Further purification and characterization of serum proteins used to detect cystic fibrosis genotypes by isoelectric focusing

Sera from cystic fibrosis (CF) homozygotes and obligate heterozygotes contain a CF factor (gamma CF factor) not found by isoelectric focusing in thin-layer polyacrylamide gels in most normal control sera. In addition, sera from most obligate heterozygotes lack another protein (bland B, C, or D) that is commonly found in sera from most normal and cystic fibrosis individuals. A standardized, biophysical assay is described that employs isoelectric focusing for the detection of both CF homozygotes and heterozygotes based on the analysis of whole serum for the presence of the gamma CF factor and bands B, C, and D. Results of analyzing sera from selected CF patients by isoelectric focusing indicated that there is a general correlation between the amount of the gamma CF factor and the clinical severity of the disease. Partial purification and characterization of the gamma CF factor and protein bands B, C, and D was accomplished by using DEAE-cellulose chromatography, Sephadex G-200 gel filtration, sequential molecular filtration through a series of Amicon Diaflo ultrafiltration membranes, affinity chromatography, and cellulose acetate electrophoresis. The gamma CF factor is a cationic protein with a pI of 8.46+/-0.05, has gamma electrophoretic mobility, a molecular weight between 3,500 and 10,000, and apparently exists in CF serum in 2 forms (free in solution and complexed to IgG). Bands B, C, and D are cationic proteins with pI values of 7.85 to 8.10, have gamma electrophoretic mobility and a molecular weight of approximately 100,000-150,000.     

Quantitative assessment of IgM antibodies towards an immunodominant B-cell epitope within the preS2 domain of HBV in the natural course and during combined prednisone/interferon alpha 2b treatment of chronic hepatitis B virus infection

A direct binding enzyme-linked immunosorbent assay (ELISA) was established for quantitative determination of serum IgM antibodies towards a synthetic peptide corresponding to a selected segment (14-21) of the preS2-gene product containing an immunodominant linear B-cell epitope. The prevalence of IgM anti-preS2 (14-21) antibody titers > 1,000 for hepatitis B e antigen (HBeAg)-positive patients with chronic hepatitis B virus (HBV) infection was 38% (22/58) and 10% (2/21) for HBeAg-negative subjects (P < 0.005). IgM anti-preS2 (14-21) reactivity was detected during the clinical course of chronic HBV infection and IgM anti-peptide antibody titers declined and disappeared before spontaneous HBe/anti-HBe seroconversion. Recombinant interferon (IFN)-alpha 2b with an antecedent short course of corticosteroids was administered to eight Chinese patients with chronic HBV infection. The IgM anti-preS2 (14-21) reactivity was monitored consecutively during treatment and patients were followed for more than 1 year. A close association between the presence of pretreatment IgM anti-preS2 (14-21) in serum and the capacity to respond favorably to the combined prednisone/IFN-alpha 2b therapy was detected. The IgM anti-preS2 (14-21) titers decreased during treatment with subsequent loss of detectable antibodies 8-16 weeks after the initiation of therapy. This decrease was concomitant with an alanine aminotransferase (ALT) augmentation preceding the disappearance of HBV-DNA and anti-HBe seroconversion. Long-term remission was not observed in treated patients who lacked detectable levels of pretreatment IgM anti-preS2 (14-21) in the circulation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of steering demand on lane keeping behaviour, self-reports, and physiology. A simulator study

In this study a driving simulator was used to determine changes in mental effort in response to manipulations of steering demand. Changes in mental effort were assessed by using subjective effort ratings, physiology, and the standard deviation of the lateral position. Steering demand was increased by exposure to narrow lane widths and high density oncoming traffic while speed was fixed in all conditions to prevent a compensatory reaction. Results indicated that both steering demand factors influence mental effort expenditure and using multiple measures contributes to effort assessment. Application of these outcomes for adaptive automation is envisaged.     

Effect of substituting fresh‐cut perennial ryegrass with fresh‐cut white clover on bovine milk fatty acid profile

BACKGROUND

Including forage legumes in dairy systems can help address increasing environmental/economic concerns about perennial ryegrass monoculture pastures. This work investigated the effect of substituting fresh‐cut grass with increasing quantities of fresh‐cut white clover (WC) on milk fatty acid (FA) profile and transfer efficiency of dietary linoleic (LA) and α‐linolenic (ALNA) acids to milk fat. Three groups of three crossbred dairy cows were used in a 3 × 3 crossover design. Dietary treatments were 0 g kg^?1 WC + 600 g kg^?1 grass, 200 g kg^?1 WC + 400 g kg^?1 grass, and 400 g kg^?1 WC + 200 g kg^?1 grass. All treatments were supplemented with 400 g kg^?1 concentrates on a dry matter basis. Cows had a 19‐day adaptation period to the experimental diet before a 6‐day measurement period in individual tie stalls.

RESULTS

Increasing dietary WC did not affect dry matter intake, milk yield or milk concentrations of fat, protein or lactose. Milk polyunsaturated FA concentrations (total n‐3, total n‐6, LA and ALNA) and transfer efficiency of LA and ALNA were increased with increasing dietary WC supply.

