An Atomistic Understanding of Allosteric Inhibition of Glutamate Racemase: a Dampening of Native Activation Dynamics

Glutamate racemases (GR) are members of the family of bacterial enzymes known as cofactor-independent racemases and epimerases and catalyze the stereoinversion of glutamate. D-amino acids are universally important for the proper construction of viable bacterial cell walls, and thus have been repeatedly validated as attractive targets for novel antimicrobial drug design. Significant aspects of the mechanism of this challenging stereoinversion remain unknown. The current study employs a combination of MD and QM/MM computational approaches to show that the GR from H. pylori must proceed via a pre-activation step, which is dependent on the enzyme's flexibility. This mechanism is starkly different from previously proposed mechanisms. These findings have immediate pharmaceutical relevance, as the H. pylori GR enzyme is a very attractive allosteric drug target. The results presented in this study offer a distinctly novel understanding of how AstraZeneca's lead series of inhibitors cripple the H. pylori GR's native motions, via prevention of this critical chemical pre-activation step. Our experimental studies, using SPR, fluorescence and NMR WaterLOGSY, show that H. pylori GR is not inhibited by the uncompetitive mechanism originally put forward by Lundqvist et al.. The current study supports a deep connection between native enzyme motions and chemical reactivity, which has strong relevance to the field of allosteric drug discovery.     

Studies on the O/C-Arylation of Cyclic β-Diketones with activated aryl fluorides

The reaction of nitrosubstituted aryl fluorides 1 with cyclic β-diketones 2 proceeds at 20–80°C in the presence of bases, such as KOH, KF or NaOEt, leading to the aryl ethers 3a – m . Depending on the base the reaction of dimedone 2a or 1,3-cyclohexanedione 2b with 2,4-dinitrofluorobenzene 1a or 4-fluoro-3-nitrobenzonitrile 1b furnishes C- and/or O-arylated products. Upon heating of 3g , e and 3m at 40–100°C in DMF/K₂CO₃, the C-arylated ketones 4a – c are formed in good yields. Starting from 3a we obtained the chromenedione 5 under the same conditions.     

Zur heterogenkatalytischen Oxydation und Ammoxydation von Isobuten. IV. Über die Beeinflussung der heterogenkatalytischen Oxydation von Isobuten zu Methacrolein durch n-Buten

On the Heterogeneous-Catalytic Oxidation and Ammoxidation of Isobutene. IV. Influence of n-Butene on the Heterogeneous-Catalytic Oxidation of Isobutene to Methacroleine The influence of n-butenes 2 on the oxidation of isobutenes 1 to methacroleine 3 has been investigated in presence of a catalyst containing Bi, Mo, P, Fe, Mg, Mn, Si and O. Addition of 2 to the gas mixture increase the selectivity and yield of 3 . This fact can be correlated by decrease of oxygen partial pressure in consequence of the dehydrogenation of 2 to butadiene 4 .     

Alkylation of Staurosporine to Derive a Kinase Probe for Fluorescence Applications

The natural product staurosporine is a high‐affinity inhibitor of nearly all mammalian protein kinases. The labelling of staurosporine has proven effective as a means of generating protein kinase research tools. Most tools have been generated by acylation of the 4′‐methylamine of the sugar moiety of staurosporine. Herein we describe the alkylation of this group as a first step to generate a fluorescently labelled staurosporine. Following alkylation, a polyethylene glycol linker was installed, allowing subsequent attachment of fluorescein. We report that this fluorescein–staurosporine conjugate binds to cAMP‐dependent protein kinase in the nanomolar range. Furthermore, its binding can be antagonised with unmodified staurosporine as well as ATP, indicating it targets the ATP binding site in a similar fashion to native staurosporine. This reagent has potential application as a screening tool for protein kinases of interest.     

A Boronic Acid Assay for the Detection of Mucin‐1 Glycoprotein from Cancer Cells

Cell surface glycoproteins are commonly aberrant in disease and act as biomarkers that facilitate diagnostics. Mucin‐1 (MUC1) is a prominent example, exhibiting truncated glycosylation in cancer. We present herein a boronic acid microplate assay for sensitive and high‐throughput detection of such glycoproteins. The immobilization of biotin–boronic acid 1 onto streptavidin plates generated a multivalent surface for glycoprotein recruitment and detection. We first validated the binding properties of 1 in solution through titrations with alizarin dye. Next, the microplate assay was explored through horseradish peroxidase (HRP) analysis as a proof‐of‐concept glycoprotein with chemiluminescence detection. Finally, this platform was applied for the detection of MUC1 directly from MCF‐7 human breast cancer cell lysates by using an HRP‐tagged antibody that targets the cancerous form of this glycoprotein. Sensitive, dose‐dependent detection of MUC1 was observed, showcasing the efficacy of this platform for detecting disease‐associated glycoproteins.     

