Polyamine and EIF5A hypusination downstream of c-Myc confers targeted therapy resistance in BRAF mutant melanoma.

BACKGROUND: BRAF inhibitors are widely employed in the treatment of melanoma with the BRAF V600E mutation. However, the development of resistance compromises their therapeutic efficacy. Diverse genomic and transcriptomic alterations are found in BRAF inhibitor resistant melanoma, posing a pressing need for convergent, druggable target that reverse therapy resistant tumor with different resistance mechanisms. METHODS: CRISPR-Cas9 screens were performed to identify novel target gene whose inhibition selectively targets A375VR, a BRAF V600E mutant cell line with acquired resistance to vemurafenib. Various in vitro and in vivo assays, including cell competition assay, water soluble tetrazolium (WST) assay, live-dead assay and xenograft assay were performed to confirm synergistic cell death. Liquid Chromatography-Mass Spectrometry analyses quantified polyamine biosynthesis and changes in proteome in vemurafenib resistant melanoma. EIF5A hypusination dependent protein translation and subsequent changes in mitochondrial biogenesis and activity were assayed by O-propargyl-puromycin labeling assay, mitotracker, mitoSOX labeling and seahorse assay. Bioinformatics analyses were used to identify the association of polyamine biosynthesis with BRAF inhibitor resistance and poor prognosis in melanoma patient cohorts. RESULTS: We elucidate the role of polyamine biosynthesis and its regulatory mechanisms in promoting BRAF inhibitor resistance. Leveraging CRISPR-Cas9 screens, we identify AMD1 (S-adenosylmethionine decarboxylase 1), a critical enzyme for polyamine biosynthesis, as a druggable target whose inhibition reduces vemurafenib resistance. Metabolomic and proteomic analyses reveal that polyamine biosynthesis is upregulated in vemurafenib-resistant cancer, resulting in enhanced EIF5A hypusination, translation of mitochondrial proteins and oxidative phosphorylation. We also identify that sustained c-Myc levels in vemurafenib-resistant cancer are responsible for elevated polyamine biosynthesis. Inhibition of polyamine biosynthesis or c-Myc reversed vemurafenib resistance both in vitro cell line models and in vivo in a xenograft model. Polyamine biosynthesis signature is associated with poor prognosis and shorter progression free survival after BRAF/MAPK inhibitor treatment in melanoma cohorts, highlighting the clinical relevance of our findings. CONCLUSIONS: Our findings delineate the molecular mechanisms involving polyamine-EIF5A hypusination-mitochondrial respiration pathway conferring BRAF inhibitor resistance in melanoma. These targets will serve as effective therapeutic targets that can maximize the therapeutic efficacy of existing BRAF inhibitors.

Discovery of N-benzylbenzamide-based allosteric inhibitors of Aurora kinase A.

Aurora kinases (AurkA/B/C) regulate the assembly of bipolar mitotic spindles and the fidelity of chromosome segregation during mitosis, and are attractive therapeutic targets for cancers. Numerous ATP-competitive AurkA inhibitors have been developed as potential anti-cancer agents. Recently, a few allosteric inhibitors have been reported that bind to the allosteric Y-pocket within AurkA kinase domain and disrupt the interaction between AurkA and its activator TPX2. Herein we report a novel allosteric AurkA inhibitor (6h) of N-benzylbenzamide backbone. Compound 6h suppressed the both catalytic activity and non-catalytic functions of AurkA. The inhibitory activity of 6h against AurkA (IC50 = 6.50 µM) was comparable to that of the most potent allosteric AurkA inhibitor AurkinA. Docking analysis against the Y-pocket revealed important pharmacophores and interactions that were coherent with structure-activity relationship. In addition, 6h suppressed DNA replication in G1-S phase, which is a feature of allosteric inhibition of AurA. Our current study may provide a useful insight in designing potent allosteric AurkA inhibitors.

Human Fibroblast-Derived Matrix Hydrogel Accelerates Regenerative Wound Remodeling Through the Interactions with Macrophages.

