Identification of an opioid kappa receptor subtype-selective N-substituent for (+)-(3R,4R)-dimethyl-4-(3-hydroxyphenyl)piperidine

A three-component library of compounds was prepared in parallel using multiple simultaneous solution-phase synthetic methodology. The compounds were biased toward opioid receptor antagonist activity by incorporating (+)-(3R,4R)-dimethyl-4-(3-hydroxyphenyl)piperidine (a potent, nonselective opioid pure antagonist) as one of the monomers. The other two monomers, which included N-substituted or unsubstituted Boc-protected amino acids and a range of substituted aryl carboxylic acids, were selected to add chemical diversity. Screening of these compounds in competitive binding experiments with the kappa opioid receptor selective ligand [3H]U69,593 led to the discovery of a novel kappa opioid receptor selective ligand, N-?(2'S)-[3-(4-hydroxyphenyl)propanamido]-3'-methylbutyl?-(3R, 4R)-dimethyl-4-(3-hydroxyphenyl)piperidine (8, RTI-5989-29). Additional structure-activity relationship studies suggested that 8 possesses lipophilic and hydrogen-bonding sites that are important to its opioid receptor potency and selectivity. These sites appear to exist predominantly within the kappa receptor since the selectivity arises from a 530-fold loss of affinity of 8 for the mu receptor and an 18-fold increase in affinity for the kappa receptor relative to the mu-selective ligand, (+)-N-[trans-4-phenyl-2-butenyl]-(3R, 4R)-dimethyl-4-(3-hydroxyphenyl)piperidine (5a). The degree of selectivity observed in the radioligand binding experiments was not observed in the functional assay. According to its ability to inhibit agonist stimulated binding of [35S]GTPgammaS at all three opioid receptors, compound 8 behaves as a mu/kappa opioid receptor pure antagonist with negligible affinity for the delta receptor.     

TRPM7-Mediated Calcium Transport in HAT-7 Ameloblasts

TRPM7 plays an important role in cellular Ca²⁺, Zn²⁺ and Mg²⁺ homeostasis. TRPM7 channels are abundantly expressed in ameloblasts and, in the absence of TRPM7, dental enamel is hypomineralized. The potential role of TRPM7 channels in Ca²⁺ transport during amelogenesis was investigated in the HAT-7 rat ameloblast cell line. The cells showed strong TRPM7 mRNA and protein expression. Characteristic TRPM7 transmembrane currents were observed, which increased in the absence of intracellular Mg²⁺ ([Mg²⁺]_i), were reduced by elevated [Mg²⁺]_i, and were inhibited by the TRPM7 inhibitors NS8593 and FTY720. Mibefradil evoked similar currents, which were suppressed by elevated [Mg²⁺]_i, reducing extracellular pH stimulated transmembrane currents, which were inhibited by FTY720. Naltriben and mibefradil both evoked Ca²⁺ influx, which was further enhanced by the acidic intracellular conditions. The SOCE inhibitor BTP2 blocked Ca²⁺ entry induced by naltriben but not by mibefradil. Thus, in HAT-7 cells, TRPM7 may serves both as a potential modulator of Orai-dependent Ca²⁺ uptake and as an independent Ca²⁺ entry pathway sensitive to pH. Therefore, TRPM7 may contribute directly to transepithelial Ca²⁺ transport in amelogenesis.     

Mutant p53 Mediates Sensitivity to Cancer Treatment Agents in Oesophageal Adenocarcinoma Associated with MicroRNA and SLC7A11 Expression

TP53 gene mutations occur in 70% of oesophageal adenocarcinomas (OACs). Given the central role of p53 in controlling cellular response to therapy we investigated the role of mutant (mut-) p53 and SLC7A11 in a CRISPR-mediated JH-EsoAd1 TP53 knockout model. Response to 2 Gy irradiation, cisplatin, 5-FU, 4-hydroxytamoxifen, and endoxifen was assessed, followed by a TaqMan OpenArray qPCR screening for differences in miRNA expression. Knockout of mut-p53 resulted in increased chemo- and radioresistance (2 Gy survival fraction: 38% vs. 56%, p < 0.0001) and in altered miRNA expression levels. Target mRNA pathways analyses indicated several potential mechanisms of treatment resistance. SLC7A11 knockdown restored radiosensitivity (2 Gy SF: 46% vs. 73%; p = 0.0239), possibly via enhanced sensitivity to oxidative stress. Pathway analysis of the mRNA targets of differentially expressed miRNAs indicated potential involvement in several pathways associated with apoptosis, ribosomes, and p53 signaling pathways. The data suggest that mut-p53 in JH-EsoAd1, despite being classified as non-functional, has some function related to radio- and chemoresistance. The results also highlight the important role of SLC7A11 in cancer metabolism and redox balance and the influence of p53 on these processes. Inhibition of the SLC7A11-glutathione axis may represent a promising approach to overcome resistance associated with mut-p53.     

Frying Performance of No-<Emphasis Type="Italic">trans</Emphasis>, Low-Linolenic Acid Soybean Oils

Two extruded-expelled physically refined soybean oils with reduced contents of linolenic acid, ultra-low- linolenic acid (ULL, 1.5%) and low-linolenic acid (LL, 2.6%), and a extruded-expelled physically refined control oil (control, 5.3% linolenic acid) were evaluated by frying French fries in a commercial-like setting for 6 h day⁻¹ during 23 days. The oils became darker, increased in yellow color at the beginning, and became redder and less green throughout the process. Free fatty acids levels were not different among the oils until day 14, after which, ULL was different from the control for the remainder of frying. The conjugated dienoic acid values were greatest in the control. Generally, ULL and LL oils had lower percentages of polar compounds than did the control, providing a frying life 2 days longer than the control and ~30% increase in frying time. A trained sensory panel evaluated the French fries on days 2, 5, and 6. Buttery and potato flavors decreased, and rancid and painty flavors increased over frying time for all products. Rancid flavor was highest in the fries from the control oil. Overall, the ULL and LL oils performed better than did the control oil and ULL tended to perform better than the LL.     

