Acid Analysis Research Articles

Progression-Free Survival Analysis of Denosumab Vs Zoledronic Acid in Intent to Transplant Multiple Myeloma Patients Based on Treatment Regimen and Baseline Characteristics

Introduction: Denosumab is a monoclonal antibody targeting Receptor Activator of Nuclear Factor-Kappa B Ligand (RANKL) that has been shown to reduce skeletal related events associated with bone lesions in patients with multiple myeloma and solid tumors. Results from the full primary analysis of 1718 patients with newly diagnosed multiple myeloma in an international double blind, randomized, controlled phase 3 (20090482) study that assessed the efficacy of denosumab (Dmab) vs zoledronic acid (ZA) for preventing SREs met its primary end point of non-inferiority regarding time to first SRE. The analysis of the PFS exploratory endpoint showed a clinically meaningful 10.7 months median PFS benefit (HR, 0.82; 95% CI, 0.68-0.99; descriptive P= 0.036) of Dmab vs ZA. This benefit was most pronounced in patients who were stratified into the "intent to undergo Autologous Stem Cell Transplant (ASCT)" group at randomization. Thus, we present an in-depth analysis of relevant baseline characteristics, treatment regimens and PFS outcome in patients with intent to undergo transplant receiving Dmab and ZA. Methods: Adult patients with newly diagnosed multiple myeloma (NDMM) and stratified as "intent to undergo ASCT" at randomization were included in this analysis. Patients received subcutaneous denosumab (120 mg) plus intravenous placebo or intravenous zoledronic acid (4 mg) plus subcutaneous placebo in 4-week cycles. In this subgroup, the PFS outcome was examined. Baseline characteristics and treatment regimens were compared between treatment arms. Results: 54.1% of the 1718 enrolled patients were stratified into "intent to undergo ASCT" as part of their front-line therapy, and 61.8% of "intent to undergo ASCT" did receive an ASCT. In the "intent to undergo ASCT" group, 19.6% patients had disease progression in the Dmab arm compared to 28.0% in the ZA arm (HR 0.65 (0.49-0.85)) (Figure 1). No imbalance in terms of triplet therapy use between the two study arms (TABLE 1). 55.1% in Dmab vs 52.6% in ZA arm received Triplet Therapies which included Bortezomib, Cyclophosphamide, Dexamethasone (VCD), Bortezomib, Thalidomide, Dexamethasone (VTD), Cyclophosphamide, Thalidomide, Dexamethasone (CTD), or Bortezomib, Lenalidomide, Dexamethasone (VRD). The percentage of triplet therapies used in the "intent to undergo ASCT"patients was higher than in patients with no intent to undergo ASCT. Percentage of patients with ECOG performance status 2 was 19.4% in the Dmab group vs 18.6% in the ZA group. 26.2% of patients in the Dmab arm and 25% in the ZA arm had Multiple Myeloma ISS stage III upon diagnosis. Among intent to transplant patients there was no imbalance in terms of age, performance status, ISS stage, risk status, weight, bone marrow plasma cell % between the ZA and the Dmab arm Conclusion: Results from this post-hoc subgroup analysis suggest a more profound PFS benefit in the "intent to undergo ASCT" patient subgroup. Multiple myeloma treatment received in the intent to undergo transplant subjects was similar between the denosumab and zoledronic acid arms. No significant imbalance in demographics or baseline disease characteristics was observed between the two treatment arms. Disclosures Terpos: Janssen: Honoraria, Other: Travel expenses, Research Funding; Amgen: Honoraria, Research Funding; Takeda: Honoraria, Other: Travel expenses, Research Funding; Medison: Honoraria; Genesis: Honoraria, Other: Travel expenses, Research Funding; Celgene: Honoraria. Shimizu:Medical Biological Laboratory: Consultancy; Takeda: Speakers Bureau; Daiichi: Consultancy; Amgen: Consultancy; Fujimoto: Consultancy. Willenbacher:Abbvie: Consultancy, Honoraria; Sandoz: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Merck: Consultancy, Membership on an entity's Board of Directors or advisory committees; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; oncotyrol: Employment, Research Funding; Gilead Science: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; Roche: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; European Commission: Research Funding; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Novartis: Consultancy, Membership on an entity's Board of Directors or advisory committees, Research Funding; Bristol-Myers Squibb: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Tirol Program: Research Funding; Fujimoto: Consultancy, Honoraria; Myelom- und Lymphomselbsthilfe Österreich: Consultancy, Honoraria; Sanofi: Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees; IQVIA: Membership on an entity's Board of Directors or advisory committees. Glennane:amgen: Employment, Equity Ownership. Dai:Amgen: Employment, Equity Ownership. Pasteiner:Amgen: Employment, Equity Ownership. Raje:Amgen Inc.: Consultancy; Bristol-Myers Squibb: Consultancy; Celgene Corporation: Consultancy; Takeda: Consultancy; Janssen: Consultancy; Merck: Consultancy.

