The Topology,in Model Membranes,of the Core Peptide Derived from the T‐Cell Receptor Transmembrane Domain

The T‐cell receptor–CD3 complex (TCR–CD3) serves a critical role in protecting organisms from infectious agents. The TCR is a heterodimer composed of α‐ and β‐chains, which are responsible for antigen recognition. Within the transmembrane domain of the α‐subunit, a region has been identified to be crucial for the assembly and function of the TCR. This region, termed core peptide (CP), consists of nine amino acids (GLRILLLKV), two of which are charged (lysine and arginine) and are crucial for the interaction with CD3. Earlier studies have shown that a synthetic peptide corresponding to the CP sequence can suppress the immune response in animal models of T‐cell‐mediated inflammation, by disrupting proper assembly of the TCR. As a step towards the understanding of the source of the CP activity, we focused on CP in egg phosphatidylcholine/cholesterol (9:1, mol/mol) model membranes and determined its secondary structure, oligomerization state, and orientation with respect to the membrane. To achieve this goal, 15‐residue segments of TCRα, containing the CP, were synthesized and spin‐labeled at different locations with a nitroxide derivative. Electron spin‐echo envelope modulation spectroscopy was used to probe the position and orientation of the peptides within the membrane, and double electron–electron resonance measurements were used to probe its conformation and oligomerization state. We found that the peptide is predominantly helical in a membrane environment and tends to form oligomers (mostly dimers) that are parallel to the membrane plane.     

The impact of anthocyanin-rich red raspberry extract (ARRE) on the properties of edible soy protein isolate (SPI) films

To modify the properties of edible soy protein isolate (SPI) films, 0.5% anthocyanin-rich red raspberry (Rubus strigosus) extract (ARRE) (0.5 g raspberry powder in 95% ethyl alcohol/water/85% lactic acid [80:19:1. v/v/v]) was incorporated into film-forming solutions. ARRE resulted in an SPI film having significantly enhanced tensile strength (P < 0.05) and % elongation at break (P < 0.05), as well as increased water swelling ratio (P < 0.05) and in vitro pepsin digestibility (P < 0.05). The resultant films also showed significantly decreased water solubility and water vapor permeability (P < 0.05). In addition, ARRE increased darkness, redness, and yellowness film appearance as evidenced by a lower L* (P < 0.05), greater positive a* (P < 0.05), and a higher b* (P < 0.05) than the control film. Scanning electron microscopy images revealed that extract-added films had denser and more compact cross-section microstructure. Fourier transform infrared spectra illustrated that ARRE-created hydrogen bonding involved conformational changes of soy protein without destroying its backbone structure. SDS-PAGE electrophoretograms revealed that the extract induced intermolecular interaction of the soy protein monomers. Natural plant extracts would be a promising ingredient to make SPI films with different physicochemical properties and applications. PRACTICAL APPLICATION: This study characterizes the potential physicochemical changes of SPI film with incorporated raspberry extract. Upon the above modification, the resultant film was found to enhance the applications of pure SPI film in food packaging. For example, SPI-ARRE film could prolong the usage life of SPI film due to increased strength, or could be useful as a desiccant (drying agent) such as a water-absorbing sheet for preserving dried foods due to its increased hydrophilic surface and water-swelling ratio. SPI-ARRE film could also be alternately used as a food wrap with unique color.     

Detection of Campylobacter jejuni in naturally contaminated chicken skin by melting peak analysis of amplicons in real-time PCR

Contamination of poultry by Campylobacter spp. is a significant source of human diarrheal diseases. Traditional methods currently used to detect Campylobacter in foods are time-consuming and labor-intensive. In this study, primers designed for the Campylobacter jejuni cadF gene sequence were used in a SYBR Green I real-time PCR assay as an alternative to a conventional bacteriological method for the rapid detection of C. jejuni from poultry. Twelve portions of chicken purchased from two local grocery stores and 39 portions obtained from a commercial processing plant were examined. Samples of the skin were enriched in Bolton broth at 37 degrees C for 3 h and then at 42 degrees C for 9, 21, or 45 h under microaerobic conditions. DNA was extracted from 1-ml aliquots of the enrichment cultures using 1% Triton X-100. The DNA was used as the template in a real-time polymerase chain reaction (PCR) assay. After 24 h of enrichment, C. jejuni was isolated from 13 samples and all of the positive cultures were also detected by the real-time PCR procedure. C. jejuni was detected by both methods from samples artificially contaminated with 1 or 10 CFU of C. jejuni per 10 g, after 24 h of enrichment. The real-time PCR method was found to be sensitive and specific. It significantly reduced the time required for the detection of C. jejuni in poultry following enrichment of samples.     

Use of dairy proteins in lean poultry meat batters – a comparative study

The use of dry whole milk, skimmed milk, caseinate, regular and modified whey, at 2% level (w/w) and with 2% additional protein level was studied in a chicken breast meat system with 51% water addition. At the 2% (w/w) level, all dairy proteins significantly reduced cooking loss compared with the control, with caseinate showing the best results. When compared on an equal protein level (2% total protein), the best performing ingredients were the whole milk and modified whey. A similar observation was made in their effect on the products’ hardness and fracturability. A cost analysis revealed that modified whey provided the most economical ingredient even when used in quantities three times greater than that of as caseinate. Microscopy results showed the formation of larger fine‐protein‐matrix regions in the treatments that provided higher fracturability values.     

