LF Samples Research Articles

Effect of selective fermentation on nutritional parameters and techno-functional characteristics of fermented millet-based probiotic dairy product

The primary goal of this study was to assess the effect of selective fermentation on the nutritional and techno-functional characteristics of fermented millet-skim milk-based product. The product was made with HHB-311 biofortified pearl millet (PM) flour, skim milk powder, and isolated cultures (either alone or in combination) of Limosilactobacillus fermentum MS005 (LF) and Lactobacillus rhamnosus GG 347 (LGG). To optimize fermentation time, time intervals 8, 16, and 24 h were explored, while the temperature was kept 37 °C. Results of protein digestibility showed that LF (16 h) and LGG (24 h) fermented samples had significantly higher (P < 0.05) protein digestibility of 90.75 ± 1.6% and 93.76 ± 3.4%, respectively, than that of control (62.60 ± 2.6%). Further, 16 h fermentation with LF showed enhanced iron (39%) and zinc (14%) bioavailability. The results suggested that LF with 16 h fermentation is most suitable for making millet-based fermented products with superior techno-functional attributes and micronutrient bioavailability.

Exposure to hull cleaning wastewater induces mortality through oxidative stress and cholinergic disturbance in the marine polychaete Perinereis aibuhitensis

While wastewater and paint particles discharged from the in-water cleaning process of ship hulls are consistently released into benthic ecosystems, their hazardous effects on non-target animals remain largely unclear. In this study, we provide evidence on acute harmful effects of hull cleaning wastewater in marine polychaete Perinereis aibuhitensis by analyzing physiological and biochemical parameters such as survival, burrowing activity, and oxidative status. Raw wastewater samples were collected during ship hull cleaning processes in the field. Two wastewater samples for the exposure experiment were prepared in the laboratory: 1) mechanically filtered in the in-water cleaning system (MF) and 2) additionally filtered with a 0.45 μm filter in the laboratory (LF). These wastewater samples contained high concentrations of metals (zinc and copper) and metal–based booster biocides (copper pyrithione and zinc pyrithione) compared to those analyzed in seawater. Polycheates were exposed to different concentrations of the two wastewater samples for 96 h. Higher mortality was observed in response to MF compared to LF–exposed polychaetes. Both wastewater samples dose-dependently decreased burrowing activity and AChE activity. Drastic oxidative stress was observed in response to the two wastewater samples. MDA levels were significantly increased by MF and LF samples. Significant GSH depletion was observed with MF exposure, while increased and decreased GSH contents were observed in LF-exposed polychaetes. Enzymatic activities of antioxidant components, catalase, superoxide dismutase, and glutathione S-transferase were significantly modulated by both wastewater samples. These results indicate that even filtered hull cleaning wastewater can have deleterious effects on the health status of polychaetes.

Research Note: Analysis of microbial diversity on the shell surface of eggs collected from geographically distinct farms in China

Micro-organisms on the eggshell surface of affect the quality of the egg. Sometimes, these microbes even pose a serious threat to the health of the egg's consumer. Bacterial 16S rDNA and fungal internal transcribed spacer region were sequenced to analyze the microbial diversity on the shell surface of the eggs collected from 4 distinct regions of China: Guyuan (GY; 1.5 million hens), Langfang (LF; 0.1 million hens), Beihai (BH; 1.2 million hens), and Dongguan (DG; 0.2 million hens). The results showed a higher bacterial and fungal abundance on the eggs collected from the northern and southern China farms, respectively. The dominant bacterial phylum detected across all egg samples was Firmicutes. In addition, the shell surfaces of the DG and LF samples harbored abundant levels of Proteobacteria. The dominant fungal phyla detected across all egg samples were Ascomycota and Basidiomycota. The bacterial compositions on eggshell surfaces differed significantly across all geographic regions, and the fungal composition differed significantly between samples collected from the southern and northern farms (P < 0.05). The abundance and composition of microbial colonies on the eggshell surface varied based on their geographical location (climate and environment) and farming scale (management). Our findings provide an important reference for optimizing the cleaning and disinfection methods for fresh eggs collected from different sources.