Role of intestinal microbiota changes in cardiovascular diseases pathogenesis

Purpose of this article is to clarify the role of intestinal microbiota changes in cardiovascular diseases pathogenesis. In the analysis of intestinal microbiota changes in patients with cardiac pathology, a diversity and lack of research on this issue are notable. Intestinal microflora performs immune, metabolic, synthetic, enzymatic, antitoxic and other functions, providing intestinal colonization resistance. The intestinal microbiota effects on the human body are primarily due to toxic metabolites which can be provisionally divided into five groups: trimethylamine and its derivatives, short-chain fatty acids, endotoxin and bacterial wall products, bile acids and uremic toxins. It is interesting to further study their role in the pathogenesis of heart diseases. For example, bacterial role in trimethylamine, choline metabolism and their effects on the pathogenesis of atherosclerosis. The possibilities of their use as diagnostic markers are studied. However, studies of trimethylamines are usually associated with difficulties, due to their strong dependence on the patient's diet. Trimethylamines are the most studied group of intestinal metabolites, although there are still no reliable ways to correct them. Analysis of short-chain fatty acids requires the identification of each fatty acid individual role, and comparing their content in peripheral blood and stool. Endotoxin is a marker of microbial activity in the intestine. It is an important component of the bacterial cell wall. A study of bile acids metabolism changes influenced by intestinal microbiota also can help to understand disorders of lipid metabolism. An increase in the intestinal microflora uremic toxins (p-cresol, indoxyl sulfate) synthesis is associated with chronic kidney disease. It is promising to develop further ways for correction of the intestinal microbiota composition, which requires a better understanding of cardio-vascular pathology pathogenetic mechanisms.Conclusions. The study of intestinal microbiota composition is appropriate taking into account synthetic activity of various bacteria. Reviewing the effect of microbial metabolites on the human body and studying the mechanisms of their synthesis, it is possible to understand their role as diagnostic markers and suggest ways of correction.

Zastosowanie termograwimetrii oraz TLC w połączeniu z densytometrią w analizie kwasu nikotynowego i jego pochodnych

Zastosowanie termograwimetrii oraz TLC w połączeniu z densytometrią w analizie kwasu nikotynowego i jego pochodnych

Rapid and comprehensive evaluation of microalgal fatty acids via untargeted gas chromatography and time-of-flight mass spectrometry.

Due to their high triacylglyceride content, microalgae are intensively investigated for bio‐economy and food applications. However, lipid analysis is a laborious task incorporating extraction, transesterification and typically gas chromatographic measurement. Co‐elution induces a significant risk of fatty acid misidentification and thus, additional purification steps like thin layer chromatography are needed. Contrary to database matching approaches, solely targeted analysis is facilitated as compound identification is driven by matching retention times or indices with standard substances. In this context, a rapid workflow for the analysis of algal fatty acids is presented. In‐situ transesterification was used to simplify sample preparation and conditions were optimized towards fast processing. If results are needed at the very day of sampling, direct processing without a preceding drying step is feasible to obtain a rough estimate about the occurrence of the major compounds. Coupling gas chromatography and time‐of‐flight mass spectrometry enables untargeted analysis. Unknown compounds may be identified by structural reconstruction of their respective fragmentation patterns and by database matching to routinely avoid mismatches by co‐elution of disturbing agents. The developed workflow was successfully applied to derive the exact stereochemistry of all fatty acids from Chlorella vulgaris and a systematic shift depending on physiological state of the cells was confirmed.

