The acid hydrolysis of various selected saccharide- and polysaccharide-based pharmaceutical excipients under acid hydrolysis and the formation of degradation compounds were studied. New degradation products formed from these excipients were discovered. Liquid chromatography-mass spectrometry and nuclear magnetic resonance techniques were employed to identify and fully characterize these unknown compounds. The degradation products were identified as [(5-formylfuran-2-yl)methoxy]acetic acid, 5-[(propan-2-yloxy)methyl]furan-2-carbaldehyde, along with the previously identified 5-(methoxymethyl)furan-2-carbaldehyde. On the basis of the identification of these degradation products, a reasonable mechanism for their formation can be proposed. Temperature and pH affect the hydrolysis rates of saccharides and polysaccharides, which in turn affects the rate of formation of furfural compounds.
- MeSH
- Furaldehyde chemical synthesis MeSH
- Mass Spectrometry MeSH
- Hydrolysis MeSH
- Kinetics MeSH
- Hydrogen-Ion Concentration MeSH
- Pharmaceutical Preparations chemistry MeSH
- Limit of Detection MeSH
- Magnetic Resonance Spectroscopy MeSH
- Polysaccharides chemistry MeSH
- Excipients MeSH
- Reproducibility of Results MeSH
- Carbohydrates chemistry MeSH
- Temperature MeSH
- Chromatography, High Pressure Liquid MeSH
- Publication type
- Journal Article MeSH
- Validation Study MeSH
In this study, a GC-MS method was developed for the quantification of saccharides in complex mixtures such as bio-oils and bio-oil aqueous phases produced by ablative pyrolysis of lignocellulosic biomass. The samples were first treated using N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) and the trimethylsilylated (more volatile) derivatives were analyzed by GC-MS. The method offers advantages of great separation capability and simultaneous identification of unknown peaks by comparison of the mass spectra and retention indices with extensive libraries available online. However, even with these tools at hand, the identification of several saccharide-resembling compounds can be challenging especially in such highly complex samples as pyrolysis bio-oils. For this reason, we devised a novel procedure, which eliminates certain saccharides depending on their specific chemical properties before subjecting the samples to the GC-MS analysis. The procedure was based on the combination of aniline treatment (elimination of reducing aldoses), and hydrolysis (elimination of anhydrosugars, glycosides, disaccharides and oligosaccharides). Based on the differences in chromatograms before and after the procedure, the unknown compounds were assigned into groups based on their susceptibility to each treatment. The combination of all methods above has allowed more accurate identification and quantification of saccharides, some of which were not as of today found in bio-oils.
Methods used for the isolation, separation and characterization of boar seminal plasma proteins are discussed, as well as techniques applied to study their binding properties. Attention is paid to interactions of these proteins with different types of saccharides and glycoconjugates, with membrane phospholipids, and to interactions between proteins. Boar seminal plasma contains different types of proteins: spermadhesins of the AQN and AWN families; DQH and PSP proteins belong to the most abundant. Some of these proteins are bound to the sperm surface during ejaculation and thus protein-coating layers of sperm are formed. Sperms coated with proteins participate in different types of interactions occurring in the course of the reproduction process, e.g. formation of the oviductal sperm reservoir, sperm capacitation, oocyte recognition and sperm binding to the oocyte.
- MeSH
- Chromatography, Affinity MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Financing, Organized MeSH
- Chromatography, Gel MeSH
- Sperm Capacitation MeSH
- Swine MeSH
- Seminal Plasma Proteins analysis isolation & purification metabolism MeSH
- Reproducibility of Results MeSH
- Protein Binding MeSH
- Chromatography, High Pressure Liquid MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Animals MeSH
The removal of algal organic matter (AOM) is a growing concern for the water treatment industry worldwide. The current study investigates coagulation of non-proteinaceous AOM (AOM after protein separation), which has been minimally explored compared with proteinaceous fractions. Jar tests with either aluminum sulphate (alum) or polyaluminium chloride (PACl) were performed at doses of 0.2-3.0 mg Al per 1 mg of dissolved organic carbon in the pH range 3.0-10.5. Additionally, non-proteinaceous matter was characterized in terms of charge, molecular weight and carbohydrate content to assess the treatability of its different fractions. Results showed that only up to 25% of non-proteinaceous AOM can be removed by coagulation under optimized conditions. The optimal coagulation pH (6.6-8.0 for alum and 7.5-9.0 for PACl) and low surface charge of the removed fraction indicated that the prevailing coagulation mechanism was adsorption of non-proteinaceous matter onto aluminum hydroxide precipitates. The lowest residual Al concentrations were achieved in very narrow pH ranges, especially in the case of PACl. High-molecular weight saccharide-like organics were amenable to coagulation compared to low-molecular weight (<3 kDa) substances. Their high content in non-proteinaceous matter (about 67%) was the reason for its low removal. Comparison with our previous studies implies that proteinaceous and non-proteinaceous matter is coagulated under different conditions due to the employment of diverse coagulation mechanisms. The study suggests that further research should focus on the removal of low-molecular weight AOM, reluctant to coagulate, with other treatment processes to minimize its detrimental effect on water safety.
