Mass Spectrometric Analysis Research Articles

Quality assessment and characterization of unregulated antiviral drugs for feline infectious peritonitis: implications for treatment, safety, and efficacy.

Feline infectious peritonitis is fatal, and due to lack of approved treatments, unregulated antiviral drugs are used to treat this disease. This study set out to determine the purity of various batches of these drugs from several companies, characterize them, and note any impurities or other unusual characteristics. We also developed a method to qualitatively assess the primary components before administration. We tested 30 vials from 17 brands of GS-441524 and 5 vials from 1 brand of GC376. We compared the GS-441524 to a control standard from Ambeed and the GC376 to a standard from Cayman Chemical. We recorded physical appearance, pH, absorbance, HPLC retention times, and thin-layer chromatography retention factors for all of the samples. Some samples were used for nuclear magnetic resonance and mass spectrometric analysis. Some of the GS-441524 vials were 10% to 25% more concentrated than advertised, but most of the GS-441524 samples tested were similar in purity and composition, both between batches and between brands. We also tested 5 vials of GC376 and found that 1 of the 5 vials contained GS-441524 rather than GC376 and the other 4 vials contained molnupiravir. While all of the GS-441524 vials contained GS-441524, none of the GC376 vials tested contained GC376. GC376 is used in cats that are unresponsive to GS-441524, and use of the wrong antiviral can cause serious side effects. We provide suggested methods for distinguishing one drug from the other in new batches.

Plasma Lipidomics Profiling to Identify the Biomarkers of Diagnosis and Radiotherapy Response for Advanced Non-Small-Cell Lung Cancer Patients.

Advanced lung cancer that contributes to a heavy burden on medical institutions is the leading cause of cancer-related death and is often accompanied by metabolic disorders. In this study, we aimed to explore the biomarkers of diagnosis and radiotherapy response in non-small-cell lung cancer (NSCLC) patients by plasma lipidomics analysis. Using triple-quadrupole mass spectrometer analysis, our research characterized the plasma lipid metabolomics profile of 25 healthy controls and 31 advanced NSCLC patients in each of three different radiotherapy phases. The results showed altered lipid elements and concentrations among NSCLC patients with different radiotherapy phases, NSCLC subtypes, and different radiotherapeutic responses. We found that compared to the healthy controls, myelin-associated glycoprotein (MAG), phosphatidylinositol (PI), and phosphatidylserine (PS) were mainly and significantly altered lipid elements (> twofold, and p < 0.05) among NSCLC patients with different radiotherapy phases. Through comparison of lipid elements between bad and good responses of NSCLC patients with radiotherapy, the obviously declined phosphatidylglycerol (PG 18 : 0/14 : 0, 18 : 1/18 : 3, and 18 : 0/20 : 1) or markedly elevated PI (20 : 0/22 : 5 and 18 : 2/22 : 4) and phosphatidic acid (PA 14 : 0/20 : 4, 14 : 0/20 : 3, and 18 : 2/22 : 4) could indicate poor therapeutic response for NSCLC patients. The results of ROC curve analysis suggested that PG (18 : 0/20 : 1 and 18 : 0/14 : 0) could clearly predict the radiotherapeutic response for NSCLC patients, and PS (18 : 0/20 : 0) and cholesterol were the first two lipid components with the most potential for the diagnosis of advanced NSCLC. Our results indicated that plasma lipidomics profiling might have a vital value to uncover the heterogeneity of lipid metabolism in healthy people and advanced NSCLC patients with different radiotherapy phase, and further to screen out radiotherapeutic response-specific biomarkers.

