KDM4 Involvement in Breast Cancer and Possible Therapeutic Approaches

Breast cancer (BC) is the second leading cause of cancer death in women, although recent scientific and technological achievements have led to significant improvements in progression-free disease and overall survival of patients. Genetic mutations and epigenetic modifications play a critical role in deregulating gene expression, leading to uncontrolled cell proliferation and cancer progression. Aberrant histone modifications are one of the most frequent epigenetic mechanisms occurring in cancer. In particular, methylation and demethylation of specific lysine residues alter gene accessibility via histone lysine methyltransferases (KMTs) and histone lysine demethylases (KDMs). The KDM family includes more than 30 members, grouped into six subfamilies and two classes based on their sequency homology and catalytic mechanisms, respectively. Specifically, the KDM4 gene family comprises six members, KDM4A-F, which are associated with oncogene activation, tumor suppressor silencing, alteration of hormone receptor downstream signaling, and chromosomal instability. Blocking the activity of KDM4 enzymes renders them “druggable” targets with therapeutic effects. Several KDM4 inhibitors have already been identified as anticancer drugs in vitro in BC cells. However, no KDM4 inhibitors have as yet entered clinical trials due to a number of issues, including structural similarities between KDM4 members and conservation of the active domain, which makes the discovery of selective inhibitors challenging. Here, we summarize our current knowledge of the molecular functions of KDM4 members in BC, describe currently available KDM4 inhibitors, and discuss their potential use in BC therapy

Green Processing of Nanoporous Biodegradable Carriers of Bioactive Agents for Pharmaceutical and Biomedical Applications

Pharmaceutical and biomedical industries demand simple, safe and reproducible processing methods thus urging the development of novel straightforward manufacturing approaches. The product manufacturing by the green processing of admixtures and end-product would avoid long and costly purification (downstream) steps. In this work, the green supercritical fluid technology is used for the processing of nanoporous carriers (aerogels) for bioactive agents [1,2]. Aerogels in the form of one micron-sized particles were processed and loaded with a model bioactive compound (ketoprofen). Results show that the carrier has excellent textural properties (specific surface area of 200 m2/g) and a high loading capacity (7 wt.%) of the bioactive compound in the amorphous form. Release profile tests show the capacity of the carrier to modulate the drug release to the medium (PBS pH 7.4). The resulting material can be incorporated in the formulation of several pharmaceutical and biomedical products

Seismic Loss Estimation in Pre-1970 Residential RC Buildings: The Role of Infills and Services in Low–Mid-Rise Case Studies

The lessons learned after recent earthquakes have highlighted the key role played by infills and services in damage and loss of Reinforced Concrete (RC) buildings. Their influence in seismic performance and loss estimation of selected RC building case studies is thoroughly analyzed here. The case study selection aims to be representative of existing buildings built in Italy before 1970, and covers a different number of stories and design typologies. The seismic responses of the case-study buildings are numerically analyzed by means of non-linear static pushover analysis (PO) considering a lumped plasticity approach with a quadri-linear flexural response for beam/column elements (properly calibrated for RC elements reinforced with plain bars) and a tri-linear compressive-only axial response with diagonal concentric struts for infill panels (empirically derived from experimental data on hollow clay masonry walls). Economic loss estimation is carried out via a component-based methodology that relies on the main repairing activities and resultant costs required for the refurbishment of infills and services for different damage levels. Accordingly, a damage analysis is performed herein, given the intensity measure, based on a comparison between Interstory drift demand from PO analysis and drift-based fragility functions specific for masonry infills. Loss curves, relating the total building repair cost to peak ground acceleration (PGA), are presented and compared for the analyzed case study buildings to show their trends and quantify the incidence of infills and services with respect to the reconstruction cost. A comparison between these outcomes and those recently found in the literature emphasizes the robustness of the considered approach and the reliability of the hypotheses about damage and loss assessment

Clinical pathways of epileptic seizures and status epilepticus: results from a survey in Italy

