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In the study, by using 5-amino-4-arylazo-3-methyl-1H-pyrazole derivatives and 2-hydroxy-5(phenyldiazenyl) benzaldehyde; eight novel heterocyclic disperse disazo-azomethine dyes were synthesized, their chemical structures were characterized via FT-IR and 1 H NMR studies. Synthesized compounds were also investigated computationally by performing various techniques. In the computational part of the study, geometry optimizations, frequency analyses, frontier molecular orbital (FMO) calculations, molecular electrostatic potential (MEP) map calculations, FT-IR and NMR spectral analyses were performed on the compounds. Additionally, to reveal their potentials against SARS-CoV-2 main protease (SARS-CoV-2 M-pro), acetylcholine esterase (AChE), heat shock protein (HSP90 alpha) and nicotinamide N-methyl transferase (NNMT), molecular docking calculations were also performed on the synthesized compounds. Molecular dynamics (MD) simulations were carried out on the top-scoring ligand-receptor complexes to evaluate the stability of the complexes and the interactions between ligands and receptors in more detail. Results showed that synthesized compounds can interact with all these four receptors effectively and can be promising structures for further studies. (C) 2021 Elsevier B.V. All rights reserved.

The application of Peptide Nucleic Acids (PNAs), mimics of DNA lacking the sugar-phosphate backbone, for antisense/anti-gene therapy and gene editing is limited by their low uptake by cells. Currently, no simple and efficient delivery systems and methods are available to solve this open issue. One of the most promising approach is the modification of the PNA structure through the covalent linkage of poliarginine tails, but this means that every PNA intended to be internalized must be modified. Herein we report the results relative to the delivery ability of a macrocyclic multivalent tetraargininocalix[4]arene (1) used as non-covalent vector for anti-miR-221-3p PNAs. High delivery efficiency, low cytotoxicity, maintenance of the PNA biological activity and ease preparation of the transfection formulation, simply attained by mixing PNA and calixarene, candidate this vector as universal delivery system for this class of nucleic acid analogues.

The implications of lysine-specific demethylase-1 (LSD1) in tumorigenesis have urged scientists to develop diagnostic tools in order to explore the function of this enzyme. In this work, we present our efforts on the development of tranylcypromine (TCP)-based functionalized probes for activity-based protein profiling (ABPP) of LSD1 activity. Biotinylated forms of selected compounds enabled dose-dependent enzyme labeling of recombinant LSD1. However, treatment with LSD1 inhibitors did not clearly reduce the LSD1 labeling efficiency thus indicating that labeling using these probes is not activity dependent. This calls for alternative strategies to develop probes for ABPP of the enzyme LSD1.

Hydrogels are three-dimensional polymeric networks capable of absorbing large amounts of water or biological fluids with the properties resembling natural living tissues. Herein, polyvinyl alcohol (PVA)/N-succinyl chitosan (NSCS)/lincomycin hydrogels for wound dressing were prepared by the freezing/thawing method, then characterized by FTIR, SEM, and TGA. The compression strength, swelling behavior, water retention capacity, antibacterial activity, drug release and cytotoxicity were systematically investigated. The results showed that the introduction of NSCS remarkably enhanced the swelling capacity, leading to the maximum swelling ratio of 19.68 g/g in deionized water. The optimal compression strength of 0.75 MPa was achieved with 30 % NSCS content.Additionally, the incorporation of lincomycin brought a remarkable antibacterial activity against both Escherichia coli and Staphylococcus aureus. Specifically, 77.71 % of Staphylococcus aureus was inhibited with 75 μg/mL lincomycin, while the MTT assay demonstrated the nontoxic nature of the composite hydrogels. In summary, this PVA/NSCS/lincomycin hydrogel showed promising potential for wound dressing. ispartof: CARBOHYDRATE POLYMERS vol:261 ispartof: location:England status: published

