Candida biotypes isolated from clinical specimens in Malaysia

The distribution of Candida species was examined using 1114 yeasts isolated from various clinical specimens. The isolates were identified by germ tube test, hyphal/pseudohyphae and chlamydoconidia production and carbohydrate assimilation test using ten carbohydrates (glucose, sucrose, trehalose, cellobiose, arabinose, galactose, mannitol, raffinose, lactose and maltose). Among the 1114 isolates studied, 9 species of Candida were identified and the relative frequency of isolation was C. albicans (44.2%), C. parapsilosis (26.0%), C. tropicalis (17.7%), C. glabrata (9.6%), C. krusei (1.2%), C. rugosa (0.6%), C. guilliermondii (0.2%), C. lusitaniae (0.08%) and C. kefyr (0.08%). Non- C. albicans was the most common Candida species isolated from blood, respiratory system, urine and skin. The isolate from vaginal swabs was predominantly C. albicans. 82.2% of C. glabrata and 64.2% of C. krusei isolated in this study were from vaginal swabs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of Substrate on the Fatty Acid Composition of Hydrocarbon-utilizing Microorganisms

The fatty acid pattern in three hydrocarbon-utilizing bacteria during growth on various substrates was examined. The predominant fatty acids in acetate-grown cells were C(16), C(16:1), C(18:1), and Br-C(19) and the major fatty acids in propane-grown cells were C(15), C(17), C(17:1), C(18:1), and Br-C(18). When one organism (Mycobacterium sp. strain OFS) was grown on the n-alkanes from C(13) to C(17), the major fatty acid in the cells was of the same chain length as the substrate. Studies on the incorporation of acetate into the cellular fatty acids of microorganisms growing on C(15) and C(17)n-alkanes suggest that the oxidative products of the substrate are incorporated into the cellular fatty acids without degradation to acetate.     

Defining the drug-binding site in the human multidrug resistance P-glycoprotein using a methanethiosulfonate analog of verapamil, MTS-verapamil

Defining the residues involved in the binding of a substrate provides insight into how the human multidrug resistance P-glycoprotein (P-gp) can transport a wide range of structurally diverse compounds out of the cell. Because verapamil is the most potent stimulator of P-gp ATPase activity, we synthesized a thiol-reactive analog of verapamil (MTS-verapamil) and used it with cysteine-scanning mutagenesis to identify the reactive residues within the drug-binding domain of P-gp. MTS-verapamil stimulated the ATPase activity of Cys-less P-gp and had a K(m) value (25 microM) that was similar to that of verapamil. 252 P-gp mutants containing a single cysteine within the predicted transmembrane (TM) segments were expressed in HEK 293 cells and purified by nickel-chelate chromatography and assayed for inhibition by MTS-verapamil. The activities of 15 mutants, Y118C (TM2), V125C (TM2), S222C (TM4), L339C (TM6), A342C (TM6), A729C (TM7), A841C (TM9), N842C (TM9), I868C (TM10), A871C (TM10), F942C (TM11), T945C (TM11), V982C (TM12), G984C (TM12), and A985C (TM12), were inhibited by MTS-verapamil. Four mutants, S222C (TM4), L339C (TM6), A342C (TM6), and G984C (TM12), were significantly protected from inhibition by MTS-verapamil by pretreatment with verapamil. Less protection was observed in mutants I868C (TM10), F942C (TM11) and T945C (TM11). These results indicate that residues in TMs 4, 6, 10, 11, and 12 must contribute to the binding of verapamil.     