Herein, a novel extracellular matrix (ECM) hydrogel is proposed fabricated solely from decellularized, human fibroblast-derived matrix (FDM) toward advanced wound healing. This FDM-gel is physically very stable and viscoelastic, while preserving the natural ECM diversity and various bioactive factors. Subcutaneously transplanted FDM-gel provided a permissive environment for innate immune cells infiltration. Compared to collagen hydrogel, excellent wound healing indications of FDM-gel treated in the full-thickness wounds are noticed, particularly hair follicle formation via highly upregulated ß-catenin. Sequential analysis of the regenerated wound tissues disclosed that FDM-gel significantly alleviated pro-inflammatory cytokine and promoted M2-like macrophages, along with significantly elevated vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) level. A mechanistic study demonstrated that macrophages-FDM interactions through cell surface integrins α5ß1 and α1ß1 resulted in significant production of VEGF and bFGF, increased Akt phosphorylation, and upregulated matrix metalloproteinase-9 activity. Interestingly, blocking such interactions using specific inhibitors (ATN161 for α5ß1 and obtustatin for α1ß1) negatively affected those pro-healing growth factors secretion. Macrophages depletion animal model significantly attenuated the healing effect of FDM-gel. This study demonstrates that the FDM-gel is an excellent immunomodulatory material that is permissive for host cells infiltration, resorbable with time, and interactive with macrophages, where it thus enables regenerative matrix remodeling toward a complete wound healing.

Identification of novel class inhibitors of NSD3 methyltransferase showing a unique, bivalent binding mode in the SET domain.

NSD3/WHSC1L1 lysine methyltransferase promotes the transcription of target genes through di- or tri-methylation at histone H3K36 using SAM as a cofactor. Genetic alterations such as amplification and gain-of-function mutation of NSD3 act as oncogenic drivers in several cancers including squamous cell lung cancer and breast cancer. NSD3 is an important therapeutic target for cancers, but the reported NSD3 inhibitors targeting the catalytic SET domain are very rare and show a poor activity. Herein, from a virtual library screening and the subsequent medicinal chemistry optimization, we identified a novel class of NSD3 inhibitors. Our docking analysis and pulldown result suggested that the most potent analogue 13i shows a unique, bivalent binding mode interacting with both SAM-binding site and BT3-bindig site within the SET domain. We found 13i inhibits NSD3 activity with IC50 = 287 µM in vitro and suppresses the proliferation of JIMT1 breast cancer cells with GI50 = 36.5 µM, which express a high level of NSD3. Also, 13i downregulated the levels of H3K36me2/3 in a dose-dependent manner. Our study could provide an insight in designing high-affinity NSD3 inhibitors. Also, as the acrylamide group of 13i was predicted to position near Cys1265 in the BT3-binding site, further optimization would lead to a discovery of novel irreversible NSD3 inhibitors.

Quantitative proteomic analysis of human serum using tandem mass tags to predict cardiovascular risks in patients with psoriasis.

Although biomarker candidates associated with psoriasis have been suggested, those for predicting the risk of cardiovascular disease (CVD) early in patients with psoriasis are lacking. We aimed to identify candidate biomarkers that can predict the occurrence of CVD in psoriasis patients. We pursued quantitative proteomic analysis of serum samples composed of three groups: psoriasis patients with and those without CVD risk factors, and healthy controls. Age/Sex-matched serum samples were selected and labeled with 16-plex tandem mass tag (TMT) and analyzed using liquid chromatography-mass spectrometry and subsequent verification with ELISA. Of the 184 proteins that showed statistical significance (P-value < 0.05) among the three groups according to TMT-based quantitative analysis, 98 proteins showed significant differences (> 2.0-fold) between the psoriasis groups with and without CVD risk factors. Verification by ELISA revealed that caldesmon (CALD1), myeloid cell nuclear differentiation antigen (MNDA), and zyxin (ZYX) levels were significantly increased in the psoriasis group with CVD risk factors. Further network analysis identified pathways including integrin signaling, which could be related to platelet aggregation, and actin cytoskeleton signaling. Three novel candidates (MNDA, ZYX, and CALD1) could be potential biomarkers for predicting CVD risks in psoriasis patients. We expect these biomarker candidates can be used to predict CVD risk in psoriasis patients in clinical settings although further studies including large validation are needed.