Pre-freezing selection of Holstein bull semen with the BoviPure colloid as double- or single-layer centrifugation improves the post-thawing quality

Artificial insemination (AI) is critical for breeding in the dairy industry. High-merit bulls can present low freezability, hampering genetic dissemination. Thawed semen can be improved using density gradient centrifugation (DGC) with colloids, but little information deals with the pre-freezing application. Thus, the BoviPure colloid (optimized for bull spermatozoa) was tested for pre-freezing application as the usual double-layer (DLC) versus single-layer (SLC, quick and economical). Semen from twelve Holstein-Friesian bulls was extended with OPTIXcell extender, frozen (Control), or processed by SLC or DLC and frozen. Sperm were assessed pre-freezing for motility and viability and post-thawing (directly and after 4 h 38 °C) for apoptosis, capacitation status, acrosomal damage, mitochondrial activity, cytoplasmic and mitochondrial reactive oxygen species (ROS), and chromatin status (SCSA for DNA fragmentation and chromatin compaction and monobromobimane, mBBr, for disulfide bridges evaluation). The DGC improved parameters post-thawing (e.g., 57.5%±10.1 motility vs. control 53.3% ± 11.2) at the cost of sperm loss (sperm recovery of DGC 14.4% ± 2.5 and SLC 17.4% ± 2.5). DNA fragmentation (%DFI) decreased (0.21% ± 0.53 vs. control 1.30% ± 0.10), and SLC reduced chromatin compaction. A clustering procedure separated lesser (LF) and greater freezability (GF) bulls. LF samples were especially benefited by DGC, with SLC providing better post-thawing results for this group. In conclusion, pre-freezing DGC improved sperm parameters post-thawing, potentially improving the cryopreservation of low-freezability semen from high-merit bulls. SLC, quicker and economical, would be preferable since it showed similar or higher performance than DLC.

The impact of freezing methods on the quality, moisture distribution, microstructure, and flavor profile of hand-grabbed mutton during long-term frozen storage

The aim of present study was to investigate the influence of conventional freezing (CF, −18 °C), low-temperaturefreezing (LF, −40 °C), and ultra-low-temperature freezing (ULF, −80 °C) on the quality, moisture distribution, microstructure, and flavor profile of hand-grabbed mutton (HGM) during frozen storage (0, 30, 60, 90, 120, 150 and 180 days). The TPC, TVB-N, and TBARS values increased significantly with prolonged storage, while the moisture content decreased (P < 0.05). Additionally, the concentrations of aldehydes, alcohols, ketones, acids, and alkenes decreased significantly as the storage duration increased. However, the concentrations of esters and heterocyclics increased (P < 0.05). Notably, at 30–180 days of storage, the TBARS and TVB-N values in ULF samples were significantly lower than those in CF and LF samples, while the moisture content was significantly higher (P < 0.05). Low field-nuclear magnetic resonance (LF-NMR) analysis showed that ULF decreased water migration and maintained the original texture characteristics of HGM during frozen storage. The ULF and LF groups had significantly higher levels of volatiles than the CF group (P < 0.05). The findings show that ULF, with its relatively rapid freezing rates, can still maintain the high quality of HGM after 180 days of frozen storage, contributing to quality control.

The Effects of Programmed Cell Death of Mesenchymal Stem Cells on the Development of Liver Fibrosis.

