Direct Cell-Cell Communication via Membrane Pores,Gap Junction Channels,and Tunneling Nanotubes: Medical Relevance of Mitochondrial Exchange

The history of direct cell-cell communication has evolved in several small steps. First discovered in the 1930s in invertebrate nervous systems, it was thought at first to be an exception to the “cell theory”, restricted to invertebrates. Surprisingly, however, in the 1950s, electrical cell-cell communication was also reported in vertebrates. Once more, it was thought to be an exception restricted to excitable cells. In contrast, in the mid-1960s, two startling publications proved that virtually all cells freely exchange small neutral and charged molecules. Soon after, cell-cell communication by gap junction channels was reported. While gap junctions are the major means of cell-cell communication, in the early 1980s, evidence surfaced that some cells might also communicate via membrane pores. Questions were raised about the possible artifactual nature of the pores. However, early in this century, we learned that communication via membrane pores exists and plays a major role in medicine, as the structures involved, “tunneling nanotubes”, can rescue diseased cells by directly transferring healthy mitochondria into compromised cells and tissues. On the other hand, pathogens/cancer could also use these communication systems to amplify pathogenesis. Here, we describe the evolution of the discovery of these new communication systems and the potential therapeutic impact on several uncurable diseases.     

Thermal Stability of Ge<Subscript>2</Subscript>Sb<Subscript>2</Subscript>Te<Subscript>5</Subscript> in Contact with Ti and TiN

The thermal stability of a Ge₂Sb₂Te₅ chalcogenide layer in contact with titanium and titanium nitride metallic thin films has been investigated mainly using x-ray diffraction and elastic nuclear backscattering techniques. Without breaking vacuum, Ti and TiN have been deposited on Ge₂Sb₂Te₅ material using magnetron sputtering. Thermal treatments have been performed in a 10⁻⁷ mbar vacuum furnace. On annealing up to 450°C, the TiN metallic film does not interact with the chalcogenide film, but at the same time adhesion problems and instabilities in contact resistance arise. To improve the adhesion and eventually stabilize the contact resistance, an interfacial Ti layer has been considered. At 300°C, a TiTe₂ compound is formed by interacting with Te segregated from the Ge₂Sb₂Te₅ layer. At higher temperatures, the Ti layer decomposes the chalcogenide film, forming several compounds tentatively identified as GeTe, Ge₃Ti₅, Ge₅Ti₆, TiTe_2,, and Sb₂Te₃. It has been found that the properties of the Ge₂Sb₂Te₅ film can be retained by controlling the decomposition rate of the chalcogenide layer, which is achieved by providing a limited supply of Ti and/or by depositing a Te-rich Ge₂Sb₂Te₅ film.     

Impact of Ge-Sb-Te compound engineering on the set operation performance in phase-change memories

The phase-change memory (PCM) technology is considered as one of the most attractive non-volatile memory concepts for next generation data storage. It relies on the ability of a chalcogenide material belonging to the Ge-Sb-Te compound system to reversibly change its phase between two stable states, namely the poly-crystalline low-resistive state and the amorphous high-resistive state, allowing the storage of the logical bit. A careful study of the phase-change material properties in terms of the set operation performance, the program window and the electrical switching parameters as a function of composition is very attractive in order to enlarge the possible PCM application spectrum. Concerning the set performance, a crystallization kinetics based interpretation of the observed behavior measured on different Ge-Sb-Te compounds is provided, allowing a physics-based comprehension of the reset-to-set transition.     