Secretome analysis reveals reduced expression of COL4A2 in hypoxic cancer-associated fibroblasts with a tumor-promoting function in gastric cancer.

BACKGROUND: Cancer-associated fibroblasts (CAFs) are major components of the tumor microenvironment (TME). Hypoxic TME is known to promote tumor progression. However, how a hypoxic condition regulates CAFs remains elusive. METHODS: To investigate the underlying mechanism involved in the regulation of gastric cancer (GC) progression by hypoxic CAFs, we performed secretome profiling. Normoxic or hypoxic CAFs conditioned media (CM) were filter-concentrated and in-gel trypsin digested. Resulting peptides were analyzed with LC-MS/MS. RESULTS: We observed that CM derived from hypoxic CAFs could promote migration of a panel of GC cell lines (AGS, SNU668, SNU638). Mass spectrometry analysis of hypoxic or normoxic CAFs CM identified 1595 proteins, of which 19 proteins (10 upregulated and 9 downregulated) were differentially expressed in the hypoxic secretome. We focused on COL4A2, whose expression was significantly decreased in hypoxic CAFs in HIF-1α-independent manner. Silencing of COL4A2 expression in normoxic CAFs phenocopied the effect of hypoxic CAFs in promoting GC cell migration. CONCLUSIONS: The reduced expression of COL4A2 in a hypoxic environment might be associated with the tumor-promoting role of hypoxic CAFs in GC.

Novel 2,6,9-Trisubstituted Purines as Potent CDK Inhibitors Alleviating Trastuzumab-Resistance of HER2-Positive Breast Cancers.

HER2-positive (HER2+) breast cancer is defined by HER2 oncogene amplification on chromosome 17q12 and accounts for 15−20% population of breast-cancer patients. Therapeutic anti-HER2 antibody such as trastuzumab is used as the first-line therapy for HER2-positive breast cancers. However, more than 50% of the patients respond poorly to trastuzumab, illustrating that novel therapy is warranted to overcome the resistance. We previously reported that in the majority of HER2+ breast-cancer patients, CDK12 is co-amplified on 17q12 and involved in developing tumors and trastuzumab resistance, proposing CDK12 as a potential drug target for HER2+ breast cancers. Here, we designed and synthesized novel 2,6,9-trisubstituted purines as potent CDK12 inhibitors showing strong, equipotent antiproliferative activity against trastuzumab-sensitive HER2+ SK-Br3 cells and trastuzumab-resistant HER2+ HCC1954 cells (GI50 values < 50 nM) both of which express a high level of CDK12. Two potent analogue 30d and 30e at 40, 200 nM greatly downregulated the levels of cyclinK and Pol II p-CTD (Ser2), as well as the expression of CDK12 downstream genes (IRS1 and WNT1) in a dose-dependent manner. We also observed structure-property relationship for a subset of potent analogues, and found that 30e is highly stable in liver microsomes with lack of CYP inhibition. In addition, 30d exhibited a synergy with trastuzumab in the both cells, suggesting that our inhibitors could be applied to alleviate trastuzumab-resistance of HER2+ breast cancers and escalate the efficacy of trastuzumab as well. Our study may provide insight into developing a novel therapy for HER2+ breast cancers.

The Kynurenine Pathway and Mediating Role of Stress in Addictive Disorders: A Focus on Alcohol Use Disorder and Internet Gaming Disorder.