Oxidative stability of soybean oils with altered fatty acid compositions

The oxidative stabilities of one canola oil and six soybean oils of various fatty acid compositions were compared in terms of peroxide values, conjugated dienoic acid values and sensory evaluations. Two of the soybean oils (Hardin and BSR 101) were from common commercial varieties. The other four soybean oils were from experimental lines developed in a mutation breeding program at Iowa State University that included A17 with 1.5% linolenate and 15.2% palmitate; A16 with 2% linolenate and 10.8% palmitate; A87-191039 with 2% linolenate and 29.6% oleate; and A6 with 27.5% stearate. Seed from the soybean genotypes was cold pressed. Crude canola oil was obtained without additives. All oils were refined, bleached and deodorized under laboratory conditions with no additives and stored at 60°C for 15 days. The A17, A16, A87-191039 and A6 oils were generally more stable to oxidation than the commercial soybean varieties and canola oil as evaluated by chemical and sensory tests. Canola oil was much less stable than Hardin and BSR 101 oils by both chemical and sensory tests. The peroxide values and flavor scores of oils were highly correlated with the initial amounts of linolenate (r=0.95, P=0.001). Flavor quality and flavor intensity had negative correlations with linolenate, (r=−0.89, P=0.007) and (r=−0.86, P=0.013), respectively.     

Laboratory study of submerged metal-mine tailings 1: Effect of solid-liquid contact time and aeration on contaminant concentrations

The dissolution of metals from various metal-mine tailings has been studied through a series of submerged tailings batch tests at U.S. Bureau of Mines laboratories. Tailings selected for this study included both reactive (as determined by column leaching studies) and benign tailings. Metal mobilization in aerated (97 pct saturated with oxygen) samples appeared to be consistently greater than metal dissolution from stagnant (79 pct or 70 pct saturated with oxygen) waters. In the early weeks of the study, sulfate concentrations increased with solid-liquid contact time, while calcium dissolution remained nearly constant. Metal concentrations varied in relation to solid-liquid contact time and aeration. Leachate pH appeared to be nearly independent of solid-liquid contact time for basic tailings. Reactive tailings having both a submerged and a surface component demonstrated increased metal mobility.     

The effect of adding devulcanized rubber on the thermomechanical properties of recycled polypropylene

This work aims to investigate the effect of adding vulcanized or partially devulcanized rubbers on recycled polypropylene (PPr), considering thermomechanical and morphological properties. The study proposes to better understand how structural changes underwent by rubber (after the devulcanization) contributed to improving the mechanical properties of the PPr. The PPr/rubber blends were prepared by a co-rotating twin-screw extruder and then were injected. The blends composed of the most devulcanized rubbers by microwaves with refined microstructure showed higher values of elongation at break and toughness. Data showed that the devulcanization process applied to the rubber interfered positively in its adhesion to the PPr. Data from dynamic mechanical analysis and atomic force microscopy indicated that the most devulcanized rubbers presented an interface more connected to PPr. These chemical interactions possibly impacted the mechanical properties of the PPr. Moreover, dilatation processes favored the fracture mechanisms of the PPr when rubber was added to it.     

The rotator cuff-deficient arthritic shoulder: diagnosis and surgical management

The symptomatic rotator cuff-deficient, arthritic glenohumeral joint poses a complex problem for the orthopaedic surgeon. Surgical management can be facilitated by classifying the disorder in one of three diagnostic categories: (1) rotator cuff-tear arthropathy, (2) rheumatoid arthritic shoulder with cuff deficiency, or (3) degenerative arthritic (osteoarthritic) shoulder with cuff deficiency. If it is not possible to repair the cuff defect, surgical management may include prosthetic arthroplasty, with the recognition that only limited goals are attainable, particularly with respect to strength and active motion. Glenohumeral arthrodesis is a salvage procedure when other surgical measures have failed. Arthrodesis is also indicated in patients with deltoid muscle deficiency. Humeral hemiarthroplasty avoids the complications of glenoid loosening and is an attractive alternative to arthrodesis, resection arthroplasty, and total shoulder arthroplasty. The functionally intact coracoacromial arch should be preserved to reduce the risk of anterosuperior subluxation. Care should be taken not to "overstuff" the gleno-humeral joint with a prosthetic component. In cases of significant internal rotation contracture, subscapularis lengthening is necessary to restore anterior and posterior rotator cuff balance. If the less stringent criteria of Neer's "limited goals" rehabilitation are followed, approximately 80% to 90% of patients treated with humeral hemiarthroplasty can have satisfactory results.     

Automatic eye fixations identification based on analysis of variance and covariance

Eye movement is the simplest and repetitive movement that enables humans to interact with the environment. The common daily activities, such as reading a book or watching television, involve this natural activity, which consists of rapidly shifting our gaze from one region to another. In clinical application, the identification of the main components of eye movement during visual exploration, such as fixations and saccades, is the objective of the analysis of eye movements: however, in patients affected by motor control disorder the identification of fixation is not banal. This work presents a new fixation identification algorithm based on the analysis of variance and covariance: the main idea was to use bivariate statistical analysis to compare variance over x and y to identify fixation. We describe the new algorithm, and we compare it with the common fixations algorithm based on dispersion. To demonstrate the performance of our approach, we tested the algorithm in a group of healthy subjects and patients affected by motor control disorder.