Novel Insights into Autophagy and Prostate Cancer: A Comprehensive Review

Autophagy is a complex process involved in several cell activities, including tissue growth, differentiation, metabolic modulation, and cancer development. In prostate cancer, autophagy has a pivotal role in the regulation of apoptosis and disease progression. Several molecular pathways are involved, including PI3K/AKT/mTOR. However, depending on the cellular context, autophagy may play either a detrimental or a protective role in prostate cancer. For this purpose, current evidence has investigated how autophagy interacts within these complex interactions. In this article, we discuss novel findings about autophagic machinery in order to better understand the therapeutic response and the chemotherapy resistance of prostate cancer. Autophagic-modulation drugs have been employed in clinical trials to regulate autophagy, aiming to improve the response to chemotherapy or to anti-cancer treatments. Furthermore, the genetic signature of autophagy has been found to have a potential means to stratify prostate cancer aggressiveness. Unfortunately, stronger evidence is needed to better understand this field, and the application of these findings in clinical practice still remains poorly feasible.     

α–ω Alkenyl-bis-S-Guanidine Thiourea Dihydrobromide Affects HeLa Cell Growth Hampering Tubulin Polymerization

Cancer is going to be the first cause of mortality worldwide in the 21th century. It is considered a multifactorial disease that results from the combined influence of many genetic aberrations, leading to abnormal cell proliferation. As microtubules are strongly implicated in cellular growth, they represent an important target for cancer treatment. The well-known microtubule-targeting agents (MTAs) including paclitaxel, colchicine and vinca alkaloids are commonly used in the treatment of various cancers. However, adverse effects and drug resistance are major limitations in their clinical use. To find new candidates able to induce microtubule alteration with reduced toxic effects or drug resistance, we studied a small new series of derivatives that present imidazolinic, guanidinic, thioureidic and hydrazinic groups ( 1 – 9 ). All the compounds were tested for their antitumor activity against a panel of six tumoral cell models. In particular, compound 8 (nonane-1,9-diyl-bis-S-amidinothiourea dihydrobromide) showed the lowest IC₅₀ value against HeLa cells, together with a low cytotoxicity for normal cells. This compound was able to induce the apoptotic mitochondrial pathway and inhibited tubulin polymerization with a similar efficacy to vinblastine and nocodazole. Taken together, these promising biological properties make compound 8 useful for the development of novel therapeutic approaches in cancer treatment.     

Discovery of New Antiproliferative Imidazopyrazole Acylhydrazones Able To Interact with Microtubule Systems

Even though immunotherapy has radically changed the search for anticancer therapies, there are still many different pathways that are open to intervention with traditional small molecules. To expand our investigation in the anticancer field, we report here a new series of compounds in which our previous pyrazole and imidazopyrazole scaffolds are linked to a differently decorated phenyl ring through an acylhydrazone linker. Preliminary tests on the library were performed at the National Cancer Institute (USA) against the full NCI 60 cell panel. The best compounds among the imidazopyrazole series were then tested by immunofluorescence staining for their inhibition of cell proliferation, apoptosis induction, and their effect on the cell cycle and on microtubules. Two compounds, in particular 4-benzyloxy-3-methoxybenzyliden imidazopyrazole-7-carbohydrazide showed good growth inhibition, with IC₅₀ values in the low-micromolar range, and induced apoptosis. Both compounds altered the cell-cycle phases with the appearance of polyploid cells. Immunofluorescence analysis evidenced microtubules alterations; tubulin polymerization assays and docking studies suggested the tubulin system to be the possible, although not exclusive, target of the new acylhydrazone series reported here.     

Identification of freshwater fish commercially labelled “perch” by isoelectric focusing and two-dimensional electrophoresis

Isoelectric focusing (IEF) and two-dimensional electrophoresis (2-DE) were used to distinguish four freshwater fish species which are sold under the generic label of “perch”: Perca fluviatilis (European perch), Lates niloticus (Nile perch), Stizostedion lucioperca (European pikeperch) and Morone chrysops x saxatilis (sunshine bass). These species have different commercial values but are easily “interchangeable” because they are sold already filleted, in view of the numerous bones of the whole fish. IEF of the water-soluble proteins extracted from fish muscle resolved in species-specific patterns. Intra-species polymorphism was low, and did not concern the bands identified as characteristic of the species. As well, 2-DE maps showed numerous species-specific protein spots. Interestingly, while none of the IEF bands was common to all four species, several major 2-DE spots were similar. Therefore, IEF of water soluble sarcoplasmic proteins is sufficient to unambiguously discriminate among the four species considered. Analysis by 2-DE, which has a higher resolution power but it is more expensive and time consuming, may be applied to obtain further knowledge of the proteome of poorly characterized species.     