The adsorption of dragon fruit (Hylocereus polyrhizus) pigments on chitin for food colorants

AbstractBACKGROUNDGrowing concern over the potential adverse health effects of synthetic colorants has led to a rising utilization of plant‐derived pigments within the food industry. Purple betacyanins from red dragon fruits (Hylocereus polyrhizus) have been subjected to an environmentally sustainable purification process for natural colorants.RESULTSThe violet‐hued pigments were absorbed onto preconditioned chitin material, a biodegraded polymer derived from shrimp shells. Liquid chromatographic–electrospray ionization–tandem mass spectrometric analysis was employed to analyze the betalain pigment composition in the raw fruit extract, making comparisons between the adsorbed supernatant and desorbed solutions. Fourier transform infrared analysis, optical microscopy, and scanning electron microscopy were conducted on the resin pre‐ and post‐adsorption stages, revealing distinctive infrared bands (RNHR, CO, CO, OH) indicative of betacyanin functional groups in the pigmented materials. The uniform adsorption of the colorant onto the adsorbents causes the intense purple color of the resin, with a maximum removal efficiency of 0.29 mg g−1. Adsorption data were subjected to various kinetic models, with the pseudo‐second‐order model demonstrating the optimal fit to the kinetic data (R2 = 0.97), while the Freundlich model proved most suitable for describing the sorption isotherm (R2 = 0.98). The adsorption energy, as determined by the Dubinin–Radushkevich equation, was 0.23 kJ mol−1, implying a physical adsorption mechanism on a heterogeneous surface.CONCLUSIONBetacyanins derived from red dragon fruits could be recovered for food colorants through chitin adsorption. This recovery mechanism is attributed to electrostatic interactions among the functional groups, including hydroxyl and carboxyl groups, and hydrogen bonds. © 2024 Society of Chemical Industry (SCI).

Endocannabinoid regulation in the cervix during pregnancy: insights into molecular mechanisms of premature labor.

The cervix plays a crucial role not only in the maintenance of pregnancy but also during delivery, when it undergoes extensive changes. This study highlights the involvement of the endocannabinoidome in cervical remodeling, emphasizing its relevance in the shift from a nonpregnant to pregnant state and its potential contribution to preterm delivery in inflammatory contexts. During pregnancy, the main role of the cervix is to isolate the fetus from outside pathogens and maintain the relatively closed system of uterine gestation. Conversely, toward the end of pregnancy, the cervix must be remodeled to increase flexibility and allow the delivery. This process is called cervical remodeling and dysregulation of the process plays a role in premature delivery. The endocannabinoidome plays an important role in several reproductive events; however, its function on cervical tissue throughout pregnancy is poorly understood. The goal of this study was to evaluate the presence and participation of the endocannabinoidome in lipopolysaccharide (LPS)-induced cervical changes. Therefore, we evaluated key components of the endocannabinoidome in cervical tissue from nonpregnant mice and pregnant mice with and without LPS treatment. Using mass spectrometric analysis, we found an increase in anandamide and 2-arachidonoylglycerol in the cervix of pregnant mice when compared to nonpregnant mice. We have also found a reduction in FAAH protein expression in these tissues. Furthermore, when treated with LPS, we observed a reduction in the cervical immunostaining with anti-CB1 and anti-CB2 antibodies. Likewise, using cervix explants from pregnant mice, we found that LPS significantly increased cervical metalloprotease activity and cyclooxygenase 2, which were subsequently modulated by cannabinoid receptor antagonists. Collectively, our findings suggest that an LPS-induced imbalance of cervix endocannabinoidome likely contributes to premature cervical remodeling, which is part of the key components that contribute to premature delivery.

Exploring the Potential of Bioactive Peptides: From Natural Sources to Therapeutics.

Bioactive peptides, specific protein fragments with positive health effects, are gaining traction in drug development for advantages like enhanced penetration, low toxicity, and rapid clearance. This comprehensive review navigates the intricate landscape of peptide science, covering discovery to functional characterization. Beginning with a peptidomic exploration of natural sources, the review emphasizes the search for novel peptides. Extraction approaches, including enzymatic hydrolysis, microbial fermentation, and specialized methods for disulfide-linked peptides, are extensively covered. Mass spectrometric analysis techniques for data acquisition and identification, such as liquid chromatography, capillary electrophoresis, untargeted peptide analysis, and bioinformatics, are thoroughly outlined. The exploration of peptide bioactivity incorporates various methodologies, from in vitro assays to in silico techniques, including advanced approaches like phage display and cell-based assays. The review also discusses the structure-activity relationship in the context of antimicrobial peptides (AMPs), ACE-inhibitory peptides (ACEs), and antioxidative peptides (AOPs). Concluding with key findings and future research directions, this interdisciplinary review serves as a comprehensive reference, offering a holistic understanding of peptides and their potential therapeutic applications.

