Transcript abundance of the pig stearoyl-CoA desaturase gene has no effect on fatty acid composition in muscle and fat tissues, but its polymorphism within the putative microRNA target site is associated with daily body weight gain and feed conversion ratio.

Fatty acid composition in porcine intramuscular fat affects the dietetic value and technological properties of meat. The stearoyl-CoA desaturase (SCD) gene is a strong positional and functional candidate for fatty acid composition. Our sequence analysis in 4 breeds (Duroc, Pietrain, Polish Landrace, and Polish Large White) revealed a novel SNP in the 5'-flanking sequence and 9 novel SNP and 2 novel indels in the 3' untranslated region (UTR). Transcript level of the SCD in subcutaneous fat was significantly greater than in muscle tissue (n=83; P<0.001) and the interbreed comparison revealed a greater transcript level in the fat tissue of Polish Landrace (P<0.01). We found no association between the abundance of the SCD transcript and fatty acid composition in any of the tissues. We performed an association analysis between 4 SNP (c.-353C>T, c.-233T>C, c.*164A>G, and c.*928G>C), 1 indel (c.*2574_2576delGTC), and production traits in Polish Large White (n=185) and synthetic line 990 (n=243). The most pronounced associations were observed for the c.*928G>C polymorphism, which occurs within a predicted target site for 2 microRNA (ssc-miR-185 and ssc-miR-491). In line 990, this polymorphism was significantly associated with daily BW gain (P<0.04 under the general model) and feed conversion ratio (P<0.0004) but not with fatness traits. The same tendency, but not significant, was observed in the Polish Large White breed. When both breeds were analyzed together, these associations were again highly significant (daily BW gain P<0.003; feed conversion ratio P<0.0001). We conclude that c.*928G>C is a promising marker for both porcine traits.

Apolipoprotein E4 allele is associated with substantial changes in the plasma lipids and hyaluronic acid content in patients with nonalcoholic fatty liver disease.

Fat may affect progression of liver damage in patients with non-alcoholic fatty liver disease (NAFLD). In this study we characterize the state of lipid metabolism in 22 patients with NAFLD and different Apo-E variants. Total concentration of plasma total fatty acids was quantified by gas chromatography, while their derivatives by liquid chromatography/tandem mass spectrometry (LC ESI MS/MS). The ratio of plasma saturated fatty acid to monounsaturated fatty acid increased, whereas the ratio of polyunsaturated fatty acids to saturated fatty acids was reduced in Apo-E4 carriers. Simultaneously, the levels of individual plasma linoleic, arachidonic, and alpha linolenic acids significantly increased in subjects with the Apo-E4 allele. The 15-lipoxygenase metabolite, 13-hydroxyoctadecadienoic acid, was significantly higher in Apo-E3 carriers (p<0.006). 5-oxo-6,8,11,14-eicosatetraenoic acid was significantly elevated in Apo-E4 carriers (p<0.009). A significant difference in hyaluronic acid concentration (p<0.0016) as well as predicted advanced fibrosis (using the BARD scoring system) was found in Apo-E4 carriers (p<0.01). We suggest that a distinct mechanism of fibrosis between Apo E alleles. In Apo-E4 carriers, an elevation in 5-oxo-6,8,11,14-eicosatetraenoic acid synthesis and fatty acid dysfunction may induce fibrosis, while an inflammatory process may be the main cause of fibrosis in Apo-E3 carriers.

[Interest of modern imagery for conservative management of a placenta percreta]. / Prise en charge conservatrice d'un placenta percreta. Apport des nouvelles techniques d'imagerie.