Correction: Effects of dietary n-6: n-3 polyunsaturated fatty acid ratios on meat quality, carcass characteristics, tissue fatty acid profiles, and expression of lipogenic genes in growing goats

[This corrects the article DOI: 10.1371/journal.pone.0188369.].

Evaluation of fatty acids in groomed fingerprint by gas chromatographic analysis using various extraction solvents and treatment methods

Extremely small amounts of fatty acids detected in latent fingerprints are important for studying fingerprint visualization and age determination through changes in composition over time. However, methods for efficiently extracting or recovering fatty acids from fingerprints have not been extensively studied. If accurate and stable quantitative estimations are established, age estimates will be possible through a better understanding of the fatty acid composition. The extraction solvent and treatment method are essential factors for achieving a reliable analysis of fatty acids. There have been few previous studies that efficiently compared fatty acids. In this study, fatty acids from sebaceous fingerprint residues were quantified with various extraction solvents and treatment methods and were evaluated with gas chromatography flame ionization detection (GC-FID). All data were analyzed using a statistical method.

Pentacylic triterpenes from Lavandula coronopifolia: structure related inhibitory activity on α-glucosidase

Ten pentacyclic triterpenes (1-10) were isolated from Lavandula coronopifolia. We evaluated their α-glucosidase inhibitory activity, and found that the aglycones, 1, 2, 3, 4, 7 and 10 showed superior IC50 values to the positive control. In order to explain the structural requirements for α-glucosidase inhibitory activity, eleven derivatives were prepared, including one new compound, 2-formyl-(A)1–19α-hydroxy-1-norursane-2, 12-dien-28-oic acid 10c. The results demonstrated that a free hydroxyl at ring-A and a free carboxylic group at position 28 are key structural features for the α-glucosidase inhibitory activity, also that an ursane skeleton is optimum for the activity. Additionally, enzyme kinetic analysis of pomolic acid 2, the most potent compound, revealed that it inhibited α-glucosidase in a mixed-type manner. The molecular docking simulation validated this type of inhibition and highlighted the role of the C-3 hydroxyl and C-28 carboxylic groups in interaction with the enzyme in silico.

Influence of Repeated Application of Wetting Agents on Soil Water Repellency and Microbial Community

Wetting agents are the primary tool used to control soil water repellency (SWR) and localized dry spot (LDS), especially on sand-based soils. However, the effect of repeated applications of wetting agents on soil microbial populations is unknown. This two-year field experiment investigated six wetting agents representing different chemistry effects on a creeping bentgrass (Agrostis stolonifera L.) putting green with existing SWR. Four out of the six wetting agents improved soil volumetric water content in the second growing season, while others showed no effect. This result was negatively correlated to the development of LDS, and positively correlated to occurrence of an air-borne turf disease. Soil microbial populations, determined by soil phospholipid fatty acid (PLFA) analysis, found that none of the treatments applied caused a shift in microbial populations between fungi and bacteria, or gram-positive and gram-negative bacteria. The stress indicators such as saturated to mono-unsaturated fatty acids were not affected by the wetting agents applied as well. However, the wetting agent that contains alkyl block polymers (ABP; Matador) with proven capability for removal of soil organic coatings showed inhibition of microbial populations at one evaluation timing. This result suggested a temporary restriction in soil carbon availability for soil microorganisms following repeated ABP application, which likely contributed to the elevated LDS development observed. Another wetting agent, a combined product of a nonionic surfactant plus acidifiers (NIS; pHAcid), which is designed to reduce inorganic carbonates while enhancing wetting, elevated all soil microbial populations tested at the end of the experiment, indicating a desirable improvement in soil health. However, repeated application of NIS did not reduce SWR at the conclusion of this experiment, which, in combination with a previous report, suggested a minimal disturbance of soil organic coatings of the hydrophobic sand. Overall, this experiment suggested that soil microbial populations can be affected by wetting agents which may further influence SWR, yet the actual effect on soil microorganisms varies depending on the chemistry of the wetting agents.

