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Herein, we disclose a facile synthetic strategy to access an important class of drug molecules that contain chiral 1,2-amino alcohol functionality utilizing highly effective ruthenium-catalyzed asymmetric transfer hydrogenation of unprotected a-ketoamines. Recently, the COVID-19 pandemic has caused a crisis of shortage of many important drugs, especially norepinephrine and epinephrine, for the treatment of anaphylaxis and hypotension because of the increased demand. Unfortunately, the existing technologies are not fulfilling the worldwide requirement due to the existing lengthy synthetic protocols that require additional protection and deprotection steps. We identified a facile synthetic protocol via a highly enantioselective one-step process for epinephrine and a two-step process for norepinephrine starting from unprotected a-ketoamines 1band 1a, respectively. This newly developed enantioselective ruthenium-catalyzed asymmetric transfer hydrogenation was extended to the synthesis of many 1,2-amino alcohol-containing drug molecules such as phenylephrine, denopamine, norbudrine, and levisoprenaline, with enantioselectivities of >99% ee and high isolated yields.

Purpose: To construct a pain classification model using binary logistic regression to calculate pain probability and monitor pain based on heart rate variability (HRV) and photoplethysmography (PPG) parameters. Methods: Heat stimulation was used to simulate pain for modeling the pain generation process, and electrocardiography and PPG signals were recorded simultaneously. After signal analysis, statistical analysis was performed using SPSS to determine the parameters that were significant for pain. Thereafter, a pain classification model with HRV and PPG parameters was established using binary logistic regression. Results: The sensitivity and specificity of the pain classification model were 60.0% and 72.0%, respectively. When pain occurred, the probability calculated using the pain classification model increased from < 50% to > 50%. When the pain was relieved, the probability decreased to < 50%. The probability of pain was consistent with the numeric rating scale value, which indicated that the model can correctly determine the presence of pain. Conclusion: This pain classification model has sufficient robustness and adaptability to be applied to different healthy people for classification and monitoring. This model is helpful in establishing a real-time pain monitoring system to improve pain management for patients in the postoperative intensive care unit and patient-controlled analgesia and provide a reference for doctors regarding medication.

Arginine methyltransferases critically regulate cellular homeostasis by modulating the functional outcome of their substrates. The protein arginine methyltransferase 5 (PRMT5) is an enzyme involved in growth and survival pathways promoting tumorigenesis. However, little is known about the biologic function of PRMT5 and its therapeutic potential in multiple myeloma (MM). In the present study, we identified and validated PRMT5 as a new therapeutic target in MM. PRMT5 is overexpressed in patient MM cells and associated with decreased progression-free survival and overall survival. Either genetic knockdown or pharmacological inhibition of PRMT5 with the inhibitor EPZ015666 significantly inhibited growth of both cell lines and patient MM cells. Furthermore, PRMT5 inhibition abrogated NF-κB signaling. Interestingly, mass spectrometry identified a tripartite motif-containing protein 21 TRIM21 as a new PRMT5-partner, and we delineated a TRIM21-dependent mechanism of NF-κB inhibition. Importantly, oral administration of EPZ015666 significantly decreased MM growth in a humanized murine model of MM. These data both demonstrate the oncogenic role and prognostic relevance of PRMT5 in MM pathogenesis, and provide the rationale for novel therapies targeting PRMT5 to improve patient outcome.

Despite the development of novel drugs, alkylating agents remain an important component of therapy in multiple myeloma (MM). DNA repair processes contribute towards sensitivity to alkylating agents and therefore we here evaluate the role of nucleotide excision repair (NER), which is involved in the removal of bulky adducts and DNA crosslinks in MM. We first evaluated NER activity using a novel functional assay and observed a heterogeneous NER efficiency in MM cell lines and patient samples. Using next-generation sequencing data, we identified that expression of the canonical NER gene, excision repair cross-complementation group 3 (ERCC3), significantly impacted the outcome in newly diagnosed MM patients treated with alkylating agents. Next, using small RNA interference, stable knockdown and overexpression, and small-molecule inhibitors targeting xeroderma pigmentosum complementation group B (XPB), the DNA helicase encoded by ERCC3, we demonstrate that NER inhibition significantly increases sensitivity and overcomes resistance to alkylating agents in MM. Moreover, inhibiting XPB leads to the dual inhibition of NER and transcription and is particularly efficient in myeloma cells. Altogether, we show that NER impacts alkylating agents sensitivity in myeloma cells and identify ERCC3 as a potential therapeutic target in MM.

