Positional isomers of unsaturated fatty acids in rat liver lipids

The fatty acids of liver lipids from rats raised on a fat free diet from the 30th to the 90th day after birth were analyzed with special regard to the detection of positional isomers of mono-, di-, tri-, and tetraenoic fatty acids. The methyl esters obtained after transesterification of total lipids were separated by argentation chromatography into five fractions: I saturated, II monoenoic, III dienoic, IV dienoic nonmethylene interrupted, V triand tetraenoic fatty acid esters. After hydroxylation of the double bonds with osmium tetroxide, the analysis of the poly-O-trimethylsilyl derivatives by gas liquid chromatography on S.C.O.T. columns combined with mass spectrometry revealed the presence of 19 monoenoic, 15 dienoic, and 9 trienoic as well as 3 tetraenoic fatty acid isomers including the normally occurring representatives of the (n−3), (n−6), (n−7), and (n−9) fatty acid families. The majority of the identified isomers can be coordinated to one of these families like 7–16∶1; 11–20∶1; 6,9–18∶2; 8,11–20∶2; 5,11–20∶2; 5,8,11–20∶3; 7,10,13–22∶3 to the (n−9) family, 11–18∶1; 13–20∶1; 5,11–18∶2; 7,13–20∶2; 6,11–18∶2; 6,9–16∶2; 8, 11–18∶2; 10,13–20∶2; 5,8,11–18∶3; 7,10,13–20∶3; 4,7,10,13–20∶4 to the (n−7) family and 11,14–20∶2; 5,11,14–20∶3; 6,9,12–18∶3; 8,11,14–20∶3; 5,8,11,14–20∶4; 7,10,13,16–22∶4 to the (n−6) family. All these naturally occuring isomers can be placed into a network of desaturation and chain elongation steps which allows certain conclusions about the substrate specificity of the Δ6-, Δ5-and Δ4-desaturase systems. The great number of isomers found in the (n−7) family indicates that the members of this family are actively metabolized in partial essential fatty acid deficiency.     

II. Yeast sequencing reports. Sequence analysis of a 78·6 kb segment of the left end of Saccharomyces cerevisiae chromosome II

We report the sequence analysis of a 78,601 bp DNA segment on the left arm of chromosome II of Saccharomyces cerevisiae. This 78·6 kb segment spans the region from the start of a subtelomeric Y′ element up to the ILS1 gene. It contains 49 open reading frames (ORFs) with more than 100 amino acids length including 14 internal and five overlapping ORFs. The gene density, excluding the internal ORFs, was calculated as one ORF per 2·2 kb. Eight ORFs (PKC1, TyA, TyB, ATP1, ROX3, RPL17a, PET112 and ILS1) correspond to previously characterized genes. ORF YBL0718 was identified as CDC27; YBL0706 as TEL1. Four other ORFs show strong similarities to already known genes. The gene product of YBL0838 is 60% identical to the ribosomal protein RPL32 from rat, mouse and man. YBL0701 encodes a protein with significant similarity to the initiation factor eIF2 associated p67 glycoprotein from rat. Eight ORFs were disrupted and the resulting yeast strains analysed with respect to their phenotype. The sequence has been deposited in the EMBL Nucleotide Sequence Database under the Accession Number X79489.     

A two-dimensional high-performance thin-layer chromatographic screening method for sulphonamides in animal tissues

Summary A procedure for the identification of sulphonamides in edible animal tissues by two-dimensional high-performance thin-layer chromatography is described. Sixteen sulphonamides can be detected at an absolute level of 10 ng. The absolute detection limit of the sulphonamide standards is 1 ng. After extraction of the sulphonamides with chloroform/acetone, the acidified extract is concentrated and purified using a cation-exchange solid phase extraction column. The column is treated with ammonia vapour and the sulphonamides are then eluted with methanol. Blank samples of edible animal tissues spiked with sulphonamides (frequently used in Belgium) result in very good separation.

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Identifizierung von sulfonamiden in geweben von schlachttieren mittels zweidimensionaler hochleistungsdünnschichtchromatographie

Zusammenfassung Eine Methode zur Identifizierung von Sulfonamiden in Geweben von Schlachttieren mittels zweidimensionaler Hochleistungsdünnschichtchromatographie wird beschrieben. 16 Sulfonamide können identifiziert werden, wobei die absolute Empfindlichkeitsgrenze bei 10 ng liegt. Die absolute Nachweisgrenze für die Referenzsubstanzen liegt bei 1 ng. Nach Extraktion der Sulfonamide mit Chloroform-Aceton wird die angesäuerte Extraktlösung auf einer Kationenaustausch-Extraktionssäule eingeengt und gereinigt. Die Säule wird mit Ammoniakdämpfen behandelt und die Sulfonamide werden anschliessend mit Methanol eluiert. Die den Gewebeextrakten zugefügten in Belgien häufig verwendeten Sulfonamide konnten sehr gut voneinander getrennt werden.

