Glucose and other reducing sugars react with proteins by a nonenzymatic, post-translational modification process called nonenzymatic glycosylation or glycation. The sugar-derived carbonyl group adds to a free amine, forming a reversible adduct which over time rearranges to produce a class of products termed advanced-glycation end-products (AGEs). These remain irreversibly bound to macromolecules and can covalently crosslink proximate amino groups. The formation of AGEs on long-lived connective tissue and matrix components accounts largely for the increase in collagen crosslinking that accompanies normal ageing and which occurs at an accelerated rate in diabetes. AGEs can activate cellular receptors and initiate a variety of pathophysiological responses. They modify an appreciable fraction of circulating low-density lipoproteins preventing uptake of these particles by their high-affinity tissue receptors. Advanced glycation has also been implicated in the pathology of Alzheimer's disease. Because AGEs may form by a pathway involving reactive alpha-dicarbonyl intermediates, we investigated a potential pharmacological strategy for selectively cleaving the resultant glucose-derived protein crosslinks. We now describe a prototypic AGE crosslink 'breaker', N-phenacylthiazolium bromide (PTB), which reacts with and cleaves covalent, AGE-derived protein crosslinks. The ability of PTB to break AGE crosslinks in vivo points to the importance of an alpha-dicarbonyl intermediate in the advanced glycation pathway and offers a potential therapeutic approach for the removal of established AGE crosslinks.

The microreactor technology is an efficient tool for kilogramscale syntheses in continuous mode and is particularly effective for hazardous reactions that do not allow scale-up in conventional reactors. Applications to several classes of reactions including highly exothermic reactions, high-temperature reactions, reactions with unstable intermediates, and reactions involving hazardous reagents are described herein.

SummaryAn overproduction ofproinflammatory cytokines by activated macrophages/monocytes mediates the injurious sequelae of inflammation, septic shock, tissue injury, and cachexia. We recently synthesized a tetravalent gnanylhydrazone compound (CNI-1493) that inhibits cytokineinducible arginine transport and nitric oxide (NO) production in macrophages, and protects mice against lethal endotoxemia and carrageenan-induced inflammation. During these investigations we noticed that CNI-1493 effectively prevented lipopolysaccharide (LPS)-induced NO production, even when added in concentrations 10-fold less than required to competitively inhibit L-arginine uptake, suggesting that the suppressive effects of this guanylhydrazone compound might extend to other LPS-induced responses. Here, we report that CNI-1493 suppressed the LPS-stimulated production of proinflammatory cytokines (tumor necrosis factor [TNF], interleukins 1[3 and 6, macrophage inflammatory proteins lcx and 1 [3) from human peripheral blood mononuclear cells. Cytokine suppression was specific, in that CNI-1493 did not inhibit either the constitutive synthesis of transforming growth factor [3 or the upregnlation of major histocompatibility complex class II by interferon ~/ (IFN-~/). In contrast to the macrophage suppressive actions of dexamethasone, which are overridden in the presence of IFN-% CNI-1493 retained its suppressive effects even in the presence of IFN-% The mechanism of cytokine-suppressive action by CNI-1493 was independent of extracellular L-arginine content and NO production and is not restricted to induction by LPS. As a selective inhibitor of macrophage activation that prevents TNF production, this tetravalent guanylhydrazone could be useful in the development ofcytokine-suppressive agents for the treatment of diseases mediated by overproduction of cytokines.

The standard method for preparing carboxylic acid hydrazides is hydrazinolysis of esters in alcoholic solutions. However, when applied to alpha,beta-unsaturated esters, the main product typically is the pyrazolidinone resulting from an undesired Michael-type cyclization. Other alternative methodologies reported for direct preparation of hydrazides from acids are inefficient. We developed an efficient and general process, involving preforming activated esters and/or amides followed by reaction with hydrazine, for the preparation of hydrazides including those of alpha,beta-unsaturated acids. This process gives the desired hydrazides in excellent yield and purity under mild conditions.

This study aimed to examine the effects of single-session anodal high-definition transcranial direct current stimulation (HD-tDCS) on the strength of intrinsic foot muscles, passive ankle kinesthesia, and static balance. Methods: In this double-blinded self-controlled study, 14 healthy younger adults were asked to complete assessments of foot muscle strength, passive ankle kinesthesia, and static balance before and after a 20-minute session of either HD-tDCS or sham stimulation (i.e., control) at two visits separated by one week. Two-way repeated-measures analysis of variance was used to examine the effects of HD-tDCS on metatarsophalangeal joint flexor strength, toe flexor strength, the passive kinesthesia threshold of ankle joint, and the average sway velocity of the center of gravity. Results: All participants completed all study procedures and no side effects nor risk events were reported. Blinding was shown to be successful, with an overall accuracy of 35.7% in the guess of stimulation type (p = 0.347). No main effects of intervention, time, or their interaction were observed for foot muscle strength (p > 0.05). The average percent change in first-toe flexor strength following anodal HD-tDCS was 12.8 ± 24.2%, with 11 out of 14 participants showing an increase in strength, while the change following sham stimulation was 0.7 ± 17.3%, with 8 out of 14 participants showing an increase in strength. A main effect of time on the passive kinesthesia threshold of ankle inversion, dorsiflexion, and anteroposterior and medial-lateral average sway velocity of the center of gravity in one-leg standing with eyes closed was observed; these outcomes were reduced from pre to post stimulation (p < 0.05). No significant differences were observed for other variables between the two stimulation types. Conclusion: The results of this pilot study suggested that single-session HD-tDCS may improve the flexor strength of the first toe, although no statistically significant differences were observed between the anodal HD-tDCS and sham procedure groups. Additionally, passive ankle kinesthesia and static standing balance performance were improved from pre to post stimulation, but no significant differences were observed between the HD-tDCS and sham procedure groups. This may be potentially due to ceiling effects in this healthy cohort of a small sample size. Nevertheless, these preliminary findings may provide critical knowledge of optimal stimulation parameters, effect size, and power estimation of HD-tDCS for future trials aiming to confirm and expand the findings of this pilot study.