Distribution of Candida species in different clinical sources in Delhi, India, and proteinase and phospholipase activity of Candida albicans isolates

Eighty-five isolates of Candida recovered from three hundred and fifty diverse clinical sources, viz. respiratory tract (sputum, bronchial washing,bronchoalveolar lavage, tracheal aspirate), blood, urine, high vaginal swab, skin and plastic devices, were studied in detail for their morphological and biochemical characters. Seven species of Candida were identified, viz., C. albicans (45.8%), C. tropicalis (24.7%), C. parapsilosis (10.5%), C. krusei (7.0%), C. kefyr (7.0%), C. guilliermondii (3.5%), and C. glabrata (1.1%). C. albicans was the predominant species isolated from all clinical specimens, except blood from which C. krusei was most frequently (38.4%) recovered. Out of 39 isolates of C. albicans, 26 (66.6%) and 19 (48.7%) exhibited strong proteinase and phospholipase activity respectively. There was a higher prevalence of proteinase producing strains amongst the vaginal and skin isolates than that in urinary and respiratory isolates. Also a greater number of phospholipase producing strains was observed in the vaginal and urinary isolates than that in the respiratory and skin isolates.     

Measuring the heat loss in horses in different seasons by infrared thermography

It is necessary to consider breed and cold tolerance in the housing and caring of horses. This study demonstrates differences in heat loss between horse types at low temperatures and examines rate of loss in different types during different seasons. Eighteen horses participated. Groups by type were light (L), warmblood (W), coldblood (C), and pony (P). A camera filmed thermographic images at 15 degrees C, 2 degrees C (all types), -8 degrees C (L, W, C), and -12 degrees C (P). The study calculated loss from the neck, trunk, and inner surfaces of front and hind legs. Loss was similar in all types at 15 degrees C. L, W, and C dissipated more heat at 2 degrees C than at 15 degrees C (p < .001) and from neck and trunk at -8 degrees C than at 2 degrees C (p < .05). P dissipated heat similarly at 2 degrees C and -12 degrees C. At 2 degrees C, loss was less from neck and trunk in C and P compared with L (p < .05). At -8 degrees C, loss in L and W was greater than in C (p < .05).     

种植模式和施肥对黄土旱塬农田土壤团聚体及其碳分布的影响

本研究以长武黄土高原农业生态试验站33年长期定位试验处理为研究对象,选取撂荒(R)、小麦连作(CK/W)、小麦玉米轮作(L),小麦连作选取单施氮肥(N)、单施磷肥(P)、施氮磷肥(NP)、单施有机肥(M)、氮肥配施有机肥(NM)、磷肥配施有机肥(PM)、氮磷肥配施有机肥(NPM)共10种不同种植模式和施肥田间处理,运用...     

Centrifugation and addition of glycerol at 22 degres C instead of 4 degrees C improve post-thaw motility and fertility of stallion spermatozoa

The aims of this study were to evaluate the effects of cooling rate to 4 degrees C and temperature at the time of centrifugation/glycerol-addition (freezing extender: INRA82 + 2% egg yolk + 2.5% glycerol) on postcentrifugation recovery rate, post-thaw motility and per-cycle fertility. When centrifugation/glycerol-addition was performed at 4 degrees C (14 ejaculates), a moderate cooling rate (37 degrees C to 4 degrees C in I h) resulted in higher post-thaw motility (45%) than when using a slow cooling rate (37 degrees C to 4 degrees C in 4 h) (39%; P<0.05). When centrifugation/glycerol-addition was performed at 22 degrees C (37 degrees C to 22 degrees C in 10 min) (10 ejaculates), post-thaw motility was lower when spermatozoa were frozen directly from 22 degrees C (23%) than when spermatozoa were cooled to 4 degrees C (22 degrees C to 4 degrees C in 1 h) before freezing (47%; P<0.0001). When centrifugation/glycerol-addition was performed at 22 degrees C (before cooling at a moderate rate), as opposed to 4 degrees C (after cooling at a moderate rate), a significant improvement of 1) recovery of spermatozoa after centrifugation (P<0,0001), 2) post-thaw motility of spermatozoa at thawing (40% vs 36% (n < or = 291 ejaculates/group), P<0.0001) and 3) per-cycle fertility (56% vs 42% (n > or = 190 cycles/group), P<0.01) was observed. In conclusion, centrifugation/glycerol-addition at 22 degrees C followed by cooling to 4 degrees C at a moderate rate results in an improvement of post-thaw motility, spermatozoa recovery rate and per cycle fertility.     