     

Regeneration of anion exchange resins by calcium carbonate and carbon dioxide

Partial demineralization of water can be achieved by bicarbonate from anion exchange resins. The exchangers can be converted into bicarbonate form by carbon dioxide and calcium carbonate. The advantage of this method lies in these chemicals not being detrimental to health and not polluting the environment. The equilibrium of the CO₂ and CaCO₃ treatment has been investigated and compared with selectivity measurements. It is obvious from the results that the regeneration efficiency will be small but sufficient for technical purposes     

Effect of clofibric acid on the turnover of the fatty acid-binding protein identified in cultured endothelial cells from bovine aorta

Several types of fatty acid-binding proteins are found in mammalian cells. Cultured endothelial cells from bovine aorta were shown to contain exclusively the cardiac-type fatty acid-binding protein (cFABP) with a mean concentration of 90 ng cFABP/mg extract protein. Only small variations were observed from passage to passage. In pulse-chase labeling experiments with L-[³⁵S]methionine, a half-life of 4.0 d was measured for cFABP which is about two times longer than the average half-life of the extracted proteins. These data imply that in aortic endothelial cells cFABP is not subject to short-term regulation. However, addition of clofibric acid to the culture medium led to a shortening of the half-life of cFABP, which was compensated for by an increase in its biosynthesis. The turnover of the bulk of extract proteins remained unchanged when the cells were challenged with clofibric acid.     

Arbutin in marjoram and oregano

Arbutin is a hydroquinone derivative that has been found in species of several plant families. Within the genus Origanum the formation of arbutin is polymorphic, with arbutin present in considerable amounts (O. dubium 20.8 ± 15.3 mg/g; wild O. majorana 51.3 ± 15.4 mg/g, cultivated O. majorana 40.6 ± 11.2 mg/g), minor amounts (O. microphyllum 0.1 ± 0.1 mg/g, wild O. onites 0.3 ± 0.1 mg/g, cultivated O. onites 0.1 ± 0.1 mg/g, O. saccatum 0.1 ± 0.1 mg/g, O. solymicum 0.4 ± 1.0 mg/g) or completely absent (O. husnucan-baseri, O. syriacum, O. vulgare). Whereas the most important commercial oregano species (O. onites and O. vulgare) contain no or only minor amounts of arbutin, marjoram (O. majorana) has considerably high amounts. The high variability of arbutin in O. majorana would allow a selection into cultivars with high arbutin content and low arbutin varieties. In a segregating F₂-generation of a species crossing between O. majorana (high content of arbutin) and O. vulgare ssp. vulgare (free of arbutin), the presence of arbutin followed a Mendelian segregation of 3:1, indicating that only one gene is responsible for the polymorphism of arbutin in the genus Origanum. The absence of arbutin in O. vulgare ssp. vulgare or O. syriacum would even enable the breeding of marjoram with no arbutin at all.     

Domestic refrigeration practices with emphasis on hygiene: analysis of a survey and consumer recommendations

A Web-enabled survey was conducted to improve knowledge of home refrigeration practices of French consumers (n = 809), with an emphasis on hygiene, and this information was used to establish recommendations. The survey targeted a convenience sample of working people. Analysis of the survey responses revealed that efforts should be directed toward improvement of microbiological control measures. Only 37% of respondents made sure the temperature in their refrigerator was 4 degrees C or below. Only 37% of respondents reported that they systematically wrapped food. Sponges, known to be frequently highly contaminated, were used by 89% of the respondents to clean their refrigerator, which indicates the need to recommend disinfection of sponges before they are used for cleaning. Twenty-seven percent of respondents used sodium hypochlorite (bleach), but it was applied without previous cleaning (21% of the users) or in the commercial concentrated form (7% of the users). The permanent presence of water condensation on the shelves was noted by 2% of respondents, suggesting imperfect closure of the door, with a consequence of higher energy consumption and water available for microbial circulation and growth. Thus, an important recommendation is to check the door gaskets and to ensure the tight closure of the door. Seventy percent of the respondents declared that they never put warm or hot food in the refrigerator. However, many people, when orally questioned, acknowledged that they leave dishes at ambient temperature overnight before putting them in the refrigerator. It therefore is essential to recommend that perishable food not be left for more than 2 h at ambient temperature.     

Genetic characterization of commercial Saccharomyces cerevisiae isolates recovered from vineyard environments

One hundred isolates of the commercial Saccharomyces cerevisiae strain Zymaflore VL1 were recovered from spontaneous fermentations carried out with grapes collected from vineyards located close to wineries in the Vinho Verde wine region of Portugal. Isolates were differentiated based on their mitochondrial DNA restriction patterns and the evaluation of genetic polymorphisms was carried out by microsatellite analysis, interdelta sequence typing and pulsed-field gel electrophoresis (PFGE). Genetic patterns were compared to those obtained for 30 isolates of the original commercialized Zymaflore VL1 strain. Among the 100 recovered isolates we found a high percentage of chromosomal size variations, most evident for the smaller chromosomes III and VI. Complete loss of heterozygosity was observed for two isolates that had also lost chromosomal heteromorphism; their growth and fermentative capacity in a synthetic must medium was also affected. A considerably higher number of variant patterns for interdelta sequence amplifications was obtained for grape-derived strains compared to the original VL1 isolates. Our data show that the long-term presence of strain VL1 in natural grapevine environments induced genetic changes that can be detected using different fingerprinting methods. The observed genetic changes may reflect adaptive mechanisms to changed environmental conditions that yeast cells encounter during their existence in nature.