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This review focuses on the acceleration of the bactericidal and fungicidal effects by uniform, adhesive Cu-based nanocomposites on textile and thin polymer surfaces. The acceleration of the bacterial inactivation kinetics mediated by Cu, TiO2/Cu and ZrO2-TiO2-Cu films in aerobic and anaerobic media is discussed for Gram negative, Gram positive bacteria and fungi. The Cu induced bacterial inactivation kinetics in the dark was observed to occur within the minute range for diverse bacteria. The characterization of the innovative Cu-films microstructure by several surface science techniques is described. This allows the correlation of the surface-reactivity features of the Cu-composite films mediating bacterial inactivation. The redox reactions on the Cu-films during the bacterial inactivation are documented by X-ray-photoelectron spectroscopy (XPS). Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy was used to monitor the changes in the functional groups leading to bacterial inactivation as a function of the bacterial inactivation time. The dependence of the inactivation kinetics on the Cu-loaded surface roughness is accounted for in qualitative way by atomic force microscopy (AFM). A mechanism for the bacterial inactivation induced by the Cu-nanoparticles by way of reactive oxygen species (ROS) in aerobic and anaerobic processes is discussed. The interfacial charge transfer (IFCT) mechanism under light irradiation between Cu and TiO2 or for the Cu with the binary oxide ZrO2-TiO2 is suggested. The relative potential band-position in the Cu-sputtered semiconductors responsible for the charge transfer is discussed. Evidence was found for the presence in anaerobic media of highly oxidative valence band holes (h+)vb at −0.4 eV generated by Cu2O. The variation in the interface surface potential (Eigenvalues) of the Cu-films during the bacterial inactivation was monitored and allowed to suggest the bacterial mineralization mechanism. Details for the reactions of the Cu-surfaces with Escherichia coli (E. coli) and Methicillin Resistant Staphylococcus Aureus (MRSA) inactivation are described. Evidence for the synergism between Cu and the TiO2 accelerating the bacterial inactivation kinetics is presented in this review.

Airborne bacterial communities are subject to conditions ill-suited to microbial activity and growth. In spite of this, air is an important transfer medium for bacteria, with the bacteria in indoor air having potentially major consequences for the health of a building's occupants. A major example is the decreased diversity and altered composition of indoor airborne microbial communities as a proposed explanation for the increasing prevalence of asthma and allergies worldwide. Previous research has shown that living on a farm confers protection against development of asthma and allergies, with airborne bacteria suggested as playing a role in this protective effect. However, the composition of this beneficial microbial community has still not been identified. We sampled settled airborne dust using a passive dust sampler from Danish pig stables, associated farmers' homes, and from suburban homes (267 samples in total) and carried out quantitative PCR measurements of bacterial abundance and MiSeq sequencing of the V3-V4 region of bacterial 16S rRNA genes found in these samples. Airborne bacteria had a greater diversity and were significantly more abundant in pig stables and farmers' homes than suburban homes (Wilcoxon rank sum test P < 0.05). Moreover, bacterial taxa previously suggested to contribute to a protective effect had significantly higher relative and absolute abundance in pig stables and farmers' homes than in suburban homes (ALDEx2 with P < 0.05), including Firmicutes, Peptostreptococcaceae, Prevotellaceae, Lachnospiraceae, Ruminococcaceae, Ruminiclostridium, and Lactobacillus. Pig stables had significantly lower airborne bacterial diversity than farmers' homes, and there was no discernable direct transfer of airborne bacteria from stable to home. This study identifies differences in indoor airborne bacterial communities that may be an important component of this putative protective effect, while showing that pig stables themselves do not appear to directly contribute to the airborne bacterial communities in the homes of farmers. These findings improve our understanding of the role of airborne bacteria in the increasing prevalence of asthma and allergy.

The intervertebral disc (ND) is the most avascular and acellular tissue in the body and therefore prone to degeneration. During IVD degeneration, the balance between anabolic and catabolic processes in the disc is deregulated, amongst others leading to alteration of extracellular matrix production, abnormal enzyme activities and production of pro-inflammatory substances like cytokines. The established treatment strategy for ND degeneration consists of physiotherapy, pain medication by drug therapy and if necessary surgery. This approach, however, has shown limited success. Alternative strategies to increase and prolong the effects of bioactive agents and to reverse the process of IVD degeneration include the use of delivery systems for drugs, proteins, cells and genes. In view of the specific anatomy and physiology of the ND and depending on the strategy of the therapy, different delivery systems have been developed which are reviewed in this article. (C) 2014 Elsevier B.V. All rights reserved.

