Characterization of expression of phosphofructokinase isoforms in isolated rat pancreatic islets and purified beta cells and cloning and expression of the rat phosphofructokinase-A isoform

We prepared polymers having a phospholipid polar group, poly [omega-methacryloyloxyalkyl phosphorylcholine (MAPC)-co-n-butyl methacrylate(BMA)], as new biomedical materials and evaluated their blood compatibility with attention to protein adsorption and platelet adhesion. The total amount of proteins adsorbed on the polymer surface from human plasma was determined, and the distribution of adsorbed proteins on the plasma-contacting surface was analyzed. The amount of proteins adsorbed on every poly (MAPC-co-BMA) was small compared with that observed on polymers without the phospholipid polar group. However, there was no significant difference in the amount of adsorbed proteins on the poly(MAPC-co-BMA) even when the methylene chain length between the phospholipid polar group and the backbone in the MAPC moiety was altered. Platelet adhesion on the polymer surface from a platelet suspension in a buffered solution was evaluated with and without plasma treatment on the surface. When a rabbit platelet suspension was brought into contact with the poly(BMA) surface after treatment with plasma, many platelets adhered and aggregated. However, a reduced amount of platelet adhered on the poly(BMA) was found in the case of direct contact with the platelet suspension. On the other hand, the poly(MAPC-co-BMA)s could inhibit platelet adhesion under both conditions. Based on these results, it can be concluded that the proteins adsorbed on the surface play an important role in determining the platelet adhesion and suppression of the protein adsorption on the surface, which is one of the most significant ways of inhibiting platelet adhesion.     

On the mechanism of human stefin B folding: II. Folding from GuHCl unfolded, TFE denatured, acid denatured, and acid intermediate states

In this report, we demonstrate the utility of interleukin-2 (IL-2) stimulation of spleen cell cultures and bivariate flow cytometry in the analysis and purification of the C57BL/6J mouse Y Chromosome. We determined that the DNA content of the C57BL/6J Y Chromosome is approximately 94.7 Mb, making it similar in size to human Chromosome 16 and significantly larger than previous estimates. In addition, we describe the bulk isolation of mouse Y Chromosomes and demonstrate enrichment of the isolated material using a fluorescence in situ hybridization strategy. We detail the construction of two small insert Y Chromosome-specific libraries, ideal for sampling Y Chromosome sequences. From these libraries 1566 clones were analyzed. We provide a detailed characterization of 103 clones, generating nearly 50 kb of sequence. For 30 of these clones, we identify regions of homology to known Y chromosomal sequences, confirming the enrichment of the sorted DNA. From the remaining characterized clones, we describe the development of 15 male-specific PCR assays and 19 male-female PCR assays potentially originating from the pseudoautosomal region or other areas of X-Y or autosome-Y homology.     

Dimensions of the average pore,the number of pores,and the surface area of hardened Portland cement paste

In the course of their small angle x-ray scattering work, Winslow and Diamond also calculated the radius of gyration of the pores. Using its value for a paste, Diamond adopted two models for the average pore: a sphere and a cylinder of equal height and diameter. This model leads to absurd values for the surface area of the paste. Using a cylindrical model, and the radius of gyration plus the hydraulic radius, the present authors calculated for a similar paste the dimensions of the average pore and obtained the values: $\text{diameter}\text{= 47.2}\text{A}\text{?}\text{,}\text{length}\text{= 466}\text{A}\text{?}$. The number of pores per gram of paste was 2.26 × 10¹⁷. The paper also discusses the surface area of hardened paste, and points out the extremely important contributions of Winslow and Diamond to the subject.     

The effects of non-insulin-dependent diabetes mellitus on the kinetics of onset of insulin action in hepatic and extrahepatic tissues

The mechanism(s) of insulin resistance in non-insulin-dependent diabetes mellitus remains ill defined. The current studies sought to determine whether non-insulin-dependent diabetes mellitus is associated with (a) a delay in the rate of onset of insulin action, (b) impaired hepatic and extrahepatic kinetic responses to insulin, and (c) an alteration in the contribution of gluconeogenesis to hepatic glucose release. To answer these questions, glucose disappearance, glucose release, and the rate of incorporation of 14CO2 into glucose were measured during 0.5 and 1.0 mU/kg-1 per min-1 insulin infusions while glucose was clamped at approximately 95 mg/dl in diabetic and nondiabetic subjects. The absolute rate of disappearance was lower (P < 0.05) and the rate of increase slower (P < 0.05) in diabetic than nondiabetic subjects during both insulin infusions. In contrast, the rate of suppression of glucose release in response to a change in insulin did not differ in the diabetic and nondiabetic subjects during either the low (slope 30-240 min:0.02 +/- 0.01 vs 0.02 +/- 0.01) or high (0.02 +/- 0.00 vs 0.02 +/- 0.00) insulin infusions. However, the hepatic response to insulin was not entirely normal in the diabetic subjects. Both glucose release and the proportion of systemic glucose being derived from 14CO2 (an index of gluconeogenesis) was inappropriately high for the prevailing insulin concentration in the diabetic subjects. Thus non-insulin-dependent diabetes mellitus slows the rate-limiting step in insulin action in muscle but not liver and alters the relative contribution of gluconeogenesis and glycogenolysis to hepatic glucose release.     

