Transcriptomic Changes in Cisplatin-Resistant MCF-7 Cells.

Breast cancer is a leading cause of cancer-related deaths among women. Cisplatin is used for treatment, but the development of resistance in cancer cells is a significant concern. This study aimed to investigate changes in the transcriptomes of cisplatin-resistant MCF7 cells. We conducted RNA sequencing of cisplatin-resistant MCF7 cells, followed by differential expression analysis and bioinformatic investigations to identify changes in gene expression and modified signal transduction pathways. We examined the size and quantity of extracellular vesicles. A total of 724 genes exhibited differential expression, predominantly consisting of protein-coding RNAs. Notably, two long non-coding RNAs (lncRNAs), NEAT1 and MALAT, were found to be dysregulated. Bioinformatic analysis unveiled dysregulation in processes related to DNA synthesis and repair, cell cycle regulation, immune response, and cellular communication. Additionally, modifications were observed in events associated with extracellular vesicles. Conditioned media from resistant cells conferred resistance to wild-type cells in vitro. Furthermore, there was an increase in the number of vesicles in cisplatin-resistant cells. Cisplatin-resistant MCF7 cells displayed differential RNA expression, including the dysregulation of NEAT1 and MALAT long non-coding RNAs. Key processes related to DNA and extracellular vesicles were found to be altered. The increased number of extracellular vesicles in resistant cells may contribute to acquired resistance in wild-type cells.

The reduction of Cr(VI) in Salvinia minima, possible involvement of an h-type thioredoxin.

Hexavalent chromium [Cr(VI)] is extremely toxic to plant cells and has been recognized to possess a high redox potential. Tolerant plant species have shown the ability to reduce Cr(VI), but the operating mechanism involved in this process is not elucidated. Thus, the aim of this study was to investigate the possible involvement of thiolic and phenolic compounds and thioredoxin expression during Cr(VI) reduction in S. minima. In addition, a probable enzymatic reduction of Cr(VI) was investigated. Plants were exposed to 20 mg L-1 Cr(VI) concentration during 7 days under controlled conditions. The amount of metal accumulated in lacinias (root-like submerged leaves) and fronds (floating leaves) indicated that a low percentage of absorbed Cr(VI) was mobilized from lacinias to fronds. X-ray absorption near-edge structure (XANES) analysis revealed that Cr(III) was the only chromium species occurring in S. minima plants. Thiols and phenolics of lacinias and fronds were increased significantly by Cr(VI) treatment, but accumulation patterns were different. The expression of an h-type thioredoxin (Trx h) was demonstrated for the first time in Cr-exposed lacinias. Enzymatic reduction showed a low contribution to the Cr(VI) reduction. Data of this study provide evidences on the involvement of thiols, thioredoxin, and phenolics in the reduction of Cr(VI) to Cr(III) in S. minima tissues.

Chronic wound healing by controlled release of chitosan hydrogels loaded with silver nanoparticles and calendula extract.

As the skin is the main protective organ of the body, it is exposed to wounds or injuries which carry out a healing process during a period of approximately 15 days depending on the severity of the injury. In the present research, the development of chitosan-based hydrogels loaded with silver nanoparticles and calendula extract (Ch-AgNPs-Ce) was proposed. This can be used to fulfill the hemostatic, anti-infective, antibacterial, healing and anti-inflammatory functions through controlled release of the nanoparticles and calendula extract in substitution of commonly used drugs. The physical properties of the silver nanoparticles were analyzed by UV-visible spectroscopy, scanning and transmission electron microscopy, showing a size between 50 and 100 nm. The antibacterial properties were evaluated by the agar well diffusion method. Antimicrobial testing of the hydrogels showed that the inclusion of silver nanoparticles provides concentration-dependent antibacterial behavior against E. coli and S. aureus. The healing properties of the system were tested in two diabetic patients to whom said hydrogels were placed, obtaining a positive curative result after a few weeks. Therefore, it can be concluded that Ch-AgNPs-Ce hydrogels can achieve healing in chronic or exposed wounds after a period of time which can be used in alternative treatments in patients with poor healing capacity.

