Degradation of connective tissue matrices by macrophages. III. Morphological and biochemical studies on extracellular, pericellular, and intracellular events in matrix proteolysis by macrophages in culture.

We have shown that macrophages in culture degrade the glycoproteins and amorphous elastin of insoluble extracellular matrices. Ultrastructural observation of the macrophage-matrix interaction revealed that connective tissue macromolecules were solubilized from the matrix extracellularly. At least part of the matrix breakdown was localized to the immediate vicinity of the cells, as shown by morphological and biochemical studies, although the rate of degradation correlated closely with the secretion of proteinases by various inflammatory stimuli in vivo, by glucocorticoids, prostaglandin E2 or colchicine, or by phagocytosis of latex, zymosan, or cholesterol-albumin complexes in culture was reflected in altered rates of glycoprotein and elastin degradation by the macrophages. Alteration of endocytosis and lysosomal digestion by cytochalasin B, NH4Cl, and proteinase inhibitors did not decrease the overall rate of matrix solubilization, but reduced the processing of the matrix fragments to peptides. Therefore, extracellular, pericellular, and lysosomal events each contribute to degradation of extracellular matrix macromolecules by inflammatory macrophages

Empirical forecasting practices of a British university

This article is based on a single case study aimed at examining behavioral issues of forecasting, in particular the role and practice of forecasting in a British university settings. Key variables were identified in establishing associations between the variables that provide suitable criteria for the purpose of this study. Data collection was based on questionnaires distributed to people involved and interviews which were held with prominent staff of the University. Fisher-exact tests were performed to identify significant associations between variables. Results indicated the various levels of perceptions and practices of forecasting produced by the people involved at the University. The study implies that useful insights can be gathered through forecasting from a different perspective of the non-profit making service industry

Development of allergy in children. I. Association with virus infections.

Children born into allergic families, with two allergic parents, are at high risk of developing allergy within the first 5 years of life. In order to observe possible external factors in the sensitization process, a prospective study of 13 such children was done, in which serial clinical and immunologic observations were made at 3- to 6-month intervals over a period of 1 to 4 yr. Eleven of these children are now clinically allergic; 5 have asthma. Immunologic evidence for allergic sensitization was observed in these 11 children by RAST, antigen-induced leukocyte histamine release, lymphoblastogenesis, and rise in serum IgE. Upper respiratory infections (URI) occurred in these 11 allergic children 1 to 2 months prior to the onset of allergic sensitization. In 10 of these 11 URI children, complement-fixing antibodies to viruses (parainfluenza, RSV, CMV) increased in the same blood samples in which immunologic allergic sensitization was first evidenced. This coincidence suggests that certain viruses may contribute to the allergic sensitization process

Profile-Based Optimal Matchings in the Student-Project Allocation Problem

In the Student/Project Allocation problem (spa) we seek to assign students to individual or group projects offered by lecturers. Students provide a list of projects they find acceptable in order of preference. Each student can be assigned to at most one project and there are constraints on the maximum number of students that can be assigned to each project and lecturer. We seek matchings of students to projects that are optimal with respect to profile, which is a vector whose rth component indicates how many students have their rth-choice project. We present an efficient algorithm for finding agreedy maximum matching in the spa context – this is a maximum matching whose profile is lexicographically maximum. We then show how to adapt this algorithm to find a generous maximum matching – this is a matching whose reverse profile is lexicographically minimum. Our algorithms involve finding optimal flows in networks. We demonstrate how this approach can allow for additional constraints, such as lecturer lower quotas, to be handled flexibly

An introduction to the Cambridge advanced modeller

Complex products and their development processes may be viewed as systems, whose different aspects can be modelled as networks of interactions between elements in different domains. Many approaches have been proposed to explore, support or improve engineering processes by building such models. Developing these approaches, and applying them to problems of realistic complexity, often requires specialised computer software suitable for manipulating large data sets. However, creating suitable tools can be difficult–because software development is time-consuming and requires skills that many researchers and practitioners do not possess. We developed an approach which aims to address this problem by recognising the iterative nature of modelling research and its often tight coupling with prototype software development, and by reducing the effort of software prototyping and revision within this process. The approach is enabled by, and embodied in, the Cambridge Advanced Modeller (CAM)–a configurable software platform we have developed, refined and applied over several years and through a number of research projects

Physiology of renal glucose handling via SGLT1, SGLT2 and GLUT2.

The concentration of glucose in plasma is held within narrow limits (4-10 mmol/l), primarily to ensure fuel supply to the brain. Kidneys play a role in glucose homeostasis in the body by ensuring that glucose is not lost in the urine. Three membrane proteins are responsible for glucose reabsorption from the glomerular filtrate in the proximal tubule: sodium-glucose cotransporters SGLT1 and SGLT2, in the apical membrane, and GLUT2, a uniporter in the basolateral membrane. 'Knockout' of these transporters in mice and men results in the excretion of filtered glucose in the urine. In humans, intravenous injection of the plant glucoside phlorizin also results in excretion of the full filtered glucose load. This outcome and the finding that, in an animal model, phlorizin reversed the symptoms of diabetes, has stimulated the development and successful introduction of SGLT2 inhibitors, gliflozins, in the treatment of type 2 diabetes mellitus. Here we summarise the current state of our knowledge about the physiology of renal glucose handling and provide background to the development of SGLT2 inhibitors for type 2 diabetes treatment

Rapid redistribution of clathrin onto macrophage plasma membranes in response to Fc receptor-ligand interaction during frustrated phagocytosis.