Subfossil freshwater bivalve mollusk shells present data on stratigraphy of Dniester and Prut riverine deposits

The article provides information on the outcrops of the Neogene-Quaternary riverine sediments of the North-Western Black Sea coastal area. A description of five outcrops of fluvial deposits located on the territory of the modern basins of the Dniester and Prut rivers is given. Based on the granulometric composition of the sediments and the presence in them of the fossil shells of freshwater bivalve mollusks (Bivalvia: Unionoida), an assumption has been made about the characteristics of the ancient river ecosystems of the Dniester and Prut on different Pliocene and Pleistocene time sections. A review of the data in the body of literature on the geological age of the studied outcrops was undertaken. The localities considered provide information on the sedimentation conditions in this region from the Pliocene to the Late Pleistocene. Previously, various approaches were used for dating such rocks and for determining the stratigraphic position of fossil material found in them. The article considers the method of amino acids racemization as one of the approaches. Racemization allows to obtain new data on the stratigraphy of Neogene-Quaternary riverine sediments and to solve the problem of their dating. The sampling principles of carbonate material for the analysis of amino acids from the mollusk shells and features of sample preparation are described. Five amino acids were used: aspartic acid (Asp), glutamic acid (Glu), leucine (Leu), phenylalanine (Phe) and isoleucine (Ile). The D/L ratios were analyzed (amino acid racemization (or AAR) is the interconversion of amino acids from one chiral form (the L – (laevo) amino acids which are the building blocks of proteins) to a mixture of L- and D- (dextro) forms. The extent of racemization is measured by the ratio of D/L isomers (it increases as a function of time and temperature and can be used for geochronology) from the same group of fossils (genera), which were preserved under similar environmental conditions, inorganic geochemistry and thermal histories. Based on the obtained values, it can be concluded that the analysis of the racemization of amino acids is useful for determining the geological age of fossil shells of freshwater bivalve mollusks, but there are limitations regarding age. In the shells of the Pliocene sites, the values of some amino acids are close to one, indicating that the racemization took place. D/L values in shells from Pleistocene localities allowed the determination of their stratigraphic position.

FAIR data in meta-omics research: Using the MOD-CO schema to describe structural and operational elements of workflows from field to publication

Nucleic acid and protein sequencing-based analyses are increasingly applied to determine origin, identity and traits of environmental (biological) objects and organisms. In this context, the need for corresponding data structures has become evident. As existing schemas and community standards in the domains of biodiversity and molecular biological research are comparatively limited with regard to the number of generic and specific elements, previous schemas for describing the physical and digital objects need to be replaced or expanded by new elements for covering the requirements from meta-omics techniques and operational details. On the one hand, schemas and standards are hitherto mostly focussed on elements, descriptors, or concepts that are relevant for data exchange and publication, on the other hand, detailed operational aspects regarding origin context and laboratory processing, as well as data management details, like the documentation of physical and digital object identifiers, are rather neglected. The conceptual schema for Meta-omics Data and Collection Objects (MOD-CO; https://www.mod-co.net/) has been set up recently Rambold et al. 2019. It includes design elements (descriptors or concepts), describing structural and operational details along the work- and dataflow from gathering environmental samples to the various transformation, transaction, and measurement steps in the laboratory up to sample and data publication and archiving. The concepts are named according to a multipartite naming structure, describing internal hierarchies and are arranged in concept (sub-)collections. By supporting various kinds of data record relationships, the schema allows for the concatenation of individual records of the operational segments along a workflow (Fig. 1). Thus, it may serve as a logical and structural backbone for laboratory information management systems. The concept structure in version 1.0 comprises 653 descriptors (concepts) and 1,810 predefined descriptor states, organised in 37 concept (sub-)collections. The published version 1.0 is available as various schema representations of identical content (https://www.mod-co.net/wiki/Schema_Representations). A normative XSD (= XML Schema Definition) for the schema version 1.0 is available under http://schema.mod-o.net/MOD-CO_1.0.xsd. The MOD-CO concepts might be integrated as descriptor/element structures in the relational database DiversityDescriptions (DWB-DD) an open-source and freely available software of the Diversity Workbench (DWB; https://diversityworkbench.net/Portal/DiversityDescriptions; https://diversityworkbench.net). Currently, DWB-DD is installed at the Data Center of the Bavarian Natural History Collections (SNSB) to build an instance of its own for storing and maintaining MOD-CO-structured meta-omics research data packages and enrich them with ‘metadata’ elements from the community standards Ecological Markup Language (EML), Minimum Information about any (x) Sequence (MIxS), Darwin Core (DwC) and Access to Biological Collection Data (ABCD). These activities are achieved in the context of ongoing FAIR ('Findable, Accessible, Interoperable and Reuseable') biodiversity research data publishing via the German Federation for Biological Data (GFBio) network (https://www.gfbio.org/). Version 1.1 of the schema with extended collections of structural and operational design concepts is scheduled for 2020.

