Aerobic Iron‐Based Cross‐Dehydrogenative Coupling Enables Efficient Diversity‐Oriented Synthesis of Coumestrol‐Based Selective Estrogen Receptor Modulators

An iron‐based cross‐dehydrogenative coupling (CDC) approach was applied for the diversity‐oriented synthesis of coumestrol‐based selective estrogen receptor modulators (SERMs), representing the first application of CDC chemistry in natural product synthesis. The first stage of the two‐step synthesis of coumestrol involved a modified aerobic oxidative cross‐coupling between ethyl 2‐(2,4‐dimethoxybenzoyl)acetate and 3‐methoxyphenol, with FeCl₃ (10 mol %) as the catalyst. The benzofuran coupling product was then subjected to sequential deprotection and lactonization steps, affording the natural product in 59 % overall yield. Based on this new methodology other coumestrol analogues were prepared, and their effects on the proliferation of the estrogen receptor (ER)‐dependent MCF‐7 and of the ER‐independent MDA‐MB‐231 breast cancer cells were tested. As a result, new types of estrogen receptor ligands having an acetamide group instead of the 9‐hydroxyl group of coumestrol were discovered. Both 9‐acetamido‐coumestrol and 8‐acetamidocoumestrol were found more active than the natural product against estrogen‐dependent MCF‐7 breast cancer cells, with IC₅₀ values of 30 and 9 nM , respectively.     

Intrinsic fluorescence polarization of amniotic fluid: II. Toward a noninvasive method for the determination of fetal lung maturity

The present study compares two methods for the determination of fetal lung maturity: the novel intrinsic fluorescence polarization ratio (IFPR) and the commercial TDx-FLMII. Amniotic fluid (AF) samples were collected from 69 women during the second and third trimesters of singleton pregnancies. Thirty-three samples were tested for IFPR only after centrifugation, and the rest were examined both before and after centrifugation. Of the latter 33 samples, 29 were assessed for lung maturity with the TDx-FLMII method as well. The results showed that IFPR values decreased with the advance in gestational age (r = 0.77, p < 0.05, n = 69). A significant correlation was found between IFPR of centrifuged and noncentrifuged samples (r = 0.94, p < 0.05, n = 36). A significant correlation was demonstrated between IFPR and TDx-FLMII values of centrifuged (r = 0.75, p < 0.05, n = 29) and noncentrifuged (r = 0.63, p < 0.05, n = 29) samples and moreover, samples considered mature by TDx-FLMII had low values of IFPR (n = 10). It can be concluded that the IFPR method can utilize noncentrifuged AF, thus suggested as a potential noninvasive method.     

Reduced Endocannabinoid Tone in Saliva of Chronic Orofacial Pain Patients

Background: the endocannabinoid system (ECS) participates in many physiological and pathological processes including pain generation, modulation, and sensation. Its involvement in chronic orofacial pain (OFP) in general, and the reflection of its involvement in OFP in salivary endocannabinoid (eCBs) levels in particular, has not been examined. Objectives: to evaluate the association between salivary (eCBs) levels and chronic OFP. Methods: salivary levels of 2 eCBs, anandamide (AEA), 2-arachidonoylglycerol (2-AG), 2 endocannabinoid-like compoundsN-palmitoylethanolamine (PEA), N-oleoylethanolamine (OEA), and their endogenous precursor and breakdown product, arachidonic acid (AA), were analyzed using liquid chromatography/tandem mass spectrometry in 83 chronic OFP patients and 43 pain-free controls. The chronic OFP patients were divided according to diagnosis into musculoskeletal, neurovascular/migraine, and neuropathic pain types. Results: chronic OFP patients had lower levels of OEA (p = 0.02) and 2-AG (p = 0.01). Analyzing specific pain types revealed lower levels of AEA and OEA in the neurovascular group (p = 0.04, 0.02, respectively), and 2-AG in the neuropathic group compared to controls (p = 0.05). No significant differences were found between the musculoskeletal pain group and controls. Higher pain intensity was accompanied by lower levels of AA (p = 0.028), in neuropathic group. Conclusions: lower levels of eCBs were found in the saliva of chronic OFP patients compared to controls, specifically those with neurovascular/migraine, and neuropathic pain. The detection of changes in salivary endocannabinoids levels related to OFP adds a new dimension to our understanding of OFP mechanisms, and may have diagnostic as well as therapeutic implications for pain.     

