A novel inhibitor of the mitochondrial respiratory complex I with uncoupling properties exerts potent antitumor activity.

Cancer cells are highly dependent on bioenergetic processes to support their growth and survival. Disruption of metabolic pathways, particularly by targeting the mitochondrial electron transport chain complexes (ETC-I to V) has become an attractive therapeutic strategy. As a result, the search for clinically effective new respiratory chain inhibitors with minimized adverse effects is a major goal. Here, we characterize a new OXPHOS inhibitor compound called MS-L6, which behaves as an inhibitor of ETC-I, combining inhibition of NADH oxidation and uncoupling effect. MS-L6 is effective on both intact and sub-mitochondrial particles, indicating that its efficacy does not depend on its accumulation within the mitochondria. MS-L6 reduces ATP synthesis and induces a metabolic shift with increased glucose consumption and lactate production in cancer cell lines. MS-L6 either dose-dependently inhibits cell proliferation or induces cell death in a variety of cancer cell lines, including B-cell and T-cell lymphomas as well as pediatric sarcoma. Ectopic expression of Saccharomyces cerevisiae NADH dehydrogenase (NDI-1) partially restores the viability of B-lymphoma cells treated with MS-L6, demonstrating that the inhibition of NADH oxidation is functionally linked to its cytotoxic effect. Furthermore, MS-L6 administration induces robust inhibition of lymphoma tumor growth in two murine xenograft models without toxicity. Thus, our data present MS-L6 as an inhibitor of OXPHOS, with a dual mechanism of action on the respiratory chain and with potent antitumor properties in preclinical models, positioning it as the pioneering member of a promising drug class to be evaluated for cancer therapy. MS-L6 exerts dual mitochondrial effects: ETC-I inhibition and uncoupling of OXPHOS. In cancer cells, MS-L6 inhibited ETC-I at least 5 times more than in isolated rat hepatocytes. These mitochondrial effects lead to energy collapse in cancer cells, resulting in proliferation arrest and cell death. In contrast, hepatocytes which completely and rapidly inactivated this molecule, restored their energy status and survived exposure to MS-L6 without apparent toxicity.

Isocitrate dehydrogenase wt and IDHmut adult-type diffuse gliomas display distinct alterations in ribosome biogenesis and 2'O-methylation of ribosomal RNA.

BACKGROUND: High-grade adult-type diffuse gliomas (HGGs) constitute a heterogeneous group of aggressive tumors that are mostly incurable. Recent advances highlighting the contribution of ribosomes to cancer development have offered new clinical perspectives. Here, we uncovered that isocitrate dehydrogenase (IDH)wt and IDHmut HGGs display distinct alterations of ribosome biology, in terms of rRNA epitranscriptomics and ribosome biogenesis, which could constitute novel hallmarks that can be exploited for the management of these pathologies. METHODS: We analyzed (1) the ribosomal RNA 2'O-ribose methylation (rRNA 2'Ome) using RiboMethSeq and in-house developed bioinformatics tools (https://github.com/RibosomeCRCL/ribomethseq-nfandrRMSAnalyzer) on 3 independent cohorts compiling 71 HGGs (IDHwt n = 30, IDHmut n = 41) and 9 non-neoplastic samples, (2) the expression of ribosome biogenesis factors using medium throughput RT-qPCR as a readout of ribosome biogenesis, and (3) the sensitivity of 5 HGG cell lines to RNA Pol I inhibitors (CX5461, BMH-21). RESULTS: Unsupervised analysis demonstrated that HGGs could be distinguished based on their rRNA 2'Ome epitranscriptomic profile, with IDHwt glioblastomas displaying the most significant alterations of rRNA 2'Ome at specific sites. In contrast, IDHmut HGGs are largely characterized by an overexpression of ribosome biogenesis factors compared to non-neoplastic tissues or IDHwt glioblastomas. Finally, IDHmut HGG-derived spheroids display higher cytotoxicity to CX5461 than IDHwt glioblastoma, while all HGG spheroids display a similar cytotoxicity to BMH-21. CONCLUSIONS: In HGGs, IDH mutational status is associated with specific alterations of the ribosome biology and with distinct sensitivities to RNA Pol I inhibitors.