Mesenchymal stem cells have shown noticeable potential for unlimited self-renewal. They can differentiate into specific somatic cells, integrate into target tissues via cell-cell contact, paracrine effects, exosomes, and other processes and then regulate the target cells and tissues. Studies have demonstrated that transplantation of MSCs could decrease the expression and concentration of collagen in the liver, thereby reducing liver fibrosis. A growing body of evidence indicates that apoptotic MSCs could inhibit harmful immune responses and reduce inflammatory responses more effectively than viable MSCs. Accumulating evidence suggests that mitochondrial transfer from MSCs is a novel strategy for the regeneration of various damaged cells via the rescue of their respiratory activities. This study is aimed at reviewing the functions of MSCs and the related roles of the programmed cell death of MSCs, including autophagy, apoptosis, pyroptosis, and ferroptosis, as well as the regulatory pathogenic mechanisms of MSCs in liver fibrosis. Research has demonstrated that the miR-200B-3p gene is differentially expressed gene between LF and normal liver samples, and that the miR-200B-3p gene expression is positively correlated with the degree of liver fibrosis, suggesting that MSCs could inhibit liver fibrosis through pyroptosis. It was confirmed that circulating monocytes could deliver MSC-derived immunomodulatory molecules to different sites by phagocytosis of apoptotic MSCs, thereby achieving systemic immunosuppression. Accordingly, it was suggested that characterization of the programmed cell death-mediated immunomodulatory signaling pathways in MSCs should be a focus of research.

Composition and assembly of the bacterial community in the overlying waters of the coral reef of China's Xisha Islands.

Coral reef ecosystems are one of the most diverse and productive habitats on Earth. Microbes in the reef-overlying waters are key players in maintaining this ecosystem through regulating biogeochemical and ecological processes. However, the composition structure and assembly mechanism of microbial community in the reef-overlying waters remain largely unknown. In the present study, the bacterial communities from the overlying waters of atolls and fringing reefs as well as the surface waters of the adjacent open ocean of the Xisha Islands in the South China Sea were investigated using 16S rRNA high-throughput sequencing combined with a size-fractionation strategy. The results showed that environments of all sampling stations were similar, characterized by an almost complete lack of inorganic nutrients such as nitrogen and phosphorus. Proteobacteria, Cyanobacteria and Bacteroidetes were the dominant phyla, and Synechococcus was most abundant at the genus level in both large fraction (LF; 1.6-200 μm) and small fraction (SF; 0.2-1.6 μm) communities. Only a slight difference in community composition between LF and SF samples was observed. The bacterial communities among the three habitat types showed noticeable differences, and the bacterial composition among the atoll reefs was more varied than that among the fringing reefs. The similarity of bacterial communities significantly declined with the increasing geographic distance, and stochastic processes were more important than deterministic processes in bacterial community assembly. This study sheds lights on the bacterial biodiversity of coral reefs and the importance of stochastic process in structuring bacterial communities.

Investigation of fungal contamination in medicinal and edible Lycii Fructus through DNA metabarcoding

Lycii Fructus (LF) is considered as a 'superfood' due to its health benefits and delicious tastes, which has gained popularity worldwide. However, LF is also a proper host for fungal growth due to its abundant nutrients. Fungal contamination seriously affects the quality and safety of LF and poses threats to consumer health. In this study, a total of 15 LF samples were collected from five provinces in China, and were divided into five groups based on the collection areas. Fungal contamination in LF was investigated by targeting the internal transcribed spacer 2 region using Illumina Miseq PE300 platform, and the differences of fungal community in groups based on collection areas were compared. Results showed that the fungal contamination was detected in all the 15 LF samples. Ascomycota, Dothideomycetes, Pleosporales and Pleosporaceae were dominant at the phylum, class, order and family levels, respectively. At the genus level, Alternaria, Cladosporium and Fusarium were the three dominant genera. In all, 24 fungal species were identified. Among which, two species, namely Penicillium oxalicum and Trichothecium roseum, were potentially toxigenic. All 15 LF samples were detected with fungal contamination. The differences of fungal community in LF samples collected from different areas were observed. DNA metabarcoding was demonstrated as an efficient method to monitor the fungal contamination in LF. This work comprehensively reveals the fungal diversity and composition in LF and provides early warning for potential mycotoxin contamination.