Objective: Patients with seizures or status epilepticus (SE) access the hospital through emergency departments and may be admitted into different wards according to the level of care required. Clinicians with different expertise are in charge of taking critical therapeutic decisions. To date, very few studies have investigated the stage at which these patients are referred to neurologists or epileptologists and how guideline recommendations are applied in clinical practice. Methods: A survey was used to investigate how patients with epileptic seizures or SE are managed in emergency and in subsequent hospital pathways in Italy. Results: One hundred and seventy-seven physicians (mainly neurologists) from all parts of Italy filled in a questionnaire. Less than half of the participants (35%) answered that, in their hospital, patients with epilepsy were managed by epileptologists. The percentages were lower for patients presenting with acute seizures (21%) or SE (16%). Diagnostic, therapeutic, and assistance pathways (PDTA) for patients presenting with seizure(s) or SE were available for both conditions in about 50% of cases, while, in the rest of the hospitals, participants indicated informal agreements (about 25% of cases) or lack of any agreement (about 25% of cases) between clinicians. Professionals more often involved in PDTA were epileptologists/neurologists, emergency physicians, and intensivists. More than half ot the participants (55%) thought that organizational issues are the most important criticalities for such patients and need to be improved (61%). Significance: There is a high variability in hospital clinical pathways for epilepsy in Italy

Frictional melting of peridotite and seismic slip

The evolution of the frictional strength along a fault at seismic slip rates (about 1 m/s) is a key factor controlling earthquake mechanics. At mantle depths, friction-induced melting and melt lubrication may influence earthquake slip and seismological data. We report on laboratory experiments designed to investigate dynamic fault strength and frictional melting processes in mantle rocks. We performed 20 experiments with Balmuccia peridotite in a high-velocity rotary shear apparatus and cylindrical samples (21.8 mm in diameter) over a wide range of normal stresses (5.4–16.1 MPa), slip rates (0.23–1.14 m/s), and displacements (1.5–71 m). During the experiments, shear stress evolved with cumulative displacement in five main stages (stages 1–5). In stage 1 (first strengthening), the coefficient of friction m increased up to 0.4–0.7 (first peak in friction). In stage 2 (abrupt firstweakening), m decreased to about 0.25–0.40. In stage 3 (gradual second strengthening), shear stress increased toward a second peak in friction (m = 0.30–0.40). In stage 4 (gradual second weakening), the shear stress decreased toward a steady state value (stage 5) with m = 0.15. Stages 1 and 2 are of too short duration to be investigated in detail with the current experimental configuration. By interrupting the experiments during stages 3, 4, and 5, microstructural (Field Emission Scanning Electron Microscope) and geochemical (Electron Probe Micro-Analyzer and Energy Dispersive X-Ray Spectroscopy) analysis of the slipping zone suggest that second strengthening (stage 3) is associated with the production of a grain-supported melt-poor layer, while second weakening (stage 4) and steady state (stage 5) are associated with the formation of a continuous melt-rich layer with an estimated temperature up to 1780 C. Microstructures formed during the experiments were very similar to those found in natural ultramafic pseudotachylytes. By performing experiments at different normal stresses and slip rates, (1) the ‘‘thermal’’ (as it includes the thermally activated first and second weakening) slip distance to achieve steady state from the first peak in strength decreased with increasing normal stress and slip rate and (2) the steady state shear stress slightly increased with increasing normal stress and, for a given normal stress, decreased with increasing slip rate. The ratio of shear stress versus normal stress was about 0.15, well below the typical friction coefficient of rocks (0.6–0.8). The dependence of steady state shear stress with normal stress was described by means of a constitutive equation for melt lubrication. The presence of microstructures similar to those found in natural pseudotachylytes and the determination of a constitutive equation that describes the experimental data allows extrapolation of the experimental observations to natural conditions and to the study of rupture dynamics in mantle rocks

Unveiling the importance of diffusion on the deterioration of cellulose acetate artefacts: The profile of plasticiser loss as assessed by infrared microscopy

Cellulose acetate (CA) artefacts are one of the most valued plastic items in museum collections and are known to present stability issues, with the loss of plasticiser being among the main degradation processes. This study investigates the concentration distribution of diethyl phthalate (DEP) plasticiser throughout the dimensions of CA using infrared microscopy for the first time. Artificial ageing experiments using reference and historic CA plasticised with DEP were performed to assess the change in the concentration profiles as a function of ageing time. Our analysis indicates that the plasticiser loss from CA artefacts is likely controlled by its diffusion, resulting in a concentration gradient in which lower plasticiser contents are observed at the external layers of the material