Fire Behavior of Thermally Thin Materials in Cone Calorimeter.

In this study, a representative set of thermally thin materials including various lignocellulosic and synthetic fabrics, dense wood, and polypropylene sheets were tested using a cone calorimeter at different heat fluxes. Time-to-ignition, critical heat flux, and peak of heat release rate (pHRR) were the main parameters considered. It appears that the flammability is firstly monitored by the sample weight. Especially, while the burning rate of thermally-thin materials does never reach a steady state in cone calorimeter, their pHRR appears to be mainly driven by the fire load (i.e., the product of sample weight and effective heat of combustion) with no or negligible influence of textile structure. A simple phenomenological model was proposed to calculate the pHRR taking into account only three parameters, namely heat flux, sample weight, and effective heat of combustion. The model allows predicting easily the peak of heat release rate, which is often considered as the main single property informing about the fire hazard. It also allows drawing some conclusions about the flame retardant strategies to reduce the pHRR..

Far-Infrared Emission Properties and Thermogravimetric Analysis of Ceramic-Embedded Polyurethane Films.

The far-infrared ray (FIR) is one kind of electromagnetic wave employed for numerous bio-interactive applications such as body thermoregulation, infrared therapy, etc. Tuning the FIR-emitting property of the functional textile surface can initiate a new horizon to utilize this property in sportswear or even smart textiles. Ceramic particles were studied for their unique ability to constantly emit FIR rays. The purpose of this research is to characterize the FIR emission properties and the thermogravimetric analysis of ceramic-embedded polyurethane films. For this purpose, ceramic particles such as aluminum oxide, silicon dioxide, and titanium dioxide were incorporated (individually) with water-based polyurethane (WPU) binder by a sonication technique to make a thin layer of film. Significant improvement in FIR emissive property of the films was found when using different ceramic particles into the polyurethane films. Reflection and transmission at the FIR range were measured with a gold integrating sphere by Fourier-transform infrared (FTIR) spectrometer. The samples were also characterized by thermogravimetric analysis (TGA). Different physical tests, such as tensile strength and contact angle measurements, were performed to illustrate the mechanical properties of the films. The study suggested that the mechanical properties of the polyurethane films were significantly influenced by the addition of ceramic particles.

Supersaturable self-microemulsifying delivery systems: an approach to enhance oral bioavailability of benzimidazole anticancer drugs.

This study explored the design of supersaturable self-microemulsifying drug delivery systems (S-SMEDDS) to address poor solubility and oral bioavailability of a novel benzimidazole derivative anticancer drug (BI). Firstly, self-microemulsifying drug delivery systems SMEDDS made of Miglyol® 812, Kolliphor® RH40, Transcutol® HP, and ethanol were prepared and loaded with the BI drug. Upon dispersion, the systems formed neutrally charged droplets of around 20 nm. However, drug precipitation was observed following incubation with simulated gastric fluid (pH 1.2). Aiming at reducing this precipitation and enhancing drug payload, supersaturable systems were then prepared by adding 1% hydroxypropyl cellulose as precipitation inhibitor. Supersaturable systems maintained a higher amount of drug in a supersaturated state in gastric medium compared with conventional formulations and were stable in simulated intestinal medium (pH 6.8). In vitro cell studies using Caco-2 cell line showed that these formulations reduced in a transient manner the transepithelial electrical resistance of the monolayers without toxicity. Accordingly, confocal images revealed that the systems accumulated at tight junctions after a 2 h exposure. In vivo pharmacokinetic studies carried out following oral administration of BI-loaded S-SMEDDS, SMEDDS, and free drug to healthy mice showed that supersaturable systems promoted drug absorption compared with the other formulations. Overall, these data highlight the potential of using the supersaturable approach as an alternative to conventional SMEDDS for improving oral systemic absorption of lipophilic drugs.

Development of Novel Polyamide 11 Multifilaments and Fabric Structures Based on Industrial Lignin and Zinc Phosphinate as Flame Retardants.

