Chromophore‐Linked Substrate (CLS405): Probing Metallo‐β‐Lactamase Activity and Inhibition

Serine‐ and metallo‐β‐lactamases present a threat to the clinical use of nearly all β‐lactam antibiotics, including penicillins, cephalosporins, and carbapenems. Efforts to develop metallo‐β‐lactamase (MBL) inhibitors require suitable screening platforms to allow the rapid determination of β‐lactamase activity and efficient inhibition. Unfortunately, the platforms currently available are not ideal for this purpose. Further progress in MBL inhibitor identification requires inexpensive and widely applicable assays. Herein the identification of an inexpensive and stable chromogenic substrate suitable for use in assays of clinically relevant MBLs is described. (6R,7R)‐3‐((4‐Nitrophenoxy)methyl)‐8‐oxo‐7‐(2‐phenylacetamido)‐5‐thia‐1‐azabicyclo[4.2.0]oct‐2‐ene‐2‐carboxylic acid 5,5‐dioxide (CLS405) was synthesised in a three‐step protocol. CLS405 was then characterised spectroscopically, and its stability and kinetic properties evaluated. With a Δλ_max value of 100 nm between the parent and hydrolysis product, a higher analytical accuracy is possible with CLS405 than with commonly used chromogenic substrates. The use of CLS405 in assays was validated by MBL activity measurements and inhibitor screening that resulted in the identification of N‐hydroxythiazoles as new inhibitor scaffolds for MBLs. Further evaluation of the identified N‐hydroxythiazoles against a panel of clinically relevant MBLs showed that they possess inhibitory activities in the mid‐ to low‐micromolar range. The findings of this study provide both a useful tool compound for further inhibitor identification, and novel scaffolds for the design of improved MBL inhibitors with potential as antibiotics against resistant strains of bacteria.     

Alanine Scan of the Peptide Antibiotic Feglymycin: Assessment of Amino Acid Side Chains Contributing to Antimicrobial Activity

The antibiotic feglymycin is a linear 13‐mer peptide synthesized by the bacterium Streptomyces sp. DSM 11171. It mainly consists of the nonproteinogenic amino acids 4‐hydroxyphenylglycine and 3,5‐dihydroxyphenylglycine. An alanine scan of feglymycin was performed by solution‐phase peptide synthesis in order to assess the significance of individual amino acid side chains for biological activity. Hence, 13 peptides were synthesized from di‐ and tripeptide building blocks, and subsequently tested for antibacterial activity against Staphylococcus aureus strains. Furthermore we tested the inhibition of peptidoglycan biosynthesis enzymes MurA and MurC, which are inhibited by feglymycin. Whereas the antibacterial activity is significantly based on the three amino acids D ‐Hpg1, L ‐Hpg5, and L ‐Phe12, the inhibitory activity against MurA and MurC depends mainly on L ‐Asp13. The difference in the position dependence for antibacterial activity and enzyme inhibition suggests multiple molecular targets in the modes of action of feglymycin.     

Enhancement of pro‐degradant performance in polyethylene/starch blends as a function of distribution

The effect of pro‐degradant distribution in polyethylene (PE)/starch blends on ultraviolet (UV) photo‐oxidative degradation was investigated. Two kinds of pro‐degradants, Fe and Co‐based, were used in this study. The distribution of pro‐degradants in the different phases was varied by a dual step process using a side‐feed on a reactive extruder. The variation in mechanical properties and evaluation of carbonyl groups by FTIR were conducted to investigate the effect of degradation following exposure to UV photo‐oxidative degradation. It was found that the variation in mechanical properties was higher when the pro‐degradants were distributed in the PE phase. The concentration of carbonyl groups increased as a function of UV exposure, and the concentration of carbonyl groups was higher when the pro‐degradants were distributed in the PE phase. Micro‐cracking was observed on the interface between starch and PE after adding the pro‐degradants. When the pro‐degradants were distributed in high‐density polyethylene (HDPE) phase, the micro‐cracks mainly appeared in HDPE matrix, and the density of micro‐crack was higher. In general, the function of the pro‐degradants in PE/starch blends was enhanced when their distribution was varied within HDPE phase. © 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Influence of precursors on the formation of 3‐MCPD and glycidyl esters in a model oil under simulated deodorization conditions

To find new ways for reducing the potential of palm oil to form 3‐monochloropropane‐1,2‐diol (3‐MCPD) and glycidyl esters during refining it is helpful to know more about the influence of different precursors like diacylglycerols (DAGs) and monoacylglycerols (MAGs), lecithin, and chlorine containing compounds. After adding increasing amounts of the different precursors to a model oil obtained by removal of polar compounds from crude palm oil and heating the mixture under standardized conditions to 240°C for 2 h the contents of 3‐MCPD and glycidyl esters were analyzed according to the standard procedure of DGF C‐VI 18 (10). DAGs and MAGs were found to increase the potential of palm oil to form 3‐MCPD and glycidyl esters, but refined lecithin showed no influence. Sodium chloride as well as tetra‐n‐butylammoniumchloride (TBAC) led to higher contents of the esters. Whereas the addition of TBAC raised the amount of glycidyl esters as well as 3‐MCPD esters, sodium chloride largely raised the amount of 3‐MCPD esters. An addition of 5 mmol of sodium carbonate/kg model oil spiked with sodium chloride reduced the amount of glycidyl esters almost completely; the 3‐MCPD esters were reduced by 50%. About 1 mmol sodium hydrogen carbonate/kg oil reduced both 3‐MCPD and glycidyl esters almost completely. Practical applications : For the mitigation of the formation of 3‐MCPD esters and related compounds in refined edible oils, it is helpful to know more about the effect of different possible precursors. Using a broader data basis, it is possible to adopt the oil processing but especially the choice of the raw material to the demands of the market for lower contents of the esters in the refined products.     