Spaltung von 1,2,4-Thiadiazol-3-onen mit Triphenylphosphan: Triphenylphosphonio-thioimidazolate und ihre Folgereaktionen

Fission of 1,2,4-Thiadiazol-3-ones by Triphenylphosphane: gewidinet Triphenylphosphonio Thioimidazolates and Their Consecutive Reactions Treatment of benzimidazo[1,2-d][1,2,4]thiadiazol-3(2H)-ones ( 1 ) and imidazo[1,2-d]-[1,2,4]thiadiazol-3(2H)-ones ( 17 ) with triphenylphosphan leads to the liberation of isocyanate and the formation of triphenylphosphonio-thiobenzimidazolat ( 4 ) and triphenylphosphonio-thioimidazolat ( 18 ), respectively. 2,4-Diphenyl-5-phenylimino-1,2,4-thiadiazol-3-one ( 23 ) is desulfurized with Ph₃P and decomposed into phenyl isocyanate and diphenylcarbodiimide whereas the N-unsubstituted imino-thiadiazol-3-one ( 25 ) is attacked at the exocyclic imino group by Ph₃P to give the N-phosphoranylidene thiourea ( 26 ). The structure of 4 can be rationalized as a dynamic system between polar and apolar valence isomeres. Reactions of 1 and 17 with cyanates, isocyanates, carbon disulfide, acetic anhydride, amines, benzyl chloride, grignard compounds, and CH acidic compounds are described.     

The Impact of Early Saliva Interaction on Dental Implants and Biomaterials for Oral Regeneration: An Overview

The presence of saliva in the oral environment is relevant for several essential health processes. However, the noncontrolled early saliva interaction with biomaterials manufactured for oral rehabilitation may generate alterations in the superficial properties causing negative biological outcomes. Therefore, the present review aimed to provide a compilation of all possible physical–chemical–biological changes caused by the early saliva interaction in dental implants and materials for oral regeneration. Dental implants, bone substitutes and membranes in dentistry possess different properties focused on improving the healing process when in contact with oral tissues. The early saliva interaction was shown to impair some positive features present in biomaterials related to quick cellular adhesion and proliferation, such as surface hydrophilicity, cellular viability and antibacterial properties. Moreover, biomaterials that interacted with contaminated saliva containing specific bacteria demonstrated favorable conditions for increased bacterial metabolism. Additionally, the quantity of investigations associating biomaterials with early saliva interaction is still scarce in the current literature and requires clarification to prevent clinical failures. Therefore, clinically, controlling saliva exposure to sites involving the application of biomaterials must be prioritized in order to reduce impairment in important biomaterial properties developed for rapid healing.     

The Fight against Cancer by Microgravity: The Multicellular Spheroid as a Metastasis Model

Cancer is a disease exhibiting uncontrollable cell growth and spreading to other parts of the organism. It is a heavy, worldwide burden for mankind with high morbidity and mortality. Therefore, groundbreaking research and innovations are necessary. Research in space under microgravity (µg) conditions is a novel approach with the potential to fight cancer and develop future cancer therapies. Space travel is accompanied by adverse effects on our health, and there is a need to counteract these health problems. On the cellular level, studies have shown that real (r-) and simulated (s-) µg impact survival, apoptosis, proliferation, migration, and adhesion as well as the cytoskeleton, the extracellular matrix, focal adhesion, and growth factors in cancer cells. Moreover, the µg-environment induces in vitro 3D tumor models (multicellular spheroids and organoids) with a high potential for preclinical drug targeting, cancer drug development, and studying the processes of cancer progression and metastasis on a molecular level. This review focuses on the effects of r- and s-µg on different types of cells deriving from thyroid, breast, lung, skin, and prostate cancer, as well as tumors of the gastrointestinal tract. In addition, we summarize the current knowledge of the impact of µg on cancerous stem cells. The information demonstrates that µg has become an important new technology for increasing current knowledge of cancer biology.