Tuning the Photothermal Effect of Carboxylated-Coated Silver Nanoparticles through pH-Induced Reversible Aggregation.

The photothermal response of mercaptoundecanoic acid (MUA)-coated Ag nanoparticles (Ag@MUA NPs) in both aqueous dispersions and paper substrates was determined as a function of pH when irradiated with a green laser or a blue LED source. Aqueous dispersions of Ag@MUA NPs showed an aggregation behavior by acidification that was used for the formation of NPs clusters of variable sizes. Aggregation was induced by changing the pH across the apparent pKa of the acid, higher than the pKa of the free acid. Formation of these aggregates was completely reversible allowing the return to the well-dispersed initial state by simply increasing the pH by the addition of a base. Aggregation produced a shift of the plasmon band that changed the spectra of the dispersions and their ability to be remotely heated when irradiated with visible light. These aggregates could be transferred to paper by simple impregnation of the substrates with the dispersion. On the solid substrate, a higher photothermal response than in the liquid medium was observed. A high local increase of up to 75 °C could be recorded on paper after only 30 s of irradiation with a green laser, whereas a blue LED array was enough for inducing the melting of a solid paraffin (Tm = 36-38 °C) deposited on it. This work demonstrates that photothermal heating can be controlled by the reversible aggregation of NPs to induce different thermal responses in liquid and solid media.

Unexpected compositional and structural modification of CoPt3 nanoparticles by extensive surface purification.

We combined synchrotron small angle X-ray scattering, X-ray fluorescence and extended X-ray absorption fine structure spectroscopy to probe the structure of chemically synthesized CoPt3 nanoparticles (NPs) after ligand removal via the commonly accepted solvent/nonsolvent approach. We showed that the improved catalytic activity of extensively purified NPs could not be explained only in terms of a "cleaner" surface. We found that extensive surface purification results in the substantial leaching of the Co atoms from the chemically synthesized CoPt3 NPs transforming them into CoPt3/Pt core/shell structures with an unexpectedly thick (∼0.5 nm) Pt shell. We indicated that the improved catalytic activity of extensively purified NPs in octyne hydrogenation reaction can be explained by the formation of CoPt3/Pt core/shell structures. Also, we demonstrated that drastic compositional and structural transformation of water transferred CoPt3 NPs was rather a result of extensive removal of native ligands via a solvent/nonsolvent approach than leaching of cobalt atoms in aqueous media. We expect that these findings can be relevant to other transition metal based multicomponent NPs.

EXAFS and DFT study of the cadmium and lead adsorption on modified silica nanoparticles.

Silica nanoparticles of 7 nm diameter were modified with (3-aminopropyl) triethoxysilane (APTES) and characterized by CP-MAS (13)C and (29)Si NMR, FTIR, zeta potential measurements, and thermogravimetry. The particles were shown to sorb successfully divalent lead and cadmium ions from aqueous solution. Lead complexation with these silica nanoparticles was clearly confirmed by EXAFS (Extended X-ray Absorption Fine Structure) with synchrotron light measurements. Predicted Pb-N and Pb-C distances obtained from quantum-chemical calculations are in very good agreement with the EXAFS determinations. The calculations also support the higher APTES affinity for Pb(2+) compared to Cd(2+).

Angiogenic effects of ionic dissolution products released from a boron-doped 45S5 bioactive glass.