Photosensitizer–Antibiotic Conjugates: A Novel Class of Antibacterial Molecules

Bacterial resistance to a variety of antibiotics has led to intensive research into the effect of photosensitizers as a cytotoxic agent against bacterial cells. In this study, we synthesized the following conjugates with or without a linker: rose bengal-penicillanic acid (RBPA), rose bengal-linker-penicillanic acid (RBLPA) and rose bengal-linker-kanamycin (RBLKAN). The antibacterial activity of these conjugates was examined on Staphylococcus aureus and Escherichia coli. Exposure of the cultures to 100 J cm⁻² showed that the minimum inhibitory concentration (MIC) of RBPA, RBLPA and RBLKAN on S. aureus was 0.195, 0.156 and 0.004 μm, respectively. The MIC of RBPA, RBLPA and RBLKAN on E. coli was 1.56, 2.5 and 0.156 μm, respectively. In dark control experiments, the MIC of these conjugates was not detected until a concentration that was 16-fold that of the MIC found in the light experiments. RBPA and RBLPA as well as RBLKAN are bactericidal for both bacterial cells. Total eradication of S. aureus and E. coli was observed with RBLKAN (0.078 and 20 μm 16 J cm⁻², respectively). Under these conditions, scanning electron microscopic analysis showed significant damage to these bacteria. However, the photosensitizer and antibiotics individually were not effective.     

The contribution of the accreditation process to public administration: promotion of vital organizational changes

Accumulative evidence suggests that the implementation of international standard operating procedures, induce, by their virtues, major organizational changes. These changes are both cultural and behavioral including changes in decision-making processes, organizational norms, and values. The decision of any organization to adopt quality assurance, specifically via accreditation, should be regarded as a strategic one. As such, it prerequisites top management commitment, budgeting the process and promoting active inter-collaboration of all organizational members. The accreditation process according to ISO/IEC 17025:1999 of the Haifa Public Health Laboratory, will be used as a case study. A number of practical benefits of such organizational changes will be demonstrated and discussed, among them improved quality customer service and handling customer's complaints, establishing multichannel communication, enhanced interlaboratory collaboration and coordination. It is important to note that the accreditation process was accompaniment by extensive training of both management and employees on-site and off-site.     

Large-scale analysis of posttranslational modifications in the hippocampus of patients with Alzheimer’s disease using pI shift and label-free quantification without enrichment

Posttranslational modifications modulate protein function in cells. Global analysis of multiple posttranslational modifications can provide insight into physiology and disease, but presents formidable challenges. In the present study, we used a technique that does not require target enrichment to analyze alterations in the phosphorylation and ubiquitination of proteins from patients with Alzheimer’s disease (AD). Guided by our previous findings, we applied three strategies to further our understanding of the dysregulation of posttranslationally modified proteins. We first identified phosphorylation sites by determining peptide pI shifts using OFFGEL. Second, using tandem mass spectrometry, we determined the ubiquitination status of the proteins using an assay for a trypsin digestion remnant of ubiquitination (Gly-Gly). Third, for large-scale discovery, we quantified the global differences in protein expression. Of the proteins expressed in AD tissue at levels of 2.0 or greater compared with controls, 60 were phosphorylated and 56 were ubiquitinated. Of the proteins expressed at levels of 0.5 or lower compared with controls, 81 were phosphorylated and 56 were ubiquitinated. Approximately 98 % of the phosphopeptides exhibited a pI shift. We identified 112 new phosphorylation sites (51.38 %), and 92 new ubiquitination sites (96.84 %). Taken together, our findings suggest that analysis of the alterations in posttranslationally modified proteins may contribute to understanding the pathogenesis of AD and other diseases.     