The present study investigated the effects of the combined treatment of two peptide nucleic acids (PNAs), directed against microRNAs involved in caspase‑3 mRNA regulation (miR‑155‑5p and miR‑221‑3p) in the temozolomide (TMZ)‑resistant T98G glioma cell line. These PNAs were conjugated with an octaarginine tail in order to obtain an efficient delivery to treated cells. The effects of singularly administered PNAs or a combined treatment with both PNAs were examined on apoptosis, with the aim to determine whether reversion of the drug‑resistance phenotype was obtained. Specificity of the PNA‑mediated effects was analyzed by reverse transcription‑quantitative polymerase‑chain reaction, which demonstrated that the effects of R8‑PNA‑a155 and R8-PNA-a221 anti‑miR PNAs were specific. Furthermore, the results obtained confirmed that both PNAs induced apoptosis when used on the temozolomide‑resistant T98G glioma cell line. Notably, co‑administration of both anti‑miR‑155 and anti‑miR‑221 PNAs was associated with an increased proapoptotic activity. In addition, TMZ further increased the induction of apoptosis in T98G cells co‑treated with anti‑miR‑155 and anti‑miR‑221 PNAs.

Sen, Ali/0000-0002-2144-5741, WOS: 000503393100009, In this study, we report anti-inflammatory and antioxidant activities of two acylated isoscutellarein glucosides isolated from ethyl acetate extract of Stachys subnuda aerial part. 4'-O-methylisoscutellarein-7-O-2 ''-O-(6 ''' acetyl-beta-D-allopyranosyl)-beta-D-glucopyranoside (SS1) and isoscutellarein-7-O-2 ''-O-(6 '''-O-acetyl-beta-D allopyranosyl)-beta-D -glucopyranoside (SS2) were isolated as major compounds from ethyl acetate extract (SSEA). Also, 2 hydroxycinnamic acid derivatives, and 5 isoscutellarein glucoside derivatives in the SSEA were identified using LC-MS/MS. SS1 with IC50 values of 2.35 and 1.98 mu g/mL and SS2 with IC50 values 13.94 and 12.76 mu g/mL showed fairly strong antioxidant activity against DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid) radicals, respectively. SS1 and SS2 inhibited 5-lipoxygenase (5-LOX) activity with IC(50 )values of 47.23 and 41.60 mu g/mL, respectively. the results demonstrated that SS1 and SS2 have significant anti-inflammatory and antioxidant potential. Acylated flavonoid glycosides have been first reported for Stachys subnuda. Also, the activities of SS1 and SS2 have been investigated for the first time in this study., Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [1919B011603877, 2209-A], This work was supported by a grant from the Scientific and Technological Research Council of Turkey (TUBITAK), under the project 1919B011603877 (Code: 2209-A).

Five copper(II) complexes 1-5 with aromatic nitrogen-containing heterocycles, pyrimidine (pm, 1), pyrazine (pz, 2), quinazoline (qz, 3 and 4) and phthalazine (phtz, 5) have been synthesized and structurally characterized by spectroscopic and single-crystal X-ray diffraction techniques. The crystallographic results show that, dependent on the ligand structure, complexes 1-5 are of different nuclearity. The antimicrobial efficiency of complexes 1-5 has been evaluated against three clinically relevant microorganisms and none of the complexes showed significant growth inhibiting activity, with values of minimum inhibitory concentrations (MIC) in the mM range. Since in many bacteria, pathogenicity and virulence are regulated by intercellular communication processes, quorum sensing (QS), the effect of the copper(II) complexes on bacterial QS has also been examined. The results indicate that the investigated complexes inhibit violacein production in Chromobacterium violaceum CV026, suggesting an anti-QS activity. In order to differentiate, which of the QS pathways was affected by the copper(II) complexes, three biosensor strains were used: the PAO1 Delta rhlIpKD-rhlA and the PA14-R3 Delta lasIPrsaI lux strain to directly measure the levels of C4-HSL (N-butanoyl-homoserine lactone) and 3OC12-HSL (N-3-oxo-dodecanoyl- homoserine lactone), respectively, and PAO1 Delta pqsA mini-CTX luxPpqsA for the detection of AHQs (2-alkyl-4-quinolones). Complexes 1-5 were shown to be efficient inhibitors of biofilm formation of the human opportunistic pathogen Pseudomonas aeruginosa PAO1, with the qz-containing complex 3 being the most active. Finally, the most anti-QS-active complexes 1 and 3 showed synergistic activity against a multi-drug resistant clinical isolate of P. aeruginosa, when supplied in combination with the known antibiotics piperacillin and ceftazidime.

