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Acid Analysis (acid + analysis)
Kinds of Acid Analysis Selected AbstractsThe Electrochemical Properties of Co(TPP), Tetraphenylborate Modified Glassy Carbon Electrode: Application to Dopamine and Uric Acid AnalysisELECTROANALYSIS, Issue 5 2006Yunlong Zeng Abstract We report the combination of the charge repelling property of tetraphenyl-borate (TPB) anion and the electrooxidation catalytic effect of cobalt(II) tetrakisphenylporphyrin (CoTPP) embedded in a sol gel ceramic film to develop a modified glassy carbon electrode (CoTPP-TPB-SGGCE) for the simultaneous determination of dopamine (DA) and uric acid (UA). The optimized CoTPP-TPB-SGGCE shows excellent sensitivity and selectivity for the DA and UA analysis. As high as 2000 fold acceptable tolerance of ascorbic acid (AA) for the determination of trace DA and UA is reached. In the presence of 0.10,mM AA, the linear concentration range for DA is from 6.0×10,8 to 2.5×10,5,M, and the detection limit is 2.0×10,8,M. For UA, the linear concentration range is from 1.0×10,7 to 3.5×10,5,M, and the detection limit is 7.0×10,8,M. Our study has also demonstrated that the novel CoTPP-TPB-SGGCE shows high stability and reliability. For 6.00,,M DA and UA, a total of 12,measurements were taken in one week, and the relative standard deviation is 2.05% and 2.68% respectively. No obvious shift of peak current and peak potential is observed over a three-month lifetime test. The response of the sensor is very quick and response time is approximately 1,s. Satisfactory results are also achieved when the CoTPP-TPB-SGGCEs being used to detect the DA and UA in human urine samples. [source] Fatty acid analyses reveal high degrees of omnivory and dietary plasticity in pond-dwelling tadpolesFRESHWATER BIOLOGY, Issue 7 2010MATT R. WHILES Summary 1.,Understanding the trophic relationships of consumers is central to ecology, but constructing meaningful food webs is often difficult because of a lack of detailed information on consumption versus assimilation and high degrees of omnivory. 2.,We used fatty acid analyses to examine the trophic relationships of three common larval anurans (Pseudacris crucifer, Lithobates catesbeianus and Lithobates clamitans) that are often classified as grazers or detritivores. Tadpoles and potential food sources were sampled in four ponds in southern Illinois and analysed for fatty acid composition. Single linkage cluster analysis was then used to compare fatty acid profiles among tadpole gut contents, tadpole muscle tissues and available food resources. 3.,Diets varied among species and within species among ponds, but organic sediments consistently contributed most to the fatty acid composition of the gut contents of all species. Fatty acid profiles also indicated that larval insects and phytoplankton were consumed by both L. catesbeianus and L. clamitans in one pond, while L. clamitans and P. crucifer consumed mainly periphyton along with sediments in another pond, and these diet differences appeared linked to physical differences among ponds, with periphyton and/or phytoplankton contributing more to tadpole diets in less shaded ponds. 4.,The fatty acid composition of muscle tissues of L. clamitans, the dominant tadpole in these systems, indicated that plant detritus and bacteria, which were the dominant components of organic sediments in the ponds, were common components of the assimilatory diet. 5.,Results demonstrate the utility of fatty acid analyses for assessing both consumption and assimilation. The tadpole assemblages we examined derive much of their energy from heterotrophic and allochthonous sources and exhibit high dietary plasticity. This information will allow for more accurate and comprehensive assessments of trophic interactions in freshwater habitats, as well as aid in amphibian conservation, management and captive propagation efforts. [source] Purification and characterization of two bacteriocins produced by lactic acid bacteria isolated from Mongolian airagJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2006B. Batdorj Abstract Aims:, The aim of this study was to isolate and identify bacteriocin-producing lactic acid bacteria (LAB) issued from Mongolian airag (traditional fermented mare's milk), and to purify and characterize bacteriocins produced by these LAB. Methods and Results:, Identification of the bacteria (Enterococcus durans) was carried out on the basis of its morphological, biochemical characteristics and carbohydrate fermentation profile and by API50CH kit and 16S rDNA analyses. The pH-neutral cell-free supernatant of this bacterium inhibited the growth of several Lactobacillus spp. and food-borne pathogens including Escherichia coli, Staphylococcus aureus and Listeria innocua. The antimicrobial agent (enterocin A5-11) was heat stable and was not sensitive to acid and alkaline conditions (pH 2,10), but was sensitive to several proteolytic enzymes. Its inhibitory activity was completely eliminated after treatment with proteinase K and , -chymotrypsin. The activity was however not completely inactivated by other proteases including trypsin and pepsin. Three-step purification procedure with high recovery yields was developed to separate two bacteriocins. The applied procedure allowed the recovery of 16% and 64% of enterocins A5-11A and A5-11B, respectively, present in the culture supernatant with purity higher than 99%. SDS-PAGE analyses revealed that enterocin A5-11 has a molecular mass of 5000 Da and mass spectrometry analyses demonstrates molecular masses of 5206 and 5218 Da for fractions A and B, respectively. Amino acid analyses of both enterocins indicated significant quantitative difference in their contents in threonine, alanine, isoleucine and leucine. Their N -termini were blocked hampering straightforward Edman degradation. Conclusions:, Bacteriocins A5-11A and B from Ent. durans belong to the class II of bacteriocins. Significance and Impact of the Study:, Judging from molecular masses, amino acid composition and