Putative association between a new polymorphism in exon 3 (Arg109Cys) of the pancreatic colipase gene and type 2 diabetes mellitus in two independent Caucasian study populations

The protein encoded by the pancreatic colipase (CLPS) gene is an essential cofactor needed by pancreatic triglyceride lipase (PNLIP) for efficient dietary lipid hydrolysis. Since the inhibition of lipase activity was shown to reduce the incidence of type 2 diabetes mellitus, we tested the hypothesis that genetic variations in the CLPS and PNLIP genes are associated with type 2 diabetes; 47 unrelated subjects were screened for polymorphisms of the CLPS and PNLIP genes. A nested-case control study of 192 incident type 2 diabetes subjects and 384 sex- and age-matched controls taken from the European Prospective Investigation into Cancer and Nutrition Potsdam Cohort (EPIC) was employed for association studies. The Metabolic Intervention Cohort Kiel (MICK) consisting of 716 males was used for verification. A novel putative functional polymorphism (Arg109Cys) was identified in the CLPS gene. The frequencies of the Arg/Cys genotype were 2.6% in EPIC and 2.2% in MICK study subjects. No homozygotes for the Cys/Cys genotype were found in either study population. Logistic regression analysis showed a statistically significant association of the Arg/Cys genotype with an increased risk of type 2 diabetes. The odds ratios estimated by the model were 3.75 (95%CI = 1.13-12.49, p = 0.03) in EPIC and 4.86 (95%CI = 1.13-20.95, p = 0.03) in MICK. No comparable associations were found with other traits of the insulin-resistance syndrome (e. g.; body mass index, waist to hip ratio). In conclusion, we obtained evidence in two German Caucasian study populations that the variant of the rare CLPS Arg109Cys polymorphism might contribute to increased susceptibility of type 2 diabetes.     

Rapid and Sensitive Detection of Staphylococcus aureus in Food Using Selective Enrichment and Real-Time PCR Targeting a New Gene Marker

Staphylococcus aureus is a bacterial pathogen considered a principal etiological agent of food poisoning. The aim of this study was to develop and evaluate a rapid and sensitive method for the detection of S. aureus in food by using selective enrichment and a new species-specific real-time polymerase chain reaction (PCR). Specific primers and a TaqMan probe targeted to specific S. aureus gene encoding for acriflavine resistance protein were designed. The real-time PCR was highly specific for S. aureus with 100% inclusivity and 100% exclusivity determined using 83 S. aureus strains and 64 non-S.-aureus strains. PCR detection limit of 6.8 × 10¹ and 3.4 × 10¹ CFU ml⁻¹ were obtained with 100% and 70% detection probability, respectively. The single selective enrichment based on the study of different enrichment conditions was selected and a lysis by boiling was used to obtain bacterial DNA. Out of 112 food samples analyzed, 61 were positive by the PCR-based method and 53 by the standard method. Out of ten food matrices artificially contaminated at a level of 10° CFU g⁻¹, ten and six were positive by the respective methods. Moreover, 10° CFU 10 g⁻¹ was detected in all ten artificially contaminated samples after a large-scale enrichment using PCR-based detection, in contrast to seven false negative by standard detection. The developed method facilitated the detection of S. aureus on the next day after the sample reception. This method can be used for S. aureus detection as a faster, highly specific, and more sensitive alternative to microbiological method with the potential for providing of improved food-processing hygiene control.     

Determination of biogenic amines in foods using ultra-performance liquid chromatography (UPLC)

A rapid ultra-performance liquid chromatographic (UPLC) method for the determination of biogenic amines (putrescine, cadaverine, spermidine, spermine, phenylethylamine, histamine, tyramine and tryptamine) in selected food samples is described. The eight biogenic amines, which are the most important to be determined in food samples, were derivatized with dansyl chloride prior to UPLC separation. The dansylated amines were separated on an Agilent Zorbax Eclipse XDB – C18 column (50 × 4.6 mm ID, 1.8 μm) using gradient elution with a binary system of acetonitrile–water, a flow rate of 1.0 ml/min and UV detection at 225 nm. The analysis is very fast, all amines are well resolved and are eluted from the column in less than 6 min. The average repeatability of the method ranged between 1.02% and 2.14%. Limits of detection (LODs) for considered amines ranged between 0.032 and 0.098 μg/l; calibration curves showed very good linearity (r = 0.9994–1.0000). The method was applied to the analysis of amines in pork, beef, chicken and fish meat, cheese and edible mushrooms.     