Discrimination and Prediction of Lonicerae japonicae Flos and Lonicerae Flos and Their Related Prescriptions by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy Combined with Multivariate Statistical Analysis

LJF and LF are commonly used in Chinese patent drugs. In the Chinese Pharmacopoeia, LJF and LF once belonged to the same source. However, since 2005, the two species have been listed separately. Therefore, they are often misused, and medicinal materials are indiscriminately put in their related prescriptions in China. In this work, firstly, we established a model for discriminating LJF and LF using ATR-FTIR combined with multivariate statistical analysis. The spectra data were further preprocessed and combined with spectral filter transformations and normalization methods. These pretreated data were used to establish pattern recognition models with PLS-DA, RF, and SVM. Results demonstrated that the RF model was the optimal model, and the overall classification accuracy for LJF and LF samples reached 98.86%. Then, the established model was applied in the discrimination of their related prescriptions. Interestingly, the results show good accuracy and applicability. The RF model for discriminating the related prescriptions containing LJF or LF had an accuracy of 100%. Our results suggest that this method is a rapid and effective tool for the successful discrimination of LJF and LF and their related prescriptions.

Effect of Lactobacillus plantarum and flavourzyme on physicochemical and safety properties of grass carp during fermentation.

The present study aimed to investigate the change in physicochemical and safety properties of grass carp during fermentation with flavourzyme and Lactobacillus plantarum (FLF). The natural fermentation (NF) and fermentation with Lactobacillus plantarum (LF) samples were used as control. The results showed that with increasing fermentation time, the pH and water activity (Aw) in each fermented grass carp sample gradually decreased, while the a-amino nitrogen (ANN), thiobarbituric acid reactive substance (TBARS), total volatile base nitrogen (TVB-N), biogenic amines (BAs), and harmful microbial gradually increased. Besides, compared with NF samples at each fermentation time, significantly lower pH, Aw, TBARS, TVB-N, BAs, and harmful microbial presented in LF and FLF samples. However, FLF samples have a higher AAN content than that of NF and LF samples during fermentation. Overall, the fermentation with Lactobacillus plantarum and flavourzyme could contribute to fermented grass carp products with better physicochemical and safety properties.

A multi-fidelity Bayesian optimization approach based on the expected further improvement

Sampling efficiency is important for simulation-based design optimization. While Bayesian optimization (BO) has been successfully applied in engineering problems, the cost associated with large-scale simulations has not been fully addressed. Extending the standard BO approaches to multi-fidelity optimization can utilize the information of low-fidelity models to further reduce the optimization cost. In this work, a multi-fidelity Bayesian optimization approach is proposed, in which hierarchical Kriging is used for constructing the multi-fidelity metamodel. The proposed approach quantifies the effect of HF and LF samples in multi-fidelity optimization based on a new concept of expected further improvement. A novel acquisition function is proposed to determine both the location and fidelity level of the next sample simultaneously, with the consideration of balance between the value of information provided by the new sample and the associated sampling cost. The proposed approach is compared with some state-of-the-art methods for multi-fidelity global optimization with numerical examples and an engineering case. The results show that the proposed approach can obtain global optimal solutions with reduced computational costs.

Biological activities of commercial bovine lactoferrin sources.

Lactoferrin (Lf) samples from several manufacturers were evaluated in vitro. The purity and protein form of each Lf were examined by SDS-PAGE, Western blot, and proteomics analysis. Assays were conducted to evaluate uptake of Lfs and iron from Lfs by enterocytes as well as Lf bioactivities, including effects on intestinal cell proliferation and differentiation, IL-18 secretion, TGF-β1 transcription, and growth of enteropathogenic Escherichia coli (EPEC). Composition of the Lfs varies; some only contain a major Lf band (∼80kDa), and some also contain minor forms. All Lfs and iron from the Lfs were absorbed by Caco-2 cells, with various efficiencies. The bioactivities of the Lfs varied considerably, but there was no consistent trend. All Lfs promoted intestinal cell proliferation, secretion of IL-18, and transcription of TGF-β1. Some Lfs exhibited pro-differentiation effects on Caco-2 cells. Effects of pasteurization (62.5°C for 30min, 72°C for 15s, or 121°C for 5min) on integrity, uptake, and bioactivities were examined using Dicofarm, Tatua, and native bovine Lfs. Results show that pasteurization did not affect protein integrity, but variously affected uptake of Lf and its effects on intestinal proliferation, differentiation, and EPEC growth. To choose a Lf source for a clinical trial, assessment of bioactivities is recommended.