The ability of peforelin (l-GnRH-III) to stimulate follicular growth, FSH release, and estrus in gilts after altrenogest treatment and in sows after weaning was investigated. In three farrow-to-wean herds, with at least 600 sows and average production performance, 216 gilts, 335 primiparous, and 1299 pluriparous sows were randomly allocated to three treatments: peforelin (M group: Maprelin), eCG (F group: Folligon), and physiological saline solution (C group). Animals were treated 48 hours after their last altrenogest treatment (gilts) or 24 hours after weaning (sows). The weaning-to-estrus interval, estrus duration, estrus rate (ER), pregnancy rate, and total born (TB), live born, and stillborn (SB) numbers were recorded and compared between treatments for the different parity groups (gilts and primiparous and pluriparous sows). Follicle sizes were measured in representative animals from each group on the occasion of their last altrenogest treatment or at weaning, and also on the occasions of their first (FS1) and second (FS2) attempted inseminations. Blood samples were taken to determine FSH concentrations at weaning and 2 hours after injection, and progesterone concentrations 10 days after the first insemination attempt. The relative change in FSH concentrations was calculated. Significant differences were found for ER within 7 days of weaning in pluriparous sows (95%, 91%, and 90% for the M, F, and C groups, respectively, P = 0.005). Gilts in the F-group had high TB numbers, and pluriparous sows in the M group had high SB numbers (TB gilts = 13.6, 15.4, and 14.9 [P = 0.02] and SB pluriparous sows = 1.8, 1.4, and 1.7 [P = 0.05] for the M, F, and C groups, respectively). The M group had the highest FS1 (for gilts) and FS2 (for pluriparous sows) values: FS1 = 5.4, 4.9, and 4.9 mm [P = 0.02] and FS2 = 6.8, 5.3, and 6.3 mm [P = 0.03] for the M, F, and C groups, respectively. There were no significant differences between the different treatments within each parity group with respect to any of the other variables. Overall, peforelin treatment had small but positive effects on the ER and follicle growth in certain parity groups but did not seem to affect litter sizes or FSH and progesterone levels in sows on the occasions of the corresponding examinations.

8-Quinolinecarboxaldehyde selenosemicarbazone (H8qasesc) and its octahedral Co(III) complex were characterized by single crystal X-ray diffraction analysis, spectroscopy methods and cyclic voltammetry. The antineoplastic activity of the ligand and the complex has been assessed on an acute monocytic leukemia cell line (THP-1) and AsPC-1 cancer stem cell (CSC) line derived from a patient suffering from pancreatic adenocarcinoma, with cisplatin (CDDP) as a reference compound. Evaluation involved determination of pro-apoptotic activity, changes in cell cycle distribution, the role of caspase activation in the process of cell death, and the ability of the investigated compounds to challenge reprogramming of the CSC phenotype. Compared to CDDP, treatment with H8qasesc induced a higher apoptotic response in both investigated cell lines. Apoptosis triggered by H8qasesc was highly caspase-dependent but did not include activation of either caspase-8 or -9. According to cell cycle changes H8qasesc delayed the transition of cells during DNA replication but in a manner different from that of CDDP. The ligand did not show nuclease activity on pUC19 plasmid, while docking studies disclosed that it does not have intercalating properties. Treatment of THP-1 cells with the Co(III) complex resulted in a strong toxic response, whereas cell death in the treated AsPC-1 line was not achieved for 24 h. Additionally, the complex concentration-dependently digested plasmid DNA which might be the cause of its cytotoxic activity. Finally, H8qasesc successfully initiated reprogramming of the CSC phenotype in the AsPC-1 cell line. This is peer-reviewed version of the following article: Filipović, N. R.; Bjelogrlić, S.; Portalone, G.; Pelliccia, S.; Silvestri, R.; Klisurić, O.; Senćanski, M.; Stanković, D.; Todorović, T. R.; Muller, C. D. Pro-Apoptotic and pro-Differentiation Induction by 8-Quinolinecarboxaldehyde Selenosemicarbazone and Its Co(III) Complex in Human Cancer Cell Lines. MedChemComm 2016, 7 (8), 1604–1616. [https://doi.org/10.1039/c6md00199h] Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3599]

