Supplementary data for the article: Nikolic, S.; Rangasamy, L.; Gligorijevic, N.; Arandelovic, S.; Radulovic, S.; Gasser, G.; Grguric-Sipka, S. Synthesis, Characterization and Biological Evaluation of Novel Ru(II)-Arene Complexes Containing Intercalating Ligands. J. Inorg. Biochem. 2016, 160, 156–165. https://doi.org/10.1016/j.jinorgbio.2016.01.005

Supplementary material for: [https://doi.org/10.1016/j.jinorgbio.2016.01.005]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2272]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3638

Effects of fluoxetine on functional outcomes after acute stroke (FOCUS): a pragmatic, double-blind, randomised, controlled trial

Background Results of small trials indicate that fluoxetine might improve functional outcomes after stroke. The FOCUS trial aimed to provide a precise estimate of these effects. Methods FOCUS was a pragmatic, multicentre, parallel group, double-blind, randomised, placebo-controlled trial done at 103 hospitals in the UK. Patients were eligible if they were aged 18 years or older, had a clinical stroke diagnosis, were enrolled and randomly assigned between 2 days and 15 days after onset, and had focal neurological deficits. Patients were randomly allocated fluoxetine 20 mg or matching placebo orally once daily for 6 months via a web-based system by use of a minimisation algorithm. The primary outcome was functional status, measured with the modified Rankin Scale (mRS), at 6 months. Patients, carers, health-care staff, and the trial team were masked to treatment allocation. Functional status was assessed at 6 months and 12 months after randomisation. Patients were analysed according to their treatment allocation. This trial is registered with the ISRCTN registry, number ISRCTN83290762. Findings Between Sept 10, 2012, and March 31, 2017, 3127 patients were recruited. 1564 patients were allocated fluoxetine and 1563 allocated placebo. mRS data at 6 months were available for 1553 (99·3%) patients in each treatment group. The distribution across mRS categories at 6 months was similar in the fluoxetine and placebo groups (common odds ratio adjusted for minimisation variables 0·951 [95% CI 0·839–1·079]; p=0·439). Patients allocated fluoxetine were less likely than those allocated placebo to develop new depression by 6 months (210 [13·43%] patients vs 269 [17·21%]; difference 3·78% [95% CI 1·26–6·30]; p=0·0033), but they had more bone fractures (45 [2·88%] vs 23 [1·47%]; difference 1·41% [95% CI 0·38–2·43]; p=0·0070). There were no significant differences in any other event at 6 or 12 months. Interpretation Fluoxetine 20 mg given daily for 6 months after acute stroke does not seem to improve functional outcomes. Although the treatment reduced the occurrence of depression, it increased the frequency of bone fractures. These results do not support the routine use of fluoxetine either for the prevention of post-stroke depression or to promote recovery of function. Funding UK Stroke Association and NIHR Health Technology Assessment Programme

A Perspective on Newly Emerging Proteolysis-Targeting Strategies in Antimicrobial Drug Discovery

Targeted protein degradation is a new aspect in the field of drug discovery. Traditionally, developing an antibiotic includes tedious and expensive processes, such as drug screening, lead optimization, and formulation. Proteolysis-targeting chimeras (PROTACs) are new-generation drugs that use the proteolytic mechanism to selectively degrade and eliminate proteins involved in human diseases. The application of PROTACs is explored immensely in the field of cancer, and various PROTACs are in clinical trials. Thus, researchers have a profound interest in pursuing PROTAC technology as a new weapon to fight pathogenic viruses and bacteria. This review highlights the importance of antimicrobial PROTACs and other similar “PROTAC-like” techniques to degrade pathogenic target proteins (i.e., viral/bacterial proteins). These techniques can perform specific protein degradation of the pathogenic protein to avoid resistance caused by mutations or abnormal expression of the pathogenic protein. PROTAC-based antimicrobial therapeutics have the advantage of high specificity and the ability to degrade “undruggable” proteins, such as nonenzymatic and structural proteins