Unambiguous Analysis and Systematic Mapping of Oxygenated Aromatic Compounds in Atmospheric Aerosols Using Ultrahigh-Resolution Mass Spectrometry.

Compositional analysis of organic aerosols (OAs) at the molecular level has been a long-standing challenge in field and laboratory studies. In this work, we applied different extraction protocols to aerosol samples collected from the ambient atmosphere and biomass burning sources, followed by Orbitrap mass spectrometric analysis with a soft electrospray ionization source operating in both positive and negative ionization modes. To systematically map the distribution of mono- and dioxygenated aromatic compounds (referred to as aromatic CHO1 and CHO2 formulas) in OA, we developed a unique two-dimensional Kendrick mass defect (2D KMD) framework. Our analysis unveiled a total of (76, 64, 70) aromatic CHO1 formulas and (103, 110, 106) CHO2 formulas, corresponding to samples obtained from ambient air, rice straw burning, and sugarcane leaf burning, respectively. These results reveal a significant number of additional distinct formulas exclusively present in ambient samples, suggesting a significant chemical transformation of OAs in the atmosphere. The analytical approach can be further extended to incorporate multiple layers of 2D KMD, enabling systematic mapping of the unexplored chemical space for complex environmental samples.

NMR characterization of uniformly 13C- and/or 15N-labeled, unsulfated chondroitins with high molecular weights.

Solution nuclear magnetic resonance (NMR) analysis of polysaccharides can provide valuable information not only on their primary structures but also on their conformation, dynamics, and interactions under physiological conditions. One of the main problems is that non-anomeric 1H signals typically overlap, and this often hinders detailed NMR analysis. Isotope enrichment, such as with 13C and 15N, will add a new dimension to the NMR spectra of polysaccharides, and spectral analysis can be performed with enhanced sensitivity using isolated peaks. For this purpose, here we have prepared uniformly 13C- and/or 15N-labeled chondroitin polysaccharides -4)-β-D-glucuronopyranosyl-(1-3)-2-acetamido-2-deoxy-β-D-galactopyranosyl-(1- with molecular weights in the range from 310 to 460 k by bacterial fermentation. The enrichment ratios for 13C and 15N were 98.9 and 99.8%, respectively, based on the mass spectrometric analysis of the constituent chondroitin disaccharides. 1H and 13C NMR signals were assigned mainly based on HSQC and 13C-detection experiments including INADEQUATE, HETCOR, and HETCOR-TOCSY. The carbonyl carbon signal of the N-acetyl-β-D-galactosamine residue was unambiguously distinguished from the C6 carbon of the β-D-glucuronic acid residue by the observation of 13C peak splitting due to 1JCN coupling in 13C- and 15N-labeled chondroitin. The T2* and T1 were measured and indicate that both rigid and mobile sites are present in the long sequence of chondroitin. The conformation, dynamics, and interactions of chondroitin and its derivatives will be further analyzed based on the results obtained in this study.

Abstract B085: K17-induced pyrimidine biosynthesis drives chemoresistance in PDAC

