Combined high pressure and temperature induced lethal and sublethal injury of Lactococcus lactis--application of multivariate statistical analysis

It was the aim of this work to determine the combined effects of pressure, temperature, and co-solutes on Lactococcus lactis, and to detect correlations between culture-dependent and culture-independent methods for assessment of cellular viability and sublethal injury. Therefore, the pressure induced inactivation of L. lactis MG 1363 was investigated in buffer and in buffer with 1.5 M sucrose or 4 M NaCl at a pressure range of 0.1 to 500 MPa and a temperature range of 5 to 50 degrees C. The inactivation was characterised by viable cell counts, stress resistant cell counts, membrane integrity, metabolic activity, and the activity of the multi-drug-resistance transport enzyme LmrP. L. lactis was most resistant to pressure application at 20-30 degrees C. Sucrose protected towards inactivation at any temperature, NaCl provided protection at high temperatures only. By using Principal Component Analysis, correlations were detected between viable cell counts and metabolic activity as well as stress resistant cell counts and LmrP activity. In conclusion, the pressure-inactivation of L. lactis is strongly temperature dependent, baroprotection by sucrose occurs at any temperature but the baroprotective effects of NaCl is temperature dependent. Further on, a combination of two experimental methods fully describe lethal and sublethal injury of pressure treated cells. These simplification of data acquisition and model development facilitates the establishment of pressure processes in food technology.     

Surface-plasmon microscopy with a two-piece solid immersion lens: bright and dark fields

We report bright-field and dark-field surface-plasmon imaging using a modified solid immersion lens and a commercial objective of moderate NA in the epi configuration. The contrast and resolution are extremely good, giving well-resolved images of protein monolayers both in air and in water. We also describe a two-part solid immersion lens that allows the sample to be moved without degrading the image quality in any observable way. The merits of the two-part lens are discussed and compared to commercially available microscope objectives. Finally, we introduce a simple Green's function model that illustrates the key features of both bright-field and dark-field surface-plasmon imaging.     

Semi-parametric estimation in the compositional modeling of multicomponent systems from Raman spectroscopic data

Identification and quantification of molecular species are central applications of molecular spectroscopy. In complex multicomponent systems like tissue samples, linear parametric models are often used to estimate the relative concentrations of the biochemical components of the sample. In situations where not all of the components of the sample are known or modeled, such parametric models can suffer from omitted variable bias and result in skewed estimates of component concentrations. We propose a semi-parametric approach that tries to avoid this omitted variable bias by effectively including unknown covariates as a non-parametric term in the regression equation. Constituent concentrations estimated with such partial linear models should outperform strict parametric linear models when the user has limited information on the composition of a multi-constituent system.     

Supplementation with extruded linseed cake affects concentrations of conjugated linoleic acid and vaccenic acid in goat milk

The aim of this research was to determine the effect of adding extruded linseed cake to the dry diet of goats on the concentrations of conjugated linoleic acid (CLA) and vaccenic acid (VA) in milk fat. Thirty crossbreed dairy goats were divided into 3 groups. Their diet was supplemented with 0% (control group), 5% (low group), or 10% (high group) of extruded linseed cake (ELC), which supplied 0, 16, and 32 g/d of linseed fat, respectively. The milk fat percentage (overall mean 3.5%) and yield did not differ with the different diets, but fatty acid composition was affected by the ELC supplements. The inclusion of ELC in the diets did not influence the concentration of fatty acids from C6:0 to C12:0. The concentrations of C14:0 and C16:0 decreased as the quantity of ELC supplements increased. The concentrations (mg/100 mg of total fatty acid methyl esters) of VA (0.70, 1.23, and 1.39 in control, low, and high groups respectively) and cis-9,trans-11 CLA (0.63, 0.96, and 1.05 in control, low, and high groups, respectively) were increased by ELC supplements. The milk fat content of VA and cis- 9,trans-11 CLA were closely correlated (R² = 0.82). Desaturation of VA in the mammary gland to produce cis-9,trans-11 CLA was higher in the control group than in the groups with ELC diets. Extruded linseed cake supplementation to lactating goats may enhance the nutritional profile of milk lipids.     

Effect of irradiation of frozen meat/fat trimmings on microbiological and physicochemical quality attributes of dry fermented sausages