Pattern Recognition Receptor-reactivity Screening of Liver Transplant Patients: Potential for Personalized and Precise Organ Matching to Reduce Risks of Ischemia-reperfusion Injury.

Pattern recognition receptors (PRRs) on immune and parenchymal cells can detect danger-associated molecular patterns (DAMPs) released from cells damaged during ischemia-reperfusion injury (IRI), in heart attack or stroke settings, but also as an unavoidable consequence of solid organ transplantation. Despite IRI being a significant clinical problem across all solid organ transplants, there are limited therapeutics and patient-specific diagnostics currently available. We screened portal blood samples obtained from 67 human liver transplant recipients both pre- [portal vein (PV) sample] and post-(liver flush; LF) reperfusion for their ability to activate a panel of PRRs, and analyzed this reactivity in relation to biopsy-proven IRI. PV samples from IRI+ orthotopic liver transplantation (OLT) patients (n = 35) decreased activation of hTLR4- and hTLR9-transfected cells, whereas PV from IRI- patients (n = 32) primarily increased hTLR7 and hNOD2 activation. LF samples from OLT-IRI patients significantly increased activation of hTLR4 and hTLR9 over IRI- LF. In addition, the change from baseline reactivity to hTLR4/9/NOD2 was significantly higher in IRI+ than IRI- OLT patients. These results demonstrate that TLR4/7/9 and NOD2 are involved in either promoting or attenuating hepatic IRI, and suggest a diagnostic screening of portal blood for reactivity to these PRRs might prove useful for prediction and/or therapeutic intervention in OLT patients before transplantation.

The role of vascularization on changes in ligamentum flavum mechanical properties and development of hypertrophy in patients with lumbar spinal stenosis

BACKGROUND CONTEXTLigamentum flavum (LF) induced lumbar spinal stenosis (LSS) is conditioned not only by its "gathering" but especially by hypertrophy. Previous studies have examined the pathophysiology and biochemical changes that cause the hypertrophy. Some studies have described a link between chronic LF inflammation and neovascularization but others have reported highly hypovascular LF tissue in LSS patients. Currently, there is no practical application for our knowledge of the pathophysiology of the LF hypertrophy. Considerations for future treatment include influencing this hypertrophy at the level of tissue mediators, which may slow the development of LSS. To our knowledge, there is no study of micromechanical properties of native LF to date. PURPOSE(1) To clarify the changes in vascularization, chondroid metaplasia, and the presence of inflammatory cell infiltration in LF associated with LSS. (2) To quantify changes in the micromechanical properties associated with LF degenerative processes. STUDY DESIGN/SETTINGVascular density analysis of degenerated and healthy human LF combined with measurement of micromechanical properties. METHODSThe study involved 35 patients who underwent surgery between November 1, 2015 and October 1, 2016. The LSS group consisted of 20 patients and the control group consisted of 15 patients. LF samples were obtained during the operation and were used for histopathological and nanoindentation examinations. Sample vascularization was examined as microvascular density (Lv), which was morphometrically evaluated using semiautomatic detection in conjunction with NIS-Elements AR image analysis software. Samples were also histologically examined for the presence of chondroid metaplasia and inflammation. Mechanical properties of native LF samples were analyzed using the Hysitron TI 950 TriboIndenter nanomechanical testing system. RESULTSVascular density was significantly lower in the LSS group. However, after excluding the effect of age, the difference was not significant. There was high association between Lv and age. With each increasing year of age, Lv decreased by 11.5 mm2. Vascular density decreased up to the age of 50. Over the age of 50, changes were no longer significant and Lv appeared to stabilize. No correlation was observed between Lv and the presence of inflammation or metaplasia; however, LSS patients had a significantly increased incidence of chondroid metaplasia and inflammatory signs. The mechanical properties of control group samples showed significantly higher stiffness than those samples obtained from the LSS group. ConclusionThis study showed that Lv changes were not dependent on LSS but were age-dependent. Vascular density was found to decrease up to the age of 50. A significantly higher incidence of chondroid metaplasia and inflammation was observed in LSS patients. The mechanical property values measured by nanoindentation showed high microstructural heterogeneity of the tested ligaments. Our results showed that healthy ligaments were significantly stiffer than LSS ligaments. CLINICAL SIGNIFICANCEPrevention of the loss of LF vascularization during aging may influence stiffness of LF which in turn may slow down the LF degenerative processes and delay onset of LSS.