spectrum of activities, bacteriocins A5-11A and B from Ent. durans show high degree of similarity with enterocins L50A and L50B isolated from Enterococcus faecium (Cintas et al. 1998, 2000) and with enterocin I produced by Ent. faecium 6T1a, a strain originally isolated from a Spanish-style green olive fermentation (Floriano et al. 1998). [source] ANTIOXIDANT ACTIVITY AND PHENOLIC ACID CONSTITUENTS OF CHESTNUT (CASTANIA SATIVA MILL.) HONEY AND PROPOLISJOURNAL OF FOOD BIOCHEMISTRY, Issue 4 2009ALI OSMAN SARIKAYA ABSTRACT This study describes the constituents of phenolic acids and antioxidant activities of chestnut (Castania sativa Mill.) honeys and propolis in Turkey. Antioxidant activity of the chestnut honeys and propolis were examined by three different methods, namely scavenging of free radical 2, 2-diphenyl-1-picrylhydrazyl, FRAP, and cupric reducing antioxidant power. Total phenolic contents were determined by using Folin,Ciocalteu reagent as GA equivalent. The phenolic constituents were also determined by HPLC. The antioxidant activities were compared with standard antioxidants such as catechin, BHT and Trolox. The antioxidant activities of all the samples were found high and related to the sample concentrations. The ethanolic propolis extracts showed the highest antioxidant activity. The major phenolic acids of the chestnut honeys and propolis identified by HPLC with PDA detection were coumaric acid, FA, cinnamic acid, CA and ChA. PRACTICAL APPLICATIONS In this study, some phenolic acid components and antioxidant capacity of chestnut (Castania sativa Mill.) honey and propolis were measured. The comparative findings from antioxidant activities and phenolic acid analyses of honey and propolis samples of chestnut origin provide important criteria for considering their nutritional and nutraceutical potentials. Comparison of our results with literature data also ranks the chestnut honey and propolis as better sources of antioxidants among those from other floral origins. [source] Recent advances in amino acid analysis by CEELECTROPHORESIS, Issue 1 2008Véréna Poinsot Abstract This paper describes a number of articles that have been published on amino acid (AA) analysis using CE during the period from June 2005 to May 2007. This review article follows the format of the previous articles of Smith (Electrophoresis 1999, 20, 3078,3083), Prata et al.. (Electrophoresis 2001, 22, 4129,4138), and Poinsot et al.. (Electrophoresis 2003, 24, 4047,4062 and Electrophoresis 2006, 27, 176,194). Several new developments in AA analysis with CE are reported describing the use of laser-emitting diodes for LIF, MS, and chips. In addition, we describe articles concerning clinical studies and neuroclinical applications. [source] Determination of amino acids by micellar EKC: Recent advances in method development and novel applications to different matricesELECTROPHORESIS, Issue 1 2008Paolo Iadarola Professor Abstract The extensive use of CE for the analysis of amino acids has been well documented in a series of research articles and reviews. Aim of this report is to address the attention of the reader on the recent advances of micellar electrokinetic chromatography for the separation and determination of these analytes. Enhancements in selectivity of this technique through the use of pseudostationary phases containing mixed micelles, polymers, and chiral selectors are presented. Selected applications concerning separation and quantitation of even minute amounts of amino acids in: (i) biological fluids; (ii) microdialysates; (iii) plant cells; (iv) food stuff; and (v) pharmaceutical formulations have also been covered. Advantages of MEKC over other techniques for the amino acid analysis have been underlined. [source] Recent advances in amino acid analysis by capillary electrophoresisELECTROPHORESIS, Issue 22-23 2003Véréna Poinsot Abstract Amino acids are studied extensively using capillary electrophoresis. In a previous article, we reviewed applications reported in the period 1999 , early 2001 (Prata, C., Bonnafous, P., Fraysse, N., Treilhou, M., Poinsot, V., Couderc, F., Electrophoresis 2001, 22, 4129,4138). In this article we follow on with this review for the period end of 2001 , beginning of 2003. We will report the developments of detection methods, separations of enantiomers, the new medical applications, and amino acids in food and plants. This review shows that CE is more and more important for the amino acid analysis. [source] Valproic Acid-Induced Hyperammonemic Encephalopathy with Triphasic WavesEPILEPSIA, Issue 7 2000Akira Kifune Summary: Purpose: To examine a patient with valproic acid (VPA)-induced hyperammonemic encephalopathy accompanied by triphasic waves. Methods: A 61-year-old male patient with epilepsy experienced disturbance of consciousness after VPA dose was increased because of poor seizure control. The electroencephalogram (EEG) taken on admission revealed triphasic waves and high-amplitude ,-activity with frontal predominance. Although serum hepatic enzymes, such as AST and ALT, were normal, serum ammonium level was high at 96 ,g/dl (normal range, 3,47 ,g/dl). Serum amino acid analysis showed multiple minor abnormalities. Administration of VPA was discontinued immediately after admission, while other anticonvulsants were continued. Results: The patient's condition was improved on the fourth day of admission. An EEG, serum ammonium level, and amino acid profile were normal on the eighth day. Based on VPA administration, serum ammonium levels, and results of amino acid analysis, this patient had VPA-induced hyperammonemic encephalopathy. Conclusions: Our case indicates that caution is required if triphasic waves appear in VPA-induced hyperammonemic encephalopathy. [source] First laboratory confirmation of Xylophilus ampelinus in Slovenia,EPPO BULLETIN, Issue 1 2005T. Dreo Bacterial blight of grapevine is caused by a slow-growing bacterium Xylophilus ampelinus. It has been suspected to occur in Slovenia on the basis of visual observation of characteristic