The placenta percreta is a rare form of anomaly of placental insertion threatening the maternal and foetal lives. The incidence of abnormal placental adhesion is correlated to frequency of caesarean sections and advanced maternal age. Patients who are at high risk should be identified during pregnancy by ultrasound examination with Color Doppler looking for characteristic features. The practice of MRI in case of echographic suspicion may be useful, particularly in posterior placentas. A late diagnosis, in an emergency context, leads generally to hysterectomy and even to partial resection of neighbour organs. Thus, we report the case of a conservative management associating uterine embolisation following an elective caesarean delivery at 36 weeks gestation. In postpartum, the placental involution was followed clinically and by imagery. In our case, a total abdominal hysterectomy was performed on the fifth postoperative week because of a severe antibiotics resistant infection. No blood transfusion was required and the postoperative period was uneventful. This clinical case aims to show, through an analysis of the recent data of the literature, the interest of modern imagery to select patients with suspected placenta percreta that would be suitable candidates for conservative management.

Main compounds responsible for off-odour of strawberries infected by Phytophthora cactorum.

AIMS: Volatile compounds present in strawberries infected with Phytophthora cactorum, especially those responsible for the characteristic off-odour of such fruits were the subject of this study. METHODS AND RESULTS: Six strawberry varieties (Redgauntlet, Selva, Korona, Tenira, Real, Pegasus) inoculated with P. cactorum strain (PC-5), isolated from naturally infected fruit and one variety inoculated with 15 strains of P. cactorum in the laboratory were analysed. All the samples had a distinct, to a various degree, off-odour reminiscent of watercolour paint with phenolic notes. Volatile compounds were isolated by solid phase microextraction and simultaneous distillation extraction methods. To detect compounds responsible for the characteristic off-odour, gas chromatography-olfactometry was used. Two compounds were found to be responsible for the characteristic off-odour of strawberries infected by P. cactorum: 4-ethyl phenol and 4-ethyl-2-metoxy phenol (4-ethyl guaiacol). The content of these compounds in infected varieties ranged from 1.12 to 22.56 mg kg(-1) and 0.14-1.05 mg kg(-1) respectively. Other volatile compounds, not detected in noninoculated sound strawberries, were also identified: camphene, 1-octene-3-ol, 3-octanone, o-cymene, phenyl methanol, cis-linaloloxide, nonanal, phenyl ethyl alcohol, 2-undecanone and alpha-muurolene. SIGNIFICANCE AND IMPACT OF THE STUDY: Volatile compounds responsible for the characteristic off-odour of strawberries infected with P. cactorum were identified. Also compounds produced as a result of P. cactorum growth on strawberry fruit were characterized.

Use of solid phase microextraction (SPME) for profiling fungal volatile metabolites.

AIMS: The influence of isolation methods: solid phase microextraction (SPME) with different fibres and simultaneous distillation extraction (SDE) on the profile of isolated fungal volatile metabolites was investigated. METHODS AND RESULTS: Four SPME fibre types: Polydimethylsiloxane, Polyacrylate, Carboxen/PDMS and Carboxen/Divinylbenzene/PDMS were evaluated in terms of their efficiency in extracting volatile metabolites emitted by Penicillium roqueforti grown on wheat kernel medium. All fibres showed varied efficiency and selectivity in extracting volatile compounds. Sesquiterpene hydrocarbons were the predominant fraction of volatile compounds isolated by all fibres, and ranged from 55.4 to 93.7% of all volatiles depending on the type of fibre used. Alcohols and ketones ranged from 2.7 to 20.5%, esters from 1.2 to 12.8%, and monoterpene hydrocarbons from 1.2 to 5.4%. Profile of volatile compounds obtained by SDE differed from SPME and the oxygenated sesquiterpenes formed the predominant fraction of volatiles isolated using SDE. SIGNIFICANCE AND IMPACT OF THE STUDY: The data in this study show that analysed profile of volatile compounds emitted by fungi is highly dependent on the extraction method.

Influence of octanoic acid addition to medium on some volatile compounds and PR-toxin biosynthesis by Penicillium roqueforti.