In regenerative medicine of vascularized tissues, there is a great interest in the use of biomaterials that are able to stimulate angiogenesis, a process necessary for rapid revascularization to allow the transport and exchange of oxygen, nutrients, growth factors and cells that take part in tissue repair and/or regeneration. An increasing number of publications have shown that bioactive glasses stimulate angiogenesis. Because it has been established that boron (B) may play a role in angiogenesis, the aim of this study was to assess the in vivo angiogenic effects of the ionic dissolution products that from a bioactive glass (BG) in the 45S5 system doped with 2 wt% B2O3 (45S5.2B). The pro-angiogenic capacity of 45S5.2B BG was assessed on the vasculature of the embryonic quail chorioallantoic membrane (CAM). Ionic dissolution products from 45S5.2B BG increased angiogenesis. This is quantitatively evidenced by the greater expression of integrin αvß3 and higher vascular density in the embryonic quail CAM. The response observed at 2 and 5 days post-treatment was equivalent to that achieved by applying 10 µg mL-1 of basic fibroblast growth factor. These results show that the ionic dissolution products released from the bioactive glass 45S5.2B stimulate angiogenesis in vivo. The effects observed are attributed to the presence the ionic dissolution products, which contained 160 ± 10 µM borate.

In vitro endothelial cell response to ionic dissolution products from boron-doped bioactive glass in the SiO2-CaO-P2O5-Na2O system.

As it has been established that boron (B) may perform functions in angiogenesis and osteogenesis, the controlled and localized release of B ions from bioactive glasses (BGs) is expected to provide a promising therapeutic alternative for regenerative medicine of vascularized tissues, such as bone. The aim of this study was to assess the in vitro angiogenic effects of the ionic dissolution products (IDPs) from BGs in the SiO2-CaO-Na2O-P2O5 (45S5) system and of those from 45S5 BG doped with 2 wt% B2O3 (45S5.2B). The results show, for the first time, the IDPs from 45S5.2B BG stimulated human umbilical vein endothelial cell (HUVEC) proliferation and migration that were associated with phosphorylation of extracellular signal-related kinase (ERK) 1/2, focal adhesion kinase (FAK) and p38 protein. It was also shown that IDPs from 45S5.2B BG could enhance in vitro HUVEC tubule formation and secretion of interleukin 6 (IL6) and the basic fibroblast growth factor (bFGF). The effects observed are attributed to the presence of B in the IDPs. These findings are relevant to bone tissue engineering and regeneration because the IDPs from 45S5.2B BG may act as inexpensive inorganic angiogenic agents providing a convenient alternative to the application of conventional angiogenic growth factors.

Resistance and inactivation kinetics of bacterial strains isolated from the non-chlorinated and chlorinated effluents of a WWTP.

The microbiological quality of water from a wastewater treatment plant that uses sodium hypochlorite as a disinfectant was assessed. Mesophilic aerobic bacteria were not removed efficiently. This fact allowed for the isolation of several bacterial strains from the effluents. Molecular identification indicated that the strains were related to Aeromonas hydrophila, Escherichia coli (three strains), Enterobacter cloacae, Kluyvera cryocrescens (three strains), Kluyvera intermedia, Citrobacter freundii (two strains), Bacillus sp. and Enterobacter sp. The first five strains, which were isolated from the non-chlorinated effluent, were used to test resistance to chlorine disinfection using three sets of variables: disinfectant concentration (8, 20 and 30 mg·L(-1)), contact time (0, 15 and 30 min) and water temperature (20, 25 and 30 °C). The results demonstrated that the strains have independent responses to experimental conditions and that the most efficient treatment was an 8 mg·L(-1) dose of disinfectant at a temperature of 20 °C for 30 min. The other eight strains, which were isolated from the chlorinated effluent, were used to analyze inactivation kinetics using the disinfectant at a dose of 15 mg·L(-1) with various retention times (0, 10, 20, 30, 60 and 90 min). The results indicated that during the inactivation process, there was no relationship between removal percentage and retention time and that the strains have no common response to the treatments.

Shape changes of Pt nanoparticles induced by deposition on mesoporous silica.