Carbohydrate, glycolipid, and lipid components from the photobiont (Scytonema sp.) of the lichen, Dictyomema glabratum

The photobiont of the lichen, Dictyonema glabratum (Scytonema sp.), was isolated and cultivated in a soil-extract medium and submitted to chemical analysis. Successive extractions with CHCl3-MeOH, aqueous MeOH, and H2O gave rise to solutions of lipids (25%), low-molecular-weight carbohydrates (22%), and polysaccharides (4%), respectively. TLC of the lipid extract showed the presence of glycolipids, which were further purified and examined by NMR spectroscopy and GC-MS. Monogalactosyldiacylglycerol (1%), digalactosyldiacylglycerol (0.8%), trigalactosyldiacylglycerol (0.4%), and sulfoquinovosyldiacylglycerol (0.5%) were identified. The most abundant fatty acid ester in each fraction was palmitic (C16:0), but a great variation of the ester composition from one to another was found. Others present were those of C12:0, C14:0, C15:0, C16:1, C17:0, C18:0, C18:1, C18:2, C18:3, C22:0, C22:2, and C24:0. The lipid extract was also subjected to acid methanolysis, which gave rise to dodecane, 2-Me-heptadecane, 2,6-Me2-octadecane, and 8-Me-octadecane, methyl esters of C14:0, C15:0, C16:0, C16:1, C17:0, C18:0, C18:1, C18:2, C20:0, and C24:0 fatty acids, and the dimethyl ester of decanedioic acid. The polysaccharide had mainly Glc, Gal, and Man, with small amounts of 3-O-methylrhamnose and 2-O-methylxylose, both found in plants, and unexpectedly, some of the units were beta-galactofuranose, typical of fungal, but not cyanobacterial polysaccharides. The low-molecular-weight carbohydrates showed mannose as the main free reducing sugar, which differs from Nostoc sp. and Trebouxia sp. photobionts.     

Structural characterization of eight cyclic lipopeptides produced by Bacillus subtilis HSO121

Lipopeptides are amphiphilic compounds which contain both hydrophobic fatty acid moieties and amphiphilic peptide moieties. From the cell-free broth of Bacillus subtilis HSO121, eight cyclic lipopeptides were isolated by reversed-phase high performance liquid chromatography (RP-HPLC). The peptide part of each lipopeptide was elucidated according to electrospray ionization quadruple-time-of-flight mass spectrometry (ESI Q-TOF MS) and the fatty acid part was analyzed by electroionization gas chromatography/mass spectrometry (EI GC/MS). It showed that fractions 1-8 had molecular masses of 1007, 1021, 1021, 1035, 1035, 1035, 1063, and 1049, respectively. Analysis of hydrolyzed lipopeptides revealed that they had invariant amino acid compositions. The differences in molecular weights represent changes in the number of methylene groups and different types of branched chains in fatty acids. Peptide sequences of two of the eight lipopeptides appeared to be N-Asp-Leu-Leu-Val-Glu-Leu-Leu-C, which was different from previously reported lipopeptides. The remaining six had an identical peptide sequence of N-Glu-Leu-Leu-Val-Asp-Leu-Leu-C. The fatty acid parts were found to be mixtures of iso C(12), iso C(13), anteiso C(13), iso C(14), n C(14), iso C(15), anteiso C(15), n C(15), anteiso C(16) and anteiso C(17) beta-hydroxy fatty acids. The structure of each lipopeptide was determined to be the beta-hydroxy fatty acid bonded to the peptide chain.     