The scaffold protein Spinophilin (Spinophilin, PPP1R9B) is one of the regulatory subunits of phosphatase-1 (PP1), directing it to distinct subcellular locations and targets. The loss of Spinophilin reduces PP1 targeting to pRb, thereby maintaining higher levels of phosphorylated pRb. Spinophilin is absent or reduced in approximately 40% of human lung tumors, correlating with the malignant grade. However, little is known about the relevance of the coordinated activity or presence of Spinophilin and its reported catalytic partners in the prognosis of lung cancer. In the present work, we show that the downregulation of Spinophilin, either by protein or mRNA, is related to a worse prognosis in lung tumors. This effect is more relevant in squamous cell carcinoma, SCC, than in adenocarcinoma. Downregulation of Spinophilin is related to a decrease in the levels of its partners PPP1CA/B/C, the catalytic subunits of PP1. A decrease in these subunits is also related to prognosis in SCC and, in combination with a decrease in Spinophilin, are markers of a poor prognosis in these tumors. The analysis of the genes that correlate to Spinophilin in lung tumors showed clear enrichment in ATP biosynthesis and protein degradation GO pathways. The analysis of the response to several common and pathway-related drugs indicates a direct correlation between the Spinophilin/PPP1Cs ratio and the response to oxaliplatin and bortezomib. This finding indicates that this ratio may be a good predictive biomarker for the activity of the drugs in these tumors with a poor prognosis., This work was funded by grants from the Spanish Ministry of Economy and Competitivity, Plan Estatal de I+D+I 2013-2016, ISCIII (Fis: PI15/00045) and CIBER de Cáncer (CB16/12/00275, CB16/12/00443, CB16/12/00442), co-funded by FEDER from Regional Development European Funds (European Union), Consejeria de Ciencia e Innovacion (CTS-1848) and Consejeria de Salud of the Junta de Andalucia (PI-0096-2014). Especial thanks to the AECC Foundation for also supporting this work. This work has also been made possible by the Fundacion BBVA.

The main aim of this study is to assess the safety and antitumor efficacy of a palladium(II) (Pd)-saccharinate complex with terpyridine. To characterize the Pd(II) complex in vitro, its cytotoxicity was evaluated using a water-soluble tetrazolium salt cell viability assay and the mechanism of cell death was assessed by DNA fragmentation/condensation and live cell imaging analyses. The antitumor efficacy and safety of the Pd(II) complex in-vivo were examined by analyzing reduction in tumor size, changes in body and organ weight, histopathological analysis of liver, kidney, and tumor sections, and biochemical analysis of serum in C57BL/6 mice. Our results showed that the Pd(II) complex was more cytotoxic to cancer cells than noncancer cell lines and caused cell death through apoptotic pathways. The treatment of the Pd(II) complex in tumor-bearing mice effectively reduced the tumor size at half the dose used for cisplatin. The Pd(II) complex appeared to exert less liver damage than the cisplatin-based complex on changes in the hepatic enzymes levels in the serum. Hence, the complex appears to be a potential chemotherapeutic drug with high antitumor efficacy and fewer hepatotoxic complications, providing an avenue for further studies. Anti-Cancer Drugs 28: 898-910 Copyright (C) 2017 Wolters Kluwer Health, Inc. All rights reserved. Scientific and Technological Research Institute of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) This work was supported by the internal funding of The Scientific and Technological Research Institute of Turkey (TUBITAK). WOS:000408162400009 28657910 Q3

The aim of the present study is to investigate DNA binding interactions of two highly cytotoxic palladium(II) saccharinate (sac) complexes with 2,2':6',2 ''-terpyridine (terpy), [Pd(sac)(terpy)](sac)center dot 4H(2)O (1) and [PdCl (terpy)](sac)center dot 2H(2)O (2). UV-Vis spectroscopy, fluorescence spectroscopy, thermal denaturation, viscosity, and gel electrophoresis measurements were employed to study binding properties of the two complexes with Fish Sperm (FS) DNA. The interaction of the palladium(II) complexes with FS-DNA was monitored by hypochromism in the UV-Vis spectra. The binding constants (K-b) obtained from absorption titrations were estimated to be 1.0 (+/- 0.05) x 10(5) M-1 for 1 and 2.0 (+/- 0.22) x 10(5) M-1 for 2. Competitive binding studies with ethidium bromide have shown that both complexes strongly bind to FS-DNA, and the apparent binding constant (K-app) calculated from emission studies was 2.0 x 10(6) M-1 for each complex. Fluorescence Scatchard plots demonstrated that the palladium(II) complexes act as dual-function metallointercalators and bind to FS-DNA by both intercalation and coordination. On addition of the two complexes, significant increases in both viscosity and melting temperature of FS-DNA solutions further support the binding mode of these complexes. Gel electrophoresis assay demonstrates that both complexes efficiently cleavage the supercoiled plasmid 8454: pCMV-VSV-G DNA to its open circular form.

