydroxyethyl) ammonium arylchalcogenylacetates, "protatranes" exert an effect on the growth of hydrocarbon-oxidizing strains of Rhodococcus erythropolis and Pseudomonas fluorescens, isolated from natural oil seepage on Lake Baikal. It has been found that "protatranes", at microconcentrations, increase the growth rate of R. erythropolis bacteria by 2-16 times. It has been established that compounds slightly effect the growth of P. fluorescens. The positive effect of "protatranes" compounds on the growth rate of hydrocarbon-oxidizing microorganisms at low positive temperatures can be used for the development of environmentally benign methods for the restoration of natural objects after their contamination with oil.

Increase in perfused boundary region of endothelial glycocalyx is associated with higher prevalence of ischemic heart disease and lesions of microcirculation and vascular wall.

OBJECTIVE: PBR characterizes penetration of red blood cells inside glycocalyx and its thickness can have profound impact on microcirculation and other vascular parameters. The goal of our study was to reliably quantify PBR and assess its potential use as a new marker of cardiovascular pathology. METHODS: The study included 208 patients (123 men and 85 women from 40 to 65 years of age) with various grades of cardiovascular SCORE risk index and IHD. PBR was quantified by sidestream dark field capillaroscopy with green light excitation. Cutaneous microcirculation was evaluated with laser Doppler fluorometry. RESULTS: Elevated PBR values over 2 mm were associated with morphological and functional lesions of arterial wall and microcirculation and lowered levels of ApoA1 lipoprotein. Moreover, elevated PBR values were associated with 2.07-fold increase in prevalence of cerebral atherosclerosis (P = .015) and 2.42-fold increase in prevalence of IHD (P = .024). Increase in PBR was associated with elevated systolic blood pressure. CONCLUSIONS: Thus, PBR can be considered a new highly reproducible and promising marker candidate for non-invasive diagnostics of IHD and cerebral atherosclerosis suggesting important role of microcirculation in development and progression of cardiovascular diseases.

Correction: Predictive chromatography of peptides and proteins as a complementary tool for proteomics.

Correction for 'Predictive chromatography of peptides and proteins as a complementary tool for proteomics' by Irina A. Tarasova et al., Analyst, 2016, 141, 4816-4832.

Accumulation of petroleum hydrocarbons in intracellular lipid bodies of the freshwater diatom Synedra acus subsp. radians.

The accumulation of hydrophobic compounds by phytoplankton plays a crucial role in the biogeochemical cycle of persistent organic pollutants (POPs) in aquatic environments. We studied the accumulation of polycyclic aromatic hydrocarbons (PAHs) in the freshwater diatom Synedra acus subsp. radians during its cultivation with crude oil hydrocarbons, using epifluorescent and laser confocal microscopy as well as gas chromatography-mass spectrometry (GC/MS) analysis. Our results revealed that in the presence of crude oil or an extract of a crude oil/n-hexane solution (light oil), S. acus subsp. radians accumulated PAHs in its lipid bodies. During cultivation in the presence of a crude oil/n-hexane solution, the cells selectively accumulated C12-C18 alkanes, with a preference for C15 and C16 homologues. The length of n-alkane hydrocarbon chains accumulated in cells was similar to the acyl chains of fatty acids of the diatom. We therefore suggest that the insertion of n-alkanes into the membrane lipid bilayer promotes the transmembrane transport of PAH in diatoms. Our results confirm the hypothesis that diatoms play a role in the elimination of hydrophobic hydrocarbons from aquatic systems.

Predictive chromatography of peptides and proteins as a complementary tool for proteomics.

In the last couple of decades, considerable effort has been focused on developing methods for quantitative and qualitative proteome characterization. The method of choice in this characterization is mass spectrometry used in combination with sample separation. One of the most widely used separation techniques at the front end of a mass spectrometer is high performance liquid chromatography (HPLC). A unique feature of HPLC is its specificity to the amino acid sequence of separated peptides and proteins. This specificity may provide additional information about the peptides or proteins under study which is complementary to the mass spectrometry data. The value of this information for proteomics has been recognized in the past few decades, which has stimulated significant effort in the development and implementation of computational and theoretical models for the prediction of peptide retention time for a given sequence. Here we review the advances in this area and the utility of predicted retention times for proteomic applications.

