Autoantibodies to the insulin receptor. Effect on the insulin-receptor interaction in IM-9 lymphocytes

The serum of some patients with insulin-resistant "diabetes" contains antibodies that bind to and block the cell membrane receptors for insulin. In this report, we have characterized the effects of the antireceptor antibodies on the interaction of (125)I-insulin with its receptor on the human lymphoblastoid cell line IM-9. Up to 95% of specific insulin binding can be inhibited by pretreatment of the cells with these immunoglobulins. The onset of the inhibitory effect is time- and temperature-dependent, and the effect is reversed extremely slowly if the cells are suspended in a large excess of antibody-free buffer. These features of antibody binding can be easily distinguished from those for insulin binding to its receptor. The inhibitory effect of the antibodies can be reversed by exposure of the cells to conditions known to elute surface immunoglobulins. The three antireceptor sera studied appear to alter the insulin-receptor interaction in different ways. Two antisera markedly reduce receptor affinity through combined effects on the insulin association and dissociation rates, and, additionally, have smaller effects on available receptor number. A third antiserum primarily affects available receptor number and has little effect on receptor affinity. All three antisera inhibit the capacity of insulin to promote negatively cooperative site-site interactions among insulin receptors. The data suggest that these autoantibodies to the insulin receptor bind to different determinants on the receptor and may therefore be useful as unique probes of insulin receptor structure and function.     

Experimental automobile-pedestrian injuries

Fifteen fresh cadaver impacts were conducted in simulation of pedestrian-automobile accidents. The test sled configuration simulated an automobile bumper and hood in "standard" and "nosedive" situations as well as "hard" and "soft" impact surfaces. Instrumentation and film demonstrated large axial compression forces in the struck leg and considerable angular velocity of the torso in all modes. Dissection revealed primary injury at the impact site at low velocities with added remote injury at high velocities. Fracture of the lateral tibial plateau was most common. Lowering the bumper height offered the greatest protection against injury at moderate impact velocities.     

Thermal Boundary Resistance in GaN Films Measured by Time Domain Thermoreflectance with Robust Monte Carlo Uncertainty Estimation

In this work, we investigate the thermal boundary resistance and thermal conductivity of GaN layers grown on Si with 100 nm AlN transition layers using time domain thermoreflectance (TDTR). The GaN layers ranged from 0.31 to 1.27 μm. Due to the challenges in determining the thermal boundary resistance of the buried interfaces found in this architecture, a new data reduction scheme for TDTR that utilizes a Monte Carlo fitting method is introduced and found to dramatically reduce the uncertainty in certain model parameters. The results show that the GaN thermal conductivity does not change significantly with layer thickness, whereas the resistance of the AlN layer decreases slightly with GaN thickness.     

Australia's housing choices: Retrospect and prospect

This article identifies issues around the Auckland Region's strategy for urban intensification and discusses the ensuing policy implications. Medium density housing forms an important element of the strategy for growth management. Implications for its success are primarily drawn from research into a major housing development in Waitakere City. It is argued that for ideological, political and technical reasons the strategy could be easily undermined. The article also suggests that something of a paradigm shift is underway in urban planning.     

Mathematical similarities in engineering network analysis

This paper examines the proposition that structural trusses and pipe networks for water supply are closely related in a mathematical sense. It shows that in terms of representation, physical quantities, constitutive equations, analysis problem formulations and methods of analysis they both belong to the same class of nonlinear potentiated networks, the truss being a special linear case for problems of analysis. The analysis of nonlinear potentiated networks may therefore be efficiently performed by linearization of the relevant nonlinear quantities and the iterative use of well-known truss analysis methods. The commonality established between structural trusses and water supply systems has potential for exploitation in respect of other network-related problems of optimum design, reliability evaluation and design for resilience.     