A highly efficient synthesis of C3-arylbutenolides by Suzuki-Miyaura coupling is described. We developed a simple protocol to accomplish scaffolds of C3-arylbutenolides which include antifungal agent (−)-incrustoporin and twelve analogues by Pd-catalyzed Suzuki-Miyaura cross coupling reaction between chiral 3-iodobutenolide and various aryl boronic acids under aerobic conditions in good to quantitative yields.

In this study we report the anticancer properties of RN5-Me, an α1-adrenergic receptor ligand. Biological screening and circular dichroism data indicate that it acts as a DNA intercalator. Docking studies, confirming this behaviour, suggest that RN5-Me possesses great selectivity for alternating AT nucleobases upon GC ones. In the cytotoxicity assay, it shows IC50 values in the range of 4.2–25.0 nM towards the cancer cell lines HN6, HN13, HeLa, SK-Me1-103, PC3, and MCF7. It is noteworthy that RN5-Me shows a higher selectivity than Cisplatin for HN6 and HN13 over NOK-SI non-cancerous cells.

The copper(I) complexes containing phosphorus donor ligands such as diazadiphosphetidine, cis-{(o-OCH2C5H4N)P(mu-(NBu)-Bu-t)}(2) (1) and aminobis(phosphonite), C6H5N{P(OC6H3(OMe-o)(C3H5-p))(2)}(2) (2, PNP), have been synthesized. Treatment of 1 with copper iodide afforded the 1D coordination polymer [{Cu(mu-I)}(2){(o-OCH2C5H4N)P(mu-(NBu)-Bu-t)}(2)](n) (3). Treatment of 3 with 2,2'-bipyridine (bpy) and 1,10-phen-anthroline (phen) produced mixed-ligand complexes [(L)(2)Cu-2{(o-OCH2C5H4N)P(mu-(NBu)-Bu-t)}(2)][I](2) (4 L = bpy; 5 L = phen) in good yields. The reaction of 2 with copper iodide yielded a rare tetranuclear copper complex [(CuI)(2)C6H5N(PR2)(2)](2) (6), which on subsequent treatment with various pyridyl ligands produced binuclear complexes [{Cu(mu-I)(py)}(2)(mu-PNP)] (7), [Cu-2(mu-I)(bpy)(2)(mu-PNP)]I (8), [Cu-2(mu-I)I(bpy)(mu-PNP)] (9), [Cu-2(phen)(bpy)(mu-PNP)](OTf)(2) (10), [Cu-2(mu-I)I(phen)(mu-PNP)] (11) and [Cu-2(mu-I(phen)(2)(mu-PNP)]I (12), in an almost quantitative yield. The new copper(I) complexes (4, 5 and 7-12) were tested for anti-cancer activity against three human tumor cell lines. Compounds 5, 10 and 12 showed in vitro antitumor activity 5-7 fold higher than cisplatin, the most used anticancer drug. These three most potent compounds (5, 10 and 12) were chosen for detailed study to understand their mechanism of action. The copper(I) compounds studied in the present investigation were found to inhibit tumor cell growth by arresting cells at the S-phase of the cell cycle. The characteristic nuclear morphology of treated cells showed signs of DNA damage. The experimental evidence clearly indicated that these compounds initiated apoptosis, which is mediated through the p53 pathway.