棉籽浓缩蛋白替代饲料中豆粕对草鱼生长性能、健康状况及肌肉品质的影响

研究旨在探讨以棉籽浓缩蛋白(Cottonseed protein concentrate, CPC)替代饲料中豆粕对草鱼(Ctenopharyngodon idella)生长性能、健康状况及肌肉品质的影响。设计了6种等氮(29%)等脂(5%)饲料,分别用棉籽浓缩蛋白替代草鱼饲料中0 (C0)、15%(C15)、30%(C30)、45%(C45)、75%(C75)和100%(C100)的豆粕,饲喂养殖于池塘网箱中平均体重为(502.46±0.40) g的草鱼(Ctenopharyngodon Idella)90d。结果显示:(1)在生长性能方面,草鱼的特定生长率和饲料系数在各组之间均无显著性差异(P>0.05)。与C0组相比, C45组草鱼的脏体比显著升高(P<0.05)。(2)在健康状况方面, C100组血清白球比显著低于C0组(P<0.05), C15组血清超氧化物歧化酶(SOD)活性显著高于C0组(P<0.05);各组草鱼肝脏、肌肉组织未发生病变。(3)在肌肉品质方面,与C0组相比, C75组草鱼肌肉粗蛋白含量和C30组异亮氨酸含量显著升高(P<0.0...     

Evaluation of a High Throughput Method for the Detection of Mutations Associated with Thrombosis and Hereditary Hemochromatosis in Brazilian Blood Donors

BackgroundThe aim of this study was to evaluate the OpenArray platform for genetic testing of blood donors and to assess the genotype frequencies of nucleotide-polymorphisms (SNPs) associated with venous thrombosis (G1691A and G20210A), hyperhomocysteinemia (C677T, A1298C), and hereditary hemochromatosis (C282Y, H63D and S65C) in blood donors from Sao Paulo, Brazil.MethodsWe examined 400 blood donor samples collected from October to November 2011. The SNPs were detected using OpenArray technology. The blood samples were also examined using a real-time PCR–FRET system to compare the results and determine the accuracy of the OpenArray method.ResultsWe observed 100% agreement in all assays tested, except HFE C282Y, which showed 99.75% agreement. The HFE C282Y assay was further confirmed through direct sequencing, and the results showed that OpenArray analysis was accurate. The calculated frequencies of each SNP were FV G1691A 98.8% (G/G), 1.2% (G/A); FII G2021A 99.5% (G/G), 0.5% (G/A); MTHFR C677T 45.5% (C/C), 44.8% (C/T), 9.8% (T/T); MTHFR A1298C 60.3% (A/A), 33.6% (A/C), 6.1% (C/C); HFE C282Y 96%(G/G), 4%(G/A), HFE H63D 78.1%(C/C), 20.3% (C/G), 1.6% (G/G); and HFE S65C 98.1% (A/A), 1.9% (A/T).ConclusionTaken together, these results describe the frequencies of SNPs associated with diseases and are important to enhance our current knowledge of the genetic profiles of Brazilian blood donors, although a larger study is needed for a more accurate determination of the frequency of the alleles. Furthermore, the OpenArray platform showed a high concordance rate with standard FRET RT-PCR.     

4'-Acetylvitexin-2'-O-rhamnoside, isoorientin, orientin, and 8-methoxykaempferol-3-O-glucoside as markers for the differentiation of Crataegus monogyna and Crataegus pentagyna from Crataegus laevigata (Rosaceae)

In our chemotaxonomic investigation of pharmaceutically relevant Crataegus species, the qualitative and quantitative flavonoid fingerprint of Crataegus monogyna and C. pentagyna is presented. Six flavonoids were identified as vitexin-2'-O-rhamnoside (1), vitexin (2), isovitexin (3), rutin (4), hyperoside (5), and isoquercitrin (6). Besides the verification of the main compounds isoorientin (7) and orientin (8) in C. pentagyna, further four flavonoids were isolated and identified as isoorientin-2'-O-rhamnoside (9), orientin-2'-O-rhamnoside (10), isovitexin-2'-O-rhamnoside (11), and 8-methoxykaempferol-3-O-glucoside (12) by means of 1D- and 2D-NMR, MS, and UV analyses. Compound 12 was isolated for the first time from C. pentagyna. In contrast to C. pentagyna, C. monogyna samples were predominated by 4'-acetylvitexin-2'-O-rhamnoside (13), which was missing in C. pentagyna. Hence, 13 represents an interesting compound for chemotaxonomy of C. monogyna, whereas the main flavonoids 7, 8, and 12 could be proposed as markers for C. pentagyna. The absence of 7, 8, 12, and 13 in C. laevigata offers an appropriate tool for additional differentiation from C. monogyna and C. pentagyna, and for sample identification and quality control of the three main Crataegus species used in European phytotherapy.     