Epigenetic signaling pathways are implicated in tumorigenesis and therefore histone deacetylases (HDACs) represent novel therapeutic targets for cancers, including multiple myeloma (MM). Although non-selective HDAC inhibitors show anti-MM activities, unfavorable side effects limit their clinical efficacy. Isoform- and/or class-selective HDAC inhibition offers the possibility to maintain clinical activity while avoiding adverse events attendant to broad non-selective HDAC inhibition. We have previously reported that HDAC3 inhibition, either by genetic knockdown or selective inhibitor BG45, abrogates MM cell proliferation. Here we show that knockdown of HDAC3, but not HDAC1or HDAC2, as well as BG45, downregulate expression of DNA methyltransferase 1 (DNMT1) mediating MM cell proliferation. DNMT1expression is regulated by c-Myc, and HDAC3 inhibition triggers degradation of c-Myc protein. Moreover, HDAC3 inhibition results in hyperacetylation of DNMT1, thereby reducing the stability of DNMT1 protein. Combined inhibition of HDAC3 and DNMT1 with BG45 and DNMT1 inhibitor 5-azacytidine (AZA), respectively, triggers synergistic downregulation of DNMT1, growth inhibition and apoptosis in both MM cell lines and patient MM cells. Efficacy of this combination treatment is confirmed in a murine xenograft MM model. Our results therefore provide the rationale for combination treatment using HDAC3 inhibitor with DNMT1 inhibitor to improve patient outcome in MM.

Recent studies have delineated cancer-type-specific roles of histone 3 lysine 27 (H3K27) demethylase KDM6B/JMJD3 depending on its H3K27 demethylase activity. Here we show that KDM6B is expressed in multiple myeloma (MM) cells; and that shRNA-mediated knockdown and CRISPR-mediated knockout of KDM6Babrogate MM cell growth and survival. Tumor necrosis factor-α or bone marrow stromal cell culture supernatants induce KDM6B, which is blocked by IKKβ inhibitor MLN120B, suggesting that KDM6Bis regulated by NF-κB signaling in MM cells. RNA-seq and subsequent ChIP-qPCR analyses reveal that KDM6B is recruited to the loci of genes encoding components of MAPK signaling pathway including ELK1and FOS, and upregulates expression of these genes without affecting H3K27 methylation level. Overexpression of catalytically inactive KDM6Bactivates expression of MAPK pathway-related genes, confirming its function independent of demethylase activity. We further demonstrate that downstream targets of KDM6B, ELK1and FOS,confer MM cell growth. Our study therefore delineates KDM6B function that links NF-κB and MAPK signaling pathway mediating MM cell growth and survival, and validates KDM6B as a novel therapeutic target in MM.

Proteasome inhibition is an effective therapy for multiple myeloma (MM) patients; however, the emergence of drug resistance is common. Novel therapeutic strategies to overcome proteasome inhibitor resistance are needed. In this study, we examined whether targeting deubiquitylating (DUB) enzymes upstream of 20S proteasome overcomes proteasome inhibitor resistance. Gene expression analysis, immunohistochemical studies of MM patient bone marrow, reverse transcription–PCR and protein analysis show that Rpn11/POH1, a DUB enzyme upstream of 20S proteasome, is more highly expressed in patient MM cells than in normal plasma cells. Importantly, Rpn11 expression directly correlates with poor patient survival. Loss-of-function studies show that Rpn11-siRNA knockdown decreases MM cell viability. Pharmacological inhibition of Rpn11 with O-phenanthroline (OPA) blocks cellular proteasome function, induces apoptosis in MM cells and overcomes resistance to proteasome inhibitor bortezomib. Mechanistically, Rpn11 inhibition in MM cells activates caspase cascade and endoplasmic stress response signaling. Human MM xenograft model studies demonstrate that OPA treatment reduces progression of tumor growth and prolongs survival in mice. Finally, blockade of Rpn11 increases the cytotoxic activity of anti-MM agents lenalidomide, pomalidomide or dexamethasone. Overall, our preclinical data provide the rationale for targeting DUB enzyme Rpn11 upstream of 20S proteasome to enhance cytotoxicity and overcome proteasome inhibitor resistance in MM.