Differential Leukocyte Counting via Fluorescent Detection and Image Processing on a Centrifugal Microfluidic Platform.

Centrifugal microfluidics has received much attention in the last decade for the automation of blood testing at the point-of-care, specifically for the detection of chemistries, proteins, and nucleic acids. However, the detection of common blood cells on-disc, particularly leukocytes, remains a challenge. In this paper, we present two analytical methods for enumerating leukocytes on a centrifugal platform using a custom-built fluorescent microscope, acridine orange nuclear staining, and image processing techniques. In the first method, cell analysis is performed in glass capillary tubes; in the second, acrylic chips are used. A bulk-cell analysis approach is implemented in both cases where the pixel areas of fractionated lymphocyte/monocyte and granulocyte layers are correlated with cell counts. Generating standard curves using porcine blood sample controls, we observed strong linear fits to measured cell counts using both methods. Analyzing the pixel intensities of the fluorescing white cell region, we are able to differentiate lymphocytes from monocytes via pixel clustering, demonstrating the capacity to perform a 3-part differential. Finally, a discussion of pros and cons of the bulk-cell analysis approach concludes the paper.

Application of Statistical Thermodynamics To Predict the Adsorption Properties of Polypeptides in Reversed-Phase HPLC.

The theory of critical chromatography for biomacromolecules (BioLCCC) describes polypeptide retention in reversed-phase HPLC using the basic principles of statistical thermodynamics. However, whether this theory correctly depicts a variety of empirical observations and laws introduced for peptide chromatography over the last decades remains to be determined. In this study, by comparing theoretical results with experimental data, we demonstrate that the BioLCCC: (1) fits the empirical dependence of the polypeptide retention on the amino acid sequence length with R(2) > 0.99 and allows in silico determination of the linear regression coefficients of the log-length correction in the additive model for arbitrary sequences and lengths and (2) predicts the distribution coefficients of polypeptides with an accuracy from 0.98 to 0.99 R(2). The latter enables direct calculation of the retention factors for given solvent compositions and modeling of the migration dynamics of polypeptides separated under isocratic or gradient conditions. The obtained results demonstrate that the suggested theory correctly relates the main aspects of polypeptide separation in reversed-phase HPLC.

Superheavy element flerovium (element 114) is a volatile metal.

The electron shell structure of superheavy elements, i.e., elements with atomic number Z ≥ 104, is influenced by strong relativistic effects caused by the high Z. Early atomic calculations on element 112 (copernicium, Cn) and element 114 (flerovium, Fl) having closed and quasi-closed electron shell configurations of 6d(10)7s(2) and 6d(10)7s(2)7p1/2(2), respectively, predicted them to be noble-gas-like due to very strong relativistic effects on the 7s and 7p1/2 valence orbitals. Recent fully relativistic calculations studying Cn and Fl in different environments suggest them to be less reactive compared to their lighter homologues in the groups, but still exhibiting a metallic character. Experimental gas-solid chromatography studies on Cn have, indeed, revealed a metal-metal bond formation with Au. In contrast to this, for Fl, the formation of a weak bond upon physisorption on a Au surface was inferred from first experiments. Here, we report on a gas-solid chromatography study of the adsorption of Fl on a Au surface. Fl was produced in the nuclear fusion reaction (244)Pu((48)Ca, 3-4n)(288,289)Fl and was isolated in-flight from the primary (48)Ca beam in a physical recoil separator. The adsorption behavior of Fl, its nuclear α-decay product Cn, their lighter homologues in groups 14 and 12, i.e., Pb and Hg, and the noble gas Rn were studied simultaneously by isothermal gas chromatography and thermochromatography. Two Fl atoms were detected. They adsorbed on a Au surface at room temperature in the first, isothermal part, but not as readily as Pb and Hg. The observed adsorption behavior of Fl points to a higher inertness compared to its nearest homologue in the group, Pb. However, the measured lower limit for the adsorption enthalpy of Fl on a Au surface points to the formation of a metal-metal bond of Fl with Au. Fl is the least reactive element in the group, but still a metal.