Chemical properties of lipopolysaccharide-like substance (LLS) extracted from Leptospira interrogans serovar canicola strain Moulton

The aqueous layer was isolated from Leptospira interrogans serovar canicola strain Moulton by the hot phenol-water method. After ultracentrifugation, the precipitate was designated as lipopolysaccharide-like substance (LLS) fraction and the chemical composition was compared with that of bacterial LPS. The LLS fraction consists of 35.2% carbohydrate, 3.8% amino sugar, 36.4% lipid, 15.2% protein, and 0.3% phosphorus. Neutral sugars were detected as rhamnose, arabinose, xylose, 4-O-methylmannose, mannose, galactose, and a small amount of erythrose, fucose and glucose by gas-liquid chromatography (GLC), but 2-keto-3-deoxyoctonic acid was not detected in the LLS by thiobarbituric acid test and high voltage paper electrophoresis. Fatty acids detected by GLC were decanoic acid (C10: 0), dodecanoic acid (C12: 0), dodecenoic acid (C12: 1), tridecenoic acid (C13: 1), tetradecanoic acid (C14: 0), hexadecanoic acid (C16: 0), hexadecenoic acid (C16: 1), and octadecenoic acid (C18: 1). With SDS-polyacrylamide gel electrophoresis, bacterial LPS showed many orderly bands, while the banding pattern of the leptospiral LLS was very simple. These findings demonstrate that the physicochemical properties and chemical composition of LLS fraction from Leptospira are different from those of LPS extracted from gram-negative bacteria such as Enterobacteriaceae, and suggesting that Leptospira has no typical LPS.     

Etiologic agents of fungemia in hospitalized patients]

The aim of performed examinations was the analysis of fungi as etiological agents of blood infections in patients hospitalized in surgical wards, internal medicine wards and intensive care units of the Medical Academy Central Clinical Hospital in Warsaw. Blood samples from patients hospitalized in 1997 were examined. Peripheral blood samples were incubated in BacT/Alert system (Organon Teknika, USA). Positive blood samples were inoculated on Sabouraud medium with chloramphenicol (bioMerieux, France or Oxoid, England). The time of cultivation was from 48 hours to 7 days at 30 degrees C. Fungal strains were identified by standard mycological procedures with the use of chromogenic medium BBL CHROMagar Candida (Becton Dickinson, USA) and biochemical test ID 32 C (bioMerieux, France). Susceptibility of strains to antifungal agents was determined by ATB FUNGUS method (bioMerieux, France). The total number of positive blood cultures in 1997 was 1380. Forty-two fungal strains were isolated from blood samples (3%). Strains belonged to the following species: C. albicans (17 isolates), C. parapsilosis (15), C. glabrata (3), melibiosica (2), C. pelliculosa (2), C. guilliermondii (1), C. tropicalis (1) and T. beigelii (1). Among fungi cultured from patients hospitalized in operative wards dominated C. parapsilosis (11) and C. albicans (10) strains, whereas from patients hospitalized in conservative wards most often C. albicans (6) strains were isolated. Candida strains were mostly susceptible to antifungal agents tested. It was interesting to culture Trichosporon beigelii (T. cutaneum) strain as an etiological agent of fungemia. This strain was multidrug-resistant.     