Abstract 3978: Direct analysis of exosome-bound proteins and nucleic acids using the ExoVerita™ Flex platform

Abstract Exosomes carry signals between cells and may influence cancer development, metastasis, and drug resistance. Unfortunately, current tools for exosome isolation and analysis have been limited to examining a specific class of markers, such as DNA, RNA, and proteins. We have developed a microelectrode array chip that uses a novel Alternating Current Electrokinetic method to perform rapid, affinity-free isolation of exosomes. The chip runs on our proprietary ExoVeritaTM Flex platform. Isolated exosomes can then be analyzed in situ or can be lysed to obtain DNA and RNA for use in downstream assays, such as sequencing. In this study, we used the ExoVerita Flex to isolate exosomes directly from plasma specimens obtained from cancer subjects with lung, colorectal, or pancreatic cancers as well as healthy donors. Isolated exosomes were analyzed in situ for exosome-bound proteins and nucleic acids, and then purified following isolation to examine their nucleic acid expression profiles. Using the ExoVerita Flex with plasma specimens and antibodies targeting the exosome-bound proteins CEA, PD-L1, GPC-1 and CD63, we observed differences in protein expression levels among the different cancer types (N = 35) and healthy donors (N = 20), such as increased PD-L1 expression in non-small cell lung cancer subject samples and elevated levels of CEA in colorectal cancer subject samples (N = 10). Furthermore, we co-stained samples with antibodies and DNA- or RNA-selective dyes to simultaneously measure exosome-bound proteins and circulating nucleic acid levels in the same sample at the same time. RNA sequencing analysis was performed by extracting RNA from the isolated exosomes using a proprietary buffer, and then performing RT-ddPCR and miRNA-Sequencing. RT-ddPCR analysis of the PGK-1 housekeeping gene RNA transcript confirmed the presence of viable RNA following extraction. miRNA profiles from isolated exosomes were generated using a targeted RNA sequencing assay and used for repeatability and differential expression studies. The ExoVerita Flex platform offers a novel, unbiased method for isolation and characterization of exosome-bound proteins and circulating nucleic acids while being compatible with standard downstream analysis. The ability to perform two-dimensional protein & nucleic acid analysis in the same sample run, at the same time, is both unique and unprecedented and will enable promising future clinical studies in cancer looking at diverse classes of biomarkers directly from blood. Citation Format: Jean M. Lewis, David Searson, Alfred Kinana, Heath Balcer, Debrah Thompson, David Bodkin, Juan P. Hinestrosa, Rajaram Krishnan. Direct analysis of exosome-bound proteins and nucleic acids using the ExoVerita™ Flex platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3978.