Nanoparticle Enhancement of Natural Killer (NK) Cell-Based Immunotherapy

Natural killer (NK) cells are one of the first lines of defense against infections and malignancies. NK cell-based immunotherapies are emerging as an alternative to T cell-based immunotherapies. Preclinical and clinical studies of NK cell-based immunotherapies have given promising results in the past few decades for hematologic malignancies. Despite these achievements, NK cell-based immunotherapies have limitations, such as limited performance/low therapeutic efficiency in solid tumors, the short lifespan of NK cells, limited specificity of adoptive transfer and genetic modification, NK cell rejection by the patient’s immune system, insignificant infiltration of NK cells into the tumor microenvironment (TME), and the expensive nature of the treatment. Nanotechnology could potentially assist with the activation, proliferation, near-real time imaging, and enhancement of NK cell cytotoxic activity by guiding their function, analyzing their performance in near-real time, and improving immunotherapeutic efficiency. This paper reviews the role of NK cells, their mechanism of action in killing tumor cells, and the receptors which could serve as potential targets for signaling. Specifically, we have reviewed five different areas of nanotechnology that could enhance immunotherapy efficiency: nanoparticle-assisted immunomodulation to enhance NK cell activity, nanoparticles enhancing homing of NK cells, nanoparticle delivery of RNAi to enhance NK cell activity, genetic modulation of NK cells based on nanoparticles, and nanoparticle activation of NKG2D, which is the master regulator of all NK cell responses

Dynamics of a Novel IVRD Pandemic Model of a Large Population over a Long Time with Efficient Numerical Methods

The model of any epidemic illness is evolved from the current susceptibility. We aim to construct a model, based on the literature, different to the conventional examinations in epidemiology, i.e., what will occur depends on the susceptible cases, which is not always the case; one must consider a model with aspects such as infections, recoveries, deaths, and vaccinated populations. Much of this information may not be available. So without artificially assuming the unknown aspects, we frame a new model known as IVRD. Apart from qualitative evaluation, numerical evaluation has been completed to aid the results. A novel approach of calculating the fundamental reproduction/transmission range is presented, with a view to estimating the largest number of aspects possible, with minimal restrictions on the spread of any disease. An additional novel aspect of this model is that we include vaccines with the actively infected cases, which is not common. A few infections such as rabies, ebola, etc., can apply this model. In general, the concept of symmetry or asymmetry will exist in every epidemic model. This model and method can be applied in scientific research in the fields of epidemic modeling, the medical sciences, virology, and other areas, particularly concerning rabies, ebola, and similar diseases, to show how immunity develops after being infected by these viruses

Can 3D-Printed Bioactive Glasses Be the Future of Bone Tissue Engineering?

According to the Global Burden of Diseases, Injuries, and Risk Factors Study, cases of bone fracture or injury have increased to 33.4% in the past two decades. Bone-related injuries affect both physical and mental health and increase the morbidity rate. Biopolymers, metals, ceramics, and various biomaterials have been used to synthesize bone implants. Among these, bioactive glasses are one of the most biomimetic materials for human bones. They provide good mechanical properties, biocompatibility, and osteointegrative properties. Owing to these properties, various composites of bioactive glasses have been FDA-approved for diverse bone-related and other applications. However, bone defects and bone injuries require customized designs and replacements. Thus, the three-dimensional (3D) printing of bioactive glass composites has the potential to provide customized bone implants. This review highlights the bottlenecks in 3D printing bioactive glass and provides an overview of different types of 3D printing methods for bioactive glass. Furthermore, this review discusses synthetic and natural bioactive glass composites. This review aims to provide information on bioactive glass biomaterials and their potential in bone tissue engineering

Synthesis, characterization and biological evaluation of novel Ru(II)–arene complexes containing intercalating ligands

