Exploitation of structural and regulatory diversity in glutamate racemases

Glutamate racemase is an enzyme essential to the bacterial cell wall biosynthesis pathway, and has therefore been considered as a target for antibacterial drug discovery. We characterized the glutamate racemases of several pathogenic bacteria using structural and biochemical approaches. Here we describe three distinct mechanisms of regulation for the family of glutamate racemases: allosteric activation by metabolic precursors, kinetic regulation through substrate inhibition, and D-glutamate recycling using a d-amino acid transaminase. In a search for selective inhibitors, we identified a series of uncompetitive inhibitors specifically targeting Helicobacter pylori glutamate racemase that bind to a cryptic allosteric site, and used these inhibitors to probe the mechanistic and dynamic features of the enzyme. These structural, kinetic and mutational studies provide insight into the physiological regulation of these essential enzymes and provide a basis for designing narrow-spectrum antimicrobial agents.     

Adrenergic reinnervation of the denervated rat urinary bladder.

In rats undergoing unilateral extirpation of the pelvic ganglion, the adrenergic innervation disappeared on the ipsilateral side of the urinary bladder. It had reappeared after 6--9 weeks.     

The effect of hexosamine inhibitors on the secretion ofγ-globulin

Zusammenfassung Hemmung von Hexosamin vermag die Sekretion von-Globulin durch antikörperproduzierende Zellen unter in vitro Bedingungen nicht zu verhindern. Dies könnte darauf hinweisen, dass unter den gewählten Bedingungen kohlehydratfreie-Globulinmoleküle sezerniert werden.     

Adrenergic reinnervation of the denervated rat urinary bladder

Summary In rats undergoing unilateral extirpation of the pelvic ganglion, the adrenergic innervation disappeared on the ipsilateral side of the urinary bladder. It had reappeared after 6–9 weeks.This work was supported by grants from the Faculty of Medicine in Lund to M.E.     

Tribological behavior of CrN-coated Cr-Mo-V steels used as die materials

DIN 1.2343 and 1.2367 steels are commonly used as die materials in aluminum extrusion, and single/duplex/multi-coatings enhance their surface properties. The design of an appropriate substrate/coating system is important for improving the tribological performance of these steels under service conditions because the load-carrying capacity of the system can be increased by decreasing the plastic deformation of the substrate. In this study, the tribological behavior of CrN-coated Cr-Mo-V steels (DIN 1.2343, 1.2367, and 1.2999 grades) was investigated using different setups and tribological pairs at room and elevated temperatures. The aim of this study was to reveal the wear resistance of a suggested system (1.2999/CrN) not yet studied and to understand both the wear and the failure characteristics of coated systems. The results showed that (i) among the steels studied, the DIN 1.2999 grade steel exhibited the lowest friction coefficient because it had the highest load-carrying capacity as a result of secondary hardening at elevated temperatures; (ii) at room temperature, both abrasive tracks and adhesive layers were observed on the worn surfaces; and (iii) a combination of chemical reactions and progressive oxidation caused aluminum adhesion on the worn surface, and the detachment of droplets and microcracking were the characteristic damage mechanisms at high temperatures.     

Determination of discontinuities in marble blocks via a nondestructive ultrasonic technique

Miners working in the marble industry have always been interested in identifying structural weaknesses in marble blocks before they are transported to marble processing plants. To achieve this difficult task, several simple methods have been developed among miners but observation-based methods do not consistently provide satisfactory results. A nondestructive method developed for testing concrete could be used for this purpose. In this study, this simple method based on differences in ultrasonic wave propagation in different materials was presented, and the test results performed both in the laboratory and a marble quarry were discussed.     

Mutations in kelch-like 3 and cullin 3 cause hypertension and electrolyte abnormalities

Hypertension affects one billion people and is a principal reversible risk factor for cardiovascular disease. Pseudohypoaldosteronism type II (PHAII), a rare Mendelian syndrome featuring hypertension, hyperkalaemia and metabolic acidosis, has revealed previously unrecognized physiology orchestrating the balance between renal salt reabsorption and K(+) and H(+) excretion. Here we used exome sequencing to identify mutations in kelch-like 3 (KLHL3) or cullin 3 (CUL3) in PHAII patients from 41 unrelated families. KLHL3 mutations are either recessive or dominant, whereas CUL3 mutations are dominant and predominantly de novo. CUL3 and BTB-domain-containing kelch proteins such as KLHL3 are components of cullin-RING E3 ligase complexes that ubiquitinate substrates bound to kelch propeller domains. Dominant KLHL3 mutations are clustered in short segments within the kelch propeller and BTB domains implicated in substrate and cullin binding, respectively. Diverse CUL3 mutations all result in skipping of exon 9, producing an in-frame deletion. Because dominant KLHL3 and CUL3 mutations both phenocopy recessive loss-of-function KLHL3 mutations, they may abrogate ubiquitination of KLHL3 substrates. Disease features are reversed by thiazide diuretics, which inhibit the Na-Cl cotransporter in the distal nephron of the kidney; KLHL3 and CUL3 are expressed in this location, suggesting a mechanistic link between KLHL3 and CUL3 mutations, increased Na-Cl reabsorption, and disease pathogenesis. These findings demonstrate the utility of exome sequencing in disease gene identification despite the combined complexities of locus heterogeneity, mixed models of transmission and frequent de novo mutation, and establish a fundamental role for KLHL3 and CUL3 in blood pressure, K(+) and pH homeostasis.