Stress plays an important role in the pathophysiology of addictive disorders. The kynurenine (KYN) pathway involved in neuroimmune and cognitive functions is activated under stress. However, the neuroimmunological-neurocognitive mechanisms in the role of stress in addictive disorders are unclear still now. Ninety-nine young adults aged 18-35 years [alcohol use disorder (AUD), N = 30; Internet gaming disorder (IGD), N = 34; healthy controls (HCs), N = 35] participated in this study. Stress levels, resilience, addiction severity, and neurocognitive functions were evaluated, and serum levels of tryptophan (TRP), 5-hydroxytryptamine (5-HT), KYN, and kynurenine acid (KYNA) were determined using liquid chromatography coupled with tandem mass spectrometry through blood samples. Both addictive disorder groups showed higher levels of stress, lower resilience, and impaired executive functions compared to the HC group. Importantly, the AUD group revealed significantly increased KYN levels and KYN/TRP ratios, as well as decreased KYNA levels and KYNA/KYN ratios compared to HCs (p < 0.001, p < 0.001, p = 0.033, and p < 0.001, respectively). The IGD group showed KYN levels and KYNA/KYN ratios intermediate between those of the AUD group and HCs. Furthermore, in the AUD group, the mediating effect of AUD on KYN through stress level was moderated by resilience [index of moderated mediation = -0.557, boot S.E = 0.331, BCa CI (-1.349, -0.081)]. Stress may induce an imbalance in downstream of KYN pathway metabolites, and the KYN/TRP ratio may play as a neuromediator between stress and behavioral changes in both addictive disorders. This study suggests that regulation of the KYN pathway is critical in the pathophysiology of addictive disorders and it may serve as an important target for future treatment modalities.

Assessment of dietary exposure to heterocyclic amines based on the Korean total diet study.

Heterocyclic amines (HCAs) are contaminants in proteinaceous foods produced by cooking at high temperatures. This study was the first assessment of exposure to HCAs using the Korean total diet study. Twelve HCAs were analysed in 1,232 pooled samples using six isotope-labelled internal standards and HPLC-MS/MS. The daily intake of HCAs in the Korean population was estimated based on the concentration of HCAs in the total diet study samples and individual food consumption data from the Korean National Health and Nutrition Examination Survey. Among HCAs, the intake of ß-carbolines, such as harman and norharman, was the highest, followed by the intake of PhIP. The primary sources of HCA intake were meat, fish, shellfish, and beverages, including alcohol. The margin of exposure to PhIP was 2,349,000 at the average level and 373,000 at the 95th percentile in the Korean population. The estimated daily intake of all HCAs in the Korean population was considered safe.

Seasonal characteristics of atmospheric water-soluble organic nitrogen in PM2.5 in Seoul, Korea: Source and atmospheric processes of free amino acids and aliphatic amines.

The seasonal characteristics of atmospheric water-soluble organic nitrogen (WSON) in particulate matter with a diameter of 2.5 µm or smaller (PM2.5) were analyzed focusing on sources and atmospheric processing. Daily collected samples over 23 h (10:00-9:00) from 7 August 2018 to 31 December 2019 on quartz filters with a high-volume sampler at the Korea Institute of Science and Technology (KIST) in Seoul were considered. The most common species in the Seoul atmosphere included Glycine (5.45 ± 9.81 ng/m3) among free amino acids (FAAs) and trimethylamine (TMA) (5.35 ± 3.80 ng/m3) among aliphatic amines (AAs). The top 10 WSON species (93.6% of all WSON species) were categorized into three groups based on correlation analysis considering meteorological data, (e.g., temperature, rainfall, relative humidity (RH), wind speed) gaseous pollutants (e.g., SO2, CO, NO2) and mass concentration of PM10 and PM2.5. Those three groups are G1 (Glycine, Alanine, and Threonine), G2 (Gln Glutamine, Lys Lysine, and Glutamic acid) and G3 (Trimethylamine (TMA), dimethylamine (DMA), and methylamine (MA)), where G1, G2 and G3 accounted for 31.1%, 8.8% and 51.1%, respectively, of the total species. Among these three groups, G1 and G3 are from combustion sources, and G2 shows secondary features generated by photochemical reactions involving ozone. Although both G1 and G3 exhibited features influenced by combustion sources, the AA species (TMA, DMA, and MA) in G3 demonstrated typical features enhanced under high-humidity conditions, suggesting not only primary sources but also secondary formation at the local scale influence to the AA in G3 group. Based on long-term measurements more than a year, our findings suggest that complex and diverse sources of atmospheric WSON are in Seoul, Korea both from primary and secondary, which may affect its environmental, climate and health.