symptoms in the 1960s. In the present study, symptoms were recorded in an infected vineyard during three consecutive years (2002/2004). Samples from this vineyard were tested by nested-PCR and isolation of bacteria on media was attempted. In the first year, angular lesions on leaves were highly expressed and an isolate morphologically similar to X. ampelinus was obtained from one sample. It was purified and identified as X. ampelinus using biochemical and nutritional tests, fatty acid analysis, immuno-fluorescence, nested PCR and partial sequencing of the 16S rRNA gene. The 16S rDNA sequence showed 99,100% homology to known sequences of X. ampelinus strains, including the type strain. Pathogenicity of the isolate was confirmed in tissue-cultured and potted grapevine plants. In the following two years, symptoms of bacterial blight were only faintly expressed. Using isolation on media and nested-PCR, 23 and 17 extracts prepared from 10 and 8 grapevines, respectively, were analysed. In 2003, no positive sample was found, but X. ampelinus was again isolated and identified by colony morphology and nested-PCR in 2004. [source] How do enamelysin and kallikrein 4 process the 32-kDa enamelin?EUROPEAN JOURNAL OF ORAL SCIENCES, Issue 2006Yasuo Yamakoshi The activities of two proteases , enamelysin (MMP-20) and kallikrein 4 (KLK4) , are necessary for dental enamel to achieve its high degree of mineralization. We hypothesize that the selected enamel protein cleavage products which accumulate in the secretory-stage enamel matrix do so because they are resistant to further cleavage by MMP-20. Later, they are degraded by KLK4. The 32-kDa enamelin is the only domain of the parent protein that accumulates in the deeper enamel. Our objective was to identify the cleavage sites of 32-kDa enamelin that are generated by proteolysis with MMP-20 and KLK4. Enamelysin, KLK4, the major amelogenin isoform (P173), and the 32-kDa enamelin were isolated from developing porcine enamel. P173 and the 32-kDa enamelin were incubated with MMP-20 or KLK4 for up to 48 h. Then, the 32-kDa enamelin digestion products were fractionated by reverse-phase high-performance liquid chromatography (RP-HPLC) and characterized by Edman sequencing, amino acid analysis, and mass spectrometry. Enamelysin cleaved the 32-kDa enamelin only after it was deglycosylated. Kallikrein 4 digestion of the 32-kDa enamelin generated nine major cleavage products, six of which were successfully characterized. After 12 h of digestion with KLK4, all of the 32-kDa enamelin had been cleaved, but some cleavage products persisted after 48 h of digestion. [source] Assignment of a single disulfide bridge in rat liver methionine adenosyltransferaseFEBS JOURNAL, Issue 1 2000María L. Martínez-Chantar Rat liver methionine adenosyltransferase incorporated 8 mol of N -ethylmaleimide per mol of subunit upon denaturation in the presence of 8 m urea, whereas 10 such groups were labelled when dithiothreitol was also included. This observation prompted a re-examination of the state of the thiol groups, which was carried out using peptide mapping, amino acid analysis and N-terminal sequencing. The results obtained revealed a disulfide bridge between Cys35 and Cys61. This disulfide did not appear to be conserved because cysteines homologous to residue 61 do not exist in methionine adenosyltransferases of other origins, therefore suggesting its importance for the differential aspects of the liver-specific enzyme. [source] Identification of phosphatidylserine decarboxylases 1 and 2 from Pichia pastorisFEMS YEAST RESEARCH, Issue 6 2009Tamara Wriessnegger Abstract Genetic manipulation of lipid biosynthetic enzymes allows modification of cellular membranes. We made use of this strategy and constructed mutants in phospholipid metabolism of Pichia pastoris, which is widely used in biotechnology for expression of heterologous proteins. Here we describe identification of two P. pastoris phosphatidylserine decarboxylases (PSDs) encoded by genes homologous to PSD1 and PSD2 from Saccharomyces cerevisiae. Using P. pastoris psd1, and psd2, mutants we investigated the contribution of the respective gene products to phosphatidylethanolamine synthesis, membrane composition and cell growth. Deletion of PSD1 caused loss of PSD activity in mitochondria, a severe growth defect on minimal media and depletion of cellular and mitochondrial phosphatidylethanolamine levels. This defect could not be compensated by Psd2p, but by supplementation with ethanolamine, which is the substrate for the cytidine diphosphate (CDP),ethanolamine pathway, the third route of phosphatidylethanolamine synthesis in yeast. Fatty acid analysis showed selectivity of both Psd1p and Psd2p in vivo for the synthesis of unsaturated phosphatidylethanolamine species. Phosphatidylethanolamine species containing palmitic acid (16:0), however, were preferentially assembled into mitochondria. In summary, this study provides first insight into membrane manipulation of P. pastoris, which may serve as a useful method to modify cell biological properties of this microorganism for biotechnological purposes. [source] Flame retardancy finish with an organophosphorus retardant on silk fabricsFIRE AND MATERIALS, Issue 6 2006Jin-Ping Guan Abstract The paper mainly deals with flame retardancy of silk fabrics treated with a commercial organophosphorus flame retardant [N-hydroxymethyl (3-dimethyl phosphono) propionamide (HDPP), also known as Pyrovatex CP], using the pad-dry-cure-wash method. The structures and properties of the treated and control sample are discussed. The Limiting Oxygen Index (LOI) value of the modified sample is above 30%. After 50 laundry cycles, it still has some flame retardancy left. HDPP and a cross-linking agent (HMM) were bound to silk fabrics which is confirmed by FT-IR spectra and amino analysis. The reaction degree of the flame retardant with silk is also high; almost all the tyrosine units have reacted, which can be confirmed by amino acid analysis. The reaction between flame retardant and silk only occurs in the amorphous region of silk fibre, which is confirmed by X-ray diffraction analysis and amino acid analysis. Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) analysis show that the flame retardant causes silk fabrics to decompose below its ignition temperature (600°C) and formed carbonaceous residue or char when exposed to fire. The char behaves as a thermal barrier to fire, so silk fabrics show good flame retardancy. The treatment has a little effect on the whiteness of the silk fabrics and the tensile strength of treated silk fabrics slightly decreased; both effects are negligible. Copyright © 2006 John Wiley & Sons, Ltd. [source] Dyskeratosis congenita with isolated neutropenia and granulocyte colony-stimulating factor treatmentINTERNATIONAL JOURNAL OF DERMATOLOGY, Issue 3 2002Kutluhan Yilmaz A 3-year-old Turkish boy with a history of chronic cough, recurrent bronchopneumonia, and a borderline sweat chloride test (40 mEq/L) was referred for further evaluation to our department. He was born at term (2100 g) to a marriage with no consanguinity. His mother and father were 40 and 46 years old, respectively. Physical examination (Fig. 1) revealed hypopigmented, atrophic, and hyperkeratotic skin lesions surrounded by reticulate hyperpigmentation on the entire body, predominantly on the face, neck, arms, shoulders, and legs, which had been noticed initially at the age of 18 months. Dystrophic toenails, sparse and thin hair, and phimosis were also observed. Laboratory tests disclosed an isolated neutropenia (white blood cell count, 1800/mm3). Bone marrow (BM) aspiration showed a decreased myelopoiesis without myelodysplastic changes, but normal erythropoiesis, megakaryopoiesis, and normal stroma. Lymphocyte subgroups containing CD4, CD5, CD6, CD8, CD19, CD23, and CD25, and immunoglobulin G (IgG), IgM, IgA, and IgE, were in the normal range; hemoglobin F (HbF), 2.8%. Spontaneous and clastogen-induced chromosome breaks were not increased. A skin biopsy showed increased pigmentation at the basal layer, dyskeratotic epidermal cells, and marked IgM deposition and cytoid bodies and mild IgA and IgG deposits at the dermo-epidermal junction. Lactate response to glucose challenge, amino acid chromatography, and urine organic acid analysis were normal. Figure 1. Hypopigmented, atrophic, and hyperkeratotic skin lesions surrounded by reticulate hyperpigmentation involving predominantly the face, neck, arms, shoulders, and legs, dystrophic toenails, and sparse and thin hair A diagnosis of dyskeratosis congenita (DC) was made with typical skin lesions, dystrophic toenails, thin and sparse hair, and neutropenia with decreased myelopoiesis in BM. Treatment with granulocyte colony-stimulating factor (G-CSF) was considered for the neutropenia. As the increase in neutrophil count at a dose of 5 µg/kg was not adequate, 10 µg/kg G-CSF was tried (Fig. 2). With 10 µg/kg once to three times a week, a 1.8,4.8-fold increase in the absolute neutrophil count (ANC) was achieved with no side-effects. Treatment was more frequent during infection (days 22,28). Figure 2. Response of absolute neutrophil count (ANC) to granulocyte colony-stimulating factor (G-CSF) administration (5 µg/kg on days 1 and 3; 10 µg/kg on days 5, 10, 16, 23, 26, 28, 34, 40, 48, 54) [source] Diversity of soil mycobacterium isolates from three sites that degrade polycyclic aromatic hydrocarbonsJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2007C.D. Miller Abstract Aims:, This paper investigates the diversity of polycyclic aromatic hydrocarbon (PAH)-degrading mycobacterium isolates from three different sites within United States: Montana, Texas and Indiana. Methods and Results:, All five mycobacterium isolates differed in chromosomal restriction enzyme-fragmentation patterns; three isolates possessed linear plasmids. The DNA sequence between the murA and rRNA genes were divergent but the sequence upstream of nidBA genes, encoding a dioxygenase involved in pyrene oxidation, was more highly conserved. Long-chain fatty acid analysis showed most similarity between three isolates from the same Montana site. All isolates were sensitive to rifampicin and isoniazid, used in tuberculosis treatment, and to syringopeptins, produced by plant-associated pseudomonads. Biofilm growth was least for isolate MCS that grew on plate medium as rough-edged colonies. The patterns of substrate utilization in Biolog plates showed clustering of the Montana isolates compared with Mycobacterium vanbaalenii and Mycobacterium gilvum. Conclusion:, The five PAH-degrading mycobacterium isolates studied differ in genetic and biochemical properties. Significance and Impact of the Study:, Different properties with respect to antibiotic susceptibility, substrate utilization and biofilm formation could influence the survival in soil of the microbe and their suitability for use in bioaugmentation. [source] Identification and serotyping of atypical Legionella pneumophila strains isolated from human and environmental sourcesJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2007J.H. Helbig Abstract Aims:, To validate identification methods for Legionella pneumophila strains that cannot be serotyped into the known serogroups and to characterize their antigenic diversity. Methods and Results:, Fifty L. pneumophila strains that could not be serogrouped, but which had been confirmed as L. pneumophila by mip gene sequencing, were further identified phenotypically. We used (i) MONOFLUO anti- Legionella Staining Reagent (Bio-Rad) (50/50), (ii) an in-house prepared immunoblot assay for the detection of L. pneumophila - specific Mip protein epitope (50/50), (iii) fatty acid analysis using the Microbial Identifications System (MIDI) (47/50) and (iv) Oxoid agglutination tests (44/50). The serological diversity was further characterized by testing with five serogroup-cross-reactive monoclonal antibodies, resulting in nine phenons. Conclusions:, The division of L. pneumophila into 15 serogroups does not reflect the serogroup heterogeneity. Results