Liver fibrosis represents a scar formation process as a response to chronic injury and a major cause of death worldwide. To date, no drug is available for this condition. Interleukin-10 (IL-10) has potent anti-inflammatory and antifibrotic properties but its short half-life in the circulation hampers its clinical use. Our aim was therefore to modify IL-10 with polyethylene glycol (PEG) to prolong its circulation time and enhance its effectivity. IL-10 was modified with 5 or 20 kDa PEG. The biological activity was preserved after PEGylation as assessed by inhibition of TNF-alpha production by macrophages. In vivo, during CCl4-induced fibrogenesis in mice, both 5PEG-IL-10 and 20PEG-IL-10 showed a longer circulation time compared to IL-10, which was associated with a significant increased liver accumulation. Immunohistochemical analysis of fibrotic livers of mice receiving treatment with IL-10 or its PEGylated forms, revealed a decrease in markers reflecting HSC and KC activation induced by 5PEG-IL10. Transcription levels of IL-6 were decreased upon treatment with IL-10 and both PEGylated forms, whereas IL-1 beta levels were only down-regulated by 5PEGIL-10 and 20PEGIL-10. We conclude that PEGylation of IL-10 is a good strategy to attenuate liver fibrosis and that 5PEGIL-10 is the most effective conjugate. (C) 2012 Elsevier B. V. All rights reserved.

Compounds of formula [Pt(diam)(Soa)]NO3, 3, and [Pt(diam)(Sob)]NO3, 4, (Soa, (methylsulfinyl)acetate; Sob, 2-(methylsulfinyl)benzoate; diam, chelating diamines: 1,2-ethylenediamine, en; (+/-)-, R,R-, S,S-, and meso-1,2-diaminocyclohexane, dach; 1,1-bis(aminomethyl)cyclohexane, damch) have been synthesized. IR, NMR, and FAB-mass spectroscopy suggest that the Soa and Sob anions are chelated to Pt through the S atom and the carboxylate group. Such a mode of coordination was confirmed for compounds 3 by the X-ray crystal structure determination of [Pt(en)(Soa)]NO3. This compound crystallizes in space group R3 (No. 146) with cell constants a = 15.139(3) angstrom and c = 12.886(2) angstrom. The structure was refined using 1001 independent reflections with I > 3sigma(I), giving a final R value of 0.018. In the complex cation Pt is in a square planar environment with en (Pt-N, 2.019(8) and 2.055(8) angstrom) and Soa (Pt-S, 2.184(2) angstrom, and Pt-O, 2.025(6) angstrom) chelated to Pt. The reactivities of 3 and 4 toward Cl-, 5'-GMP, and 9-methylguanine have been investigated by H-1 NMR spectroscopy at 40-degrees-C and complex concentration 10(-2) mol L-1 in D2O. Compounds 3 react readily with Cl- by displacement of the carboxylate group, yielding [PtCl(diam)(Soa-S)] (with monodentate S-coordinated Soa) which reacts with excess chloride to give [PtCl2-(diam)] at a very slow rate (for the en derivative formation of [PtCl2(en)] 24 h, but the concentration of the intermediates with S-coordinated monodentate Sob remains very low throughout the course of the reaction, indicating that the Sob chelate ring is more inert toward ring opening. Reactions of 3 and 4 toward 5'-GMP are rather fast: formation of [Pt(dach)(GMP)2]2- is complete in 3 and 1 h respectively (Pt/GMP = 1/2). These reactions proceed via the formation of intermediates with one N(7)-bound GMP and one monodentate Soa, coordinated to Pt via the S atom, or Sob probably bound to Pt via the O (sulfinyl) atom. Reactions with 9-methylguanine are slower and occur with a similar mechanism. The first step of the reaction of 5 with 2 mol of GMP is displacement of Cl by GMP. Formation of [Pt(en)(GMP)2]2- is complete in 75 min. Complexes 3 and 4 are moderately cytotoxic toward L1210 leukemia cells; the dach and damch derivatives are cytotoxic also against the L1210 cisplatin-resistant line. The cytotoxicities of the dach complexes depend not only on the absolute configuration of the diamine, but also on the configuration of the leaving group.