Biobased lignin represents one of the possible materials for next-generation flame retardant additives due to its sustainability, environmental benefits and comparable efficiency to other flame retardant (FR) additives. In this context, this study presents the development of FR polyamide 11 (PA11) multifilament yarns and fabric structures containing different industrial lignins (i.e., lignosulfonate lignin (LL), and Kraft lignin (KL)) and zinc phosphinate (ZnP). The combination of ZnP and lignin (KL or LL) at different weight ratios were used to prepare flame retarded PA11 blends by melt mixing using a twin-screw extruder. These blends were transformed into continuous multifilament yarns by the melt-spinning process even at a high concentration of additives as 20 wt%. The mechanical test results showed that the combination of KL and ZnP achieved higher strength and filaments showed regularity in structure as compared to the LL and ZnP filaments. Thermogravimetric (TG) analysis showed the incorporation of lignin induces the initial decomposition (T5%) at a lower temperature; at the same time, maximum decomposition (Tmax) shifts to a higher temperature region and a higher amount of char residue is reported at the end of the test. Further, the TGA-FTIR study revealed that the ternary blends (i.e., the combination of LL or KL, ZnP, and PA11) released mainly the phosphinate compound, hydrocarbon species, and a small amount of phosphinic acid during the initial decomposition stage (T5%), while hydrocarbons, carbonyls, and phenolic compounds along with CO2 are released during main decomposition stage (Tmax). The analysis of decomposition products suggests the stronger bonds formation in the condensed phase and the obtainment of a stable char layer. Cone calorimetry exploited to study the fire behavior on sheet samples (polymer bulk) showed an improvement in flame retardant properties with increasing lignin content in blends and most enhanced results were found when 10 wt% of LL and ZnP were combined such as a reduction in heat release rate (HRR) up to 64% and total heat release (THR) up to 22%. Besides, tests carried out on knitted fabric structure showed less influence on HRR and THR but the noticeable effect on postponing the time to ignition (TTI) and reduction in the maximum average rate of heat emission (MARHE) value during combustion.

Development of a Model for Chemical Screening Based on Collateral Sensitivity to Target BTK C481S Mutant.

Targeted therapies have improved the outcome of cancer, but their efficacy is intrinsically limited by the emergence of subclones with a mutation in the gene encoding the target protein. A few examples of collateral sensitivity have demonstrated that the conformational changes induced by these mutations can create unexpected sensitivity to other kinase inhibitors, but whether this concept can be generalized is unknown. Here is described the development of a model to screen a library of kinase inhibitors for collateral sensitivity drugs active on the Bruton Tyrosine Kinase (BTK) protein with the ibrutinib resistance mutation C481S. First, we demonstrate that overexpression of the constitutively active mutant of BTK harboring the E41K mutation in Ba/F3 cells creates an oncogenic addiction to BTK. Then, we have exploited this phenotype to perform a screen of a kinase inhibitor library on cells with or without the ibrutinib resistance mutation. The BTK inhibitors showed the expected sensitivity profile, but none of the drugs tested had a specific activity against the C481S mutant of BTK, suggesting that extending the collateral sensitivity paradigm to all kinases targeted by cancer therapy might not be trivial.

Bio-Functional Textiles: Combining Pharmaceutical Nanocarriers with Fibrous Materials for Innovative Dermatological Therapies.

In the field of pharmaceutical technology, significant attention has been paid on exploiting skin as a drug administration route. Considering the structural and chemical complexity of the skin barrier, many research works focused on developing an innovative way to enhance skin drug permeation. In this context, a new class of materials called bio-functional textiles has been developed. Such materials consist of the combination of advanced pharmaceutical carriers with textile materials. Therefore, they own the possibility of providing a wearable platform for continuous and controlled drug release. Notwithstanding the great potential of these materials, their large-scale application still faces some challenges. The present review provides a state-of-the-art perspective on the bio-functional textile technology analyzing the several issues involved. Firstly, the skin physiology, together with the dermatological delivery strategy, is keenly described in order to provide an overview of the problems tackled by bio-functional textiles technology. Secondly, an overview of the main dermatological nanocarriers is provided; thereafter the application of these nanomaterial to textiles is presented. Finally, the bio-functional textile technology is framed in the context of the different dermatological administration strategies; a comparative analysis that also considers how pharmaceutical regulation is conducted.