of these tests indicate that there are more serogroups. Significance and Impact of the Study:, MONOFLUO anti- Legionella Staining Reagent is the only commercially available tool for identifying atypical strains of L. pneumophila. If necessary for epidemiological purposes, the antigenic heterogeneity of these strains can be analysed by monoclonal antibodies. [source] Characterization of micro-organisms isolated from dairy industry after cleaning and fogging disinfection with alkyl amine and peracetic acidJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2005E. Bore Abstract Aims:, To characterize micro-organisms isolated from Norwegian dairy production plants after cleaning and fogging disinfection with alkyl amine/peracetic acid and to indicate reasons for survival. Methods and Results:, Microbial samples were collected from five dairy plants after cleaning and fogging disinfection. Isolates from two of these production plants, which used fogging with alkylamino acetate (plant A), and peracetic acid (plant B), were chosen for further characterization. The sequence of the 16S ribosomal DNA, fatty acid analysis and biochemical characteristics were used to identify isolates. Three isolates identified as Rhodococcus erythropolis, Methylobacterium rhodesianum and Rhodotorula mucilaginosa were isolated from plant A and one Sphingomonas sp. and two M. extorquens from plant B. Different patterns of resistance to seven disinfectants in a bactericidal suspension test and variable degree of attachment to stainless steel were found. The strains with higher disinfectant resistance showed lower degree of attachment than susceptible strains. Conclusions:, The study identifies and characterizes micro-organisms present after cleaning and fogging disinfection. Both surface attachment and resistance were shown as possible reasons for the presence of the isolates after cleaning and disinfection. Significance and Impact of the Study:, These results contribute to the awareness of disinfectant resistance as well as attachment as mechanisms of survival in dairy industry. It also strengthens the argument of frequent alternation of disinfectants in the food processing industry to avoid the establishment of resistant house strains. [source] Negative and positive ion matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and positive ion nano-electrospray ionization quadrupole ion trap mass spectrometry of peptidoglycan fragments isolated from various Bacillus speciesJOURNAL OF MASS SPECTROMETRY (INCORP BIOLOGICAL MASS SPECTROMETRY), Issue 2 2001Gerold Bacher Abstract A general approach for the detailed characterization of sodium borohydride-reduced peptidoglycan fragments (syn. muropeptides), produced by muramidase digestion of the purified sacculus isolated from Bacillus subtilis (vegetative cell form of the wild type and a dacA mutant) and Bacillus megaterium (endospore form), is outlined based on UV matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and nano-electrospray ionization (nESI) quadrupole ion trap (QIT) mass spectrometry (MS). After enzymatic digestion and reduction of the resulting muropeptides, the complex glycopeptide mixture was separated and fractionated by reversed-phase high-performance liquid chromatography. Prior to mass spectrometric analysis, the muropeptide samples were subjected to a desalting step and an aliquot was taken for amino acid analysis. Initial molecular mass determination of these peptidoglycan fragments (ranging from monomeric to tetrameric muropeptides) was performed by positive and negative ion MALDI-MS using the thin-layer technique with the matrix ,-cyano-4-hydroxycinnamic acid. The results demonstrated that for the fast molecular mass determination of large sample numbers in the 0.8,10 pmol range and with a mass accuracy of ±0.07%, negative ion MALDI-MS in the linear TOF mode is the method of choice. After this kind of muropeptide screening often a detailed primary structural analysis is required owing to ambiguous data. Structural data could be obtained from peptidoglycan monomers by post-source decay (PSD) fragment ion analysis, but not from dimers or higher oligomers and not with the necessary sensitivity. Multistage collision-induced dissociation (CID) experiments performed on an nESI-QIT instrument were found to be the superior method for structural characterization of not only monomeric but also of dimeric and trimeric muropeptides. Up to MS4 experiments were sometimes necessary to obtain unambiguous structural information. Three examples are presented: (a) CID MSn (n = 2,4) of a peptidoglycan monomer (disaccharide-tripeptide) isolated from B. subtilis (wild type, vegetative cell form), (b) CID MSn (n = 2,4) of a peptidoglycan dimer (bis-disaccharide-tetrapentapeptide) obtained from a B. subtilis mutant (vegetative cell form) and (c) CID MS2 of a peptidoglycan trimer (a linear hexasaccharide with two peptide side chains) isolated from the spore cortex of B. megaterium. All MSn experiments were performed on singly charged precursor ions and the MS2 spectra were dominated by fragments derived from interglycosidic bond cleavages. MS3 and MS4 spectra exhibited mainly peptide moiety fragment ions. In case of the bis-disaccharide-tetrapentapeptide, the peptide branching point could be determined based on MS3 and MS4 spectra. The results demonstrate the utility of nESI-QIT-MS towards the facile determination of the glycan sequence, the peptide linkage and the peptide sequence and branching of purified muropeptides (monomeric up to trimeric forms). The wealth of structural information generated by nESI-QIT-MSn is unsurpassed by any other individual technique. Copyright © 2001 John Wiley & Sons, Ltd. [source] The oxidation metabolites of endomorphin 1 and its fragments induced by free radicalsJOURNAL OF PEPTIDE SCIENCE, Issue 5 2009Pin Gong Abstract Endomorphin 1 (EM1), an endogenous µ-opioid receptor agonist, acts as a free radical scavenger in vitro and an antioxidant in vivo. The modification of EM1 by ROS and the properties