Effect of enterocin AS-48 in combination with high-intensity pulsed-electric field treatment against the spoilage bacterium Lactobacillus diolivorans in apple juice

Enterocin AS-48 was tested in apple juice against the cider-spoilage, exopolysaccharide-producing strain Lactobacillus diolivorans 29 in combination with high-intensity pulsed-electric field (HIPEF) treatment (35 kV/cm, 150 Hz, 4 μs and bipolar mode). A response surface methodology was applied to study the bactericidal effects of the combined treatment, with AS-48 concentration and HIPEF treatment time as process variables. At subinhibitory bacteriocin concentrations, microbial inactivation by the combined treatment increased as the bacteriocin concentration and the HIPEF treatment time increased (from 0.5 to 2.0 μg/ml and from 100 to 1000 μs, respectively). Highest inactivation (4.87 logs) was achieved by 1000 μs HIPEF treatment in combination with 2.0 μg/ml AS-48. While application of treatments separately did not protect juice from survivors during storage, survivors to the combined treatment were inactivated within the following 24 h of storage, and the treated samples remained free from detectable lactobacilli for at least 15 days at temperatures of 4 °C as well as 22 °C. The combined treatment could be useful for inactivation of exopolysaccharide-producing L. diolivorans in apple juice.     

Thermal denaturation of proteins in Post rigor muscle tissue as studied by differential scanning calorimetry

Post rigor bovine M. semimembranosus was analysed by differential scanning calorimetry (d.s.c.). After extractive removal of sarcoplasmic proteins, subsequent pH adjustment and manual connective tissue removal, d.s.c. yielded reproducible thermograms which permitted investigation of the individual major myofibrillar proteins in various pH and salt environments without prior isolation. The positions of two major peaks, interpreted as myosin transitions, proved to be strongly pH dependent. At pH 5.4, the peak maxima occurred at 58 and 65°C, respectively, at a heating rate of 10°C min-¹. Above pH 6.5 their order of denaturation was reversed. In the pH range 5.4–6.5 the peak ascribed to actin had its maximum near 80°C in intact muscle. Above this pH range it was displaced to lower temperature. The thermal stability of actin was studied after treatment of the muscle tissue with different salt solutions. At equal ionic strengths (μ = 0.15) at pH 5.5, calcium chloride and sodium chloride caused 6.5°C and 4°C displacement to lower temperature, respectively. The thermograms of bovine semimembranosus muscle were compared to those of two red and two white muscle types (bovine cardiac and rabbit soleus muscles, chicken breast and rabbit semimembranosus muscles, respectively) at two pH levels. Greater myosin differences were found between red and white muscles than between muscles from different animal species. All muscles gave similar actin transitions, with exception of the heart muscle where the actin peak appeared at 3 °C lower temperature. The necessity of a strict pH control in order to obtain reproducible muscle thermograms is demonstrated.     

Effect of cofermentation of grape varieties on aroma profiles of la mancha red wines

The effect of winemaking using blends of red grape varieties cultivated in La Mancha region (Spain) on the aroma profile of wines was researched by chemical characterization. Free and glycosidically bound aroma compounds were isolated by solid phase extraction using dichloromethane and ethyl acetate, respectively, as solvents in elution and then analyzed by gas chromatography-mass spectrometry. Free and bound volatile compounds were analyzed in Cencibel, Bobal, and Moravia Agria monovarietal wines, and in 3 wines obtained with the blending of grapes: Cencibel (50%) + Bobal (50%); Cencibel (50%) + Moravía Agria (50%); Cencibel (33%) + Bobal (33%) + Moravía Agria (33%). Aroma compounds were studied in terms of odor activity values (OAVs). Ninety free aroma compounds and sixty-five bound aroma compounds were identified and quantified. The odor activity values for the different compounds were classified into 7 odorant series. The fruity and sweet series contributed most strongly to the aroma profile of all wines, independently of the winemaking technique used. In general, co-winemaking wines present a more complex chemical profile than monovarietal wines. Practical Application: Some grape varieties could benefit from this process with the presence of other varieties that might have an excess of aroma compounds. In this study, the wines were elaborated by blending different grape varieties together; this process implies co-maceration and co-fermentation steps. The co-winemaking technique could benefit from additional molecules provided by the other varieties, which results in a more complex formation than in the case of monovarietal wines. This technique provides a viable alternative to traditional winemaking methods for improving and enhancing the sensory profile of elaborated wines.     