International audience; The ovine cervix relaxes at estrus allowing easier entry of spermatozoa into the uterus. The mechanism responsible for this relaxation is not fully elucidated and we hypothesized that cervical relaxation at estrus is induced by ovarian and pituitary hormones stimulating the local production of prostaglandin E-2 Via a biosynthetic pathway involving a number Of mediators including oxytocin, phospholipase A(2) (cPLA(2)), cyclooxygenase-2 (COX-2), and peroxisome proliferator-activated receptor gamma (PPAR gamma). The aim of this study was to investigate the cervical expression of estradiol receptor alpha (ER alpha), oxytocin receptor (OTR), cPLA(2), COX-2, and PPAR gamma at three stages of the estrous cycle (the luteal phase and two times during the follicular phase, just before and just after the LH surge). An experiment was conducted during the breeding season, in 25 ewes to test this hypothesis. Samples of cervical tissue were collected from groups of ewes at three stages of the estrous cycle: the luteal (N = 8), "pre-LH surge" (N = 8), and "post-LH surge" (N = 9) stages. Cervical tissue from uterine, mid, and vaginal regions of the cervix were analyzed by Western immunoblot analysis for ER alpha, OTR, cPLA(2), COX-2, and PPAR gamma. The results showed that the levels of all five proteins were lowest during the luteal phase of the estrous cycle in all regions of the cervix. The levels of all except cPLA(2), increased significantly during the "pre-LH surge" stage. The levels of cPLA(2) and ER alpha increased in the "post-LH surge" stage and those for OTR and PPARy were unchanged and those for COX-2 were lower. These data show that the cervical levels of all five of the intermediates in the synthesis of prostaglandin E-2 that were examined in this study were higher in the "pre-" and "post-LH surge" stages compared with the luteal phase of the estrous cycle and these findings are consistent with our hypothesis. (C) 2013 Elsevier Inc. All rights reserved.

Background: To overcome the hurdles of cisplatin, majorly its toxicity and resistance, there has been extensive search for alternative anti-cancer metal-based compounds. Here, three Cu(II)-complexes, Cu(Sal-Gly)(phen), Cu(Sal-Gly)(pheamine), Cu(Sal-Gly)(phepoxy) are characterized for their interaction with DNA, cytotoxicity and mechanism of action. Methods: The binding ability of the complexes to Calf-Thymus DNA was evaluated by competition fluorescence studies with thiazole-orange, UV-Vis and circular dichroism spectroscopic titrations. Cytotoxicity was evaluated by MTT analysis. The DNA damage was analyzed through cleavage of supercoiled DNA via agarose gel-electrophoresis, and 8-oxo-guanidine and gamma H2AX staining in cells. Apoptosis was detected via DNA condensation/fragmentation, mitochondrial membrane potential, Annexin V staining and caspase 3/7 activity. Formation of reactive oxygen species was determined by DCFDA- and GSSG/GSH-analysis. Results: Binding constants to DNA were evaluated as 1.7 x 10(6) (Cu(Sal-Gly)(phen)), 2.5 x 10(6) (Cu(Sal-Gly)(pheamine)) and 3.2 x 10(5) (Cu(Sal-Gly)(phepoxy)). All compounds induced DNA damage. Apoptosis was the main form of cell death. There was an increase in ROS, which is most likely responsible for the observed DNA-damage. Although the compounds were cytotoxic to all tested cancer cell lines, only Cu(Sal-Gly)(pheamine) displayed significantly lower toxicity towards non-cancer cells, its associated phenotypes differing from the other two Cu-complexes. Thus, Cu(Sal-Gly)(pheamine) was further assayed for molecular changes in response to drug treatment using a custom designed RT-qPCR array. Results showed that Harakiri was significantly upregulated. Presence of p53 was not required for apoptosis in response to Cu-complexes. Conclusions and general significance: These Cu-complexes, namely Cu(Sal-Gly)(pheamine), may be considered promising anticancer agents with activity in cancer cells even with deficient p53 status. Portuguese Foundation for Science and Technology UID/QUI/00100/2013 IST-UTL Center of the Portuguese Mass Spectrometry Networks REM2013 - RECI/QEQ-QIN/0189/2012

