Positive association of renal insufficiency with agriculture employment and unregulated alcohol consumption in Nicaragua

Endemic renal insufficiency (RI) of unknown etiology is a major public health issue with high mortality in the Pacific coastal regions of Central America. We studied RI in León and Chinandega, Nicaragua, evaluating associations with known risk factors and hypothesized exposures

A Simple Method of Hydrogen Insertion in Transition Metal Oxides to Obtain Bronzes

Reaction of $WO_3$, $MoO_3$ and $V_2O_5$ with alcohols or glycols is shown to yield hydrogen bronzes of the oxides. Cubic and tetragonal $H_XWO_3$, $H_{0.47}MoO_3$ as well as $H_{0.4}V_2O_5$ have been prepared by this method

PH modulation method to detect ligand-receptor binding

The present disclosure relates to detecting receptor-ligand binding by measuring local pH modulation using a pH-sensitive fluorophore.U

PH modulation method to detect ligand-receptor binding

The present disclosure relates to detecting receptor-ligand binding by measuring local pH modulation using a pH-sensitive fluorophore.U

Detecting Protein−Ligand Binding on Supported Bilayers by Local pH Modulation

Herein, we describe a highly sensitive technique for detecting protein−ligand binding at the liquid/solid interface. The method is based upon modulation of the interfacial pH when the protein binds. This change is detected by ortho-Texas Red DHPE, which is doped into supported phospholipid bilayers and used as a pH-sensitive dye. The dye molecule fluoresces strongly at acidic pH values but not basic ones and has an apparent pKA of 7.8 in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine membranes containing 0.5 mol % biotin-cap-PE. This method was used to detect antibiotin/biotin binding interactions as well as the binding of cholera toxin B subunits to GM1. Since these proteins are negatively charged under the conditions of the experiment the interface became slightly more acidic upon binding. In each case, the equilibrium dissociation constant was determined by following the rise in fluorescence as protein was introduced. This change is essentially linear with protein coverage under the conditions employed. For the biotin/antibiotin system it was determined that KD = 24 ± 5 nM, which is in excellent agreement with classical measurements made by total internal reflection fluorescence microscopy involving fluorophore-conjugated antibody molecules. Moreover, the limit of detection was ∼350 fM at the 99% confidence level. This corresponds to 1 part in 69 000 of the KD value. Such a finding compares favorably with surface plasmon resonance studies of similar systems and conditions. The assay could be run in imaging mode to obtain multiple simultaneous measurements using a CCD camera

Impact of Hapten Presentation on Antibody Binding at Lipid Membrane Interfaces

AbstractWe report the effects of ligand presentation on the binding of aqueous proteins to solid supported lipid bilayers. Specifically, we show that the equilibrium dissociation constant can be strongly affected by ligand lipophilicity and linker length/structure. The apparent equilibrium dissociation constants (KD) were compared for two model systems, biotin/anti-biotin and 2,4-dinitrophenyl (DNP)/anti-DNP, in bulk solution and at model membrane surfaces. The binding constants in solution were obtained from fluorescence anisotropy measurements. The surface binding constants were determined by microfluidic techniques in conjunction with total internal reflection fluorescence microscopy. The results showed that the bulk solution equilibrium dissociation constants for anti-biotin and anti-DNP were almost identical, KD(bulk)=1.7±0.2 nM vs. 2.9±0.1 nM. By contrast, the dissociation constant for anti-biotin antibody was three orders of magnitude tighter than for anti-DNP at a lipid membrane interface, KD=3.6±1.1 nM vs. 2.0±0.2 μM. We postulate that the pronounced difference in surface binding constants for these two similar antibodies is due to differences in the ligands’ relative lipophilicity, i.e., the more hydrophobic DNP molecules had a stronger interaction with the lipid bilayers, rendering them less available to incoming anti-DNP antibodies compared with the biotin/anti-biotin system. However, when membrane-bound biotin ligands were well screened by a poly(ethylene glycol) (PEG) polymer brush, the KD value for the anti-biotin antibody could also be weakened by three orders of magnitude, 2.4±1.1μM. On the other hand, the dissociation constant for anti-DNP antibodies at a lipid interface could be significantly enhanced when DNP haptens were tethered to the end of very long hydrophilic PEG lipopolymers (KD=21±10nM) rather than presented on short lipid-conjugated tethers. These results demonstrate that ligand presentation strongly influences protein interactions with membrane-bound ligands

Towards complete specifications with an error calculus

Abstract. We present an error calculus to support a novel specification mechanism for sound and/or complete safety properties that are to be given by users. With such specifications, our calculus can form a foundation for both proving program safety and/or discovering real bugs. The basis of our calculus is an algebra with a lattice domain of four abstract statuses (namely unreachability, validity, must-error and may-error) on possible program states and four operators for this domain to calculate suitable program status. We show how proof search and error localization can be supported by our calculus. Our calculus can also be extended to separation logic with support for user-defined predicates and lemmas. We have implemented our calculus in an automated verification tool for pointer-based programs. Initial experiments have confirmed that it can achieve the dual objectives, namely of safety proving and bug finding, with modest overheads.

SSD1 Is Integral to Host Defense Peptide Resistance in Candida albicans▿

Candida albicans is usually a harmless human commensal. Because inflammatory responses are not normally induced by colonization, antimicrobial peptides are likely integral to first-line host defense against invasive candidiasis. Thus, C. albicans must have mechanisms to tolerate or circumvent molecular effectors of innate immunity and thereby colonize human tissues. Prior studies demonstrated that an antimicrobial peptide-resistant strain of C. albicans, 36082R, is hypervirulent in animal models versus its susceptible counterpart (36082S). The current study aimed to identify a genetic basis for antimicrobial peptide resistance in C. albicans. Screening of a C. albicans genomic library identified SSD1 as capable of conferring peptide resistance to a susceptible surrogate, Saccharomyces cerevisiae. Sequencing confirmed that the predicted translation products of 36082S and 36082R SSD1 genes were identical. However, Northern analyses corroborated that SSD1 is expressed at higher levels in 36082R than in 36082S. In isogenic backgrounds, ssd1Δ/ssd1Δ null mutants were significantly more susceptible to antimicrobial peptides than parental strains but had equivalent susceptibilities to nonpeptide stressors. Moreover, SSD1 complementation of ssd1Δ/ssd1Δ mutants restored parental antimicrobial peptide resistance phenotypes, and overexpression of SSD1 conferred enhanced peptide resistance. Consistent with these in vitro findings, ssd1 null mutants were significantly less virulent in a murine model of disseminated candidiasis than were their parental or complemented strains. Collectively, these results indicate that SSD1 is integral to C. albicans resistance to host defense peptides, a phenotype that appears to enhance the virulence of this organism in vivo