B-cell maturation antigen (BCMA) is a highly plasma cell-selective protein that is expressed on malignant plasma cells of multiple myeloma (MM) patients and therefore is an ideal target for T-cell redirecting therapies. We developed a bispecific T-cell engager (BiTE) targeting BCMA and CD3ɛ (BI 836909) and studied its therapeutic impacts on MM. BI 836909 induced selective lysis of BCMA-positive MM cells, activation of T cells, release of cytokines and T-cell proliferation; whereas BCMA-negative cells were not affected. Activity of BI 836909 was not influenced by the presence of bone marrow stromal cells, soluble BCMA or a proliferation-inducing ligand (APRIL). In ex vivo assays, BI 836909 induced potent autologous MM cell lysis in both, newly diagnosed and relapsed/refractory patient samples. In mouse xenograft studies, BI 836909 induced tumor cell depletion in a subcutaneous NCI-H929 xenograft model and prolonged survival in an orthotopic L-363 xenograft model. In a cynomolgus monkey study, administration of BI 836909 led to depletion of BCMA-positive plasma cells in the bone marrow. Taken together, these results show that BI 836909 is a highly potent and efficacious approach to selectively deplete BCMA-positive MM cells and represents a novel immunotherapeutic for the treatment of MM.

AbstractIn today’s globally competitive environment, processes involving multiple manufacturing lines are quite common due to economies of scale considerations. Production yield measure indices Cpkand Cpuare the most popular yield-based indices and have been widely used in manufacturing industries for two-sided and one-sided specification limits, respectively. In this paper, two production yield indices for multiple lines and are considered and the approximate distributions of two natural estimators and are derived. For the convenience of industry applications, the lower confidence bounds and critical values are provided.

Proteasome inhibitor bortezomib is an effective therapy for relapsed and newly diagnosed multiple myeloma (MM); however, dose-limiting toxicities and the development of resistance can limit its long-term utility. Recent research has focused on targeting ubiquitin receptors upstream of 20S proteasome, with the aim of generating less toxic therapies. Here we show that 19S proteasome-associated ubiquitin receptor Rpn13 is more highly expressed in MM cells than in normal plasma cells. Rpn13-siRNA (small interfering RNA) decreases MM cell viability. A novel agent RA190 targets Rpn13 and inhibits proteasome function, without blocking the proteasome activity or the 19S deubiquitylating activity. CRISPR/Cas9 Rpn13-knockout demonstrates that RA190-induced activity is dependent on Rpn13. RA190 decreases viability in MM cell lines and patient MM cells, inhibits proliferation of MM cells even in the presence of bone marrow stroma and overcomes bortezomib resistance. Anti-MM activity of RA190 is associated with induction of caspase-dependent apoptosis and unfolded protein response-related apoptosis. MM xenograft model studies show that RA190 is well tolerated, inhibits tumor growth and prolongs survival. Combining RA190 with bortezomib, lenalidomide or pomalidomide induces synergistic anti-MM activity. Our preclinical data validates targeting Rpn13 to overcome bortezomib resistance, and provides the framework for clinical evaluation of Rpn13 inhibitors, alone or in combination, to improve patient outcome in MM.

The anti-CD38 monoclonal antibody SAR650984 (SAR) is showing promising clinical activity in treatment of relapsed and refractory multiple myeloma (MM). Besides effector-mediated antibody-dependent cellular cytotoxicity and complement-mediated cytotoxicity, we here define molecular mechanisms of SAR-directed MM cell death and enhanced anti-MM activity triggered by SAR with Pomalidomide (Pom). Without Fc-cross-linking agents or effector cells, SAR specifically induces homotypic aggregation (HA)-associated cell death in MM cells dependent on the level of cell surface CD38 expression, actin cytoskeleton and membrane lipid raft. SAR and its F(ab)’2 fragments trigger caspase 3/7-dependent apoptosis in MM cells highly expressing CD38, even with p53 mutation. Importantly, SAR specifically induces lysosome-dependent cell death (LCD) by enlarging lysosomes and increasing lysosomal membrane permeabilization associated with leakage of cathepsin B and LAMP-1, regardless of the presence of interleukin-6 or bone marrow stromal cells. Conversely, the lysosomal vacuolar H+-ATPase inhibitor blocks SAR-induced LCD. SAR further upregulates reactive oxygen species. Pom enhances SAR-induced direct and indirect killing even in MM cells resistant to Pom/Len. Taken together, SAR is the first therapeutic monoclonal antibody mediating direct cytotoxicity against MM cells via multiple mechanisms of action. Our data show that Pom augments both direct and effector cell-mediated MM cytotoxicity of SAR, providing the framework for combination clinical trials.