Cocaprins, β-Trefoil Fold Inhibitors of Cysteine and Aspartic Proteases from Coprinopsis cinerea

We introduce a new family of fungal protease inhibitors with β-trefoil fold from the mushroom Coprinopsis cinerea, named cocaprins, which inhibit both cysteine and aspartic proteases. Two cocaprin-encoding genes are differentially expressed in fungal tissues. One is highly transcribed in vegetative mycelium and the other in the stipes of mature fruiting bodies. Cocaprins are small proteins (15 kDa) with acidic isoelectric points that form dimers. The three-dimensional structure of cocaprin 1 showed similarity to fungal β-trefoil lectins. Cocaprins inhibit plant C1 family cysteine proteases with K_i in the micromolar range, but do not inhibit the C13 family protease legumain, which distinguishes them from mycocypins. Cocaprins also inhibit the aspartic protease pepsin with K_i in the low micromolar range. Mutagenesis revealed that the β2-β3 loop is involved in the inhibition of cysteine proteases and that the inhibitory reactive sites for aspartic and cysteine proteases are located at different positions on the protein. Their biological function is thought to be the regulation of endogenous proteolytic activities or in defense against fungal antagonists. Cocaprins are the first characterized aspartic protease inhibitors with β-trefoil fold from fungi, and demonstrate the incredible plasticity of loop functionalization in fungal proteins with β-trefoil fold.     

Harnessing real-world depth edges with multiflash imaging

A method for capturing geometric features of real-world scenes relies on a simple capture setup modification. The system might conceivably be packaged into a portable self-contained device. The multiflash imaging method bypasses 3D geometry acquisition and directly acquires depth edges from images. In the place of expensive, elaborate equipment for geometry acquisition, we use a camera with multiple strategically positioned flashes. Instead of having to estimate the full 3D coordinates of points in the scene (using, for example, 3D cameras) and then look for depth discontinuities, our technique reduces the general 3D problem of depth edge recovery to one of 2D intensity edge detection. Our method could, in fact, help improve current 3D cameras, which tend to produce incorrect results near depth discontinuities. Exploiting the imaging geometry for rendering provides a simple and inexpensive solution for creating stylized images from real scenes. We believe that our camera will be a useful tool for professional artists and photographers, and we expect that it will also let the average user easily create stylized imagery. This article is available with a short video documentary on CD-ROM.     

Observations of tropical cyclone structure from WindSat

Passive microwave (PMW) radiometric observations of clouds from multichannel imaging sensors onboard low Earth-orbiting environmental satellites are now a vital operational dataset. The first operational passive microwave sensor was the Special Sensor Microwave/Imager onboard the Defense Meteorological Satellite Program satellites, which has been gathering hydrological data records since 1987, and continued with the Tropical Rainfall Measuring Mission (TRMM) and the Advanced Microwave Scanning Radiometer onboard Aqua. These sensors view the underlying scene with an Earth incidence angle near 53/spl deg/ and with a variable azimuthal angle, depending upon the orbit direction and scan position. The WindSat sensor onboard the Coriolis satellite, launched in January 2003, is a five-channel polarimetric PMW radiometer designed to optimize ocean surface wind vector retrievals. While it does not have 85-GHz channels, an added feature is its unique fore-aft viewing capability across a portion of its fore scan swath. This provides a view of the underlying scene from two separate azimuthal directions, which provides added information on the three-dimensional (3-D) structure of clouds and their evolution. In this paper, we compare WindSat and TRMM Precipiation Radar observations of tropical cyclones (TCs) with Monte Carlo radiative transfer simulations performed on idealized 3-D convective cloud structures. The TC 3-D structure and possible tilt in the convective cloud structure are inferred from the difference between the 37-GHz equivalent blackbody brightness temperatures (T/sub B/) from the corresponding fore and aft view observations. The information gained from this analysis is important since asymmetries in the cloud vertical and horizontal structure may be an indication of upper level wind shear, which plays a major role in influencing changes of the TC intensity.     

The existence of a precursor of cathepsin D: evidence from autolysis, denaturation and activation studies

This report evidences that the single polypeptide chain of cathepsin D undergoes in vitro autolysis resulting in heavy (Mr about 30000) and light (Mr about 15000) polypeptide chains. These two chains are held together through non-covalent interaction, thus constituting a stable active conformation. Fluorescence and circular dichroism measurements demonstrate the irreversible denaturation of cathepsin D. The existence of cathepsin D precursor, cathepsinogen D, of about 50000 molecular weight was proved. Cathepsinogen D is converted to the active enzyme by intramolecular activation, releasing activation-inhibitory peptide(s).     

Applications of microcontinuum fluid mechanics

A comprehensive review of the existing theories of microcontinuum fluid mechanics has been presented by the authors in a previous paper. In this paper the published applications of these microfluid theories to real and ideal flow problems have been reviewed briefly and these short reviews are presented in tabular form for easy reference.     

Rapid isolation of cathepsin D by affinity chromatography on the immobilized synthetic inhibitor

To study the influence of tetanus antitoxins on the outcome of human tatanus, we analyzed data on 545 cases reported to the Center for Disease Control from 1965 through 1971. Patients treated with antitoxin had a significantly lower case-fatality ratio than untreated patients, and the effect of serotherapy was not modified significantly by the age or race of the subjects. Antitoxin of equine origin and human tetanus immune globulin (TIG) were equally effective. The data on the effect of different doses of TIG suggest that 500 units may be as effective as the currently recommended therapeutic dose of 3,000 to 10,000 units. The importance of possible bias introduced by unmeasured factors such as quality of supportive therapy could not be determined, but confounding by 11 recorded potentially confounding factors was minimal.