Abstract Background: The 5-year survival rate of pancreatic cancer is extremely low compared with other cancers, although it has increased to 12% in 2023 in the United States. 80% of pancreatic ductal adenocarcinoma (PDAC) patients are not eligible for surgical procedure, and unfortunately, PDAC is refractory to standard-of-care chemotherapy. Thus, developing novel and more effective treatment strategies for PDAC is urgently needed. We previously reported keratin 17 (K17) as a novel negative prognostic and predictive biomarker, whose overexpression confers the resistance to chemotherapies. Here, we investigated the mechanisms of chemoresistance and tumor-specific vulnerabilities that can be exploited for K17-mediated targeted therapies. Methods: Survival analyses, immunohistochemistry (IHC), and matrix assisted laser desorption/ionization mass spectrometry (MALDI MSI) metabolite imaging were applied in patient derived samples. The K17 stable expression cell line models (KPC and PANC-1) and K17 loss-of-function cell line model (L3.6) were generated, where the analyses were applied including western blots, qRT-PCR, mass spectrometric metabolomics analysis, immunofluorescence imaging, cell viability and proliferation assays. KPC cells with or without K17 expression were orthotopically implanted into the head of the pancreas of c57B6J mice for tumor growth and treatment study. Results: Unbiased metabolomic studies in isogenic PDAC models identified several key metabolic pathways that are upregulated in the presence of K17. We demonstrate that K17 increases pyrimidine biosynthesis, a pathway that drives chemoresistance. Patient dataset analysis revealed that K17 expression and enzymes involved in pyrimidine, but not purine, de novo biosynthesis is associated with shorter patient survival. Furthermore, K17-expressing cells were more sensitive to Brequinar, a specific inhibitor of dihydroorotate dehydrogenase (DHODH), the rate-limiting enzyme in de novo pyrimidine biosynthetic pathway. Targeting DHODH by small interfering RNA or by Brequinar with Gemcitabine synergistically inhibited the viability of K17- positive cells. Importantly, the combination of Gemcitabine and Brequinar significantly inhibited the growth of K17-expressing tumors and extended survival of mice bearing K17-positive tumors. Mechanistically, K17 increased mitochondrial DHODH translocation which made these cells more sensitive to Brequinar. We also found that the coil 2 domain of K17 was crucial for triggering metabolic reprogramming and promoting tumor aggression. Conclusions: Overall, we define a novel connection between K17, nucleotide metabolism, and chemoresistance in PDAC. We report for the first time that K17, a negative biomarker of the most aggressive PDAC subtype, drives chemoresistance by up-regulating de novo pyrimidine biosynthesis, which provides a tumor-specific vulnerability: DHODH. Further study could lead to the development of a DHODH targeted therapy for K17-expressing PDAC. Citation Format: Yinghuan Lyu, Kenneth Shroyer, Chun-Hao Pan, Robert Tseng, Md Afjalus Siraj, Girish Halemirle Rajacharya, Bo Chen, Katie L Donnelly, Michael Horowitz, Carlos Mauricio Mejia Arbelaez, Sumedha Chowdhury, Lyanne Oblein, Natalia Marchenko, Luisa Escobar-Hoyos, Pankaj K. Singh. K17-induced pyrimidine biosynthesis drives chemoresistance in PDAC [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B085.

Mass spectrometric analysis of transition metal complexes formed through contact of artificial sweat with circulating Euro coins

AbstractIt is well‐known that some metals/alloys sensitize the skin of susceptible individuals. Trace amounts of metals and chemical compounds containing metal ions can affect human health, i. e., cause allergy or inflammation. However, the details of this metal‐induced allergic reaction are not well understood. Complexes of nickel, copper, and zinc are known to be easily formed through contact of metal alloys with ubiquitously available organic compounds such as triacylglycerols, amino acids, urea, and fatty acids found in skin secretions or skin care products. Thus, an analytical technique is required to characterize and identify metal complexes formed on metallic everyday products, ideally offering a chemically specific, laterally resolved view of allergy‐causing processes. Here we present a mass‐spectrometry‐based analytical workflow to characterize metal‐containing compounds formed on the surface of 1‐euro coins when in contact with artificial sweat. This study identified different metal complexes by using tandem mass spectrometry (MS/MS). Surface imaging of 1‐Euro coins employing laser‐based mass spectrometry revealed the lateral distribution of Zn, Cu, and Ni complexes with low‐micrometer resolution.