AbstractIn this paper a system of depth-integrated equations for over-saturated debris flows on three-dimensional topography is derived. The lower layer is a saturated mixture of density preserving solid and fluid constituents, where the pore fluid is in excess, so that an upper fluid layer develops above the mixture layer. At the layer interface fluid mass exchange may exist and for this a parameterization is needed. The emphasis is on the description of the influence on the flow by the curvature of the basal surface, and not on proposing rheological models of the avalanching mass. To this end, a coordinate system fitted to the topography has been used to properly account for the geometry of the basal surface. Thus, the modeling equations have been written in terms of these coordinates, and then simplified by using (1) the depth-averaging technique and (2) ordering approximations in terms of an aspect ratio ϵ which accounts for the scale of the flowing mass. The ensuing equations have been complemented by closure relations, but any other such relations can be postulated. For a shallow two-layer debris with clean water in the upper layer, flowing on a slightly curved surface, the equilibrium free surface is shown to be horizontal.

To investigate the association between brain amyloid load in Alzheimer disease (AD) measured by 11CPIB-PET, regional cerebral glucose metabolism (rCMRGlc) measured by 18FFDG-PET, and cognition.

To evaluate the relationship between clinical improvement and in vivo synaptic dopamine (DA) release after a single oral dose of levodopa (LD) in patients with advanced Parkinson disease (PD).

Huntington disease (HD) is characterized by the progressive death of medium spiny dopamine receptor bearing striatal GABAergic neurons. In addition, microglial activation in the areas of neuronal loss has recently been described in postmortem studies. Activated microglia are known to release neurotoxic cytokines, and these may contribute to the pathologic process.

A fast combinatorial approach to access information about the immobilization behavior and kinetics of enzymes on a variation of surfaces is presented. As a test system, Candida Antarctica Lipase B was immobilized on a self-assembled monolayer bearing a gradient of surface energy. The respective immobilization behavior was monitored by Fourier transform infrared microspectroscopy. In addition, the activity of the immobilized enzyme was monitored over the entire film in real time with a specially developed fluorescence activity assay embedded into a siloxane gel. It was found that the highest amount of active protein was immobilized on the hydrophilic end of the gradient surface. This effect is associated with a higher surface roughness of this area resulting in hydrophobic microenviroments in which the enzyme gets immobilized.

Laparoscopic appendectomy has been shown to improve postoperative recovery when compared with open appendectomy. The present randomized trial was conducted to evaluate any further difference in outcome between needlescopic appendectomy (NA) and conventional laparoscopic appendectomy (CLA) in the management of acute appendicitis. Patients with the clinical diagnosis of acute appendicitis were randomized to either NA (instrument size ≤3 mm) or CLA (instrument size ≥5 mm). Standardized anesthetic technique and perioperative management were adopted. The primary end point was length of postoperative hospital stay. Other parameters such as conversion rate, postoperative pain score and analgesic requirement, return of bowel function, resumption of normal activities, complication rate, and length of the final scars were also assessed and compared. A total of 363 patients (NA: 174, CLA: 189) were recruited. Both approaches could accurately arrive at the diagnosis (NA: 98.3%; CLA: 100%). Compared with CLA, NA resulted in a significantly longer operation time (P= 0.015) and a higher conversion rate (P< 0.001). The final scars of the NA group were significantly shorter when compared with the CLA group (P< 0.001). Otherwise, there was no statistical difference between the 2 groups in terms of complication rate, postoperative pain score, length of postoperative stay, and other recovery parameters. NA resulted in a longer operation time and higher conversion rate. Except for a smaller scar, the present study was unable to demonstrate any other short-term benefits. Thus, the technique cannot be routinely recommended.

Novel mono‐ and multifunctional vinyl ester monomers containing thioether groups were synthesized via an amine‐catalyzed Michael addition reaction between vinyl acrylate and multifunctional thiols. Using photo‐differential scanning calorimetry and real‐time Fourier transform infrared (RTIR) spectroscopy, the polymerization kinetics and oxygen inhibition of the homopolymerizations of the vinyl ester monomers were investigated. The effect of the vinyl ester and thioether group on acrylate/vinyl ester and thiol/vinyl ester copolymerizations was determined using real‐time IR spectroscopy to monitor polymerization rates of acrylate, vinyl, and thiol groups simultaneously. Polymerization of the vinyl esters used was found to be relatively insensitive to oxygen inhibition. We propose that the thioether group is responsible for reducing oxygen inhibition by a series of chain transfer/oxygen‐scavenging reactions. In polymerization of a acrylate/vinyl ester mixture both in nitrogen and in air, the vinyl ester monomer significantly enhances the polymerization rates and the conversion of the acrylate double bonds via plasticization of the crosslinked matrix and reduction of inhibition by oxygen. Ultimately, the vinyl ester monomer is incorporated into the polymer network. Thiol/vinyl ester free‐radical copolymerization is much faster than either thiol/allylether copolymerization or vinyl ester homopolymerization. The electron‐rich vinyl ester double bonds ensure rapid copolymerization with thiol. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 4424–4436, 2004