AIMS: The effect of wheat kernel medium supplementation with octanoic acid on the formation of PR toxin and some volatiles by Penicillium roqueforti was investigated. METHODS AND RESULTS: Octanoic acid was added to the medium once, prior to inoculation (4.55 mg g-1), or periodically (total 27.3 mg g-1) during the 10 day course of incubation. No octanoic acid was added to the reference sample. Levels of 2-heptanone, 2-heptanol and aristolochene, a volatile intermediate in PR toxin biosynthesis, were monitored using a solid phase microextraction (SPME) technique. The contents of PR toxin and ergosterol were determined after incubation. Aristolochene was observed in the reference sample, and 10.4 mg kg-1 of PR toxin was detected after 10 days. In cultures periodically supplemented with octanoic acid, no aristolochene or PR toxin were observed. However, in samples supplemented with octanoic acid only prior to incubation, the aristolochene level was 25% that in the reference sample, and PR toxin content was 3.4 mg kg-1. SIGNIFICANCE AND IMPACT OF THE STUDY: These data suggest that a high level of octanoic acid in the medium prevents PR toxin formation by P. roqueforti.

Derivatization of the mutagen MX (3-chloro-4(dichloromethyl)-5-hydroxy-2(5H)-furanone) with butyl alcohols prior to GC-MS analysis.

An extremely potent mutagen, 3-chloro-4(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) is commonly present in chlorinated drinking water. Due to its high mutagenic activity and according to WHO guidelines its concentration should be controlled in drinking waters. Determination of MX is difficult due to the low (ppt) levels at which the compound usually exists in drinking waters. Results obtained with butanols as MX derivatization agents are shown and derivatization with sec-butanol is presented as a method which significantly lowers GC/MS detection levels of MX.

Changes in the phospholipid composition of the arterial cell can result in severe atherosclerotic lesions.

The oxysterol concentration in the plasma and the phospholipid composition of vascular tissue obtained by coronary artery bypass grafting (CABG) were compared with plasma and vascular tissue from age and sex matched controls. The plasma from CABG patients had a higher concentration of oxysterols than was present in the controls. Human endothelial cells were cultured for 72 hours in a medium containing plasma obtained from CABG patients, from controls or from the same controls to which 5 oxysterols were added to make the total oxysterol level equivalent to that in the CABG plasma and then pulsed with calcium (45Ca(2+)) for one hr. A significantly higher influx of 45Ca(2+) was noted in the endothelial cells cultured in the plasma obtained from CABG patients and from the controls with 5 added oxysterols, but not in those cultured without added oxysterols indicating that oxysterols increased calcium influx into endothelial cells. A phospholipid analysis indicated that the arterial tissue from CABG patients had 48.2% sphingomyelin in its phospholipid fraction compared to 10% in arterial tissue from umbilical cords. The saphenous vein obtained during CABG surgery from the same patient had only 24% sphingomyelin in its phospholipid fraction and unlike the coronary arteries had no atherosclerotic lesions. The higher level of oxysterol in the plasma of patients suffering from severe atherosclerosis could increase the concentration of sphingomyelin in the arterial cell membrane and thereby increase calcium influx required for producing the calcific type VII lesions in the coronary arteries.

Application of sec-butanol to the derivatization of hydroxyfuranones.

This paper reports the use of sec-butanol for the derivatization of chlorinated hydroxyfuranones. The following hydroxyfuranones were investigated: 3,4-dichloro-5-hydroxy-2(5H)-furanone, 3,4-dibromo-5-hydroxy-2(5H)-furanone, 3-chloro-4-(chloromethyl)-5-hydroxy-2(5H)-furanone, and 3-chloro-4-methyl-5-hydroxy-2(5H)-furanone. Their derivatization products were analyzed by gas chromatography/mass spectrometry. The sec-butyl derivatives of the hydroxyfuranones investigated yield ions that are less abundant than those obtained for the corresponding isopropyl derivatives. However, sec-butyl derivatives are easily detectable in especially dirty matrixes because they produce double peaks in the chromatogram. The ion intensity of the first peak in the pair is lower than that of the second peak, but both are characterized by the same spectrum. The formation of multiple peaks is related to the formation of diastereoisomers during derivatization.