Mass spectrometric analysis of unprecedented high levels of carbonaceous aerosol particles long-range transported from wildfires in the Siberian Arctic

Abstract. Wildfires in Siberia generate large amounts of aerosols, which may be transported over long distances and pose a threat to the sensitive ecosystem of the Arctic. Particulate matter (PM) of aged wildfire plumes originating from Yakutia in August 2021 was collected in Nadym and on Bely Island (both in northwestern Siberia). An advanced analysis of the chemical composition of aerosol particles was conducted through a multi-wavelength thermal–optical carbon analyzer (TOCA) coupled to resonance-enhanced multiphoton ionization time-of-flight mass spectrometry (REMPI-TOFMS) as well as through ultra-high-resolution Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS). In Nadym, concentrations of organic carbon (OC) and elemental carbon (EC) peaked at 100 and 40 µg m−3, respectively, associated with Angström absorption exponents for 405 and 808 nm (AAE405/808) between 1.5 and 3.3. The weekly average on Bely Island peaked at 8.9 µg m−3 of OC and 0.3 µg m−3 of EC and AAE405/808 close to unity. In particular, ambient aerosol in Nadym had a distinct biomass burning profile with pyrolysis products from carbohydrates, such as cellulose and hemicellulose, as well as lignin and resinoic acids. However, temporarily higher concentrations of five- and six-ring polycyclic aromatic hydrocarbons (PAHs), different from the PAH signature of biomass burning, suggest a contribution of regional gas flaring. FT-ICR MS with electrospray ionization (ESI) revealed a complex mixture of highly functionalized compounds, containing up to 20 oxygen atoms, as well as nitrogen- and sulfur-containing moieties. Concentrations of biomass burning markers on Bely Island were substantially lower than in Nadym, flanked by the appearance of unique compounds with higher oxygen content, higher molecular weight, and lower aromaticity. Back-trajectory analysis and satellite-derived aerosol optical depth suggested long-range transport of aerosol from the center of a Yakutian wildfire plume to Nadym and from the plume periphery to Bely Island. Owing to lower aerosol concentrations in the plume periphery than in its center, it is demonstrated how dilution affects the chemical plume composition during atmospheric aging.

Gas chromatography tandem mass spectrometric analysis of alkylphosphonofluoridic acids as verification targets of nerve agents

Alkylphosphonofluoridic Acids (APFA) are the major thermal degradation products of G- and A-series nerve agents and thus play a vital role in the verification analysis of Chemical Weapons Convention. Present study focuses on the development of sample clean-up, derivatization procedures and gas chromatography tandem mass spectrometric analysis of APFA in aqueous samples. APFA were found to be much more delicate than the corresponding alkylphosphonic acids and thus required subtle optimizations. Retention of analytes on silica and polymer-based anion exchangers followed by elution under alkaline conditions yielded best recoveries. Elution under acidic conditions led to partial or complete degradation of the analytes to alkylphosphonic acids. Silylation reactions, particularly with MTBSTFA were found the best in terms of chromatographic responses and resolution of the derivative peaks. Methylations with diazomethane, which requires acidic reaction media, failed to produce desired yields of the derivatives. Under optimized conditions, the analytes produced the recoveries ranging from 76.9 to 94.5% with RSD ≤9.2%. The best LOD's in the tandem mass spectrometric analysis ranged from 13 to 56 ng/ml. The applicability of the method was tested by spiking the analytes in the retained aqueous samples received for the 52nd proficiency test conducted by the Organization for the Prohibition of Chemical Weapons (OPCW).

Identification of a rare [Gγ(Aγδβ)0] -thalassemia using tandem mass spectrometry.

Thalassemias are a group of inherited monogenic disorders characterized by defects in the synthesis of one or more of the globin chain subunits of the hemoglobin tetramer. Delta-beta (δβ-) thalassemia has large deletions in the β globin gene cluster involving δ- and β-globin genes, leading to absent or reduced synthesis of both δ- and β-globin chains. Here, we used direct globin-chain analysis using tandem mass spectrometry for the diagnosis of δβ-thalassemia. Two cases from unrelated families were recruited for the study based on clinical and hematological evaluation. Peptides obtained after trypsin digestion of proteins extracted from red blood cell pellets from two affected individuals and their parents were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Mass spectrometric analysis revealed a severe reduction in δ, β, and Aγ globin proteins with increased Gγ globin protein in the affected individuals. The diagnosis of Gγ(Aγδβ)0 -thalassemia in the homozygous state in the affected individuals and in the heterozygous state in the parents was made from our results. The diagnosis was confirmed at the genetic level using multiplex ligation-dependent probe amplification (MLPA). Our findings demonstrate the utility of direct globin protein quantitation using LC-MS/MS to quantify individual globin proteins reflecting changes in globin production. This approach can be utilized for accurate and timely diagnosis of hemoglobinopathies, including rare variants, where existing diagnostic methods provide inconclusive results.