Pristine and strongly irradiated self-assembled monolayers of [1,1‘:4‘,1‘ ‘-terphenyl]-4,4‘ ‘-dimethanethiol (TPDMT) on Au have been characterized by near-edge X-ray absorption fine structure spectroscopy using partial electron yield acquisition mode. The TPDMT films were found to be extremely stable toward electron irradiation, which is explained by cross-linking between the aromatic backbones. In addition, we assume that a large delocalization and a strong relaxation of the initial electronic excitations in the densely packed film contributed to the film stability. The data analysis implies an inhomogeneous distribution of the irradiation-induced dehydrogenation and cross-linking of the terphenyl moieties in the TPDMT film, being most pronounced close to the film−ambient interface. The inhomogeneity was explained by quenching of the electronically excited C−H* states via dipole−dipole interaction with the states' image at the metal surface, which has a reduced probability with increasing separation from the metal surface. Generally, the results suggest the importance of relaxation processes for the response of self-assembled monolayers to ionizing radiation.

Abstract: For the first time, the yield strength of thin-film parylene-C is measured from membrane load-deflection experiments and surface profile analysis. To do so, the onset pressure which causes plastic deformation of the membrane is first experimentally measured. Then a new 2-step displacement model, together with the energy minimization technique [1], is developed to convert the onset pressure to the yield strength on the pre-stressed parylene membrane under a uniform pressure loading. The results depict a Yield Strength of 59 MPa (or 0.012 of strain) for thin-film parylene-C in comparison to 55 MPa reported by parylene vendor (measured from large samples) [2]. To double check with the result, the balloon model [3] is further used to compare with the stress value from our model at the center of parylene membranes and good agreements are obtained.

A nickel-based tubule electrode array was fabricated by using electroless deposition with a polycarbonate membrane serving as a template. Hypophosphite was the reducing agent in the Ni deposition, and electrodes made of a Ni−P alloy were formed. The tubule has an internal diameter of ca. 180 nm, a wall thickness of ca. 20 nm, and a length of ca. 2 μm. The electrode array provided an exposed area ca. 8.8 times as large as that of a planar electrode, suggesting a potential application of the array in sensor devices. Cyclic voltammetric measurements in KOH showed that the electrochemical response from the Ni(OH)2/NiOOH redox reaction for the electrode arrays was more than 40 times larger than that for an planar electrode. The nanoscale structure of the tubule was concluded to be responsible for this magnification of the redox response. Heat treatment combined with overcharge oxidation of the electrode arrays was found to significantly improve the charge storage capacity resulting from the redox reaction. The present work has provided important concepts capable of improving the performance of a nickel hydroxide electrode for nickel metal hydride batteries.

The adsorption and thermal decomposition behavior of azomethane on the Si(111)-7 × 7 surface has been studied by high-resolution electron energy loss spectroscopy (HREELS), synchrotron radiation photoelectron spectroscopy (SR-PES) in the soft X-ray and UV photon energy ranges, and scanning tunneling microscopy (STM). It is found that at 90 K substrate temperature, azomethane is adsorbed largely intact in both trans and cis forms. The decomposition of the molecules starts at an annealing temperature around 200 K with N&dbd;N bond breaking, and at 300 K, a mixture of remaining intact azomethane, NCHx, and Si−NHx species is present on the surface with three distinct N 1s photoemission peaks. At this temperature, the Si−N species begins to appear as nitride, but no Si−C bonding is detected. Carbon dissociates from nitrogen and reacts directly with the Si surface above 400 K with CH3 adsorption as an intermediate adspecies. The CH3 adsorption is strongly enhanced if azomethane is thermally precracked prior to the exposure. At 700 K, all surface carbon and nitrogen are bonded to Si. The remaining hydrogen is desorbed at 1000 K, and Si−C and Si−N phases are formed. After it is annealed to 1500 K, photoemission features typical for the clean Si(111)-7 × 7 surface are immediately recovered. Si−C and Si−N species are desorbed, leaving a homogeneous but disordered surface with an underlying 1 × 1 structure, which transforms back into the 7 × 7 structure by forming characteristic triangular-shaped patterns as intermediate steps in the surface reconstruction process.