Three new ruthenium(II)–arene complexes, namely [(η6-p-cymene)Ru(Me2dppz)Cl]PF6 (1), [(η6-benzene)Ru(Me2dppz)Cl]PF6 (2) and [(η6-p-cymene)Ru(aip)Cl]PF6 (3) (Me2dppz = 11,12-dimethyldipyrido[3,2-a:2′,3′-c]phenazine; aip = 2-(9-anthryl)-1H-imidazo[4,5-f] [1,10] phenanthroline) have been synthesized and characterized using different spectroscopic techniques including elemental analysis. The complexes were found to be well soluble and stable in DMSO. The biological activity of the three complexes was tested in three different human cancer cell lines (A549, MDA-MB-231 and HeLa) and in one human non-cancerous cell line (MRC-5). Complexes 1 and 3, carrying η6-p-cymene as the arene ligand, were shown to be toxic in all cell lines in the low micromolar/subnanomolar range, with complex 1 being the most cytotoxic complex of the series. Flow cytometry analysis revealed that complex 1 caused concentration- and time-dependent arrest of the cell cycle in G2-M and S phases in HeLa cells. This event is followed by the accumulation of the sub-G1 DNA content after 48 h, in levels higher than cisplatin and in the absence of phosphatidylserine externalization. Fluorescent microscopy and acridine orange/ethidium bromide staining revealed that complex 1 induced both apoptotic and necrotic cell morphology characteristics. Drug-accumulation and DNA-binding studies performed by inductively coupled plasma mass spectrometry in HeLa cells showed that the total ruthenium uptake increased in a time- and concentration-dependent manner, and that complex 1 accumulated more efficiently than cisplatin at equimolar concentrations. The introduction of a Me2dppz ligand into the ruthenium(II)-p-cymene scaffold was found to allow the discovery of a strongly cytotoxic complex with significantly higher cellular uptake and DNA-binding properties than cisplatin

Molecular Imaging Probes Based on Matrix Metalloproteinase Inhibitors (MMPIs)

Matrix metalloproteinases (MMPs) are a family of zinc- and calcium-dependent endopeptidases which are secreted or anchored in the cell membrane and are capable of degrading the multiple components of the extracellular matrix (ECM). MMPs are frequently overexpressed or highly activated in numerous human diseases. Owing to the important role of MMPs in human diseases, many MMP inhibitors (MMPIs) have been developed as novel therapeutics, and some of them have entered clinical trials. However, so far, only one MMPI (doxycycline) has been approved by the FDA. Therefore, the evaluation of the activity of a specific subset of MMPs in human diseases using clinically relevant imaging techniques would be a powerful tool for the early diagnosis and assessment of the efficacy of therapy. In recent years, numerous MMPIs labeled imaging agents have emerged. This article begins by providing an overview of the MMP subfamily and its structure and function. The latest advances in the design of subtype selective MMPIs and their biological evaluation are then summarized. Subsequently, the potential use of MMPI-labeled diagnostic agents in clinical imaging techniques are discussed, including positron emission tomography (PET), single-photon emission computed tomography (SPECT) and optical imaging (OI). Finally, this article concludes with future perspectives and clinical utility

Mixed Ligand Copper(II) Complexes of <i>N</i>,<i>N</i>-Bis(benzimidazol-2-ylmethyl)amine (BBA) with Diimine Co-Ligands: Efficient Chemical Nuclease and Protease Activities and Cytotoxicity