Calmodulin-Connexin Partnership in Gap Junction Channel Regulation-Calmodulin-Cork Gating Model

In the past four decades numerous findings have indicated that gap junction channel gating is mediated by intracellular calcium concentrations ([Ca²⁺_i]) in the high nanomolar range via calmodulin (CaM). We have proposed a CaM-based gating model based on evidence for a direct CaM role in gating. This model is based on the following: CaM inhibitors and the inhibition of CaM expression to prevent chemical gating. A CaM mutant with higher Ca²⁺ sensitivity greatly increases gating sensitivity. CaM co-localizes with connexins. Connexins have high-affinity CaM-binding sites. Connexin mutants paired to wild type connexins have a higher gating sensitivity, which is eliminated by the inhibition of CaM expression. Repeated trans-junctional voltage (Vj) pulses progressively close channels by the chemical/slow gate (CaM’s N-lobe). At the single channel level, the gate closes and opens slowly with on-off fluctuations. Internally perfused crayfish axons lose gating competency but recover it by the addition of Ca-CaM to the internal perfusion solution. X-ray diffraction data demonstrate that isolated gap junctions are gated at the cytoplasmic end by a particle of the size of a CaM lobe. We have proposed two types of CaM-driven gating: “Ca-CaM-Cork” and “CaM-Cork”. In the first, the gating involves Ca²⁺-induced CaM activation. In the second, the gating occurs without a [Ca²⁺]_i rise.     

Influence of Nano,Micro, and Macro Topography of Dental Implant Surfaces on Human Gingival Fibroblasts

Current research on dental implants has mainly focused on the influence of surface roughness on the rate of osseointegration, while studies on the development of surfaces to also improve the interaction of peri-implant soft tissues are lacking. To this end, the first purpose of this study was to evaluate the response of human gingival fibroblasts (hGDFs) to titanium implant discs (Implacil De Bortoli, Brazil) having different micro and nano-topography: machined (Ti-M) versus sandblasted/double-etched (Ti-S). The secondary aim was to investigate the effect of the macrogeometry of the discs on cells: linear-like (Ti-L) versus wave-like (Ti-W) surfaces. The atomic force microscopy (AFM) and scanning electron microscopy (SEM) analysis showed that the Ti-S surfaces were characterized by a significantly higher micro and nano roughness and showed the 3D macrotopography of Ti-L and Ti-W surfaces. For in vitro analyses, the hGDFs were seeded into titanium discs and analyzed at 1, 3, and 5 days for adhesion and morphology (SEM) viability and proliferation (Cck-8 and MTT assays). The results showed that all tested surfaces were not cytotoxic for the hGDFs, rather the nano-micro and macro topography favored their proliferation in a time-dependent manner. Especially, at 3 and 5 days, the number of cells on Ti-L was higher than on other surfaces, including Ti-W surfaces. In conclusion, although further studies are needed, our in vitro data proved that the use of implant discs with Ti-S surfaces promotes the adhesion and proliferation of gingival fibroblasts, suggesting their use for in vivo applications.     

A new data mining approach to estimate causal effects of policy interventions

This paper presents a data driven approach that enables one to obtain a measure of comparability between-groups in the presence of observational data.The main idea lies in the use of the general framework of conditional multiple correspondences analysis as a tool for investigating the dependence relationship between a set of observable categorical covariates X and an assignment-to-treatment indicator variable T, in order to obtain a global measure of comparability between-groups according to their dependence structure. Then, we propose a strategy that enables one to find treatment groups, directly comparable with respect to pre-treatment characteristics, on which estimate local causal effects.