Polyvinylpyrollidone (PVP)-capped platinum nanoparticles (NPs) are found to change shape from spherical to flat when deposited on mesoporous silica substrates (SBA-15). Transmission electron microscopy (TEM), small-angle X-ray scattering (SAXS), and extended X-ray absorption fine structure (EXAFS) analyses are used in these studies. The SAXS results indicate that, after deposition, the 2 nm NPs have an average gyration radius 22% larger than in solution, while the EXAFS measurements indicate a decrease in first neighbor co-ordination number from 9.3 to 7.4. The deformation of these small capped NPs is attributed to interactions with the surface of the SBA-15 support, as evidenced by X-ray absorption near-edge structure (XANES).

Synthesis and characterization of gold at gold(i)-thiomalate core at shell nanoparticles.

In this paper, the synthesis of gold at gold(I)-thiolate core at shell nanoparticles is described for the first time. The chemical nature and structure of these nanoparticles were characterized by a multi-technique approach. The prepared particles consist of gold metallic cores, about 1 nm in size, surrounded by stable gold(I)-thiomalate shells (Au at Au(I)-TM). These nanoparticles could be useful in medicine due to the interesting properties that gold(I)-thiomalate has against rheumatoid arthritis. Furthermore, the described results give new insights in the synthesis and characterization of metallic and core at shell nanoparticles.

Osteoconductivity of strontium-doped bioactive glass particles: a histomorphometric study in rats.

There is accumulating evidence that strontium (Sr)-containing bioceramics have positive effects on bone tissue repair. The aims of the present study were to evaluate the osteoconductivity of Sr-doped bioactive glass (BG) particles implanted in rat tibia bone marrow, and characterize the neoformed bone tissue by SEM-energy-dispersive X-ray microanalysis. Melt-derived BGs were prepared from a base 45S5 BG. Sr-doped glass (45S5.6Sr) was prepared using 6 wt % SrO as a substitute for the CaO. Histological analysis using undecalcified sections showed that new lamellar bone had formed along the surface of both 45S5 and 45S5.6Sr BG particles within 4 weeks. To evaluate osteoconductivity, affinity indices were calculated. At 30 days after implantation, 45S5 and 45S5.6Sr BGs had almost identical affinity indices (88% +/- 7% and 87% +/- 9%; p > 0.05). Strontium was not detected in the neoformed bone tissue surrounding 45S5.6Sr BG particles. These results indicate that 45S5.6Sr BG particles are osteoconductive when implanted inside the intramedullary canal of rat tibiae, and no alterations in bone mineralization, in terms of Ca/P ratio, were observed in the neoformed bone tissue around 45S5.6Sr BG particles.

Biocompatibility and bone mineralization potential of 45S5 Bioglass-derived glass-ceramic scaffolds in chick embryos.

The aim of the present study was to evaluate the biocompatibility and bone mineralization potential of 45S5 Bioglass-derived glass-ceramic scaffolds using a chick embryo shell-less (ex ovo) culture system. Chick embryos were divided into two groups: control (C) and experimental (E). Scaffolds were placed on the chorioallantoic membrane (CAM) in embryos of group E at 10 days of total incubation. The 45S5 Bioglass-derived glass-ceramic scaffolds proved to be biocompatible in terms of the absence of inflammatory response at the implant site (CAM). Moreover, no alterations in the other end-points assessed, i.e. survival, stage of embryonic development and body weight, were detected. However, body length was greater in group E embryos than in group C embryos (p0.05). A marked reduction (93%) in Ca content in the scaffolds was evidenced by energy-dispersive X-ray analysis at 5 days post-implantation. Calcium release from the scaffold implanted on the CAM might have been responsible for the restoration of the bone-like phenotype in chick embryonic skeleton of group E as detected by Alcian blue-Alizarin red double staining, as well as by histological and microchemical analyses. Conversely, the control embryos exhibited a chondrogenic phenotype.

Electrochemical preparation and delivery of melanin-iron covered gold nanoparticles.