Expression, purification and folding of native like mitochondrial carrier proteins in lipid membranes

Mitochondrial Carrier Family proteins (MCFs) are located in the mitochondrial inner membrane and play essential roles in various cellular processes. Due to the relatively low abundance of many members of the family, in vitro structure and function determination of most MCFs require over-expression and purification of recombinant versions of these proteins. In this study, we report on a new method for overexpression of MCFs in Escherichia coli (E. coli) membranes, efficient purification of native-like proteins, and their reconstitution in mitochondrial inner membrane lipid mimics. cDNAs of Uncoupling Protein 4 (UCP4), Adenine Nucleotide Translocase (ANT) and Phosphate Translocase (PiT) were subcloned into the pET26b (+) expression vector such that fusion proteins with a short N-terminal pelB leader sequence and a six-histidine tag were produced to target the proteins toward the inner membrane of E. coli and facilitate affinity purification, respectively. Utilizing a modified autoinduction method, these proteins were overexpressed and extracted from the membrane of E. coli BL21 (DE3) and two modified strains, E. coli BL21 C43 (DE3) and E. coli BL21 Lobstr (DE3), in high yields. The proteins were then purified by immobilized metal affinity chromatography as monomers. Purity, identity, and concentration of the eluted monomers were determined by semi-native SDS-PAGE, Western blotting and mass spectrometry, and a modified Lowry assay, respectively. Cleavage of the pelB leader sequence from proteins was verified by mass spectrometric analysis. The purified proteins, surrounded by a shell of bacterial membrane lipids, were then reconstituted from the mild non-denaturing octyl glucoside (OG) detergent into phospholipid liposomes. Monomeric UCP4 spontaneously self-associated to form stable tetramers in lipid membranes, which is consistent with our previous studies. However, PiT and ANT remained dominantly monomeric in both detergent and liposome milieus, as detected by a combination of spectroscopic and electrophoretic methods. Native-like helical conformations of proteins were then confirmed by circular dichroism spectroscopy. Overall, this study demonstrates that targeting mitochondrial carrier family proteins to E. coli membranes provides an effective expression system for producing this family of proteins for biophysical studies.

Preclinical evaluation of an 18F-labeled Nε-acryloyllysine piperazide for covalent targeting of transglutaminase 2.

Transglutaminase 2 (TGase2) is a multifunctional protein and has a prominent role in various (patho)physiological processes. In particular, its transamidase activity, which is rather latent under physiological conditions, gains importance in malignant cells. Thus, there is a great need of theranostic probes for targeting tumor-associated TGase 2, and targeted covalent inhibitors appear to be particularly attractive as vector molecules. Such an inhibitor, equipped with a radionuclide suitable for noninvasive imaging, would be supportive for answering the general question on the possibility for functional characterization of tumor-associated TGase 2. For this purpose, the recently developed 18F-labeled Nε-acryloyllysine piperazide [18F]7b, which is a potent and selective irreversible inhibitor of TGase2, was subject to a detailed radiopharmacological characterization herein. An alternative radiosynthesis of [18F]7b is presented, which demands less than 300µg of the respective trimethylammonio precursor per synthesis and provides [18F]7b in good radiochemical yields (17 ± 7%) and high (radio)chemical purities (≥ 99%). Ex vivo biodistribution studies in healthy mice at 5 and 60min p.i. revealed no permanent enrichment of 18F-activity in tissues with the exception of the bone tissue. In vivo pretreatment with ketoconazole and in vitro murine liver microsome studies complemented by mass spectrometric analysis demonstrated that bone uptake originates from metabolically released [18F]fluoride. Further metabolic transformations of [18F]7b include mono-hydroxylation and glucuronidation. Based on blood sampling data and liver microsome experiments, pharmacokinetic parameters such as plasma and intrinsic clearance were derived, which substantiated the apparently rapid distribution of [18F]7b in and elimination from the organisms. A TGase2-mediated uptake of [18F]7b in different tumor cell lines could not be proven. Moreover, evaluation of [18F]7b in melanoma tumor xenograft models based on A375-hS100A4 (TGase2 +) and MeWo (TGase 2 -) cells by ex vivo biodistribution and PET imaging studies were not indicative for a specific targeting. [18F]7b is a valuable radiometric tool to study TGase2 in vitro under various conditions. However, its suitability for targeting tumor-associated TGase2 is strongly limited due its unfavorable pharmacokinetic properties as demonstrated in rodents. Consequently, from a radiochemical perspective [18F]7b requires appropriate structural modifications to overcome these limitations.