Valorization of Industrial Lignin as Biobased Carbon Source in Fire Retardant System for Polyamide 11 Blends.

In this study, two different types of industrial lignin (i.e., lignosulphonate lignin (LL) and kraft lignin (DL)) were exploited as charring agents with phosphorus-based flame retardants for polyamide 11 (PA11). The effect of lignins on the thermal stability and fire behavior of PA11 combined with phosphinate additives (namely, aluminum phosphinate (AlP) and zinc phosphinate (ZnP)) has been studied by thermogravimetric analysis (TGA), UL 94 vertical flame spread, and cone calorimetry tests. Various blends of flame retarded PA11 were prepared by melt process using a twin-screw extruder. Thermogravimetric analyses showed that the LL containing ternary blends are able to provide higher thermal stability, as well as a developed char residue. The decomposition of the phosphinates led to the formation of phosphate compounds in the condensed phase, which promotes the formation of a stable char. Flammability tests showed that LL/ZnP ternary blends were able to achieve self-extinction and V-1 classification; the other formulations showed a strong melt dripping and higher burning. In addition to this, cone calorimetry results showed that the most enhanced behavior was found when 10 wt % of LL and AlP were combined, which strongly reduced PHRR (-74%) and THR (-22%), due to the interaction between LL and AlP, which not only promotes char formation but also confers the stability to char in the condensed phase.

Influence of Ammonium Polyphosphate/Lignin Ratio on Thermal and Fire Behavior of Biobased Thermoplastic: The Case of Polyamide 11.

Flame retardancy of polymers is a recurring obligation for many applications. The development trend of biobased materials is no exception to this rule, and solutions of flame retardants from agro-resources give an advantage. Lignin is produced as a waste by-product from some industries, and can be used in the intumescent formation development as a source of carbon combined with an acid source. In this study, the flame retardancy of polyamide 11 (PA) is carried out by extrusion with a kraft lignin (KL) and ammonium polyphosphate (AP). The study of the optimal ratio between the KL and the AP makes it possible to optimize the fire properties as well as to reduce the cost and facilitates the implementation of the blend by a melting process. The properties of thermal decomposition and the fire reaction have been studied by thermogravimetric analyzes, pyrolysis combustion flow calorimetry (PCFC) and vertical flame spread tests (UL94). KL permits a charring effect delaying thermal degradation and decreases by 66% the peak of heat release rate in comparison with raw PA. The fire reaction of the ternary blends is improved even if KL-AP association does not have a synergy effect. The 25/75 and 33/67 KL/AP ratios in PA give an intumescence behavior under flame exposure.

Chitosan⁻Carboxymethylcellulose-Based Polyelectrolyte Complexation and Microcapsule Shell Formulation.

Chitosan (CH)⁻carboxymethyl cellulose sodium salt (NaCMC) microcapsules containing paraffin oil were synthesized by complex formation, and crosslinked with glutaraldehyde (GTA). The electrostatic deposition of NaCMC onto the CH-coated paraffin oil emulsion droplets was demonstrated by zeta potential and optical microscopy. The optimal process conditions were identified in terms of pH of the aqueous solution (5.5) and CH/NaCMC mass ratio (1:1). Encapsulation of paraffin oil and microcapsule morphology were analyzed by ATR-FTIR and SEM, respectively. The effect of GTA crosslinking on paraffin oil latent heat was investigated by DSC and combined with the values of encapsulation efficiency and core content, supporting the compact shell formation.

Influence of process parameters on microcapsule formation from chitosan-Type B gelatin complex coacervates.

A series of chitosan/gelatin based microcapsules containing n-hexadecane was synthesized through complex phase coacervation from chitosan (CH) and type-B gelatin (GB), and crosslinked by glutaraldehyde (GTA). This research was conducted to clarify the influence of different parameters on the encapsulation process, i.e., the emulsion formation and the shell formation, using zeta potential and surface tension measurements, attenuated total reflectance (ATR), and thermal analysis such as differential scanning calorimetry (DSC). The optimal values of biopolymer ratios (TBP), crosslinker amount, emulsion time and feeding weight ratio of core/shell polymer (RCS) were identified. The stability of the emulsion was depended on the surface activity and TBP ratio, which also affected the droplet size distribution and the thickness of the shell. Furthermore, core content, encapsulation efficiency and thermal properties of the microcapsules were related to TBP and RCS; with the lowest RCS giving the best microcapsules features.