Attractive combination: Biopolymer-modified nanoparticles which combine magnetic properties with biocompatibility are prepared and delivered following a three-step strategy (see figure): i) Adsorption of thiol-capped metal nanoparticles on graphite, ii) electrochemical modification, iii) potential-induced delivery of the modified nanoparticles to the electrolyte. Thiol-capped gold nanoparticles modified with iron-melanin are attractive because they combine magnetic properties and biocompatibility. The biopolymer modified nanoparticles are prepared and delivered following a three step strategy: i) adsorption of thiol-capped metal nanoparticles on graphite, ii) electrochemical deposition of melanin-iron, iii) potential-induced delivery of the modified nanoparticles to the electrolyte.

Genetic analysis and effect of triiodothyronine and prednisone trial on bone turnover in a patient with craniotubular hyperostosis.

Craniotubular hyperostosis are a group of high bone mass disorders related to mutations in the LRP5 and SOST genes, although other causative genes remain to be identified. Little is known about the bone turnover and the response to T3 or glucocorticoids in these patients. We describe a patient with craniotubular hyperostosis, including mutation analyses of the LRP5, SOST, DKK1 and KRM1 genes. We also studied bone turnover and bone mineral density (BMD), before and after a trial with T3 (75 microg/d for 28 weeks) and T3 and prednisone (T3 100 microg/d for 2 weeks, followed by 10 weeks on prednisone 10 mg/d, and a final 2 weeks period off of medicactions, completing 3 cycles in 42 weeks. Mutation analysis of the complete coding region and flanking highly conserved sequences of SOST, evaluation of the presence of the 52-kb deletion associated with Van Buchem disease in Dutch patients and mutation analysis of exons 2-4 of LRP5, and the coding regions of DKK1 and KRM1 did not reveal any disease-causing mutations. A baseline 5 to 7 fold increase in osteocalcin and in deoxypiridinoline was detected. After 4 weeks on 75 microg/d of T3, osteocalcin decreased 36%, but at week 28, it returned to basal. Deoxypiridinoline did not change. After the first cycle on T3 and prednisone, osteocalcin decreased 72%, and at the end of the third cycle it remained 44% below basal value. Deoxypiridinoline was stable and high during the three cycles; no changes in BMD were observed. As we failed to identify any disease-causing mutations in our patient with craniotubular hyperostosis, we suggest that another gene must be involved in the pathogenesis of his condition. This study provides additional data about the high bone turnover described in craniotubular hyperostosis, and also suggests an abnormal response to T3 excess in this condition.

Electronic perturbation in a molecular nanowire of [IrCl5(NO)]- units.

The nitrosyl in [IrCl5(NO)]- is probably the most electrophilic known to date. This fact is reflected by its extremely high IR frequency in the solid state, electrochemical behavior, and remarkable reactivity in solution. PPh4[IrCl5(NO)] forms a crystal in which the [IrCl5(NO)]- anions are in a curious wire-like linear arrangement, in which the distance between the N--O moiety of one anion and the trans chloride of the upper one nearby is only 2.8 A. For the same complex [IrCl5(NO)]- but with a different counterion, Na[IrCl5(NO)], the anions are stacked one over the other in a side-by-side arrangement. In this case the electronic distribution can be depicted as the closed-shell electronic structure Ir III-NO+, as expected for any d(6) third-row transition metal complex. However, in PPh4[IrCl5(NO)] an unprecedented electronic perturbation takes place, probably due to NO*-Cl- acceptor-donor interactions among a large number of [IrCl5(NO)]- units, favoring a different electronic distribution, namely the open-shell electronic structure Ir IV-NO*. This conclusion is based on XANES experimental evidence, which demonstrates that the formal oxidation state for iridium in PPh4[IrCl5(NO)] is +4, as compared with +3 in K[IrCl5(NO)]. In agreement, solid-state DFT calculations show that the ground state for [IrCl5(NO)]- in the PPh4+ salt comprises an open-shell singlet with an electronic structure which encompasses half of the spin density mainly localized on a metal-centered orbital, and the other half on an NO-based orbital. The electronic perturbation could be seen as an electron promotion from a metal-chloride to a metal-NO orbital, due to the small HOMO-LUMO gap in PPh4[IrCl5(NO)]. This is probably induced by electrostatic interactions acting as a result of the closeness and wire-like spatial arrangement of the Ir metal centers, imposed by lattice forces due to pi-pi stacking interactions among the phenyl rings in PPh4+. Experimental and theoretical data indicate that in PPh4[IrCl5(NO)] the Ir-N-O moiety is partially bent and tilted.