Analysis of the Snake Venom Peptidome.

Snake venom peptidomes are known to be a large source of molecules with different pharmacological properties. The complexity and variability of snake venoms, the presence of proteinases, and the lack of complete species-specific genome sequences make snake venom peptidome profiling a challenging task that requires especial technical strategies for sample processing and mass spectrometric analysis. Here, we describe a method for assessing the content of snake venom peptides and highlight the importance of sampling procedures, as they substantially influence the peptidomic complexity of snake venoms.

Comparative analyses of adsorbed circulating proteins in the PMMA and PES hemodiafilters in patients on predilution online hemodiafiltration.

Acute and chronic inflammation are common in patients with end-stage kidney disease (ESKD). So, the adsorption of pro-inflammatory cytokines by the hollow fiber of the dialysis membrane has been expected to modify the inflammatory dysregulation in ESKD patients. However, it remains to be determined in detail what molecules of fiber materials can preferably adsorb proteins from the circulating circuit. We aimed this study to analyze directly the adsorbed proteins in the polymethyl methacrylate (PMMA) and polyethersulfone (PES) membranes in patients on predilution online hemodiafiltration (OL-HDF). To compare the adsorbed proteins in the PMMA and PES hemodiafilters membrane, we initially performed predilution OL-HDF using the PES (MFX-25Seco) membrane while then switched to the PMMA (PMF™-A) membrane under the same condition in three patients. We extracted proteins from the collected hemodiafilters by extraction, then SDS-PAGE of the extracted sample, protein isolation, in-gel tryptic digestion, and nano-LC MS/MS analyses. The concentrations of adsorbed proteins from the PMMA and PES membrane extracts were 35.6±7.9 μg/μL and 26.1±9.2 μg/μL. SDS-PAGE analysis revealed distinct variations of adsorbed proteins mainly in the molecular weight between 10 to 25 kDa. By tryptic gel digestion and mass spectrometric analysis, the PMMA membrane exhibited higher adsorptions of β2 microglobulin, dermcidin, retinol-binding protein-4, and lambda-1 light chain than those from the PES membrane. In contrast, amyloid A-1 protein was adsorbed more potently in the PES membrane. Western blot analyses revealed that the PMMA membrane adsorbed interleukin-6 (IL-6) approximately 5 to 118 times compared to the PES membrane. These findings suggest that PMMA-based OL-HDF therapy may be useful in controlling inflammatory status in ESKD patients.

Geoecological and geochemical assessment of the impact of NMMC mining enterprises on the environment

Study of the dynamics of natural and man-made processes and their impact on the environment in the hills of Uzbekistan; study of ecological and geochemical processes occurring in technogenic and natural landscapes of mining enterprises of the NMMC; Conducting geochemical and analytical studies to identify the contents and distribution of toxic and other associated elements in technogenic waste from NMMC mining enterprises, soils, and plants in connection with their impact on the ecology of the environment. Study of geochemical features of the productive valuable and toxic element`s behavior in space and time in industrial waste area Geological and geochemical mapping of soil, plants and water sources. Study of the chemical composition, behavior and forms of occurrence of toxic elements (in soils, water streams, dust, gases, smoke) in the water area and in the vicinity of the Navoi Mining and Metallurgical Combine - geochemical ecology. As a result of investigations of initial samples by using mass spectrometric analysis, the limits of fluctuations and average contents of not only toxic elements, but also other associated elements were revealed.