Enterobacter soli sp. nov.: A Lignin-Degrading ��-Proteobacteria Isolated from Soil

A Gram-negative bacterium that formed cream-colored colonies designated strain LF7 was isolated from soil collected in the Tambopata National Reserve in Madre de Dios, Peru. 16S rRNA sequence comparisons indicate that LF7 is a novel Enterobacter sp. closely related to E. asburiae JCM 6051(T) [AB004744] and E. aerogenes JCM 1235(T) [AB004750] based on their sequence homologies (p-distance: 1.06 and 1.19%, respectively). DNA G + C content was 52.8 mol% which is within the range reported for E. asburiae (55-57 mol%). The major cellular fatty acids present in the LF7 strain were C(16:0) (27.3%), C(16:1) ω7c and/or C(16:1) ω6c (16.3%), C(18:1) ω7c (16.1%), C(17:0) cyclo (12.4%), C(14:0) 3-OH and/or C(16:1) iso-I (8.9%), C(14:0) (7.6%), C(12:0) (3.9%), C(17:0) (2.4%), C(13:0) 3-OH and/or C(15:1) iso-H (1.7%), C(13:0) (1.1%), and C(18:2) ω6,9c and/or C(18:0) ante (0.5%). The cellular fatty acid profile, G + C content, phenotypic and biochemical characteristics were consistent with its placement in the genus Enterobacter. The name Enterobacter soli is proposed for this bacterium.     

Genetic population structure of Norwegian brown trout

Biochemical genetic variation in populations of anadromous and resident brown trout, Salmo trutta L., was studied. Altogether 50 Norwegian populations were screened for 32 enzyme loci. Genetic polymorphism was found at the following 11 loci: AAT-4 * (E.C. 2.6.1.1), CK-1 * (E.C. 2.7.3.2), G3PDH-2 * (E.C. 1.1.1.8), IDHP-2 * (E.C. 1.1.1.42), LDH-5 * (E.C. 1.1.1.27), MDH-2 * (E.C. 1.1.1.37), MDH-3/4 * (E.C. 1.1.1.37), MEP-2 * (E.C. 1.1.1.40), GPI-2 * (E.C. 5.3.1.9). GPI-5 * (E.C. 5.3.1.9) and PGM-1 * (E.C. 5.4.2.2), giving an overall polymorphism of 34%, ranging from 3.7 to 29.6% among individual populations. The average calculated heterozygosity ranged from 1.4 to 10.2% among populations. Genetic heterogeneity was observed among anadromous populations, and significant differences in allelic frequencies were found between anadromous populations in neighbouring watercourses, among resident populations and between anadromous and resident populations inhabiting the same watercourses. Significant heterogeneity was also found among 12 populations from Lake Mjøsa, with a major division between the western and eastern populations of the lake. Differences in allelic frequencies were found between wild stocks and their hatchery derivatives, and between different hatchery derivatives originating from the same wild population. In some cases release of hatchery populations into wild stocks may have influenced the genetic characteristics of wild stocks. The data support the hypothesis of eastern as well as western postglacial colonization lines for Norwegian brown trout.     

Application of solid surface fluorescence EEM spectroscopy for tracking organic matter quality of native halophyte and furrow-irrigated soils