Pulmonary rehabilitation improves depression, anxiety, dyspnea and health status in patients with COPD.

OBJECTIVE: To determine the impact of an 8-wk program of comprehensive pulmonary rehabilitation on depression, anxiety, dyspnea, and health-related quality of life in patients with chronic obstructive pulmonary disease (COPD). DESIGN: We studied 24 patients with severe COPD randomized either to pulmonary rehabilitation (PR), (n = 10; FEV1 30 +/- 9%) or control (C; n = 14; FEV1 34 +/- 11%). The PR program included disease education, energy conservation techniques, relaxation, and exercise including 20-min arm elevation with dumbbells and 20-min leg exercise sessions three times a week for 8 wks. At baseline and after completion of the program, all patients were evaluated using the Beck Depression Inventory, State Trait Anxiety Inventory (STAI), Modified Medical Research Council Scale (MRC), and St. George's Respiratory Questionnaire (SGRQ). RESULTS: After PR, there was a significant improvement in the severity of depression (P < 0.01), a decrease in symptoms (P < 0.05), an increase in daily living activities (P < 0.05), and a decrease in the total score of the SGRO (P < 0.01). Dyspnea measured by the MRC scale was significantly better in the PR group (P < 0.01). CONCLUSIONS: The present study shows that in patients with severe COPD, pulmonary rehabilitation induces important changes on depression and anxiety independent of changes in dyspnea and health-related quality-of-life.

Biological performance of boron-modified bioactive glass particles implanted in rat tibia bone marrow.

The aim of the present study was to characterize the neoformed bone tissue around boron-modified bioactive glass particles implanted in rat tibia bone marrow by histologic, histomorphometric and microchemical evaluation. Melt-derived glasses were prepared from a base 45S5 bioactive glass of nominal composition (45% SiO(2), 24.5% CaO, 24.5% Na(2)O and 6% P(2)O(5) in wt%). The glass composition was modified by adding 2% wt of boron oxide (45S5.2B). Histological and histomorphometric analyses using undecalcified sections showed that at 15 days post-implantation the area of neoformed bone tissue around the 45S5.2B particles was significantly higher than control 45S5 glass. No statistically significant differences were observed at 30 days post-implantation. The thickness of osseointegrated tissue on 45S5.2B BG particles was significantly greater than on the control at all experimental time-points evaluated. A statistically significant increase in the Ca:P ratio was observed in the neoformed bone around 45S5.2B particles 15 days post-implantation. The results of the present study provide evidence that particles of boron-modified 45S5 BG (45S5.2B) enhance bone formation more than 45S5 glass when implanted into the intramedullary canal of rat tibiae.

Reversal of hypogonadotropic hypogonadism with tamoxifen in a patient with hyperprolactinemia resistant to dopamine agonists.

OBJECTIVE: To determine the response to tamoxifen in a woman with hypogonadotropic hypogonadism induced by hyperprolactinemia resistant to dopamine agonist drugs. DESIGN: Case report. SETTING: Academic fertility center. PATIENT(S): A young woman with persistent amenorrhea, symptomatic hypogonadotropic hypogonadism, and hyperprolactinemia. INTERVENTION(S): Tamoxifen was administered in addition to bromocriptine. MAIN OUTCOME MEASURE(S): Measurements of follicle-stimulating hormone, estradiol, prolactin, and progesterone. RESULT(S): Recovery of gonadal-hypothalamic-pituitary axis. CONCLUSION(S): Tamoxifen can revert the effects of chronic hypogonadotropic hypogonadism induced by hyperprolactinemia resistant to dopamine agonists.