Characterising plasticised cellulose acetate-based historic artefacts by NMR spectroscopy: a new approach for quantifying the degree of substitution and diethyl phthalate contents

As one of the first semi-synthetic plastics produced industrially, cellulose acetate (CA)-based artefacts represent valued items in museum collections and archives which, however, present stability issues. High temperature and relative humidity conditions have long been known to promote changes in CA properties, for instance, due to the deacetylation of CA polymer chains and the loss of plasticiser from the polymer matrix. However, there is a need for improved methods for the quantification of plasticiser loss and CA deacetylation. In this context, this contribution presents a new approach for enabling the investigation of plasticiser loss and deacetylation degradation processes in historic plasticised CA-based artefacts which is based on high-resolution proton nuclear magnetic resonance spectroscopy (1H NMR). The proposed methods allow for simple and fast quantification of diethyl phthalate contents and average degree of substitution (DS), while requiring no need for extractive separation between the plasticiser and the CA polymer matrix prior to analysis. Both methods are demonstrated by their application towards a series of reference samples, historic artefacts and artificially aged plasticised CA materials. Our analysis indicates that plasticiser content and DS can be accurately quantified by using high-resolution 1H NMR and both methods have been compared to analyses performed using infrared spectroscopy

Arthropods on Mars?

As presented in this report numerous fossils like forms resembling a variety of marine arthropods including crustaceans, sea spiders, scorpions, arachnids, nematodes, annelids, tube worms, sea snakes, Kimberlla, Namacalathus, Lophotrochozoa, armored trilobites and millipedes have been found in Gale Crater (on Sols 302, 553, 753, 781, 809, 869, 880, 905, 1032), and Meridiani Planum both of which have hosted rivers, lakes, and inland seas. Similar specimens are mixed within a variety of divergent fossillike forms and are also found on distant sediment and mud stone. All specimens are distinct from underlying substrate and there are no obvious patterns or repetitions typically produced by erosion or weathering. Although without extraction and direct examination it is impossible to precisely determine the identity of all these specimens, the same problems bedevil identification of Burgess Shale fossils some of which are presented in this report for comparative analysis. The discoveries presented here and in other reports supports the theory that metazoans and other marine organisms evolved in the lakes, oceans and inland seas of Mars

Aerodynamic Properties and Drug Solubility of Dry Powders Prepared by Spray Drying: Clarithromycin Versus its Hydrochloride Salt