Effect of mixotrophic growth on the ultrastructure and fatty acid composition of the diatom Synedra acus from Lake Baikal.

BACKGROUND: Interest in studies concerning the effect of organic carbon sources on the growth of diatoms is largely aimed at subsequent physiological changes occurring in their cells. There are no data on structural changes in the cytoplasm and their relationship with changes in the composition of fatty acids in the course of mixotrophic culturing of freshwater diatoms. To elucidate the role of lipids in the growth of diatom cells in autotrophic and mixotrophic cultures, it is necessary to obtain information on the distribution of fatty acids among intracellular compartments and on possible ultrastructural changes in the cells. RESULTS: In this study, the results demonstrated that Synedra acus cells cultured in the presence of 80 mM glycerol contained lipid bodies of increased size, while cells from cultures supplemented with 40 mM glucose accumulated polysaccharide (chrysolaminarin) granules. An increase in the relative amounts of palmitic and stearic acids was revealed at the exponential growth phase of S. acus in the medium with 80 mM glycerol, which was indicative of the accumulation of fatty acids contained in triacylglycerols. CONCLUSIONS: Synedra acus subsp. radians have an ability to proliferate in the presence of high concentrations of organic substances and their transport into cells is evidence for its high adaptation potential, which, along with other factors, accounts for the dominance of this diatom in the spring-summer plankton of the oligotrophic Lake Baikal.

Limitation of predictive 2-D liquid chromatography in reducing the database search space in shotgun proteomics: in silico studies.

A two-dimensional (2-D) liquid chromatography (LC) separation of complex peptide mixtures that combines a normal phase utilizing hydrophilic interactions and a reversed phase offers reportedly the highest level of 2-D LC orthogonality by providing an even spread of peptides across multiple LC fractions. Matching experimental peptide retention times to those predicted by empirical models describing chromatographic separation in each LC dimension leads to a significant reduction in a database search space. In this work, we calculated the retention times of tryptic peptides separated in the C18 reversed phase at different separation conditions (pH 2 and pH 10) and in TSK gel Amide-80 normal phase. We show that retention times calculated for different 2-D LC separation schemes utilizing these phases start to correlate once the mass range of peptides under analysis becomes progressively narrow. This effect is explained by high degree of correlation between retention coefficients in the considered phases.

On the utility of predictive chromatography to complement mass spectrometry based intact protein identification.

The amino acid sequence determines the individual protein three-dimensional structure and its functioning in an organism. Therefore, "reading" a protein sequence and determining its changes due to mutations or post-translational modifications is one of the objectives of proteomic experiments. The commonly utilized approach is gradient high-performance liquid chromatography (HPLC) in combination with tandem mass spectrometry. While serving as a way to simplify the protein mixture, the liquid chromatography may be an additional analytical tool providing complementary information about the protein structure. Previous attempts to develop "predictive" HPLC for large biomacromolecules were limited by empirically derived equations based purely on the adsorption mechanisms of the retention and applicable to relatively small polypeptide molecules. A mechanism of the large biomacromolecule retention in reversed-phase gradient HPLC was described recently in thermodynamics terms by the analytical model of liquid chromatography at critical conditions (BioLCCC). In this work, we applied the BioLCCC model to predict retention of the intact proteins as well as their large proteolytic peptides separated under different HPLC conditions. The specific aim of these proof-of-principle studies was to demonstrate the feasibility of using "predictive" HPLC as a complementary tool to support the analysis of identified intact proteins in top-down, middle-down, and/or targeted selected reaction monitoring (SRM)-based proteomic experiments.