Bimodal neural probe for highly co-localized chemical and electrical monitoring of neural activities in vivo.

Investigation of the chemical and electrical signals of cells in vivo is critical for studying functional connectivity and brain diseases. Most previous studies have observed either the electrical signals or the chemical signals of cells because recording electrical signals and neurochemicals are done by fundamentally different methods. Herein, we present a bimodal MEMS neural probe that is monolithically integrated with an array of microelectrodes for recording electrical activity, microfluidic channels for sampling extracellular fluid, and a microfluidic interface chip for multiple drug delivery and sample isolation from the localized region at the cellular level. In this work, we successfully demonstrated the functionality of our probe by monitoring and modulating bimodal (electrical and chemical) neural activities through the delivery of chemicals in a co-localized brain region in vivo. We expect our bimodal probe to provide opportunities for a variety of in-depth studies of brain functions as well as for the investigation of neural circuits related to brain diseases.

Maintenance and differentiation of human ES cells on polyvinylidene fluoride scaffolds immobilized with a vitronectin-derived peptide.

Polyvinylidene fluoride (PVDF) is biocompatible, easy to fabricate, and has piezoelectric properties; it has been used for many biomedical applications including stem cell engineering. However, long-term cultivation of human embryonic stem cells (hESCs) and their differentiation toward cardiac lineages on PVDF have not been investigated. Herein, PVDF nanoscaled membrane scaffolds were fabricated by electrospinning; a vitronectin-derived peptide-mussel adhesive protein fusion (VNm) was immobilized on the scaffolds. hESCs cultured on the VNm-coated PVDF scaffold (VNm-PVDF scaffold) were stably expanded for more than 10 passages while maintaining the expression of pluripotency markers and genomic integrity. Under cardiac differentiation conditions, hESCs on the VNm-PVDF scaffold generated more spontaneously beating colonies and showed the upregulation of cardiac-related genes, compared with those cultured on Matrigel and VNm alone. Thus, VNm-PVDF scaffolds may be suitable for the long-term culture of hESCs and their differentiation into cardiac cells, thus expanding their application in regenerative medicine.

Analysis of saponin composition and comparison of the antioxidant activity of various parts of the quinoa plant (Chenopodium quinoa Willd.).

Quinoa plant is a valuable food crop because of its high nutritional and functional values. Total saponin content, sapogenins, polyphenol, and flavonoid contents and antioxidant activities were analyzed in various parts of quinoa plants, including sprout, seeds, bran, pericarp, leave, stem, and root. Quinoa seeds (QS) had significantly higher sapogenin content than quinoa stem (QT), quinoa leaves (QL), and quinoa roots (QR). Quinoa saponin was mainly composed of phytolaccagenic acid. Quinoa root (QR) had the highest amount of total saponin (13.39 g 100 g-1), followed by quinoa bran. The highest total phenolic content (30.96 mg GAE 100 g-1) and total flavonoid content (61.68 mg RE 100 g-1) were observed in quinoa root extract and 1-month-old sprout extract, respectively. Quinoa sprouts showed better antioxidant activity than fully grown parts of the quinoa plant. Overall, root and sprout had a higher antioxidant capacity compared to other parts of the quinoa plant, suggesting the potential use of quinoa root and sprout as a nutraceutical ingredient in the health food industry.

Validation of analytical methods for heterocyclic amines in seven food matrices using high-performance liquid chromatography-tandem mass spectrometry.

Heterocyclic amines (HCAs) are potent mutagens generated by the high temperatures of the cooking process. The purpose of this study was to develop and validate analytical methods for HCAs determination using high-performance liquid chromatography-tandem mass spectrometry in seven food matrices: corn oil, milk, 20% ethanol, pork, flat fish, sea mustard (Undaria pinnatifida), and radish. Six isotopically labelled internal standards were used for quantitation, and Chem Elut and Oasis hydrphilic-liphophilic balance cartridges were applied for sample preparation to remove interferences. Calibration curves showed good linearity (R2 > 0.99) in all matrices. The ranges of the method detection limit and method quantitation limit were 0.009-2.35 ng g-1 and 0.025-7.13 ng g-1, respectively. The recoveries ranged from 67.5% to 119.6%. The coefficients of variation ranged from 0.3% to 15.1% for intra-day and ranged from 0.8% to 19.1% for inter-day. The methods were applied to 24 total diet study samples for HCAs quantitation. These results indicate that the established methods are reliable for determining HCAs in various foods.