This study reports on new proteolytically stable, pharmacologically active endomorphin analogues, incorporating Dmt(1), Achc(2), pFPhe(4), or beta MePhe(4) unnatural amino acids. Consistent with earlier results, it was found that the analogues carrying anti and Achc2 residues displayed the highest p-opioid receptor affinities, depending upon the configuration of the incorporated Achc(2). Combination of such derivatives with pFPhe(4) or beta MePhe(4) yielded further compounds with variable binding potencies. Combined application of Dmt(1), cis-(1S,2R)Achc(2), and pFPhe(4) (compound 16) resulted in the most potent analogue. Ligand stimulated [S-35]GTP gamma S binding assays indicated that the analogues retained their agonist activities and opioid receptor specificities. NMR and molecular modeling studies of the analogues containing beta MePhe(4) or pFPhe(4) confirmed the predominance of bent structures, however, it is apparent that bent structures are energetically more favored than random/extended structures for all studied compounds.

It has recently been shown that some growth factors (GFs) have strong interactions with nonproteoglycan extracellular matrix proteins. Relevant here, the 12th-14th type three repeats of fibronectin (FN III12-14) have been shown to bind insulin-like growth factor binding-protein-3, fibroblast growth factor (FGF)-2, and vascular endothelial growth factor (VEGF)-A with high affinity. Since FN III12-14 is known to bind GFs from different families, we hypothesized that this domain could be highly promiscuous in its GF-binding capacity. We used biochemical approaches and surface plasmon resonance to investigate such interactions with recombinant FN III12-14. We found that FN III12-14 binds most of the GFs from the platelet-derived growth factor (PDGF)/VEGF and FGF families and some GFs from the transforming growth factor-β and neurotrophin families, with K(D) values in the nanomolar range, without inhibiting GF activity. Overall, 25 new binding interactions were identified. In a clinically relevant fibrin matrix, a fibrin-binding variant of FN III12-14 was highly effective as a GF delivery system. For instance, in matrices functionalized with FN III12-14, PDGF-BB-induced sprouting of human smooth muscle cell spheroids was greatly enhanced. We show that FN III12-14 is a highly promiscuous ligand for GFs and also holds great potential in clinical healing applications.

Autoimmune uveitis is a blinding disease presenting with autoantibodies against eye-specific proteins as well as autoagressive T cells invading and attacking the immune-privileged target tissue retina. The molecular events enabling T cells to invade and attack the tissue have remained elusive. Changes in membrane protein expression patterns between diseased and healthy stages are especially interesting because initiating events of disease will most likely occur at membranes. Since disease progression is accompanied with a break-down of the blood-retinal barrier, serum-derived proteins mask the potential target tissue-related changes. To overcome this limitation, we used membrane-enriched fractions derived from retinas of the only available spontaneous animal model for the disease equine recurrent uveitis, and compared expression levels by a label-free LC-MSMS-based strategy to healthy control samples. We could readily identify a total of 893 equine proteins with 57% attributed to the Gene Ontology project term "membrane." Of these, 179 proteins were found differentially expressed in equine recurrent uveitis tissue. Pathway enrichment analyses indicated an increase in proteins related to antigen processing and presentation, TNF receptor signaling, integrin cell surface interactions and focal adhesions. Additionally, loss of retina-specific proteins reflecting decrease of vision was observed as well as an increase in Müller glial cell-specific proteins indicating glial reactivity. Selected protein candidates (caveolin 1, integrin alpha 1 and focal adhesion kinase) were validated by immunohistochemistry and tissue staining pattern pointed to a significant increase of these proteins at the level of the outer limiting membrane which is part of the outer blood-retinal barrier. Taken together, the membrane enrichment in combination with LC-MSMS-based label-free quantification greatly increased the sensitivity of the comparative tissue profiling and resulted in detection of novel molecular pathways related to equine recurrent uveitis.