Abstract 3985: Dynamic changes of tumor-derived extracellular vesicles and related RNAs in blood samples of NSCLC patients

Abstract Liquid biopsy is defined as molecular analysis of rare cells or cell-free nucleic acids circulating in blood or in other biofluids. The concept of liquid biopsy aims at closely monitoring the status of a disease or treatment efficacy in a simple, fast, cost efficient way and at any point in time with minimal risk and burden for the patient. It has been suggested that the most reproducible approach to investigate “liquid” mRNAs would be to focus on cell-free extracellular vesicle (EV)-transcripts rather than on circulating cell-free mRNAs, as the former are well protected from degradation and therefore more stable. However, the volumes of available blood samples can be limiting for molecular analysis and rare cells as well as cell-free nucleic acids are normally present in low abundance. Therefore, detection of rare events or quantification of limited material requires robust, highly sensitive technologies like droplet digital PCR (ddPCR). In this pilot study, we investigated the absolute expression of 10 EV-transcripts typically involved in cancer development and chemotherapy resistance in longitudinal serum samples of 13 non-small cell lung cancer (NSCLC) patients (age range 37-76 years) treated with cisplatin at the Department of Inner Medicine II, University of Ulm, between May 2011 and August 2012. Blood samples were collected at baseline, after 3-6 months and after 9-12 months of chemotherapy from patients showing objective clinical response. Then, EV-RNAs were isolated by sequentially processing obtained serum samples with miRCURY® exosome kit and miRNeasy kit (both Qiagen, Hilden, Germany). Afterwards, the status of selected EV-transcripts was investigated via ddPCR (Biorad, Milan, Italy). After start of chemotherapy, we observed a downregulation of at least 2-fold of the following potentially cancer-related EV-transcripts : PTEN, ERBB2, FOSL1, IL-8, MET, RPS27A, SF3B1 and of the following housekeeping EV-mRNAs: ACTB, HIST1H3H and HSPA1A, in all patients. Interestingly, in none of patients' samples, the expression of EV-AKT1 and of EV-PD-L1 transcripts could be detected. Our preliminary results demonstrates significant dynamic changes of cancer-related EV-mRNA expression in serum samples of NSCLC patients during cisplatin treatment, thus suggesting a potential overall decrease of EVs in response to chemotherapy. Moreover, despite the small amounts of mRNA in extracellular vesicles from patient blood, using ddPCR differential mRNA levels could be determined in patient serum and expand the scope of biomarker analysis of EVs. This work is supported by IMI JU & EFPIA (grand no. 115749, CANCER-ID). Samples from patients and healthy volunteers, respectively, were collected under signed informed consent. Citation Format: Rita Lampignano, Martin Neumann, Vera Kloten, Nina Kessler, Anna Babayan, Laura Keller, Ines Stevic, Klaus Pantel, Thomas Krahn, Thomas Schlange, for the IMI CANCER-ID consortium. Dynamic changes of tumor-derived extracellular vesicles and related RNAs in blood samples of NSCLC patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3985.

Isolation, structure elucidation and quantitative analysis of callicarpone and oleanolic acid in the leaves of Callicarpa candicans collected from different provinces in Thailand

Isolation, structure elucidation and quantitative analysis of callicarpone and oleanolic acid in the leaves of Callicarpa candicans collected from different provinces in Thailand

A Low-Cost Time-Correlated Single Photon Counting Portable DNA Analyzer.

Photon-counting analysis of nucleic acids plays a key role in many diagnostics applications for its accurate and non-invasive nature. However, conventional photon-counting instrumentations are bulky and expensive due to the use of conventional optics and a lack of optimization of electronics. In this paper, we present a portable, low-cost time-correlated single photon-counting (TCSPC) analysis system for DNA detection. Both optical and electronic subsystems are carefully designed to provide effective emission filtering and size reduction, delivering good DNA detection and fluorescence lifetime extraction performance. DNA detection has been verified by fluorescence lifetime measurements of a V-carbazole conjugated fluorophore lifetime bioassay. The time-to-digital module of the proposed TCSPC system achieves a full width at half maximum (FWHM) timing resolution from 121 to 145 ps and a differential non-linearity (DNL) between −8.5% and +9.7% of the least significant bit (LSB) within the 500 ns full-scale range (FSR). With a detection limit of 6.25 nM and a dynamic range of 6.8 ns, the proposed TCSPC system demonstrates the enabling technology for rapid, point-of-care DNA diagnostics.