Five new strains of lactobacilli isolated from goatling's stomach were identified by molecular-biological approaches. Profiles of fermentable saccharides, Gram staining, and cell morphology were also determined. They were identified as Lactobacillus reuteri (strains KO4b, KO4m, KO5) and as Lactobacillus plantarum (strains KG1z, KG4). In DNA samples of all newly isolated L. reuteri strains as well as in L. reuteri E (Lreu E; originated from lamb), the part of gldC gene, coding large subunit of glycerol dehydratase, that is necessary for 3-hydroxypropionaldehyde (3-HPA; reuterin) production, was amplified using two designed primer sets. However, the 3-HPA production was revealed only in the strain Lreu E. It produced five- or ten-fold lower amount of 3-HPA in comparison with probiotic L. reuteri ATCC 55730 in aerobic or anaerobic conditions, respectively. Moreover, Lreu E completely lost its production ability after ca. five passages in MRS medium. The co-incubation of Lreu E, but not other L. reuteri isolates, with Escherichia coli re-induced 3-HPA production. In the case of L. reuteri ATCC 55730, the 3-HPA production increased more than four times after co-incubation with E. coli.
Předmět sdělení: Recidivující aftózní stomatitida (RAS) je jedním z nejčastějších onemocnění sliznice dutiny ústní, které se projevuje tvorbou bolestivých erozí až ulcerací. Diagnostika RAS je založena na anamnestických údajích a klinickém vzhledu lézí; neexistují žádné laboratorní testy k potvrzení diagnózy. Léčba tohoto onemocnění je pouze symptomatická a málo efektivní. Etiopatogeneze RAS dosud není známa, v literatuře je ale popsána řada rizikových faktorů, které ke vzniku a rozvoji onemocnění mohou přispívat. Kromě lokálního traumatu, potravinových alergenů, mikrobiální dysbiózy, infekčních agens, nutričních faktorů (deficitu vitaminu B12, železa a kyseliny listové), stresu a hormonálních změn, hraje roli i imunologický profil pacienta a také genetické predispozice jedince k této multifaktoriální chorobě. Vliv dědičnosti na vznik, resp. rozvoj onemocnění byl již dříve potvrzen studiemi dvojčat a rodin. Aktuálně jsou publikovány genetické asociační studie zabývající se variabilitou vybraných genů u pacientů s RAS ve srovnání se zdravými kontrolami (tzv. studie kontrol a případů) v různých populacích. Za kandidátní jsou považovány zejména ty geny, které souvisejí s funkcí imunitního systému, s reakcí organismu na oxidační stres, s metabolismem tkání sliznic, vitaminů a minerálních látek. Cílem těchto analýz bylo nalezení rizikových nebo naopak protektivních variant, a to v genech interleukinů-1 (IL) a antagonisty jejich receptoru (IL-1RN), IL-4, IL-6, IL-10, tumor nekrotizujícího faktoru α (TNFα), NOD-like receptoru 3 (NLRP3), Toll-like receptoru 4 (TLR4), E- a L-selektinů (SEL), angiotenzin konvertujícího enzymu (ACE), v genu pro středomořskou horečku (MEFV), serotoninový transportér (SLC6A4), matrix metaloproteinázu 9 (MMP9), metylentetrahydrofolát reduktázu (MTHFR) a syntázu oxidu dusnatého 2 (NOS2), které mohou v kontextu dalších faktorů ovlivňovat náchylnost jedince k rozvoji onemocnění. Závěr: V předloženém přehledovém článku jsou shrnuty a diskutovány závěry těchto genetických asociačních studií. Je pravděpodobné, že výzkum RAS na molekulární úrovni by mohl vést k alespoň částečnému pochopení etiopatogeneze tohoto onemocnění, a tím ke zlepšení prevence, diagnostiky a léčby postižených pacientů.