Variable-Fidelity Lower Confidence Bounding Approach for Engineering Optimization Problems with Expensive Simulations

Engineering design optimization with expensive simulations is usually a computationally prohibitive process. As one of the most famous efficient global optimization approaches, the lower confidence bounding (LCB) approach has been widely applied to relieve this computational burden. However, the LCB approach can be used only for design optimization problems with the single-fidelity level. In this paper, a variable-fidelity lower confidence bounding (VF-LCB) approach is developed to extend the LCB approach to engineering problems with multifidelity levels. First, a VF-LCB function is analytically derived to adaptively select LF or HF samples according to the predicted values and uncertainty from the VF metamodel. Then the coefficient of variations (CoV) of the predicted values and uncertainty of the VF model, which can reflect the degree of dispersion of the prediction values and uncertainty, respectively, are introduced to balance the global exploration and local exploitation objectively. To illustrate the effectiveness and merits of the proposed VF-LCB approach, eight numerical examples, and the design of micro-aerial vehicle (MAV) fuselage are tested. Comparative results between the proposed approach and the other five existing methods show that the average cost savings are about 25% for eight numerical examples and about 45% for the MAV problem compared with the other five existing methods.

Illumina-Based Analysis of Endophytic and Rhizosphere Bacterial Diversity of the Coastal Halophyte Messerschmidia sibirica.

Halophytes play important roles in coastal ecosystems. However, few reports have described bacterial communities related to halophytes, and the distribution patterns of these bacteria in different plant tissues have been rarely compared. This paper mainly studied the diversity and community structure of endophytic and rhizosphere (Rh) bacteria related to the halophyte Messerschmidia sibirica, a dominant species in the coastal zone of Shandong Peninsula, China. We collected leaf (Lf), stem (Sm), root (Rt), Rh, and bulk (Bl) control soil samples, and sequenced the V5–V7 region of the bacterial 16S rRNA gene using the Illumina HiSeq platform to identify bacterial communities originating from different plant habitats. We found that the bacterial richness and diversity in Rh were significantly higher than those in the leaves, Sm, and Rt, but lower than those of the Bl control soil. In total, 37 phyla and 438 genera were identified. Microbial-diversity analysis showed that Proteobacteria and Actinobacteria were the dominant phyla and that Pseudomonas, Bacillus, Sphingomonas, Streptomyces, Microbacterium, Rhizobium, and Nocardioides were the dominant genera. However, there were clear differences in community diversity and structure among the samples. Endophytic bacteria community in Lf, Sm, and Rt shared more similarity than those in Rh and Bl control soil. The numbers of operational taxonomic units exclusive to the Lf, stem, Rt, Rh, and Bl control soil samples were 51, 43, 122, 139, and 922, respectively, implying habitat-specific patterns. Principal coordinate analysis demonstrated differences were apparent in the bacterial communities associated with habitats. On the whole, M. sibirica affected bacterial diversity and structured the bacterial community. This study provides insight into the complex microbial compositions of coastal halophytes.

Changes in Gut Microbial Ecology and Immunological Responses of Mice Fed the Insoluble Fraction of Brassica rapa L. that was Fermented or Not.