of the OM attracted our attention. In vitro assays were performed via RP-HPLC, spectrophotometric measurements, EPR and amino acid analysis, Schmorl's reaction to define the formation of melanin-like compounds transformed from EM1, collectively named EM1,melanin and by solubility assay, radioligand-binding assay, NADH oxidation, superoxide anion scavenging assay to study some physical and chemical properties of EM1,melanin. Possible pathways of the formation of EM1,melanin were proposed. Copyright © 2009 European Peptide Society and John Wiley & Sons, Ltd. [source] Solid phase peptide synthesis on epoxy-bearing methacrylate monolithsJOURNAL OF PEPTIDE SCIENCE, Issue 12 2004E. Vlakh Abstract Monoliths based on a copolymer of glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) can be used directly as sorbents for affinity chromatography after solid phase peptide synthesis. The quality of the synthesized products, the amount of grown peptides on a support and the reproducibility of the process must be considered. A determination of the quantity of the introducing ,-Ala (and, consequently, the total amount of synthesized peptide) was carried out. Three peptides complementary to recombinant tissue plasminogen activator (t-PA) have been synthesized using Fmoc-chemistry on GMA-EDMA disks. The peptidyl ligands were analysed by amino acid analysis, ES-MS and HPLC methods. The affinity binding parameters were obtained from frontal elution data. The results were compared with those established for GMA-EDMA affinity sorbents formed by the immobilization of the same but separately synthesized and purified ligands. The immobilization on GMA-EDMA disks was realized using a one-step reaction between the amino groups of the synthetic ligand and the original epoxy groups of monolithic material. The affinity constants found for two kinds of sorbent did not vary significantly. Finally, the directly obtained affinity sorbents were tested for t-PA separation from a cellular supernatant. Copyright © 2004 European Peptide Society and John Wiley & Sons, Ltd. [source] Synthesis, conformation and biological activity of linear and cyclic Thr6 -bradykinin analogues containing N -benzylglycine in place of phenylalanine,JOURNAL OF PEPTIDE SCIENCE, Issue 12 2001L. Biondi Abstract Three linear Thr6 -bradykinin analogues in which either one or both the two phenylalanine residues in the peptide sequence have been substituted by N -benzylglycine (BzlGly) and their head-to-tail cyclic analogues were synthesized and tested on an isolated rat duodenum preparation. The linear (BzlGly5,Thr6 -BK, BzlGly8,Thr6 -BK and BzlGly5,8,Thr6 -BK) and the cyclic (cyclo BzlGly5,Thr6 -BK, cyclo BzlGly8,Thr6 -BK and cyclo BzlGly5,8,Thr6 -BK) peptoid-like analogues were characterized by amino acid analysis, optical rotation, analytical HPLC and MALDI-TOF mass spectroscopy. The conformational features of both the linear and cyclic derivatives were investigated by FT-IR and CD measurements. Preliminary molecular mechanics calculations were also performed on some synthetic peptides. Pharmacological screening using the relaxation of the isolated rat duodenum preparation showed that incorporation of N -benzylglycine at positions 5 and/or 8 in the linear Thr6 -BK causes a substantial decrease in potency. Comparable incorporation in cyclo Thr6 -BK, at position 8, or 5 and 8, resulted in nearly inactive analogues. However, cyclo BzlGly5,Thr6 -BK showed a potency which is of the same order of magnitude as for cyclo -BK and cyclo Thr6 -BK. Copyright © 2001 European Peptide Society and John Wiley & Sons, Ltd. [source] Nitrate modifies the assimilation pattern of ammonium and urea in wheat seedlingsJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 3 2010Maria Garnica Abstract BACKGROUND: In certain plant species, ammonium or urea nutrition can cause negative effects on plant development which can result in toxic symptoms. Some authors suggest that the presence of nitrate can alleviate these symptoms by increasing ammonium and urea assimilation, avoiding its accumulation. In order to study this hypothesis, wheat (Triticum aestivum L.) seedlings were grown with various nitrogen supplies containing the main nitrogen forms (ammonium, nitrate and urea). Amino acids content and the activity of the three main enzymes involved in nitrogen assimilation (nitrate reductase, glutamine synthetase and urease) were studied. RESULTS: The application of nitrate along with urea and/or ammonium was not associated with a time-sustained increase in the activity of glutamine synthetase and urease. Amino acid analysis revealed that nitrate induced changes in amino acid metabolism enhancing its concentration. Likewise the content of protein was also higher in nitrate-treated plants. CONCLUSION: These results suggest that the effect of nitrate is compatible with a rapid and transient increase in the activity of glutamine synthetase and urease during the first hour after the onset of treatments. Nevertheless, a possible effect of nitrate reducing ammonium accumulation through the activation of alternative metabolic pathways different from that involving glutamine synthetase cannot be ruled out. Finally, nitrate effects on amino acid concentration indicate that, whereas ammonium assimilation takes place principally in the root, urea and nitrate assimilation occurred in the shoot, under the conditions of the experiment. Copyright © 2009 Society of Chemical Industry [source] Replacement of Fish Meal with Poultry By-product Meal as a Protein Source in Pond-raised Sunshine Bass, Morone chrysops , × M. saxatlis ,, DietsJOURNAL OF THE WORLD AQUACULTURE SOCIETY, Issue 5 2008Harvey J. Pine Replacement of fish meal (FM) as a protein source with alternative sources of protein in aquaculture diets has been widely explored in aquaculture. The goal of replacement of FM in production diets is to maintain growth, lower production costs, and increase sustainability. Evaluation of the replacement of FM with poultry by-product meal (PBM) in phase II sunshine