The Escherichia coli RecA protein complements recombination defective phenotype of the Saccharomyces cerevisiae rad52 mutant cells

The Saccharomyces cerevisiae rad52 mutants are sensitive to many DNA damaging agents, mainly to those that induce DNA double-strand breaks (DSBs). In the yeast, DSBs are repaired primarily by homologous recombination (HR). Since almost all HR events are significantly reduced in the rad52 mutant cells, the Rad52 protein is believed to be a key component of HR in S. cerevisiae. Similarly to the S. cerevisiae Rad52 protein, RecA is the main HR protein in Escherichia coli. To address the question of whether the E. coli RecA protein can rescue HR defective phenotype of the rad52 mutants of S. cerevisiae, the recA gene was introduced into the wild-type and rad52 mutant cells. Cell survival and DSBs induction and repair were studied in the RecA-expressing wild-type and rad52 mutant cells after exposure to ionizing radiation (IR) and methyl methanesulphonate (MMS). Here, we show that expression of the E. coli RecA protein partially complemented sensitivity and fully complemented DSB repair defect of the rad52 mutant cells after exposure to IR and MMS. We suggest that in the absence of Rad52, when all endogenous HR mechanisms are knocked out in S. cerevisiae, the heterologous E. coli RecA protein itself presumably takes over the broken DNA.     

Changes in selected biochemical and sensory parameters as affected by the superficial inoculation of Penicillium camemberti on dry fermented sausages

An atoxigenic strain of Penicillium camemberti was superficially inoculated on fermented sausages in an attempt to improve their sensory properties. The growth of this mould on the surface of the sausages resulted in an intense proteolysis and lipolysis, which caused an increase in the concentration of free amino acids, free fatty acids (FFA) and volatile compounds. Many of these were derived from amino acid catabolism and were responsible for the "ripened flavour", i.e. branched aldehydes and the corresponding alcohols, acids and esters. The development of the fungal mycelia on the surface of the sausages also protected lipids from oxidation, resulting in both lower 2-thiobarbituric acid (TBARS) values and lipid oxidation-derived compounds, such as aliphatic aldehydes and alcohols. The sensory analysis of superficially inoculated sausages showed clear improvements in odour and flavour and, as a consequence, in the overall quality of the sausages. Therefore, this strain is proposed as a potential starter culture for dry fermented sausage production.     

Variation in fatty acid composition of ewes' colostrum and mature milk fat

Temporal content variations of approximately 70 C4–C24 fatty acids (FA) in colostrum and milk of ewes consuming winter diet were determined by gas chromatography. The content of monounsaturated, polyunsaturated and straight-chain saturated FA 14:0–16:0 in colostrum was higher whereas the content of 4:0–12:0, 18:0 and branched-chain saturated FA 15:0–17:0 was lower than that in mature milk. The effect of ewe breed on the FA profiles was not significant. The composition of FA changed most significantly 1–2 days after lambing in agreement with the time schedule of colostrum formation. Nevertheless, the content of FA further successively changed up to approximately the 6th day of lactation, whereas further changes up to the 60th day of lactation were smaller. This is consistent with the recommendation that ewes' milk is suitable for human consumption after six-to-eight days of lactation. The higher contents of palmitic and myristic acids in colostrum compared with mature milk suggest that FA composition in colostrum matches the changing needs of the growing lambs.     

Daten amtlicher Lebensmittelkontrolle in Deutschland – Analyse und Erfassung der Unterschiede

Whether or not food control data reported according to former directive 89/397 EEC in Germany are comparable and whether or not recording or aggregation differences appear between the Länder should be proved within a project conducted by the BVL. The result of a data analysis of official control data was, that indicators like “the percentage of establishments with non-conformity reports” were not comparable between the Länder. It should be analysed, whether or not this is based on structural differences in the Länder and/or based on a matter of recording or aggregation differences. An analysis of these differences was conducted with the help of the Länder in several local authorities and upper authorities. The towns and authorities were chosen at random. Structured interviews were conducted with food control experts. It could be recognized that recording differences appeared for inspection data. These refer mainly to the deepness of data recording, the total number of recorded establishments, associated controls, and additional establishment information. Aggregation differences were mainly found for the sample data. Here, two main points are important: (1) which samples were actually aggregated; this is which sampling reasons were taken into account (population)? and (2) which samples were summarised in which product group. Less concise are differences in sample planning, the analysis spectrum for one sample, the assessment of analysis results and the consequence of the sample analysis outcome (advice, sanction). Since a statistical analysis of these food control data is difficult because of the mentioned differences in data recording and data aggregation, a harmonized usage of definitions for and recording as well as aggregation of these data is worthwhile. Not till then data could be the basis for a meaningful statistic and picture the success or failure of control activities.