EFFECT OF TEMPERATURE ON THE ADSORPTION OF METHYLENE BLUE DYE ONTO SULFURIC ACID–TREATED ORANGE PEEL

The present study explains the preparation and application of sulfuric acid–treated orange peel (STOP) as a new low-cost adsorbent in the removal of methylene blue (MB) dye from its aqueous solution. The effects of temperature on the operating parameters such as solution pH, adsorbent dose, initial MB dye concentration, and contact time were investigated for the removal of MB dye using STOP. The maximum adsorption of MB dye onto STOP took place in the following experimental conditions: pH of 8.0, adsorbent dose of 0.4 g, contact time of 45 min, and temperature of 30°C. The adsorption equilibrium data were tested by applying both the Langmuir and Freundlich isotherm models. It is observed that the Freundlich isotherm model fitted better than the Langmuir isotherm model, indicating multilayer adsorption, at all studied temperatures. The adsorption kinetic results showed that the pseudo-second-order model was more suitable to explain the adsorption of MB dye onto STOP. The adsorption mechanism results showed that the adsorption process was controlled by both the internal and external diffusion of MB dye molecules. The values of free energy change (ΔG ^o) and enthalpy change (ΔH ^o) indicated the spontaneous, feasible, and exothermic nature of the adsorption process. The maximum monolayer adsorption capacity of STOP was also compared with other low-cost adsorbents, and it was found that STOP was a better adsorbent for MB dye removal.     

Dendrimers or Nanoparticles as Supports for the Design of Efficient and Recoverable Organocatalysts?

The Jørgensen–Hayashi catalyst [(S)‐α,α‐diphenylprolinol trimethylsilyl ether] was grafted onto the surface of two different supports: phosphorus dendrimers (generations 1 to 3) and magnetic, polymer‐coated cobalt/carbon (Co/C) nanobeads. These new supported catalysts displayed high activities and selectivities in the Michael additions of a wide range of aldehydes to different nitroolefins. Moreover, the dendrimer of the third generation displayed excellent recycling abilities since it could be recovered and reused in 7 consecutive runs without loss of activity.     

Processing and properties of transparent hydroxyapatite and β tricalcium phosphate obtained by HIP process

Stoichiometric β-tricalcium phosphate (β-TCP) and hydroxyapatite (HA) powders were synthesized by chemical precipitation of aqueous solutions of diammonium phosphate and calcium nitrate. After a calcination treatment and a milling step, the powders were shaped by slip casting. The sintering temperature effect on the relative density and the average grain size was investigated. By natural sintering at 1200 and 1120 °C, densities of 98% and 99% were obtained for HA and TCP, respectively. After determination of minimum temperatures to obtain only closed porosity and a pre-sintering at these temperatures, hot isostatic pressing (HIP) treatment was carried out. Transparent or translucent samples were obtained, indicating a relative density very close to the theoretical value (>99.9%). Mechanical properties (three-point bending strength, fracture toughness, Young's modulus and Vickers hardness) were measured on both materials with similar grain size (～ 1 μm). Bending strengths of 181 and 105 MPa were measured for TCP and HA, respectively.     

Associations between apolipoprotein E genotype and circulating F<Subscript>2</Subscript>-isoprostane levels in humans

Apolipoprotein E (apoE), an important determinant of plasma lipoprotein metabolism, has three common alleles (ε2, ε3, and ε4). Population studies have shown that the risk of diseases characterized by oxidative damage, such as coronary heart disease and Alzheimer’s disease, is significantly higher in ɛ4 carriers. We evaluated the association between apoE genotypes and plasma F₂-isoprostane levels, an index of lipid peroxidation, in humans. Two hundred seventy-four healthy subjects (104 males, 170 females; 46.9±13.0 yr; 200 whites, 74 blacks; 81 nonsmokers, 64 passive smokers, and 129 active smokers) recruited for a randomized clinical antioxidant intervention trial were included in this analysis. ApoE genotype was determined by PCR and restriction enzyme digestion. Free plasma F₂-isoprostane was measured by GC-MS. Genotype groups were compared using multiple regression analysis with adjustment for sex, age, race, smoking status, body mass index, plasma ascorbic acid, and β-carotene. Subjects with ε3/ε4 and ε4/ε4 genotype (ε4-carriers) and with ε2/ε3 and ε3/ε3 (non-ε4-carriers) were pooled for analysis. In subjects with high cholesterol levels (total cholesterol above 200 mg/dl), plasma F₂-isoprostane levels were 29% higher in ε4 carriers than in non-ε4-carriers (P=0.0056). High-cholesterol subjects that are ε4 carriers have significantly higher levels of lipid peroxidation as assessed by circulating F₂-isoprostane levels.