MOLECULAR WEIGHT COMPOSITION OF HUMIC ACIDS IN URBAN SOILS (THE NORTHERN ADMINISTRATIVE DISTRICT OF MOSCOW CITY TAKEN AS AN EXAMPLE)

The article provides research results and comparative analysis of humic acids in sod-podzolic soil of the Experimental Forest of the Russian State Agrarian University (protected area) and urban soils located in the Timiryazev district of Moscow. Humic acids of the zonal sod-podzolic soil of the Experimental Forest include four fractions with varying molecular weight: the 1 st fraction – ≥ 23 440 unified atomic mass units (amu), the 2 nd one – 13 340 amu, the 3 rd one – 5 500 amu and the 4 th one – 2 460 amu. The fraction with a molecular weight of 5 500 amu and a relative content of 38 % dominates among them, while the share of low-molecular fractions (< 20 000 amu) accounts for 70 % of the total mass of humic acids. When weight-average molecular mass of humic acids is 17 530 amu, the average molecular mass of the low-molecular fractions is 9 960 amu. Humic acids of urbanozems differ in molecular weight composition from humic acids of sod-podzolic soil. In most cases it consists of 5–6, less commonly of 3, fractions with molecular weight from 1 780 to ≥ 23 440 amu. The share of medium- and high-molecular fractions fluctuates from 31–37 % to 47–50 % of the total mass of humic acids. The characteristic feature of humic acids of urbanozems is the presence in their composition of low-molecular fractions with such molecular weights, which are not found in humic acids of sod-podzolic soil. Thus, humic acids of urbanozems are characterized by higher weight-average molecular mass ranging from 17 680 to 19 980 amu, as well as by higher weigh-average molecular mass of the low-molecular fractions which vary from 10 680 to 13 650 amu.

Direct quantitative LC-MS/MS analysis of amino acids in plasma

Direct quantitative LC-MS/MS analysis of amino acids in plasma

UV Method Development and Validation of Ellagic Acid for its Rapid Quantitative Estimation

Development and validation of a simple UV- Spectroscopy method was done for the quantitative analysis of Ellagic Acid (EA). The stock solution of 50μg/ml was prepared and scanned, for which absorption maxima was found to be 277nm. Further dilutions to different concentrations (1-5μg/ ml) were prepared and analyzed at 277nm. The method so developed was validated as per ICH guidelines for: linearity, robustness, precision, accuracy, limit of detection and quantification. The Lambert- Beer’s law is followed in the range (1-5μg/ml) with correlation coefficient value 0.9994. It was observed that the method is precise and accurate for EA analysis with good recovery percent of 94.47% to 106.83%. The method developed was further employed for determining the entrapment efficiency of ellagic acid and its release from its nanoparticle dosage form. The method may be utilized for determining the concentration of EA when present as formulation and in combination with other drugs.

The use of tris (tetraphenylimidodiphosphinate) of praseodymium chemical shift reagent in proton NMR for the evaluation of the argan oil fatty acids autoxidation and the analysis of the argan pulp fatty acids

<p>Proton NMR is a method of molecular investigation that has its limitations when applied to complex molecules or molecules with many nearly equivalent sites. Previous studies have resorted to the use of paramagnetic chemical shift reagents, having as formula tris (tetraphenylimidodiphosphinate) of lanthanides ln((tpip.)3.<strong> </strong>The use of reagent Pr(tpip)3 in proton RMN has allowed us to evaluate the autoxidation of fatty acids mixture (stored 6 and 12 months after oil extraction) by the dosage of saturated and unsaturated acids on the one hand, and that of oleic and linoleic acids on the other. We note between 6 and 12 months of storage at 4°C a decrease in the percentage of unsaturated acids (76% to 63%) and an increase in the percentage of saturated acids (24% to 36%). The results show that the oleic acid maintained the same percentage (35%) as it is not easily oxidized whereas, for the linoleic acid, we observe a decrease in percentage from 22.5% to 18.5% (slow autoxidation at 4°C). We also used this NMR method for the analysis of the argan pulp fatty part. The GC analysis shows that it contains very few unsaturated fatty acids and that the main fatty acids are myristic (C14:0) and palmitic (C16:0) acids. The proton NMR with Pr(tpip)3 allowed us to confirm these results. This method that does not require derivation has proven to be interesting, simple and efficient.</p>