1,4:3,6-Dianhydrohexites

It is known that derivatives of isosorbide and isomannide can impart a sweet taste, for example, dimethyl ether of isosorbide is used as an ingredient in the production of chewing gums, hard sweets [1], isosorbide dipropanoate is widely used in the production of bakery products [1]. Mixed isohexite esters and esters (especially isosorbide) are used as flavor enhancers. Currently, isosorbide is proposed to be used as a component of the mixture used for the water-base ink pigment. This component has excellent dispersion stability, which is necessary for printing [1]. All production methods leading to isohexites use the corresponding hexitol as the starting material. To obtain isohexites with a quantitative yield, and in order to avoid adverse reactions, a number of techniques have been developed. The rate of the dehydration sequence in reactions using as the starting compound: 1,4-anhydro derivatives of D-iditol, D-gulitol, D-glucitol, and D-mannitol (from which 1,4:3,6-dianhydro compounds of isoidide (5) and isomannide (4) are obtained) depends on the location of the hydroxyl group (with5 endo as in D-mannitol and D-glucitol, or exo as in D-iditol and D-glucitol). All three isohexites (3-5) were studied by mass spectrometry, especially D-isomannide and their O-deuterated isomers, and 5-nitroisosorbide [1]. The isomannide cation formed as a result of dehydration reactions was also detected in the mass spectra of some isomannide derivatives [1]. Mass spectrometric analysis is used to determine isosorbide and its 5-nitro derivative in human urine and blood plasma.

Thiol Redox Proteomics for Identifying Redox-Sensitive Cysteine Residues Within the Protein of Interest During Stress.

Redox modulation is a common posttranslational modification to regulate protein activity. The targets of oxidizing agents are cysteine residues(Cys), which have to be exposed at the surface of the proteins and are characterized by an environment that favors redox modulation. This includes their protonation state and the neighboring amino acids. The Cys redox state can be assessed experimentally by redox titrations to determine the midpoint redox potential in the protein. Exposed cysteine residues and putative intramolecular disulfide bonds can be predicted by alignments with structural data using dedicated software tools and information on conserved cysteine residues. Labeling with light and heavy reagents, such as N-ethylmaleimide (NEM), followed by mass spectrometric analysis, allows for the experimental determination of redox-responsive cysteine residues. This type of thiol redox proteomics is a powerful approach to assessing the redox state of the cell, e.g., in dependence on environmental conditions and, in particular, under abiotic stress.

Detection of several volatile organic compounds through Ar+ induced chemical ionisation using inductively coupled plasma-tandem mass spectrometry (ICP-MS/MS).

A new analytical technique for detection of organic compounds using inductively coupled plasma-tandem mass spectrometry (ICP-MS/MS) is described. Volatile organic compounds (VOCs) were introduced into the collision/reaction cell (CRC), instead of through the ICP ion source, and the molecules were ionised through an ion reaction, induced by collision with the primary ions (Ar+) produced in the ICP. The ionisation characteristics of this new approach were investigated by mass spectrometric analysis of eight VOCs (i.e., benzene, toluene, ethyl acetate, methyl butyrate, ethyl butyrate, pentyl acetate, pyridine, and 2-methylfuran). These molecules were detected as molecular ions (M+), protonated ions ([M + H]+), or deprotonated ions ([M - H]+), demonstrating that soft ionisation was achieved by the present ionisation protocol using ICP-MS/MS. In addition, a volatile selenium-containing organic compound, dimethyl diselenide (Se2(CH3)2), was also analysed to investigate the feasibility of this ionisation protocol to achieve soft and hard ionisation simultaneously. Several Se-related ions such as Se+, SeH+, Se2+, [SeCH3]+, and [Se2CH3]+, together with [Se2(CH3)2]+, were observed, suggesting that while soft ionisation was possible, ion reaction-induced-fragmentation and hard ionisation also occurred. To demonstrate the analytical capability of the present technique, volatile components released from coffee beans were subjected to the present mass spectrometric analysis. Many ion peaks originating from VOCs were detected from the coffee beans. The data obtained here demonstrated that ICP-MS equipped with a CRC can become an effective tool for analyzing both elements and molecules.