Background: Recurrent aphthous stomatitis (RAS), one of the most common diseases of the oral mucosa, is characterized by the formation of painful oral erosions or even ulcers. RAS diagnosis is based on anamnestical data and appearance of lesions; no laboratory tests to confirm the diagnosis are available. Treatment of this condition is only symptomatic and less effective. The disease etiopathogenesis is unknown, but risk factors associated with the origin and development of the disease have been described in the literature. Besides local trauma, food allergens, oral microbial dysbiosis, infectious agents, nutritious factors (deficiency of B12 vitamin, iron, and folic acid), stress and hormonal changes, the immunological profile of the patient and his/her genetic predispositions to this multifactorial disease play a role. The effect of heredity on the disease origin and development was previously confirmed by studies of twins and families. Genetic variability of the selected genes in patients with RAS compared with healthy controls (case-control study) conducted in different populations have been published. The main candidates for RAS are the genes associated with the immune system, response of the organism to oxidative stress, metabolism of mucosal tissues, vitamins, and minerals. The aim of these studies was to find risk, or on the contrary protective, gene variants in the interleukin-1 (IL) and its receptor antagonist (IL-1RN), IL-4, IL-6, IL-10, tumor necrosis factor alpha (TNFalpha), NOD-like receptor 3 (NLRP3), Toll-like receptor 4 (TLR4), E- and L-selectin (SEL), angiotensin converting enzym (ACE), gene for Mediterranean fever (MEFV), serotonin transporter (SLC6A4), matrix metalloproteinase 9 (MMP9), methylenetetrahydrofolate reductase (MTHFR) and nitric oxide syntase 2 (NOS2), which may together with other factors influence the individual susceptibility to the disease development. In the present review, we summarize and discuss findings of genetic association studies. Conclusion: We assume that further research into RAS on the molecular level may lead to better understanding of this disease etiopathogenesis and improve prevention, diagnosis and treatment of the affected patients.
- MeSH
- Stomatitis, Aphthous * diagnosis etiology genetics MeSH
- Genetic Predisposition to Disease * MeSH
- Genetic Association Studies MeSH
- Genes MeSH
- Humans MeSH
- Intercellular Signaling Peptides and Proteins MeSH
- Cell Adhesion Molecules MeSH
- Mouth Diseases MeSH
- Oxidative Stress MeSH
- Risk Factors MeSH
- Mouth Mucosa pathology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
Introduction: The prognosis of multiple myeloma is still unfavorable due to inherent characteristics of the disease and the often-delayed diagnosis due to widespread and unspecific symptoms such as back pain and fatigue. Therefore, a simple diagnostic blood test would be helpful to speed up the diagnostic procedure in such patients (pts.). Here, we evaluated the diagnostic value of plasma levels of carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) in the peripheral blood and bone marrow of pts. with plasma cell disorders and in healthy controls. Materials and Methods: Immunoreactive CEACAM6 was determined in the peripheral blood and bone marrow (n = 95/100) of pts. with monoclonal gammopathy of unknown significance (MGUS: 28/37), newly diagnosed multiple myeloma (NDMM: 42/40), and relapsed/refractory multiple myeloma (RRMM: 25/23) by sandwich ELISA. Results: Median CEACAM6 levels in the peripheral blood of pts. with plasma cell disorders were significantly higher than those of healthy controls (healthy controls: 15.2 pg/ml (12.1-17.1); MGUS: 19.0 pg/ml (16.4-22.5); NDMM: 18.0 pg/ml (13.4-21.2); and RRMM: 18.9 pg/ml (15.2-21.5); p < 0.001). Plasma levels of CEACAM6 discriminated healthy subjects from MGUS/NDMM pts. (AUC = 0.71, 95% CI: 0.6-0.8); i.e., a CEACAM6 level > 17.3 pg/ml has an 82% (95% CI: 70-90) predictive probability for the identification of MGUS or NDMM. Moreover, CEACAM6 levels in the bone marrow were significantly higher in RRMM pts. than in NDMM pts. (p = 0.04), suggesting a role of this molecule in disease progression. Conclusion: CEACAM6 plasma levels can noninvasively identify pts. with a plasma cell disorder and should be evaluated prospectively as a potential diagnostic marker. Moreover, due to high CEACAM6 levels in the bone marrow in RRMM pts., this adhesion molecule might be a therapeutic target in multiple myeloma pts.