Headspace solid-phase microextraction use for the characterization of volatile compounds in vegetable oils of different sensory quality.

Headspace solid-phase microextraction (HS-SPME) was used to isolate the volatile compounds, which are formed during peroxidation of fatty acids in vegetable oils. Isolated compounds were characterized by GC-MS and quantified using GC with FID detection. Four fibers for HS-SPME method development were tested, and the divinylbenzene/carboxene/PDMS fiber was selected as providing the best detection of analyzed compounds. Extraction curves, limits of detection, repeatability, and linearity were investigated for 14 aldehydes, ketones, hydrocarbons, and alcohols being products of fatty acids autoxidation. Limits of detection for 11 of these were below 1 microg/L. For quantitative purposes, to minimize the influence of temperature on hydroperoxide formation and the changes in the volatiles profile of the extracts, sampling was performed at 20 degrees C. For compound characterization by GC-MS, sampling temperature of 50 degrees C was applied. The developed method was applied to the analysis of refined and cold-pressed rapeseed oil stored at 60 degrees C for 10 days, and for 10 different vegetable oils of various degree of peroxidation. All samples were subjected to sensory analysis. The results of PCA sensory analysis were related to the amount of volatile compounds isolated by SPME method. In cases where the amount of compounds was highest, the samples were perceived as the worst, whereas those with low levels of volatile compounds were the most desired ones according to sensory evaluation. The relation was observed for both total volatiles, quantified C5-C9 aldehydes, and 14 compounds selected in method development. SPME revealed to be a rapid and sensitive method for the extraction and quantitation of trace volatile compounds from plant oils even at ambient temperature.

Determination of cholesterol oxidation products in milk powder and infant formulas by gas chromatography and mass spectrometry.

In this paper a method for the cholesterol oxidation products (oxysterols) determination in milk powder and infant formulas has been presented. In the sample preparation step lipids transesterification has been performed. The recoveries of oxysterols have been determined and ranged from 94.2% to 99.9% for all but 20a-hydroxy cholesterol (74.2%). Detection limits were 0.018-0.034 ppm and the relative standard deviations (RSD) values were 4.6%-18.3%. The method has been utilized for the determination of oxysterols in milk-based infant formulas and milk powder available on the market. The concentration of oxysterols was between 0.04 and 4.20 ppm of a lipid extract fraction.

Natural contamination of spring barley with group A trichothecene mycotoxins in south-eastern Poland.

Strains (10705) of microscopic fungi were isolated from spring barley heads in the region of Lublin (south-eastern Poland). Fusarium sporotrichioides Sherb was found in 418 (3.9%) of isolated strains. Group A trichothecene mycotoxins were detected in the collected barley kernels colonized by F. sporotrichioides, with Fusarium head blight symptoms. Among 24 samples analysed, 12 were T-2 toxin positive in a range of contamination from 0.02 to 2.40 micrograms/g (average 0.45), while in five samples HT-2 toxin ranged from 0.01 to 0.37 micrograms/g (average 0.23) and T-2 tetraol was detected in two samples in a range of 0.01-0.21 micrograms/g (average 0.11). Twelve samples were free of detectable amounts of the toxic metabolites analysed.

New derivatization method for determination of 3-chloro-4(dichloromethyl)-5-hydroxy-2(5H)-furanone in water.

An extremely potent mutagen, 3-chloro-4(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) is commonly present in chlorinated drinking water. Due to its high mutagenic activity and according to World Health Organization guidelines its concentration should be controlled in drinking waters. Determination of MX is difficult due to ppt levels at which the compound usually exists in drinking waters. Derivatization of MX with 2-propanol is presented as a method which significantly lowers the GC-MS detection level compared to other alcohol derivatization agents.

Production of volatile sesquiterpenes by Fusarium sambucinum strains with different abilities to synthesize trichothecenes.