bass diets, Morone chrysops × M. saxatilis, was conducted in ponds over 246 d. Four diets were formulated to be isonitrogenous (37%) and isocaloric (4 kcal/g) with different levels of FM replacement with PBM (0, 33, 67, and 100%, Diets 1,4, respectively). Twelve ponds were stocked with 400 phase II sunshine bass (mean weight 5.6 g) and randomly assigned one of the four diets. Fish were fed below satiation based on predicted growth and feed conversion, initially once daily (1700 h) and then twice daily (0700 and 1700 h) as water temperatures and feeding activity increased. Diets were evaluated based on production and performance indicators, body composition, and economic analysis. Production results revealed no significant differences in mean final individual fish weight (511 ± 21 g), net production (4257 ± 247 kg/ha), and survival (85 ± 2%). No significant differences occurred between the performance indicators: mean feed conversion ratio (2.47 ± 0.11), specific growth rate (1.84 ± 0.02), and protein conversion efficiency (23 ± 1.3%). Body composition was statistically similar for mean percent fillet weight (49 ± 0.6%) and percent intraperitoneal fat (9.8 ± 1.0%); however, the hepatosomatic index was significantly different between Diets 3 (3.7 ± 0.1%) and 4 (3.2 ± 0.1%). Mean proximate analysis of whole fish (dry weight basis) was not significantly different among treatments yielding the following: percent protein (46 ± 0.4%), lipid (47 ± 1.3%), and ash (8 ± 0.7%). Mean fillet composition (dry weight basis) also revealed no significant differences: percent protein (72 ± 0.8%), percent lipid (30 ± 1.6%), and percent ash (5 ± 0.2%). Proximate analysis was also performed on the diets and revealed a significantly lower protein content in Diet 3 (34.3 ± 0.5%) compared to the other diets (37.1 ± 0.4%). Amino acid analysis of the diets indicated a possible deficiency in methionine in Diets 3 and 4. Based on production, performance, and body composition, the results indicate that complete replacement of FM with PBM in sunshine bass diets is feasible; however, economic analysis suggests that the replacement of FM with PBM may result in reduced revenue over feed costs. [source] Role of Alicyclobacillus acidoterrestris in the development of a disinfectant taint in shelf-stable fruit juiceLETTERS IN APPLIED MICROBIOLOGY, Issue 1 2003N. Jensen Abstract Aims: This study was undertaken to identify the bacterium and metabolic products contributing to a disinfectant taint in shelf-stable fruit juice and to determine some of the growth conditions for the organism. Methods and Results: Microbiological examination of tainted and untainted fruit juice drinks detected low numbers of acid-dependent, thermotolerant, spore-forming bacteria in the tainted juices only. The presence of ,-cyclohexyl fatty acids was confirmed in two of the isolates by cell membrane fatty acid analysis. The isolates were subsequently identified as Alicyclobacillus acidoterrestris by partial 16S rDNA sequencing. Studies on the isolates showed growth at pH 2·5,6·0 and 19·5,58 °C. Gas chromatography/mass spectrometry (GC/MS) was used to identify and quantify 2,6-dibromophenol (2,6-DBP) and 2,6-dichlorophenol (2,6-DCP) in the tainted juice. Challenge studies in a mixed fruit drink inoculated with the two isolates and the type strain of A. acidoterrestris, incubated at 44,46 °C for 4 d, showed the production of both metabolites, which were confirmed and quantified by GC/MS. Conclusions: The results show that A. acidoterrestris can produce 2,6-DBP and 2,6-DCP in shelf-stable juices. Significance and Impact of the Study: This is the first report detailing experimental methodology showing that A. acidoterrestris can produce 2,6-DCP in foods. Control of storage temperatures (to <,20 °C) immediately after processing may provide an effective control measure for the fruit juice industry to prevent spoilage by A. acidoterrestris. [source] Properties and Bioapplications of Blended Cellulose and Corn Protein FilmsMACROMOLECULAR BIOSCIENCE, Issue 9 2009Quanling Yang Abstract A series of blend films have been prepared from cellulose and corn protein in a NaOH/urea solution by a simple, low cost, and ,green' pathway. Their structure and properties are characterized by amino acid analysis, X-ray diffraction, scanning electron microscopy, thermogravimetry, and tensile testing. The results reveal that a certain miscibility exists between cellulose and corn protein and their thermal stability and mechanical properties are improved significantly, compared with the protein materials, when the protein content is less than 18 wt.-%. The protein, which contains tyrosine and histidine, could remain in the blend films after being washed for ten days, which indicates the strong hydrogen bonding between the hydroxy groups of cellulose and the hydroxyphenyl of tyrosine and imidazolyl of histidine in the protein. Furthermore, they exhibit good biocompatibility capable of supporting cell adhesion and proliferation. [source] Standardization of allergen products: 1.ALLERGY, Issue 7 2009Detailed characterization of GMP-produced recombinant Bet v 1.0101 as biological reference preparation Background:, Standardization of allergen extracts requires the availability of well-characterized recombinant allergens, which can be used as reference standards provided by the European regulatory authorities. The objective of this study was the detailed physicochemical and immunological characterization of rBet v 1.0101, which shall be used in a ring trial within the framework of the Biological Standardization Programme BSP090 of the European Directorate for Quality of Medicines and Healthcare. Methods:, Recombinant Bet v 1.0101 Y0487 was produced under good manufacturing practice conditions and analysed by an array of physicochemical and immunological methods for identity, quantity, homogeneity, folding and denaturation, aggregation state and stability in solution, as well as biological activity. Results:, Batch Y0487 was shown to contain monomeric and