Lipid oxidation is a major factor in meat quality. In order to relate human perceptions of lipid oxidation, as determined by a trained taste panel, to a chemical measurement of oxidation, we studied meat from animals with a wide range of potential oxidation through differences in their PUFA composition and by displaying the meat in high oxygen modified atmosphere packs for varying lengths of time. Meat was obtained from 73 Angus- and Charolais-cross steers from different trials that had been raised on 10 different diets: grass silage (high in C18:3, n-3), cereal concentrate (high in C18:2, n-6), three diets with 3% added fat consisting of three levels of protected lipid supplement (high in C18:2, n-6 and C18:3, n-3, ratio 1:1), a control with Megalac^® (relatively saturated), three diets with three levels of inclusion of protected fish oil (high in C20:5 n-3 and C22:6 n-3) plus a constant amount of unprotected fish oil and a final diet with an unprotected fish oil control. The longissimus dorsi muscle was excised from the left carcass side, aged vacuum packaged for 10–13 days depending on the projects and frozen for less than eight months. TBARS and sensory analyses were performed on steaks displayed for 0, 4 or 9 days under simulated retail conditions, exposed to light in modified atmosphere packaging (CO₂:O₂; 25:75). Meat oxidation increased throughout display for each of the diets, as shown by a rise in TBARS values. This increase was not linear, differences between 0 and 4 days of display were smaller than between 4 and 9 days of display. The lowest TBARS and lowest increment occurred in the two control diets and the grass-fed animals, probably due to the more saturated fat of meat from animals fed the control diets and the higher content of vitamin E. Sensory attributes were also influenced by time of display. Positive attributes, such as beef flavour or overall liking, decreased throughout display, whereas negative attributes, such as abnormal and rancid flavours, increased.     

Calcium-free vitrification reduces cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes

Despite the success of embryo cyropreservation, routine oocyte freezing has proved elusive with only around 200 children born since the first reported birth in 1986. The reason for the poor efficiency is unclear, but evidence of zona pellucida hardening following oocyte freezing indicates that current protocols affect oocyte physiology. Here we report that two cryoprotectants commonly used in vitrification procedures, dimethyl sulfoxide (DMSO) and ethylene glycol, cause a large transient increase in intracellular calcium concentration in mouse metaphase II (MII) oocytes comparable to the initial increase triggered at fertilization. Removal of extracellular calcium from the medium failed to affect the response exacted by DMSO challenge, but significantly reduced the ethylene glycol-induced calcium increase. These results suggest that the source of the DMSO-induced calcium increase is solely from the internal calcium pool, as opposed to ethylene glycol that causes an influx of calcium across the plasma membrane from the external medium. By carrying out vitrification in calcium-free media, it was found that zona hardening is significantly reduced and subsequent fertilization and development to the two-cell stage significantly increased. Furthermore, such calcium-free treatment appears not to affect the embryo adversely, as shown by development rates to the blastocyst stage and cell number/allocation. Since zona hardening is one of the early activation events normally triggered by the sperm-induced calcium increases observed at fertilization, it is possible that other processes are negatively affected by the calcium rise caused by cryoprotectants used during oocyte freezing, which might explain the current poor efficiency of this technique.     

Online coupling of digital microfluidic devices with mass spectrometry detection using an eductor with electrospray ionization

MS detection coupled with digital microfluidic (DMF) devices has most commonly been demonstrated in an offline manner using matrix assisted laser desorption ionization. In this work, an eductor is demonstrated which facilitated online coupling of DMF with electrospray ionization MS detection. The eductor consisted of a transfer capillary, a standard ESI needle, and a tapered gas nozzle. As a pulse of N(2) was applied to the nozzle, a pressure differential was induced at the outlet of the ESI needle that pulled droplets from the DMF, past the ESI needle, and into the flow of gas exiting the nozzle, allowing detection by MS. Operating position, ionization potential, and N(2) pressure were optimized, with the optimum ionization potential and N(2) pressure found to be 3206 V and 80 psi, respectively. Online MS detection was demonstrated from both open and closed DMF devices using 2.5 μL and 630 nL aqueous droplets, respectively. Relative quantitation by DMF-MS was demonstrated by mixing droplets of caffeine with droplets of theophylline on an open DMF device and comparing the peak area ratio obtained to an on-chip generated calibration curve. This eductor-based method for transferring droplets has the potential for rapid, versatile, and high-throughput microfluidic analyses.     

Nanopatterned electrically conductive films of semiconductor nanocrystals

We present the first semiconductor nanocrystal films of nanoscale dimensions that are electrically conductive and crack-free. These films make it possible to study the electrical properties intrinsic to the nanocrystals unimpeded by defects such as cracking and clustering that typically exist in larger-scale films. We find that the electrical conductivity of the nanoscale films is 180 times higher than that of drop-cast, microscopic films made of the same type of nanocrystal. Our technique for forming the nanoscale films is based on electron-beam lithography and a lift-off process. The patterns have dimensions as small as 30 nm and are positioned on a surface with 30 nm precision. The method is flexible in the choice of nanocrystal core-shell materials and ligands. We demonstrate patterns with PbS, PbSe, and CdSe cores and Zn(0.5)Cd(0.5)Se-Zn(0.5)Cd(0.5)S core-shell nanocrystals with a variety of ligands. We achieve unprecedented versatility in integrating semiconductor nanocrystal films into device structures both for studying the intrinsic electrical properties of the nanocrystals and for nanoscale optoelectronic applications.