Analytical Method of Multi-Mycotoxins in Table-Ready Foods for a Total Diet Study Using Stable Isotope Dilution Liquid Chromatography-Tandem Mass Spectrometry.

Background: Determining the multi-mycotoxins present in table-ready foods is necessary for a total diet study. However, so far, most methods of analyzing multi-mycotoxins apply to raw foods. Therefore, a reliable method for analyzing multi-mycotoxins in table-ready foods is needed. Objective: We developed and validated methods of multi-mycotoxin analysis that employed stable isotope dilution with LC-tandem MS (LC-MS/MS) using two representative matrices. Methods: Samples were fortified with [13C]-labeled mycotoxins as internal standards and extracted with 50% acetonitrile in water for high-carbohydrate foods and 3% formic acid in acetonitrile for high-protein and/or high-fat foods, cleaned up with n-hexane and the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method, and followed up by LC-MS/MS. Results: Validation of the methods was performed, and the results were as follows: the correlation coefficient, R², was 0.99; method detection limit, 0.01-2.4 µg/kg; recoveries, 83.6-114%; precision, 0.8-10 (intraday) and 3.1-22% (interday); interlaboratory reproducibility, ≤15%; and uncertainty, 3.5-19%error. The applicability of the methods was evaluated by analyzing table-ready foods spiked with standards. Conclusions: These methods were successfully evaluated and deemed appropriate for determining the multi-mycotoxins in table-ready foods. Highlights: This work demonstrates that stable isotope dilution with LC-MS/MS can be effectively used to analyze multi-mycotoxins simultaneously in a total diet study.

Preparation and characterization of sericin powder extracted with deep sea water.

In this study, we developed an effective technology for the extraction of sericin from silk cocoons by deep sea water (DSW). We focused on extraction of sericin in the absence of chemical additives to obtain a safe, effective, inexpensive sericin powder. Sericin was extracted using a simple high-temperature process involving heating, condensation with Molus alba, filtering with cotton cloth, cold storage, and lyophilization. The results showed that the yield of sericin (26%) extracted by DSW was approximately 2% higher than that obtained using a chemical buffer (0.2 M Na2CO3, 24%). The marine mineral sericin M. alba (MSM) showed a size distribution of 15-250 kDa, with major peaks at 75-250 kDa with a galactose chain. Additionally, this MSM product had high antioxidant, whitening, and antibiosis effects and could be safely stored for a long time. Thus, our findings supported the use of a DSW extraction method, which was ecofriendly and yielded a proteinous, biodegradable biopolymer, for preparation of sericin.

Nitrite ion mitigates the formation of N-nitrosodimethylamine (NDMA) during chloramination of ranitidine.

Ranitidine (RNT) has been an important tertiary amine precursor of N-nitrosodimethylamine (NDMA) in chlorine-based water treatment, due to reaction with monochloramine (NH2Cl) with exceptionally high molar yields up to 90%. This study examined the effects of nitrite ions (NO2-) on the kinetics of NDMA formation during the chloramination of RNT under variable concentrations of dissolved oxygen (DO, 0.7-7.5mg/L), RNT (5-30µM), NH2Cl (5-20mM), NO2- or NO3- (0-2mM) and pH (5.6-8.6). In the absence of the NO2-, the ultimate molar yield of NDMA after 6h of reaction was primarily influenced by [DO] and pH, while marginally affected by initial [RNT] and [NH2Cl]. A kinetic model, prepared in accordance with the reaction sequence of NDMA formation, suggested that the rate determining step was accelerated with increasing [NH2Cl]0, [DO], and pH. A Kinetic study together with ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometer (UPLC-Q-TOF MS) and gas chromatography (GC)/TOF MS analyses in parallel demonstrated that the nitrite ion inhibited the nucleophilic substitution of the terminal amine on NH2Cl, and reduced the pseudo-steady state concentration of N-peroxyl radicals, significantly decreasing the ultimate yields of NDMA.