Kinetics of interaction of HIV fusion protein (gp41) with lipid membranes studied by real-time AFM imaging

One of the most important steps in the process of viral infection is a fusion between cell membrane and virus, which is mediated by the viral envelope glycoprotein. The study of activity of the glycoprotein in the post-fusion state is important for understanding the progression of infection. Here we present a first real-time kinetic study of the activity of gp41 (the viral envelope glycoprotein of human immunodeficiency virus—HIV) and its two mutants in the post-fusion state with nanometer resolution by atomic force microscopy (AFM). Tracking the changes in the phosphatidylcholine (PC) and phosphatidylcholine–phosphatidylserine (PC:PS) membrane integrity over one hour by a set of AFM images revealed differences in the interaction of the three types of protein with zwitterionic and negatively charged membranes. A quantitative analysis of the slow kinetics of hole formation in the negatively charged lipid bilayer is presented. Specifically, analysis of the rate of roughness change for the three types of proteins suggests that they exhibit different types of kinetic behavior.     

Unusual Two-Step Assembly of a Minimalistic Dipeptide-Based Functional Hypergelator

Self-assembled peptide hydrogels represent the realization of peptide nanotechnology into biomedical products. There is a continuous quest to identify the simplest building blocks and optimize their critical gelation concentration (CGC). Herein, a minimalistic, de novo dipeptide, Fmoc-Lys(Fmoc)-Asp, as an hydrogelator with the lowest CGC ever reported, almost fourfold lower as compared to that of a large hexadecapeptide previously described, is reported. The dipeptide self-assembles through an unusual and unprecedented two-step process as elucidated by solid-state NMR and molecular dynamics simulation. The hydrogel is cytocompatible and supports 2D/3D cell growth. Conductive composite gels composed of Fmoc-Lys(Fmoc)-Asp and a conductive polymer exhibit excellent DNA binding. Fmoc-Lys(Fmoc)-Asp exhibits the lowest CGC and highest mechanical properties when compared to a library of dipeptide analogues, thus validating the uniqueness of the molecular design which confers useful properties for various potential applications.     

Surface-induced dissociation of peptides and protein complexes in a quadrupole/time-of-flight mass spectrometer

A novel in-line surface-induced dissociation (SID) device was designed and implemented in a commercial QTOF instrument (Waters/Micromass QTOF II). This new setup allows efficient SID for a broad range of molecules. It also allows direct comparison with conventional collision-induced dissociation (CID) on the same instrument, taking advantage of the characteristics of QTOF instrumentation, including extended mass range, improved sensitivity, and better resolution compared with quadrupole analyzers and ion traps. Various peptides and a noncovalent protein complex have been electrosprayed and analyzed with the new SID setup. Here we present SID of leucine enkephalin, fibrinopeptide A, melittin, insulin chain-B, and a noncovalent protein complex from wheat, heat shock protein 16.9. The SID spectra were also compared to CID spectra. With the SID setup installed, ion transmission proved to be efficient. SID fragmentation patterns of peptides are, in general, similar to CID, with differences in the relative intensities of some peaks such as immonium ions, backbone cleavage b- versus y-type ions, and y- versus y-NH3 ions, suggesting enhanced accessibility to high-energy/secondary fragmentation channels with SID. Furthermore, these results demonstrate that the in-line SID setup is a valid substitute for CID, with potential advantages for activation of singly/multiply charged peptides and larger species such as noncovalent protein complexes.     

Isomorphic Representations and Well-Formedness of Engineering Systems

When representing the elements of different engineering systems as vertices and edges of a mathematical graph, the well-formedness of the topology of the graph, and hence the topology of the engineering system, can be explicitly computed. This enables checking the well-formedness of the engineering system before investing the effort needed for a complete analysis. This method is demonstrated in the paper for the following fields: trusses, dynamic mass-spring-damper oscillator systems and planetary gear systems. The approach facilitates achieving rapid design, correct first time, which is an important aim of modern design computation.     

Standford B型主动脉夹层覆膜支架腔内修复术围手术期并发症分析

目的总结覆膜支架腔内修复术治疗Standford B型主动脉夹层的经验,分析围手术期并发症的发生率及处理方法。方法２００５年１月至２０１０年１月,１２７例B型主动脉夹层患者接受覆膜支架腔内修复术治疗。B型夹层１１２例,B型主动脉穿透性溃疡１５例。术前、术后１０d行６４排CTA检查,证实手术效果。统计分析围手术期病死率、主要并发症发生率,探讨死亡及发生主要并发症的可能原因。结果手术成功率１００％,围手术期死亡３例,病死率为２．４％,主要并发症发生率为１３．４％（１７／１２７）,包括截瘫１例,逆行性A型夹层１例,脑梗死致意识障碍２例,支架相关并发症９例,严重内漏４例。结论覆膜支架腔内修复术治疗B型主动脉夹层安全、有效,但主要并发症的发生不容忽视。