Surface behavior and bulk properties of aqueous chitosan and type-B gelatin solutions for effective emulsion formulation.

The behavior of aqueous chitosan (CH), type-B gelatin (GB) and CH-GB coacervate was studied on oil-in-water emulsion formulation at various pH and concentration ratio. The coacervate was formed by phase separation at ratios CH:GB, 1:10 to 1:1 with total biopolymer concentrations of 0.55%-1.0% (w/v) at pH 4.0-5.5. Soluble complexes were formed below pH 5.0 and coacervate formation was confirmed at pH 5.0 and above by zeta potential and UV-spectroscopy measurements. The coacervate formation was found maximum at the CH-GB ratios of 1:10 and 1:5 at pH 5.5. Formulated emulsions (>10µm droplets) using 1% (w/v) chitosan and GB were found stable (+52.5mv and creaming index 86%) and unstable respectively. Emulsion stabilized by mixed CH:GB 1:5 (3%w/v) had no creaming effect. The instability was attributed to the lower surface activity (K=5.0Lg-1) of pure GB compared to CH (K=14.3Lg-1). The formulation and methods can successfully tune the stability of the emulsions.

Application of Flame-Retardant Double-Layered Shell Microcapsules to Nonwoven Polyester.

A microencapsulated flame retardant was used in order to produce a flame retardant nonwoven substrate. Melamine-formaldehyde polymer-shell microcapsules, containing Afflamit® PLF 280 (resorcinol bis(diphenyl phosphate)) as the core substance, were coated by an outer thermoplastic wall (polystyrene (PS) or poly(methyl methacrylate)), before being applied to a core/sheet-type bi-component PET/co-PET spunbond nonwoven substrate using impregnation. The outer wall of the microcapsules was heated to the softening temperature of the thermoplastic shell in order to be bonded onto the textile fibres. The thermal stability of the microcapsules was examined using thermogravimetric analysis. The textile samples were observed with a scanning electron microscope, and the flame retardancy performance was evaluated using the NF P92-504 standard. The results show that the composition of the outer polymeric shell affected the thermal stability of the microcapsules, since the particles with a PS shell are more stable. Furthermore, the microcapsules were more located at the nonwoven surface without affecting the thickness of the samples. Based on the results of the NF P92-504 test, the flame spread rate was relatively low for all of the tested formulations. Only the formulation with a low content of PS was classified M2 while the others were M3.

PLA with Intumescent System Containing Lignin and Ammonium Polyphosphate for Flame Retardant Textile.

Using bio-based polymers to replace of polymers from petrochemicals in the manufacture of textile fibers is a possible way to improve sustainable development for the textile industry. Polylactic acid (PLA) is one of the available bio-based polymers. One way to improve the fire behavior of this bio-based polymer is to add an intumescent formulation mainly composed of acid and carbon sources. In order to optimize the amount of bio-based product in the final material composition, lignin from wood waste was selected as the carbon source. Different formulations of and/or ammonium polyphosphate (AP) were prepared by melt extrusion and then hot-pressed into sheets. The thermal properties (thermogravimetric analyses (TGA) and differential scanning calorimetry (DSC)) and fire properties (UL-94) were measured. The spinnability of the various composites was evaluated. The mechanical properties and physical aspect (microscopy) of PLA multifilaments with lignin (LK) were checked. A PLA multifilament with up to 10 wt % of intumescent formulation was processed, and the fire behavior of PLA fabrics with lignin/AP formulation was studied by cone calorimeter.

Identification of new high mobility group A1 associated proteins.