We aimed to investigate the effects of feeding fermented Brassica rapa L. on ecological and immunological changes in the mouse gut using in vitro cultivation tests and in vivo experiments in normal mice. In the preliminary in vitro study, two B. rapa L. products from different fermentation periods (one d [SF] or six months [LF]) were evaluated along with non-fermented vegetables (NF). Among the components of each product, the insoluble fraction resulted in the most prominent change such as a relative increase in butyrate production during a cultivation inoculated with mouse cecum contents. Based on this result, the boiled water-insoluble fractions of B. rapa L. (SF, LF, and NF samples) were selected as test materials for the subsequent in vivo experiment. Male C57BL/6J mice were divided into four groups and fed either a control diet (CON) or control diet plus one of the insoluble fractions for two weeks. The NF and LF groups had higher relative populations of Faecalibacterium prausnitzii than the CON group. Therefore, colonic butyrate concentrations were higher in the NF and LF groups than in the CON group. The oral administration of B. rapa L. extract induced immune regulatory effects, even when mice were fed NF and SF, but not LF, as assessed by an increase in regulatory T cell numbers. Our results indicate that feeding a purified insoluble fraction from B. rapa L. affects enteric short-chain fatty acid production and immunological responses in the mouse gut in a similar manner, regardless of the fermentation status.

Anisotropy field distribution in soft magnetic Hitperm alloys submitted to different field annealing processes

The magnetic anisotropy field distribution is discussed for Hitperm alloys annealed under different field conditions leading to different induced magnetic anisotropies: zero (ZF), transversal (TF), and longitudinal (LF) field annealing and compared to that of as-quenched (AQ) melt-spun amorphous ribbon. In order to accurately use the present method, the demagnetizing factor has been obtained by analyzing the field dependence of the inverse of the field derivative of the magnetization. The coherence of the analysis is supported by testing the normalization of the complete distribution of anisotropy fields. Independently of the composition, two groups can be distinguished among the studied samples: those with mainly perpendicular anisotropy field contributions (ZF and TF samples) and those with mainly longitudinal anisotropy field contributions (LF and AQ samples). Behavior of TF samples is well reproduced using Stoner–Wohlfarth model and, in the case of as-quenched amorphous samples, the anisotropy field depends almost linearly on the thickness of the ribbon.

Evaluation of different methods for determination of the iron saturation level in bovine lactoferrin

Lactoferrin (Lf) samples with ca. 25%, 50%, 75%, 85% and 100% iron saturation were prepared for the purpose of evaluating Chromametry, Differential Scanning Calorimetry (DSC) and Circular Dichroism (CD) spectropolarimetry for their suitability in determining the iron saturation level. Numerical values for colour from Chromametry, enthalpy change of denaturation (ΔHcal) from DSC and molar ellipticities from CD were statistically analysed to evaluate their correlation with the level of iron saturation in Lf. Linear regression analysis of colour coordinates Chroma (C∗) and hue (h°) angle on percentage iron saturation level of Lf showed that the values can be used to estimate the iron saturation level. The ΔHcal for the iron saturated peak and the CD ellipticities in the 310–340nm region provided reliable data for the estimation of iron saturation level of Lf up to 75%. Mono- and di-saturated Lf displayed the same thermal stability and very similar tertiary structures.

Composition, structure and properties of POPC–triolein mixtures. Evidence of triglyceride domains in phospholipid bilayers

We have in this study investigated the composition, structure and spectroscopical properties of multilamellar vesicles composed of a phospholipid, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and up to 10mol% of triolein (TO), a triglyceride. We found in agreement with previous results that the mixtures with 10mol% TO spontaneously separate into two distinct phases, heavy (HF) and light (LF), with different densities and found this also to be the case for 2 and 5mol% TO. The compositions of the two phases were investigated by quantitative lipid mass spectrometric analysis, and with this method we found that TO had a solubility maximum of about 4mol% in the HF, whereas it was markedly up-concentrated in the LF. Electron paramagnetic resonance spectroscopy indicated POPC membranes of all tested concentrations of TO in both phases to be almost unperturbed by the presence of TO and to exist as vesicular structures containing entrapped water. Bilayer structure of the membranes was supported by small angle X-ray scattering that showed the membranes to form a lamellar phase. Fluorescence spectroscopy with the polarity sensitive dye Nile red revealed, that the LF samples with more than 5mol% TO contained pure TO domains. These observations are consistent with an earlier MD simulation study by us and our co-workers suggesting triglycerides to be located in lens shaped, blister-like domains between the two lipid bilayer leaflets (Khandelia et al. (2010) [26]).