Solid surface fluorescence excitation-emission matrix (EEM) is developed a potential method to characterize soil organic matter (SOM). Solid surface EEM spectroscopy with parallel factor analysis (PARAFAC) and hierarchical cluster analysis (HCA) is used to extract fluorescent components, to seek latent factors, and to investigate spatial distribution of SOM. Soil samples were collected from four native halophyte and two furrow-irrigated soil profiles, i.e. Comm. Salicornia europaea (CSE), Comm. Suaeda glauca (CSG), Comm. Kalidium cuspidatum (CKC), Comm. Sophora alopecuroides (CSA), corn fields (CFD), and wheat fields (WFD). SOM contained six fluorescent components: microbial/terrestrial fulvic-like fluorescent components (C1), tryptophan-like/lignin-derived phenol fluorescent components (C2), terrestrial humic-like fluorescent component (C3), lignin oxidative degradation by-products (C4 and C5), and amino acids (C6). The C 4 and C5 were the representative components of SOM within the CSE, CSG, CKC, CSA and CFD soil profiles, while the C2 and C6 were dominated within the WFD soil profile. The C4, C5, C1 and C2 were latent factors, and they could roughly distinguish SOM within the whole saline soil profiles except the CFD. A humification index (H/L) deduced from the fluorescent components, was very suitable to indicate humification levels of SOM. Humification levels of SOM within the halophyte soil profiles decreased with soil depth, but the opposite trends within the furrow-irrigated soil profiles. The H/L was closely correlated with exchangeable sodium percentage (ESP), and humification levels increased with the decreasing ESP. Soil surface EEM may not only indicate organic matter fractions of saline soils, but may be transferred to other types of landscape.     

Inhibition of phospholipases by Met-Leu-Phe-Ile-Leu-Ile-Lys-Arg-Ser-Arg-His-Phe, C terminus of middle-sized tumor antigen

The N and C terminals and tyrosine-phosphorylating site of the middle-sized tumor antigen of polyoma virus were chemically synthesized. The sequences of these peptides were Met-Asp-Arg-Val-Leu-Ser-Arg-Ala-Asp-Lys (N-MT), Met-Leu-Phe-Ile-Leu-Ile-Lys-Arg-Ser-Arg-His-Phe (C-MT), and Glu-Glu-Glu-Glu-Tyr-Met-Pro-Met-Glu (MT-Tyr), respectively. Among these peptides, the C-MT peptide inhibited phospholipase A2 (EC 3.1.1.4), phospholipase C (EC 3.1.4.3), and phospholipase D (EC 3.1.4.4). In addition, phosphatidylinositol-specific phospholipase C (EC 3.1.4.10) was also inhibited by this peptide. To study the mechanism of the inhibition, kinetic analysis was performed using phospholipase A2 from porcine pancreas. The degree of inhibition of phospholipase was dose dependent, and maximal inhibition was observed at pH 8.8. This peptide inhibited phospholipase A2 in a competitive manner for low-affinity sites of Ca2+, and in a noncompetitive manner for phospholipid substrates. When a fatty acid in the 2 position of the glycerol moiety of phosphatidylcholine was replaced by palmitic acid (C16:0), oleic acid (C18:1), linoleic acid (C18:2), eicosatrienoic acid (C20:3), or arachidonic acid (C20:4), the degree of inhibition of phosphatidylcholine hydrolysis by the C-MT peptide decreased. Inhibition of phospholipase A2 by the C-MT peptide was reversed by low concentrations of sodium deoxycholate but not by Triton X-100 or Nonidet P40, nonionic detergents. These detergents and the modification of acyl groups altered the micellar state of phospholipids. These results, taken together, suggest that the binding of the C-MT peptide near the low-affinity Ca2+ binding sites modifies the interaction of phospholipid substrates with the active center of phospholipase A2.     

Absorption and metabolism of cyanidin 3-O-beta-D-glucoside in rats.

We have clarified for the first time how cyanidin 3-O-beta-D-glucoside (C3G), which is a potent antioxidant anthocyanin, is absorbed and metabolized in vivo. Rats were orally administered C3G (0.9 mmol/kg body weight), and C3G rapidly appeared in the plasma. However, the aglycon of C3G (cyanidin; Cy) was not detected, although it was present in the jejunum. Protocatechuic acid (PC), which may be produced by degradation of Cy, was present in the plasma and the concentration was 8-fold higher than that of C3G. These results suggest that plasma PC and C3G may contribute to the antioxidant activity of the plasma. In the liver and kidney, C3G was metabolized to methylated C3G (methyl-C3G), suggesting that C3G and/or methyl-C3G act as antioxidants in the tissues.