A series of mononuclear mixed ligand copper­(II) complexes [Cu­(bba)­(diimine)]­(ClO₄)₂ <b>1</b>–<b>4</b>, where bba is <i>N</i>,<i>N</i>-bis­(benzimidazol-2-ylmethyl)­amine and diimine is 2,2′-bipyridine (bpy) (<b>1</b>), 1,10-phenanthroline (phen) (<b>2</b>), 5,6-dimethyl-1,10-phenanthroline (5,6-dmp) (<b>3</b>), or dipyrido­[3,2-<i>d</i>:2′,3′-<i>f</i>]­quinoxaline (dpq) (<b>4</b>), have been isolated and characterized by analytical and spectral methods. The coordination geometry around copper­(II) in <b>2</b> is described as square pyramidal with the two benzimidazole nitrogen atoms of the primary ligand bba and the two nitrogen atoms of phen (<b>2</b>) co-ligand constituting the equatorial plane and the amine nitrogen atom of bba occupying the apical position. In contrast, the two benzimidazole nitrogen atoms and the amine nitrogen atom of bba ligand and one of the two nitrogen atoms of 5,6-dmp constitute the equatorial plane of the trigonal bipyramidal distorted square based pyramidal (TBDSBP) coordination geometry of <b>3</b> with the other nitrogen atom of 5,6-dmp occupying the apical position. The structures of <b>1</b>–<b>4</b> have been optimized by using the density functional theory (DFT) method at the B3LYP/6-31G­(d,p) level. Absorption spectral titrations with Calf Thymus (CT) DNA reveal that the intrinsic DNA binding affinity of the complexes depends upon the diimine co-ligand, dpq (<b>4</b>) > 5,6-dmp (<b>3</b>) > phen (<b>2</b>) > bpy (<b>1</b>). The DNA binding affinity of <b>4</b> is higher than <b>2</b> revealing that the π-stacking interaction of the dpq ring in between the DNA base pairs with the two bzim moieties of the bba ligand stacked along the DNA surface is more intimate than that of phen. The complex <b>3</b> is bound to DNA more strongly than <b>1</b> and <b>2</b> through strong hydrophobic interaction of the methyl groups on 5,6-positions of the phen ring in the DNA grooves. The extent of the decrease in relative emission intensities of DNA-bound ethidium bromide (EB) upon adding the complexes parallels the trend in DNA binding affinities. The large enhancement in relative viscosity of DNA upon binding to <b>3</b> and <b>4</b> supports the DNA binding modes proposed. Interestingly, the 5,6-dmp complex <b>3</b> is selective in exhibiting a positive induced CD band (ICD) upon binding to DNA suggesting that it induces a B to A conformational change. In contrast, <b>2</b> and <b>4</b> show induced CD responses indicating their involvement in strong DNA binding. Interestingly, only the dpq complex <b>4</b>, which displays the strongest DNA binding affinity and is efficient in cleaving DNA in the absence of an activator with a rate constant of 5.8 ± 0.1 h^–1, which is higher than the uncatalyzed rate of DNA cleavage. All the complexes exhibit oxidative DNA cleavage ability, which varies as <b>4</b> > <b>2</b> > <b>3</b> > <b>1</b> (ascorbic acid) and <b>3</b> > <b>2</b> > <b>4</b> > <b>1</b> (H₂O₂). Also, the complexes cleave the protein bovine serum albumin in the presence of H₂O₂ as an activator with the cleavage ability varying in the order <b>3</b> > <b>4</b> > <b>2</b> > <b>1</b>. The highest efficiency of <b>3</b> to cleave both DNA and protein in the presence of H₂O₂ is consistent with its strong hydrophobic interaction with the biopolymers. The IC₅₀ values of <b>1</b>–<b>4</b> against cervical cancer cell lines (SiHa) are almost equal to that of cisplatin, indicating that they have the potential to act as effective anticancer drugs in a time-dependent manner. The morphological assessment data obtained by using acridine orange/ethidium bromide (AO/EB) and Hoechst 33258 staining reveal that <b>3</b> induces apoptosis much more effectively than the other complexes. Also, the alkaline single-cell gel electrophoresis study (comet assay) suggests that the same complex induces DNA fragmentation more efficiently than others

Organometallic Derivatization of the Nematocidal Drug Monepantel Leads to Promising Antiparasitic Drug Candidates

The discovery of novel drugs against animal parasites is in high demand due to drug-resistance problems encountered around the world. Herein, the synthesis and characterization of 27 organic and organometallic derivatives of the recently launched nematocidal drug monepantel (Zolvix®) are described. The compounds were isolated as racemates and were characterized by 1H, 13C, and 19F NMR spectroscopy, mass spectrometry, and IR spectroscopy, and their purity was verified by microanalysis. The molecular structures of nine compounds were confirmed by X-ray crystallography. The anthelmintic activity of the newly designed analogues was evaluated in vitro against the economically important parasites Haemonchus contortus and Trichostrongylus colubriformis. Moderate nematocidal activity was observed for nine of the 27 compounds. Three compounds were confirmed as potentiators of a known monepantel target, the ACR-23 ion channel. Production of reactive oxygen species may confer secondary activity to the organometallic analogues. Two compounds, namely, an organic precursor (3 a) and a cymantrene analogue (9 a), showed activities against microfilariae of Dirofilaria immitis in the low microgram per milliliter range