Elimination of microcystin-LR and residual Mn species using permanganate and powdered activated carbon: Oxidation products and pathways.

The oxidation of microcystin-LR (MC-LR) in deionized water (DI) and river water using potassium permanganate (KMnO4) at a neutral pH and at 23 ± 2 °C was investigated. These two aqueous systems (i.e., DI and river water) gave comparable second-order rate constants (289.9 and 285.5 M-1s-1 (r2 > 0.99), respectively), which confirmed the effectiveness of this oxidation process for the treatment of natural surface water. The presence of either humic or fulvic acid reduced the removal efficiency of MC-LR, with the latter exhibiting a greater inhibitory effect. Monitoring of MC-LR and residual Mn2+ levels with adding KMnO4 (1 mg/L) and powdered activated carbon (PAC, 5-20 mg L-1) before and during coagulation, respectively, revealed that 60 min of permanganate pre-oxidation followed by coagulant addition with PAC was the most effective approach for reducing both levels below limits stated by WHO guidelines. The MC-LR degradation products were the result of oxidation occurring at the diene and aromatic moieties of the Adda (3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid) side-chain, in addition to amine bond hydrolysis of the Mdha (N-methyldehydroalanine) moiety. Several toxic by-products with an intact Adda chain were observed during the reaction, but completely disappeared after 60 min. This further supports the conclusion that sufficient contact time with permanganate (i.e., >60 min) is essential to reducing the residual toxicity and maximizing the efficiency of MC-LR oxidation when treating raw water.

Discovery of 1-(3-(benzyloxy)pyridin-2-yl)-3-(2-(piperazin-1-yl)ethyl)urea: A new modulator for amyloid beta-induced mitochondrial dysfunction.

Herein, we report a new series of aliphatic substituted pyridyl-urea small molecules synthesized as potential modulators for amyloid beta (Aß) induced mitochondrial dysfunction. Their blocking activities against Aß-induced mitochondrial permeability transition pore (mPTP) opening were evaluated by JC-1 assay which measures the change of mitochondrial membrane potential (ΔΨm). The inhibitory activity of sixteen compounds against Aß-induced mPTP opening was superior or almost similar to that of the standard Cyclosporin A (CsA). Among them, 1-(3-(benzyloxy)pyridin-2-yl)-3-(2-(piperazin-1-yl)ethyl)urea (5x) effectively maintained mitochondrial function and cell viabilities on ATP assay, MTT assay, and ROS assay. Using CDocker algorithm, a molecular docking model presented a plausible binding mode for 5x with cyclophilin D (CypD) receptor as a major component of mPTP. Moreover, hERG and BBB-PAMPA assays presented safe cardiotoxicity and high CNS bioavailability profiles for 5x. Taken as a whole, this report presents compound 5x as a new nonpeptidyl mPTP blocker may hold a promise for further development of Alzheimer's disease (AD) therapeutics.

Synthesis of new diarylamides with pyrimidinyl pyridine scaffold and evaluation of their anti-proliferative effect on cancer cell lines.

A new series of diarylamides, having a pyrimidinyl pyridine scaffold, was designed and synthesized. The target compounds were synthesized in three steps. A selected group from the target compounds was tested over a panel of 60 cancer cell lines at a single dose concentration of 10 µM, and the most active compound, 5j, was further tested in a five-dose testing mode to determine its IC50 value over the 60 cell lines. In single-dose testing mode, compound 5j showed the highest growth inhibition against the NCI-60 cancer cell lines, while other tested compounds showed a weak to moderate inhibitory activity against a range of different cancer cell lines. In five-dose testing mode, compound 5j showed strong inhibitory activity in micro molar range against many cancer cell lines. Its major activity was against melanoma cancer cell lines. Therefore, compound 5j is a promising hit compound targeting this severe form of cancer.