Aerodynamic Properties and Drug Solubility of Dry Powders Prepared by Spray Drying: Clarithromycin Versus its Hydrochloride Salt. Russo P, Manniello M.D., Simonetti A., Petrone A.M., Porta A., Del Gaudio P., Aquino R.P. Department of Pharmacy (DIFARMA), University of Salerno, Fisciano (SA) – Italy; ([email protected]). Introduction and Objectives. The antibiotic therapy for a direct administration to the lung in cystic fibrosis patients has to provide suitable drug availability, possibly in the lower respiratory tract characterized by the presence of thick secretions. Apart from deposition, systemic or local pharmacological activity of an inhalation product depends on drug dissolution into the biological fluids lining the lung. Therefore, one of the crucial step in the therapeutic management of the respiratory disease is the drug solubilization in this site of action. Clarithromycin (CLA; fig.1) is a broad spectrum and a well know macrolide antibiotic usually prescribed particularly for the treatment of respiratory infections, interestingly showing an additional anti-inflammatory effect (Pukhalsky et al., 2004); CLA is characterized by a very poor water solubility (0.33 mg/L). One of the common strategy, for increasing drug solubility in aqueous medium is represented by the production of dry powders in amorphous form using the spray drying technique (Yonemochi et al., 1999). Moreover, CLA has a dimethylamino group, which can be salified for solubility/dissolution improvement (fig.1). Hence, the aim of the present study was to obtain respirable powders of clarithromycin, while improving drug aqueous solubility. Powders were produced with CLA or CLA hydrochloride and characterized in terms of drug content, aerodynamic properties and drug solubility. Finally, in order to assess the effect of the spray-drying process on the antibiotic activity of the engineered particles, microbiological tests were performed. Materials and Methods. Several batches of micronized particles were prepared by spray drying different feed solutions; critical process parameters were solvent composition (isopropyl alcohol/water ratio), drug concentration and pH of the liquid feeds (table 1). Saturated solubility measurements were carried out keeping an excess amount of CLA raw material (RM), CLA spray-dried suspension and hydrochloride spray-dried in phosphate buffer (0.05 M, pH 6.75) at 37°C for 72h. After filtration, the solubility was measured by HPLC method and expressed in mg/ml. The results were reported as mean of three measurements and standard deviation. Particle size distribution of Raw Material and engineered particles was determined using a light-scattering laser granulometer, while particle morphology was assessed by scanning electron microscopy (SEM). The in vitro deposition of the micronized powders was evaluate by means of a Single-Stage Glass Impinger (SSGI; apparatus A; European Pharmacopoeia 8.0), using a proper device for the aerosolization. The antibacterial assay was carried out in MHB by microdilution method using 96-well microtiter plates. Briefly, 200 µl of 1X107 CFU/ml of P. aeruginosa ATCC 27853 were incubated at 37°C with different concentration of drug (4, 6 and 8 µg/ml). Results and Discussion. Morphology and aerodynamic properties of spray-dried particles were strongly dependent on organic solvent concentration as well as on pH of the liquid feeds processed, both influencing drug solubility. Adding clarithromycin to hydroalcoholic mixtures, alkaline feeds (pH~10.5; #1, #2, tab.1) in form of suspensions were obtained.   The resulting spray-dried powders showed good process yield, but unsatisfactory aerodynamic properties, due to a high particle size (tab. 1) and the presence of residual drug crystals mixed to spherical particles (fig.2; #2). With the aim to obtain clear feeds in form of solutions, we tested clarithromycin in its hydrochloride salt form, obtained lowering pH values of feed solutions (pH 6.5). Micronized salified powders showed higher process yield and very interesting FPF values, thanks to smaller and wrinkled particles (fig.2 #4, #6; tab.1). Moreover, water solubility of spray-dried powders was strongly influenced by clarithromycin form. Powders obtained from alkaline feed suspensions showed lower solubility in a phosphate buffer 0.05 M, pH 6.75 (fig.3, # 1). A substantial increase in drug solubility was obtained, at the same conditions, with powders dried from feed solution containing clarithromycin hydrochloride (fig.3 #9-#5). To verify the ability of the produced formulations to was performed. Three different drug concentrations were tested in a multi-well plate. Clarithromycin raw material (RM, line C, figure 4) and #1 (line D, figure 4) showed a lower activity against P. aeruginosa growth compared to #4, #5, #6, containing the hydrochloride form. This different behavior against P. aeruginosa may be due to the lower CLA solubility; at higher concentration, clarithromycin precipitates in RM and #1 wells, becoming unavailable for antibiotic purpose. Conclusions. Clarithromycin inhalable powders containing the drug in its hydrochloride form showed good aerodynamic properties and higher water solubility. Thanks to a fine-tuning of the process parameters and liquid feed composition, no excipients were necessary to obtain respirable powders. The spray drying process of CLA hydrochloride not only preserved antimicrobial activity, but also, increasing drug solubility, improved drug efficacy against P. aeruginosa. References. Pukhalsky, A. L., Shmarina, G. V., Kapranov, N. I., Kokarovtseva, S. N., Pukhalskaya, D., & Kashirskaja, N. J. (2004). Anti-inflammatory and immunomodulating effects of clarithromycin in patients with cystic fibrosis lung disease. Mediators Inflamm, 13(2), 111-117. doi: 10.1080/09629350410001688495 Yonemochi, E., Kitahara, S., Maeda, S., Yamamura, S., Oguchi, T., & Yamamoto, K. (1999). Physicochemical properties of amorphous clarithromycin obtained by grinding and spray drying. Eur J Pharm Sci, 7(4), 331-338