- MeSH
- Antigens, CD blood metabolism MeSH
- GPI-Linked Proteins blood metabolism MeSH
- Bone Marrow metabolism MeSH
- Middle Aged MeSH
- Humans MeSH
- Multiple Myeloma blood MeSH
- Cell Adhesion Molecules blood metabolism MeSH
- Biomarkers, Tumor blood metabolism MeSH
- Aged MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
Markers for evaluating the establishment of cyanobacteria based on their sensitivity or resistance to antibiotics, saccharide utilization patterns and PCR generated fingerprints were developed. Four selected strains (isolates from rhizosphere soils of diverse agro-ecosystems) have shown potential as diazotrophs and exhibited plant growth promoting abilities. Different responses were obtained on screening against 40 antibiotics, which aided in developing selectable antibiotic markers for each strain. Biochemical profiles generated using standardized chromogenic identification system (including saccharide utilization tests) revealed that 53 % of the saccharides tested were not utilized by any strain, while some strains exhibited unique ability for utilization of saccharides such as melibiose, cellobiose, maltose and glucosamine. PCR based amplification profiles developed using a number of primers based on repeat sequences revealed the utility of 3 primers in providing unique fingerprints for the strains.
- Keywords
- Rhizosphere,
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Drug Resistance, Bacterial MeSH
- Disaccharides metabolism MeSH
- DNA Fingerprinting methods MeSH
- DNA Primers MeSH
- Financing, Organized MeSH
- Phylogeny MeSH
- Microbial Sensitivity Tests MeSH
- Molecular Sequence Data MeSH
- Polymerase Chain Reaction methods MeSH
- Soil Microbiology MeSH
- Cyanobacteria genetics classification metabolism drug effects MeSH
- Agriculture methods MeSH
BACKGROUND: Lectins are proteins of non-immune origin capable of binding saccharide structures with high specificity and affinity. Considering the high encoding capacity of oligosaccharides, this makes lectins important for adhesion and recognition. The present study is devoted to the PA-IIL lectin from Pseudomonas aeruginosa, an opportunistic human pathogen capable of causing lethal complications in cystic fibrosis patients. The lectin may play an important role in the process of virulence, recognizing specific saccharide structures and subsequently allowing the bacteria to adhere to the host cells. It displays high values of affinity towards monosaccharides, especially fucose--a feature caused by unusual binding mode, where two calcium ions participate in the interaction with saccharide. Investigating and understanding the nature of lectin-saccharide interactions holds a great potential of use in the field of drug design, namely the targeting and delivery of active compounds to the proper site of action. RESULTS: In vitro site-directed mutagenesis of the PA-IIL lectin yielded three single point mutants that were investigated both structurally (by X-ray crystallography) and functionally (by isothermal titration calorimetry). The mutated amino acids (22-23-24 triad) belong to the so-called specificity binding loop responsible for the monosaccharide specificity of the lectin. The mutation of the amino acids resulted in changes to the thermodynamic behaviour of the mutants and subsequently in their relative preference towards monosaccharides. Correlation of the measured data with X-ray structures provided the molecular basis for rationalizing the affinity changes. The mutations either prevent certain interactions to be formed or allow formation of new interactions--both of afore mentioned have strong effects on the saccharide preferences. CONCLUSION: Mutagenesis of amino acids forming the specificity binding loop allowed identification of one amino acid that is crucial for definition of the lectin sugar preference. Altering specificity loop amino acids causes changes in saccharide-binding preferences of lectins derived from PA-IIL, via creation or blocking possible binding interactions. This finding opens a gate towards protein engineering and subsequent protein design to refine the desired binding properties and preferences, an approach that could have strong potential for drug design.
- MeSH
- Adhesins, Bacterial genetics chemistry MeSH
- Chromatography, Affinity MeSH
- Financing, Organized MeSH
- Polymorphism, Single Nucleotide MeSH
- Protein Conformation MeSH
- Crystallography, X-Ray MeSH
- Lectins genetics chemistry MeSH
- Models, Molecular MeSH
- Monosaccharides chemistry MeSH
- Mutagenesis, Site-Directed MeSH
- Protein Engineering MeSH
- Pseudomonas aeruginosa genetics MeSH
- Ralstonia solanacearum chemistry MeSH
- Recombinant Proteins genetics isolation & purification MeSH
- Plant Lectins chemistry MeSH
- Amino Acid Substitution MeSH
- Binding Sites MeSH