Three analogues of 1α,25-dihydroxyvitamin D 3 (calcitriol), featuring a trans- fused decalin C,D-core with local S 2 -symmetry, and possessing identical side-chain and seco-B,A-ring structures, have been synthesized starting from readily available (4a R ,8a S )-octahydronaphthalene-1,5-dione ( 7 ). The very short sequences involve the simultaneous introduction of the side-chain and seco-B,A-ring fragments via Suzuki and Sonogashira coupling reactions. The analogues are devoid of relevant biological activity. Keywords: Suzuki coupling; Sonogashira reaction; Calcitriol analogues. Introduction Since the discovery that the biological action of vitamin D 3 originates from the dihydroxylated metabolite 1α,25-dihydroxyvitamin D 3 ( 1 , calcitriol) and that, next to its classical role in the regulation of calcium homeostasis, these actions also involve immunomodulation, cell differentiation and antiproliferation, there has been an intense search for structural analogues of calcitriol that might show a separation in calcemic and antiproliferative-prodifferentiating activities (Scheme 1) [1]. In this context various successful structural modifications have been introduced in each one of the three parts that one may distinguish in its structure: the rigid central C,D-ring system and the flexible parts of the molecule, comprising the side chain and the seco-B,A-ring [2].

New platinum complexes have been synthesized by the reaction of Na2PtCl4 with 2-acetylpyridine-4-cyclohexyl-thiosemicarbazone, HAc4CyclHexyl (1). The new complexes [Pt(Ac4Cyc1Hexyl)Cl] (2) and [Pt(Ac4CyclHexyl)(2)] (3) have been characterized by elemental analyses and spectroscopic studies. The crystal structure of the complex [Pt(Ac4CyclHexy1)Cl] - DMF has been solved by single-crystal X-ray diffraction. The anion of Ac4CyclHexyl coordinates in a planar conformation to the central platinum(II) through the pyridyl N, azomethine N and thiolato S atoms. The crystal packing is determined by double intermolecular hydrogen interactions, pi-pi, Pt-C and Pt-pi contacts. The cytotoxic activities of 1-3 have been evaluated for antiproliferative activity in vitro against the cells of three human cancer cell lines: MCF-7 (human breast cancer cell line), T24 (bladder cancer cell line), A-549 (non-small cell lung carcinoma) and a mouse L-929 (a fibroblast-like cell line cloned from strain L). The compounds 1-3 display IC50 values in a mu M range better than that of the antitumor drug cisplatin and are considered as agents with potential antitumor activity candidates for further stages of screening in vitro and/or in vivo. (C) 2007 Elsevier Ltd. All rights reserved. Polyhedron

The actin-ADP-ribosylating binary Clostridium botulinum C2 toxin consists of two individual proteins, the binding/translocation component C2II and the enzyme component C2I. To elicit its cytotoxic action, C2II binds to a receptor on the cell surface and mediates cell entry of C2I via receptor-mediated endocytosis. Here we report that binding of C2II to the surface of target cells requires cleavage of C2II by trypsin, Trypsin cleavage causes oligomerization of the activated C2II (C2IIa) to give SDS-stable heptameric structures, which exhibit a characteristic annular or horseshoe shape and form channels in lipid bilayer membranes. Cytosolic delivery of the enzyme component C2I is blocked by bafilomycin bat Plot by brefeldin A or nocodazole, indicating uptake from an endosomal compartment and requirement of endosomal acidification for cell entry. In the presence of C2IIa and C2I, short term acidification of the extracellular medium (pH 5.4) allows C2I to enter the cytosol directly. Our data indicate that entry of C2 toxin into cells involves (i) activation of C2II by trypsin-cleavage, (ii) oligomerization of cleaved C2IIa to heptamers, (iii) binding of the C2IIa oligomers to the carbohydrate receptor on the cell surface and assembly with C2I, (iv) receptor-mediated endocytosis of both C2 components into endosomes, and finally (v) translocation and release of C2I into the cytosol after acidification of the endosomal compartment.

We determined the in vitro cytotoxic activity of the sesquiterpene lactone endoperoxide artemisinin (1) and some chemically prepared derivatives, which have been found to display cytotoxicity to cloned murine Ehrlich ascites tumour (EAT) cells and human HeLa cells and against murine bone marrow using a clonogenic assay for committed progenitor cells of the granulocyte-monocyte lineage (CFU-GM assay). Comparing artemisinin (1) to deoxyartemisinin (2), the endoperoxide group appeared to play a role in cytotoxicity to CFU-GM cells. Dimers of dihydroartemisinin and dihydrodeoxyartemisinin revealed that the stereochemistry of the ether linkage of the dimers was a more important determinant for this cytotoxic activity. The nonsymmetrical dimer of dihydroartemisinin (3) and the corresponding endoperoxide-lacking dimer of dihydrodeoxyartemisinin (5) were equally cytotoxic to CFU-GM cells. Despite the differences between both systems, it may be stated that most compounds displayed higher cytotoxicity to CFU-GM cells than to EAT cells. Dimers of dihydroartemisinin (3, 4) were selected as potential antitumour compounds and subjected to the National Cancer Institute drug-screening programme consisting of about sixty human cancer cell lines derived from nine different tissues. Both compounds displayed the same specific cytotoxicity pattern. Throughout the screen dimer 3 was more active than 4.