We have observed increases in assembled clathrin on the plasma membrane during "frustrated phagocytosis," the spreading of macrophages on immobilized immune complexes. Resident macrophages freshly harvested from the peritoneal cavity of mice and attached to bovine serum albumin (BSA)-anti-BSA-coated surfaces at 4 degrees C had almost no clathrin basketworks on their adherent plasma membrane (less than 0.01 coated patch/micron 2), as observed by immunofluorescence, immunoperoxidase, and platinum-carbon replica techniques, although abundant assembled clathrin was observed in the perinuclear Golgi region. When the cells were warmed to 37 degrees C they started to spread by 4 min and reached their maximum extent by 20 min. Spreading preceded clathrin assembly at the plasma membrane. Clathrin-coated patches were first observed on the adherent plasma membrane at 6 min. Between 12 and 20 min assembled clathrin coats appeared on both adherent and nonadherent plasma membranes with a concomitant decrease in identifiable clathrin in the perinuclear region. A new steady state emerged by 2 h, as perinuclear clathrin began to reappear. At 20 min at 37 degrees C the adherent plasma membranes of macrophages spreading on BSA alone had 0.9 coated patch/micron 2, whereas in cells spread on immune complex-coated surfaces, the clathrin patches increased, dependent on ligand concentration, to a maximum of 2.1 coated patches/micron 2. Because frustrated phagocytosis of immune complex-coated surfaces at 37 degrees C increased the area of adherent plasma membrane, the total area coated by clathrin basket-works increased 5-fold (28 micron 2/cell) as compared with cells plated on BSA alone (5.6 micron 2/cell) and 200-fold as compared with cells adhering to immune complexes at 4 degrees C. We then determined that macrophages cultured on BSA-coated coverslips for 24 h already have abundant surface clathrin. When immune complexes were formed by the addition of anti-BSA IgG to already spread macrophages cultured on BSA-coated coverslips for 24 h, clathrin assembled at the sites of ligand-receptor interaction even at 4 degrees C, before spreading, and a 2.6-fold increase in assembled clathrin was observed on the adherent plasma membrane of cells on immune complexes as compared with cells on BSA alone. Clathrin was reversibly redistributed to the Golgi region, returning to the steady state by 2 h.(ABSTRACT TRUNCATED AT 400 WORDS

Subpopulations of liver coated vesicles resolved by preparative agarose gel electrophoresis.

Rat liver clathrin coated vesicles (CVs) were separated into several distinct subpopulations using non-sieving concentrations of agarose, which allowed the separation of species differing primarily in surface charge. Using preparative agarose electrophoresis (Kedersha, N. L., and L. H. Rome, 1986, Anal. Biochem., in press), the CVs were recovered and analyzed for differences in morphology, coat protein composition, and stripped vesicle protein composition. Coat proteins from different populations appeared identical on SDS PAGE, and triskelions stripped from the different populations showed the same mobility on the agarose gel, suggesting that the mobility differences observed in intact CVs were due to differences in the surface charge of underlying vesicles rather than to variations in their clathrin coats. Several non-coat polypeptides appeared to segregate exclusively with different populations as resolved by two-dimensional electrophoresis. Stripped CVs also exhibited considerable heterogeneity when analyzed by Western blotting: the fast-migrating population was enriched in the mannose 6-phosphate receptor, secretory acetylcholine esterase, and an Mr 195,000 glycoprotein. The slow-migrating population of CVs was enriched in the asialoglycoprotein receptor, and it appeared to contain all detectable concanavalin A-binding polypeptides as well as the bulk of detectable WGA-binding proteins. When CVs were prepared from 125I-asialoorosomucoid-perfused rat liver, ligand was found in the slow-migrating CVs, suggesting that these were endocytic in origin. Morphological differences were also observed: the fast-migrating population was enriched in smaller CVs, whereas the slow-migrating population exhibited an enrichment in larger CVs. As liver consists largely of hepatocytes, these subpopulations appear to originate from the same cell type and probably represent CVs of different intracellular origin and destination

Modelling binary alloy solidification with adaptive mesh refinement

The solidification of a binary alloy results in the formation of a porous mushy layer, within which spontaneous localisation of fluid flow can lead to the emergence of features over a range of spatial scales. We describe a finite volume method for simulating binary alloy solidification in two dimensions with local mesh refinement in space and time. The coupled heat, solute, and mass transport is described using an enthalpy method with flow described by a Darcy-Brinkman equation for flow across porous and liquid regions. The resulting equations are solved on a hierarchy of block-structured adaptive grids. A projection method is used to compute the fluid velocity, whilst the viscous and nonlinear diffusive terms are calculated using a semi-implicit scheme. A series of synchronization steps ensure that the scheme is flux-conservative and correct for errors that arise at the boundaries between different levels of refinement. We also develop a corresponding method using Darcy's law for flow in a porous medium/narrow Hele-Shaw cell. We demonstrate the accuracy and efficiency of our method using established benchmarks for solidification without flow and convection in a fixed porous medium, along with convergence tests for the fully coupled code. Finally, we demonstrate the ability of our method to simulate transient mushy layer growth with narrow liquid channels which evolve over time

Parametric Oscillation with Squeezed Vacuum Reservoirs

Employing the quantum Hamiltonian describing the interaction of two-mode light (signal-idler modes) generated by a nondegenerate parametric oscillator (NDPO) with two uncorrelated squeezed vacuum reservoirs (USVR), we derive the master equation. The corresponding Fokker-Planck equation for the Q-function is then solved employing a propagator method developed in Ref. \cite{1}. Making use of this Q-function, we calculate the quadrature fluctuations of the optical system. From these results we infer that the signal-idler modes are in squeezed states and the squeezing occurs in the first quadrature. When the NDPO operates below threshold we show that, for a large squeezing parameter, a squeezing amounting to a noise suppression approaching 100% below the vacuum level in the first quadrature can be achieved.Comment: 16 page