Electrochemical detection of ursolic acid from spruce (Picea abies) essential oils using modified amperometric microsensors

Ursolic acid (UA) is a natural pentacyclic triterpenoid carboxylic acid found in some medicinal plant species. In this paper, amperometric microsensors based on a powder which contained graphite (G) and carbon nanoparticles (CN) (G-CN) unmodified and modified with chitosan (CHIT) and tetraphenyl-porphine cobalt(II) (Co(II)TPP) are proposed for the analysis of UA plant source essential oils obtained from spruce (Picea Abies). Differential pulse voltammetry (DPV) was used to optimize the method and for the determination of ursolic acid from different types of botanical samples. The optimum working pH was 5.00 for the G-CN and (Co(II)TPP)/G-CN microsensors and pH 3.00 for CHIT/G-CN in the presence of a 0.1 mol L−1 KCl supporting electrolyte. The linear concentration ranges for ursolic acid (UA) were between 0.1 and 100 µmol L−1 for the unmodified microsensor (G-CN), 0.01, 1 µmol L−1 for the microsensor modified with chitosan (CHIT/G-CN), and 0.01 and 10 µmol L−1 for the microsensor modified with (Co(II)TPP)/G-CN). It is the first time these amperometric microsensors have been used for the reliable analysis of ursolic acid (UA) in three original botanical samples obtained from different parts of spruce (Picea abies): resin essential oil, cons essential oil, cons and sprouts essential oil, with recovery rate values up to 99.29%.

Adult hypophosphatasia with compound heterozygous p.Phe327Leu missense and c.1559delT frameshift mutations in tissue-nonspecific alkaline phosphatase gene: a case report

BackgroundHypophosphatasia is an inherited bone disease characterized by low alkaline phosphatase activity encoded by ALPL. Clinically, hypophosphatasia can be categorized as perinatal, infantile, childhood, and adult forms, as well as odonto-hypophosphatasia, according to the age at first sign or dental manifestations. Adult hypophosphatasia typically presents in middle-aged patients who appear to be in good health in early adulthood and manifests as painful feet caused by recurrent, slow-healing stress fractures of the lower limb. Because the symptoms of adult hypophosphatasia vary and are common, many patients with hypophosphatasia might be not diagnosed accurately and thus may receive inappropriate treatment.Case presentationWe report a case of a 35-year-old Japanese woman with low serum alkaline phosphatase detected at a routine medical checkup. She had mild muscle/bone pain but no history of rickets, fractures, or dental problems. Measurement of bone mineral density of the lumbar spine and the femoral neck revealed osteopenia below the expected range for age in a young adult. Abdominal ultrasonography revealed numerous microcalcifications in both kidneys. Analysis of amino acids in urine revealed that phosphoethanolamine was elevated. Low serum alkaline phosphatase activity, elevation of phosphoethanolamine, and low bone mineral density supported the diagnosis of hypophosphatasia. ALPL mutation analysis revealed two mutations: p.Phe327Leu and c.1559delT. These genetic abnormalities were previously reported in perinatal, infantile, and childhood but not adult hypophosphatasia. On the basis of the clinical presentation, laboratory and imaging findings, and genetic analyses, the patient was definitively diagnosed with adult hypophosphatasia. To the best of our knowledge, this is the first case report of adult hypophosphatasia with the compound heterozygous mutations p.Phe327Leu and c.1559delT.ConclusionsAlthough the risk of bone fracture was high in this case, treatment approaches differ between osteoporosis and hypophosphatasia. Because adult hypophosphatasia diagnosis is often difficult because of their varied symptoms, hypophosphatasia should be considered in the differential diagnosis of low serum alkaline phosphatase. Early diagnosis is important so that appropriate treatment can be initiated.