High mobility group A (HMGA) proteins (HMGA1a, HMGA1b, HMGA1c and HMGA2) are nonhistone chromosomal proteins that do not have transcriptional activity per se, but they orchestrate the assembly of multiprotein complexes involved in gene transcription, replication and chromatin structure through a complex network of protein-DNA and protein-protein interactions. To better understand their mechanisms of action, we have used a combination of coimmunoprecipitation, 1-D gel SDS-PAGE and MS to identify new potential molecular interactors. We have found 11 proteins that associate with HMGA1. These proteins belong to three different classes: mRNA processing proteins, RNA helicases and protein chaperones. Some interactions were confirmed by coimmunoprecipitation and pull-down experiments in human embryonal kidney 293 cells. These experimental data suggest that HMGA1 proteins can associate with proteins that are strictly involved in chromatin structure and in several important mRNA processing steps, supporting the idea that HMGA1 proteins can also participate in these events.

US11 of herpes simplex virus type 1 interacts with HIPK2 and antagonizes HIPK2-induced cell growth arrest.

Homeodomain-interacting protein kinase 2 (HIPK2) is a nuclear serine/threonine kinase of the subfamily of dual-specificity Yak1-related kinase proteins. HIPK2 was first described as a homeodomain-interacting protein kinase acting as a corepressor for homeodomain transcription factors. More recently, it was reported that HIPK2 plays a role in p53-mediated cellular apoptosis and could also participate in the regulation of the cell cycle. US11 protein of herpes simplex virus type 1 is a multifunctional protein involved in the regulation of several processes related to the survival of cells submitted to environmental stresses by mechanisms that are not fully elucidated. In an attempt to better understand the multiple functions of US11, we identified cellular binding partners of this protein by using the yeast two-hybrid system. We report that US11 interacts with HIPK2 through the PEST domain of HIPK2 and that this interaction occurs also in human cells. This interaction modifies the subcellular distribution of HIPK2 and protects the cell against the HIPK2-induced cell growth arrest.

Different growth properties of neointimal and medial smooth muscle cells in response to growth factors.

Different smooth muscle cell phenotypes coexist in arteries. The majority of cells cultured from a normal rat aortic media have a spindle-shaped phenotype while cells isolated from an intimal thickening 15 days after endothelial injury show a distinct epithelioid phenotype. These two phenotypes express their own specific set of genes and differ in their proliferation and migration characteristics. We studied growth factor-induced DNA synthesis in both phenotypes and investigated the potential mechanisms behind the differences in growth characteristics. Insulin-like growth factor-I (IGF-I), platelet-derived growth factor (PDGF-BB), and basic fibroblast growth factor (FGF) increased thymidine incorporation in both phenotypes, but the increase was markedly stronger in neointimal cells than in medial cells. Northern blot analysis 30 min after growth factor stimulation showed that c-FOS and c-JUN mRNAs were induced more strongly in neointimal than in medial cells. IGF-I receptor and PDGF-R beta levels were higher in neointimal cells than in medial cells, but the FGF receptor level was not different between the cell types. In summary, our results indicate that neointimal cells are more sensitive to growth factors than medial cells, likely due to a higher expression of IGF-I receptor and PDGF-R beta. Our results provide insight into the mechanism by which epithelioid cells play a primary role in vascular neointima formation.

Alteration of ribosomal protein maps in herpes simplex virus type 1 infection.

At present, the effect of herpes simplex virus infection on the entire proteomes of infected cells is very poorly documented. Following several studies performed over the past few years, the modifications of a sub-cellular fraction induced by herpes simplex virus type 1 can be documented. These studies were performed in order to characterize the virally-induced modifications of a major component of the translational apparatus, the ribosomes. The very basic nature of most of the ribosomal proteins renders them very difficult to separate using isoelectric focusing (IEF). Therefore these studies were achieved using several different but related two-dimensional electrophoretic systems which allowed several two-dimensional ribosomal protein maps to be built. Comparison of the ribosomal protein maps built from non-infected cells with those built from infected cells demonstrated that infection by herpes simplex virus type 1 (HSV-1) induces important modifications of ribosomes: (i) non-reversible phosphorylation of ribosomal protein S6; (ii) unusual phosphorylation of several proteins of the small and the large subunits; and (iii) association of viral and cellular proteins to the ribosomal fraction. An overview of these published studies is presented in this review.