A constrained binary knapsack approximation for shortest path network interdiction

A modified shortest path network interdiction model is approximated in this work by a constrained binary knapsack which uses aggregated arc maximum flow as the objective function coefficient. In the modified shortest path network interdiction problem, an attacker selects a path of highest non-detection probability on a network with multiple origins and multiple available targets. A defender allocates a limited number of resources within the geographic region of the network to reduce the maximum network non-detection probability between all origin-target pairs by reducing arc non-detection probabilities and where path non-detection probability is modeled as a product of all arc non-detection probabilities on that path. Traditional decomposition methods to solve the shortest path network interdiction problem are sensitive to problem size and network/regional complexity. The goal of this paper is to develop a method for approximating the regional allocation of defense resources that maintains accuracy while reducing both computational effort and the sensitivity of computation time to network/regional properties. Statistical and spatial analysis methods are utilized to verify approximation performance of the knapsack method in two real-world networks.     

Probability-based validation of protein identifications using a modified SEQUEST algorithm

Database-searching algorithms compatible with shotgun proteomics match a peptide tandem mass spectrum to a predicted mass spectrum for an amino acid sequence within a database. SEQUEST is one of the most common software algorithms used for the analysis of peptide tandem mass spectra by using a cross-correlation (XCorr) scoring routine to match tandem mass spectra to model spectra derived from peptide sequences. To assess a match, SEQUEST uses the difference between the first- and second-ranked sequences (ACn). This value is dependent on the database size, search parameters, and sequence homologies. In this report, we demonstrate the use of a scoring routine (SEQUEST-NORM) that normalizes XCorr values to be independent of peptide size and the database used to perform the search. This new scoring routine is used to objectively calculate the percent confidence of protein identifications and posttranslational modifications based solely on the XCorr value.     

Automated identification of amino acid sequence variations in proteins by HPLC/microspray tandem mass spectrometry

Amino acid sequence variations resulting from single-nucleotide polymorphisms (SNPs) were identified using a novel mass spectrometric method. This method obtains 99+% protein sequence coverage for human hemoglobin in a single LC-microspray tandem mass spectrometry (microLC-MS/MS) experiment. Tandem mass spectrometry data was analyzed using a modified version of the computer program SEQUEST to identify the sequence variations. Conditions of sample preparation, chromatographic separation, and data collection were optimized to correctly identify amino acid changes in six variants of human hemoglobin (Hb C, Hb E, Hb D-Los Angeles, Hb G-Philadelphia, Hb Hope, and Hb S). Hemoglobin proteins were isolated and purified, dehemed, (S)-carboxyami-domethylated, and then subjected to a combination proteolytic digestion to obtain a complex peptide mixture with multiple overlaps in sequence. Reversed-phase chromatographic separation of peptides was achieved on-line with MS utilizing a robust fritless microelectrospray interface. Tandem mass spectrometry was performed on an ion trap mass spectrometer using automated data-dependent MS/MS procedures. Tandem mass spectra were collected from the five most abundant ions in each scan using dynamic and isotopic exclusion to minimize redundancy. The spectra were analyzed by a version of the SEQUEST algorithm modified to identify amino acid substations resulting from SNPs.     

Proteome profiling-pitfalls and progress

In this review we examine the current state of analytical methods in proteomics. The conventional methodology using two-dimensional electrophoresis gels and mass spectrometry is discussed, with particular reference to the advantages and shortcomings thereof. Two recently published methods which offer an alternative approach are presented and discussed, with emphasis on how they can provide information not available via two-dimensional gel electrophoresis. These two methods are the isotope-coded affinity tags approach of Gygi et al. and the two-dimensional liquid chromatography-tandem mass spectrometry approach as presented by Link et al. We conclude that both of these new techniques represent significant advances in analytical methodology for proteome analysis. Furthermore, we believe that in the future biological research will continue to be enhanced by the continuation of such developments in proteomic analytical technology.     

Sex-and age-related differences in ceramide dihexosides of primary human brain tumors

Neutral glycolipids (NGL) are promising diagnostic markers of human gliomas, but differences in NGL with age and sex have not been examined. Previous work demonstrated that ceramide dihexosides (CDH) levels in mouse kidney are age- and sex-dependent, probably due to levels of sex hormones. We quantitated CDH in 181 human gliomas and found significant differences with sex and age, particularly menopause and male puberty. This emphasizes the importance of assessing results of studies on glycolipids in disease states with respect to age and sex in order to avoid erroneous conclusions concerning the relationship of glycolipid composition with diagnosis and pathogenesis.