Light-activated antibacterial surfaces comprise photosensitizers

Antibacterial surfaces were prepared using a base polyethylene sheet topped with a layer containing a mixed powder of poly (vinylidene fluoride) and photosensitizers (PSs). A crimpled stamp was placed on the mixed powder, and then it was passed through a heating and pressing device. The three chosen PSs were rose bengal, toluidine blue O and methylene blue. Scanning electron microscope analysis showed that the PS surface texture was coarse and highly developed. Measurement of the apparent contact angles of the droplets deposited on the PS surfaces using goniometry showed that all three surfaces were hydrophobic. Photodynamic analysis of the surfaces into which the PSs were incorporated indicated significant reactive oxygen species formation after illumination with light fluency rate of 1.46 mW cm(-2) for 30 min. Photodynamic inactivation assays performed in nutrient broth demonstrated more than 4 log reduction of the attached Escherichia coli after illumination (1.46 mW cm(-2)) for 24 h when the inoculum was 10(3) CFU mL(-1). However, more than 4 log reduction of Staphylococcus aureus occurred even when the cultures were illuminated for only 6 h. Our results provide an inexpensive, simple, state-of-the-art method for preparing antibacterial surfaces that may help prevent infections in hospital surroundings and in some medical devices.     

Nondiffracting accelerating wave packets of Maxwell's equations

We present the nondiffracting spatially accelerating solutions of the Maxwell equations. Such beams accelerate in a circular trajectory, thus generalizing the concept of Airy beams to the full domain of the wave equation. For both TE and TM polarizations, the beams exhibit shape-preserving bending which can have subwavelength features, and the Poynting vector of the main lobe displays a turn of more than 90°. We show that these accelerating beams are self-healing, analyze their properties, and find the new class of accelerating breathers: self-bending beams of periodically oscillating shapes. Finally, we emphasize that in their scalar form, these beams are the exact solutions for nondispersive accelerating wave packets of the most common wave equation describing time-harmonic waves. As such, this work has profound implications to many linear wave systems in nature, ranging from acoustic and elastic waves to surface waves in fluids and membranes.     

Solubilization of hydrophobic guest molecules in the monoolein discontinuous QL cubic mesophase and its soft nanoparticles

Hydrophobic bioactive guest molecules were solubilized in the discontinuous cubic mesophase (QL) of monoolein. Their effects on the mesophase structure and thermal behavior, and on the formation of soft nanoparticles upon dispersion of the bulk mesophase were studied. Four additives were analyzed. They were classified into two types based on their presumed location within the lipid bilayer and their influence on the phase behavior and structure. Differential scanning calorimetry (DSC), small-angle X-ray scattering (SAXS), polarized light microscopy, cryogenic-transmission electron microscopy (cryo-TEM), and dynamic light scattering (DLS) were used for the analysis. We found that carbamazepine and cholesterol (type I molecules) likely localize in the hydrophobic domains, but close to the hydrophobic-hydrophilic region. They induce strong perturbation to the mesophase packing by influencing both the order of the lipid acyl chains and interactions between lipid headgroups. This results in significant reduction of the phase transition enthalpy, and phase separation into lamellar and cubic mesophases above the maximum loading capacity. The inclusion of type I molecules in the mesophase also prevents the formation of soft nanoparticles with long-range internal order upon dispersion. In their presence, only vesicles or sponge-like nanoparticles form. Phytosterols and coenzyme Q10 (type II molecules) present only moderate effects. These molecules reside in the hydrophobic domains, where they cannot alter the lipid curvature or transform the QL mesophase into another phase. Therefore, above maximum loading, excess solubilizate precipitates in crystal forms. Moreover, when type II-loaded QL is dispersed, nanoparticles with long-range order and cubic symmetry (i.e., cubosomes) do form. A model for the growth of the ordered nanoparticles was developed from a series of intermediate structures identified by cryo-TEM. It proposes the development of the internal structure by fusion events between bilayer segments.