well-folded protein being identical with rBet v 1.0101, as determined by mass spectrometry. SDS-PAGE, isoelectric focusing, deamidation analysis and size-exclusion chromatography with light scattering revealed sample homogeneity of >99.9%. Upon storage at +4°C batch Y0487 retained the monomeric state up to 3 months. Protein quantification determined by amino acid analysis was found coinciding with half-maximal inhibition of serum IgE in ELISA. Biological activity of batch Y0487 was shown to be comparable to natural Bet v 1 by IgG and IgE immunoblotting, as well as basophil and T-cell activation. Conclusion:, Recombinant Bet v 1.0101 Y0487 was characterized extensively by physicochemical and immunological methods. It was shown highly stable, monomeric and immunologically equivalent to its natural counterpart. Thus, it represents an appropriate candidate reference standard for Bet v 1. [source] Borrelia burgdorferi membranes are the primary targets of reactive oxygen speciesMOLECULAR MICROBIOLOGY, Issue 3 2008Julie A. Boylan Summary Spirochetes living in an oxygen-rich environment or when challenged by host immune cells are exposed to reactive oxygen species (ROS). These species can harm/destroy cysteinyl residues, iron-sulphur clusters, DNA and polyunsaturated lipids, leading to inhibition of growth or cell death. Because Borrelia burgdorferi contains no intracellular iron, DNA is most likely not a major target for ROS via Fenton reaction. In support of this, growth of B. burgdorferi in the presence of 5 mM H2O2 had no effect on the DNA mutation rate (spontaneous coumermycin A1 resistance), and cells treated with 10 mM t -butyl hydroperoxide or 10 mM H2O2 show no increase in DNA damage. Unlike most bacteria, B. burgdorferi incorporates ROS-susceptible polyunsaturated fatty acids from the environment into their membranes. Analysis of lipoxidase-treated B. burgdorferi cells by Electron Microscopy showed significant irregularities indicative of membrane damage. Fatty acid analysis of cells treated with lipoxidase indicated that host-derived linoleic acid had been dramatically reduced (50-fold) in these cells, with a corresponding increase in the levels of malondialdehyde by-product (fourfold). These data suggest that B. burgdorferi membrane lipids are targets for attack by ROS encountered in the various stages of the infective cycle. [source] Phytate in foods and significance for humans: Food sources, intake, processing, bioavailability, protective role and analysisMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue S2 2009Ulrich Schlemmer Abstract The article gives an overview of phytic acid in food and of its significance for human nutrition. It summarises phytate sources in foods and discusses problems of phytic acid/phytate contents of food tables. Data on phytic acid intake are evaluated and daily phytic acid intake depending on food habits is assessed. Degradation of phytate during gastro-intestinal passage is summarised, the mechanism of phytate interacting with minerals and trace elements in the gastro-intestinal chyme described and the pathway of inositol phosphate hydrolysis in the gut presented. The present knowledge of phytate absorption is summarised and discussed. Effects of phytate on mineral and trace element bioavailability are reported and phytate degradation during processing and storage is described. Beneficial activities of dietary phytate such as its effects on calcification and kidney stone formation and on lowering blood glucose and lipids are reported. The antioxidative property of phytic acid and its potentional anticancerogenic activities are briefly surveyed. Development of the analysis of phytic acid and other inositol phosphates is described, problems of inositol phosphate determination and detection discussed and the need for standardisation of phytic acid analysis in foods argued. [source] Proximate composition, fatty acid analysis and protein digestibility-corrected amino acid score of three Mediterranean cephalopodsMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 10 2006Spiros Zlatanos Abstract Proximate composition, fatty acid analysis and protein digestibility-corrected amino acid score (PDCAAS) in three commercially important cephalopods of the Mediterranean sea (cuttlefish, octopus and squid) were determined. The results of the proximate analysis showed that these species had very high protein:fat ratios similar to lean beef. Docosahexaenoic, palmitic and eicosipentaenoic acid were the most abundant fatty acids among analyzed species. The amount of n- 3 fatty acids was higher than that of saturated, monounsaturated and n- 6 fatty acids. Despite the fact that cephalopods contain small amounts of fat they were found quite rich in n- 3 fatty acids. Finally, PDCAAS indicated that these organisms had a very good protein quality. [source] Trichothiodystrophy-like Hair Abnormalities in a Child with Keratitis Ichthyosis Deafness SyndromePEDIATRIC DERMATOLOGY, Issue 4 2008L. De Raeve M.D., Ph.D. It appears to be genetically heterogeneous and may be caused by mutations in the connexin 26 (Cx26) gene (GJB2) or in the connexin 30 gene. It is characterized by the association of ichthyosis-like skin lesions, hearing loss, and vascularizing keratitis. We report the clinical and molecular findings in a 5-year-old girl with keratitis ichthyosis deafness syndrome. DNA sequencing in our patient revealed a p.Ser17Phe mutation in GJB2. Besides the typical clinical features of keratitis ichthyosis deafness syndrome, a peculiar intriguing finding not previously described in the literature in this condition was that polarizing light microscopy of the scalp hair in our patient revealed striking bright and dark bands as seen in trichothiodystrophy. Amino acid analysis of the hair sample also disclosed a reduced cysteine index. We emphasize that it would be of great benefit to examine hair shafts in other patients with keratitis ichthyosis deafness syndrome for trichothiodystrophy-like abnormalities. [source] | ||||||||||||||||||||||||||||||||||