Background: neuropeptide S (NPS) and its receptor NPSR1 act along the hypothalamic-pituitary-adrenal axis to modulate anxiety, fear responses, nociception and inflammation. The importance of the NPS-NPSR1 signaling pathway is highlighted by the observation that, in humans, NPSR1 polymorphism associates with asthma, inflammatory bowel disease, rheumatoid arthritis, panic disorders, and intermediate phenotypes of functional gastrointestinal disorders. Because of the genetic complexity at the NPSR1 locus, however, true causative variations remain to be identified, together with their specific effects on receptor expression or function. To gain insight into the mechanisms leading to NPSR1 disease-predisposing effects, we performed a thorough functional characterization of all NPSR1 promoter and coding SNPs commonly occurring in Caucasians (minor allele frequency .0.02). Principal Findings: we identified one promoter SNP (rs2530547 [2103]) that significantly affects luciferase expression in gene reporter assays and NPSR1 mRNA levels in human leukocytes. We also detected quantitative differences in NPSinduced genome-wide transcriptional profiles and CRE-dependent luciferase activities associated with three NPSR1 nonsynonymous SNPs (rs324981 [Ile107Asn], rs34705969 [Cys197Phe], rs727162 [Arg241Ser]), with a coding variant exhibiting a loss-of-function phenotype (197Phe). Potential mechanistic explanations were sought with molecular modelling and bioinformatics, and a pilot study of 2230 IBD cases and controls provided initial support to the hypothesis that different ciscombinations of these functional SNPs variably affect disease risk. Significance: these findings represent a first step to decipher NPSR1 locus complexity and its impact on several human conditions NPS antagonists have been recently described, and our results are of potential pharmacogenetic relevance. Citation: Anedda F, Zucchelli M, Schepis D, Hellquist A, Corrado L, et al. (2011) Multiple Polymorphisms Affect Expression and Function of the Neuropeptide S

While human chorionic gonadotropin (hCG) appears to have an essential role in early pregnancy, it is controversial whether the hyperglycosylated form of hCG (hCG-h), which is the major hCG isoform during the first 4–5 weeks of pregnancy, is able to activate LH/hCG receptor (LHCGR). To address this, we utilized different extensively characterized hCG and hCGβ reference reagents, cell culture- and urine-derived hCG-h preparations, and an in vitro reporter system for LHCGR activation. The WHO hCG reference reagent (99/688) was found to activate LHCGR with an EC50-value of 3.3 ± 0.6 pmol/L (n = 9). All three studied hCG-h preparations were also able to activate LHCGR, but with a lower potency (EC50-values between 7.1 ± 0.5 and 14 ± 3 pmol/L, n = 5–11, for all P

Inspired by the preferential, allosteric binding of RAPTA-T and auranofin to the nucleosome core particle, we describe the design and synthesis of a series of heterobimetallic ruthenium(II)-gold(I) complexes with varying spacer lengths ranging from four to eight polyethylene glycol units. Evaluation of their cytotoxicity reveals IC50 values in the low micromolar range against cisplatin sensitive and resistant human ovarian carcinoma (A2780, A2780cisR) and nontumoral human embryonic kidney (HEK293) cell lines. Binding studies monitored via mass spectrometry revealed an affinity for histidine residues on a fragment of the amyloid beta-protein (residues 1-16, employed as a model system), which is in accordance with the binding sites of parent drugs, RAPTA-C and auranofin, to the nucleosome core particle.