Twenty-five strains of Fusarium sambucinum grown on wheat kernels were examined for trichothecene production and the synthesis of volatile sesquiterpenes. The volatiles were purged with air and collected on Tenax traps. Adsorbed compounds were eluted from the traps and injected into a gas chromatograph coupled with a mass spectrometer. Ten strains isolated from potato tubers produced high amounts of diacetoxyscirpenol and its derivatives. These strains were characterized by the production of high amounts of diverse sesquiterpenes. In 10 cultures, 19 compounds were detected, of which 6 were predominant and composed as much as 82% of the volatile sesquiterpene fraction (e.g., beta-farnesene, beta-chamigrene, beta-bisabolene, alpha-farnesene, trichodiene, and an unidentified compound). Fifteen strains isolated from various sources that did not produce trichothecenes produced much less volatile sesquiterpenes, with less chemical diversity. No more than six compounds were present in cultures. Two of these compounds were present in the toxigenic strains isolated from potatoes (beta-farnesene and acoradiene), but four were unique to the strains not producing trichothecenes (longifolene, isocaryophyllene, delta-elemene, and an unidentified one). The pattern of volatile sesquiterpenes was characteristic and distinctive for both toxic and nontoxic strains.

[Purification and characterization of cAMP-dependent protein kinases of yeasts in a Saccharomyces cerevisiae wild strain and selected mutants of cAMP metabolism]. / Reinigung und Charakterisierung von cAMP-abhängigen Proteinkinasen bei Hefen in einem Saccharomyces cerevisiae Wildstamm und ausgewählten Mutanten des cAMP-Stoffwechsels.

Protein kinases represent a diverse family of enzymes that play a critical role in regulation. Among nearly 100 known protein kinases, the cAMP-dependent enzyme is best understood biochemically. Unlike other protein kinases, cAMP-dependent protein kinase consists of two different types of subunits that dissociate, a regulatory subunit (R), which is the receptor for cAMP, and a catalytic subunit (C). In the absence of cAMP, the enzyme exists as an inactive tetramer, R2C2. The binding of intracellular cAMP to the R subunit decreases the affinity of the R subunit for the C subunit by approximately four orders of magnitude and, under physiological conditions, leads to dissociation of the holoenzyme into R2(cAMP)4 dimer and two free C subunits that are catalytically active. Mutants of the cAMP metabolism, adenylate cyclase and cell cycle mutants, provided further information about protein synthesis and cellular growth in Saccharomyces cerevisiae. The purified protein kinases were divided into different types according to their elution profiles from the DEAE-cellulose matrix. Two types of cAMP-dependent and two types of cAMP-independent protein kinases were isolated from the wild strain. Differences in the activities of the kinases in the mutants showed a close relationship to the locus of the respective mutations in the cell-cycle. Some properties of the protein kinases are discussed with respect to individual mutations.

Improved methods of zearalenone and moniliformin preparation.

Modified procedures of zearalenone and moniliformin preparation, using solid substrate (rice or corn kernels) has been developed. Preliminary purification of toxins by 1iquid-liquid partition was applied, followed by column chromatography on silicagel and charcoal. Final yield was about lg from 1kg of dry cultures of crystalline zearalenone and liophylized moniliformin o high purity.

Biosynthesis and preparation of T-2 Toxin, HT-2 Toxin and Neosolantol.

Biosynthesis of trichothecenes by strains ofF. sporotriahioides KF 9 6 and KF 530 was performed on rice as a medium. Three toxins in significant amounts were produced, with yield: T-2 toxin 0.7g from 600g of drv culture, being the highest of the three metabolites, HT-2 toxin 0.06g an Neosolaniol 0.015g. Toxins were extracted with methanol from ground and defatted dry culture. Liquid/liquid partition, using chloroform/water saturated solution of sodium bicarbonate as a preliminary purification was applied. Silicagel and charcoal columns were used in further purification. Toxin were separated by preparative TLC and crystallized from methanol.