CONCLUSION

Inclusion of WC in pastures may increase concentrations of nutritionally beneficial FA, without influencing milk yield and basic composition, but any implications on human health cannot be drawn. © 2018 The Authors. Journal of the Science of Food and Agriculture published by JohnWiley & Sons Ltd on behalf of Society of Chemical Industry.

     

Phenotypic and functional characterization of normal rat pleural macrophages in comparison with autologous peritoneal and alveolar macrophages

It is not well appreciated that nutritional status can modulate apoptosis, a process that eliminates unwanted or damaged cells. Choline is an essential nutrient, and its absence induces apoptosis. When PC12 cells were cultivated in a choline-free medium, apoptosis was induced (27.4% of cells apoptotic at 72 h as compared to 4.4% in control medium). In choline-free medium at 72 h, there was a 49% decrease in phosphatidylcholine concentration (P<0.01) and a 34% decrease in sphingomyelin concentration (P<0.01); however, there was no change in phosphatidylethanolamine concentration. Before detecting increased apoptosis in choline-deficient cells, we measured a significant increase in ceramide (218% control) and diacyglycerol (155% control) concentrations. The addition of a cell-permeable ceramide to cells in control medium induced apoptosis; however, adding a cell-permeable diacyglycerol did not induce apoptosis. Caspase is a common mediator of apoptosis, and choline deficiency-induced apoptosis was prevented completely by replacing choline or adding a caspase inhibitor into the medium within 48 h of initial choline deprivation. In those cells rescued by replacing choline at 36 h, the concentrations of phosphatidylcholine, sphingomyelin, ceramide, and diacyglycerol returned to levels of control cells. In those cells rescued by adding a caspase inhibitor at 36 h, the concentrations of sphingomyelin and ceramide returned to control levels, but the concentrations of phosphatidylcholine and diacyglycerol did not return to normal. We propose that availability of dietary factors (choline in this model) can modulate apoptosis. Mechanisms that we identify using this model may help us to explain why dietary choline influences brain development.     

From food structure to texture

Understanding the relationship between food texture perception and food structure is of increasing importance for companies wishing to produce texturally attractive food products. The perception of texture is a complex process involving the senses of vision, hearing, somesthesis and kinesthesis. Texture perception takes place partly during the dynamic process of food breakdown in the mouth and is affected by oral processes, such as motility, saliva production and temperature. To take account of these factors, a multidisciplinary approach is proposed for studying the relationship between food structure and texture perception, combining sensory research, physiology studies and research into food physicochemical characteristics. Recent developments in these three areas that give potential for a better understanding of texture perception and its relationship with food structure are discussed.     

Film Properties of Hydroxypropyl Starch

The present study describes the preparation of low‐substituted hydroxypropyl (HP) starches and investigation of their suitability for the development of flexible, highly transparent cast films. Changes in molecular composition due to processing conditions were identified by determining molar mass distribution. Molecular characterisation was performed both by static light scattering (SLS) and size‐exclusion chromatography with multi‐angle laser light scattering (SEC‐MALLS) detection. The weight‐average molar mass, mean square radius of gyration, second virial coefficient, and molar mass distribution were determined. Rheological characterisation of concentrated aqueous solutions included a study of flow behaviour as a function of shear rate and oscillatory measurement of viscoelastic properties. Significant differences in rheological properties due to hydroxypropylation conditions were established. Mechanical testing was carried out by measuring tensile strength and elongation and determining the elastic modulus. The tensile strength of cast films based on hydroxypropyl starches was 40–50 MPa. Elongation values were dependent on the type of starch, use of plasticiser and storage conditions.     

Da Vinci's yeast: Blastobotrys davincii f.a., sp. nov

A new species of the yeast genus Blastobotrys was discovered during a worldwide survey of culturable xerophilic fungi in house dust. Several culture-dependent and independent studies from around the world detected the same species from a wide range of substrates including indoor air, cave wall paintings, bats, mummies, and the iconic self-portrait of Leonardo da Vinci from ca 1512. However, none of these studies identified their strains, clones, or OTUs as Blastobotrys. We introduce the new species as Blastobotrys davincii f.a., sp. nov. (holotype CBS H-24879) and delineate it from other species using morphological, phylogenetic, and physiological characters. The new species of asexually (anamorphic) budding yeast is classified in Trichomonascaceae and forms a clade along with its associated sexual state genus Trichomonascus. Despite the decade-old requirement to use a single generic name for fungi, both names are still used. Selection of the preferred name awaits a formal nomenclatural proposal. We present arguments for adopting Blastobotrys over Trichomonascus and introduce four new combinations as Blastobotrys allociferrii (≡ Candida allociferrii), B. fungorum (≡ Sporothrix fungorum), B. mucifer (≡ Candida mucifera), and Blastobotrys vanleenenianus (≡ Trichomonascus vanleenenianus). We provide a nomenclatural review and an accepted species list for the 37 accepted species in the Blastobotrys/Trichomonascus clade. Finally, we discuss the identity of the DNA clones detected on the da Vinci portrait, and the importance of using appropriate media to isolate xerophilic or halophilic fungi.