Extracellular matrix synthesis and remodelling are driven by increased activity of transforming growth factor beta 1 (TGF-β1). In tendon tissue repair, increased activity of TGF-β1 leads to progressive fibrosis. Decorin (DCN) and interleukin 10 (IL-10) antagonise pathological collagen synthesis by exerting a neutralising effect via downregulation of TGF-β1. Herein, we report that the delivery of DCN and IL-10 transgenes from a collagen hydrogel system supresses the constitutive expression of TGF-β1 and a range of pro-fibrotic extracellular matrix genes.

The role of sex steroid hormones in reproductive function in women is well established. However, in the last two decades it has been shown that receptors for estrogens, progesterone and androgens are expressed in non reproductive tissue /organs (bone, brain, cardiovascular system) playing a role in their function. Therefore, it is critical to evaluate the impact of sex steroid hormones in the pathophysiology of some diseases (osteoporosis, Alzheimer, atherosclerosis). In particular, women with primary ovarian insufficiency, polycystic ovary syndrome, endometriosis and climacteric syndrome may have more health problems and therefore an hormonal treatment may be crucial for these women.

In 2006, the European Commission has established maximum levels for ochratoxin A in wine and grape products, using occurrence data up to 2001 and toxicity data up to 2006. This paper presents an up-to-date overview of the occurrence of mycotoxins in grapes and wine produced in Europe in the period 1995-2010. In addition, for the most frequently occurring mycotoxins, factors influencing the occurrence, and the toxicological effects are presented. To evaluate possible trends in occurrence, contamination data were grouped into three periods of time, i.e. 1995-1999, 2000-2006 and 2007-2010. Most of the available contamination data on mycotoxins in grapes and wine refer to ochratoxin A, but occurrence data on this toxin from 2006 onwards are very limited. The occurrence of ochratoxin A is higher in the southern European countries than in the northern countries, and higher in red and sweet wines as compared to white wines. Fumonisins occur frequently, but in low concentrations. Data on the natural occurrence of Alternaria toxins are not available. The most important factors that influence ochratoxin A contamination of grapes and wine include: temperature and relative humidity in the month before harvesting the berries, the type of wine (maceration), and the percentage of damaged berries before vinification. Applying good agricultural practices in the vineyard, including minimizing damaged berries and chemical or biological control of the fungi, are the best methods to limit mycotoxin formation in grapes and wine. Ochratoxin A, Alternaria toxins and fumonisins are toxic to animals. These toxins are of concern to human health, but clear evidence on their relationship with human disease is not available yet. Therefore, more research in this area would be desirable.

Objective: To develop a new cementoblast culture method and to detect bone sialoprotein (BSP) expression in response to high and low mechanical tensile stress in cementoblast in vitro. Materials and Methods: Cementoblasts were collected from the roots of newborn bovine teeth and were identified with cementum-derived attachment protein (CAP) antibody 3G9. Cell proliferation was evaluated by MTT [3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, and mineralization was confirmed by von Kossa staining. Mechanical tensile stress was applied in vitro to the cementoblast with the use of a uniaxial four-point bending system with 2000 or 4000 microstrains, at a frequency of 0.5 Hz for 3, 6, 12, 24, or 36 hours. BSP mRNA level was quantified by real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Results: A large amount of cementoblast was observed to be expressing CAP. Cementoblasts had a proliferation tendency similar to that of osteoblasts but different from that of periodontal ligament (PDL) cells. Cementoblasts had the ability to become mineralized between osteoblasts and PDL cells. The mechanical tensile stress significantly up-regulated BSP mRNA expression, which reached a peak at 24 hours in both 2000 and 4000 microstrain groups (P < .01) and was tenfold and sixfold higher than that of controls, respectively. BSP expression dropped toward baseline levels at 36 hours in both groups. Conclusions: Mechanical tensile stress up-regulated the expression of BSP. Low mechanical tensile stress induced earlier and more intensive up-regulation of BSP mRNA; this might represent the optimal stimuli for cementoblast activity.

Background: Over the past few decades, the field of nanotechnology has led to significant advances in healthcare, impacting both diagnosis and therapy. Systemic delivery of therapeutics via inhalation route has also advanced with the use of sub micron particles as colloidal drug carriers. Use of inhalable nanocarriers for delivering drugs systemically offers additional degree of control and manipulation, thereby maximizing the alveolar deposition and minimizing clearance. The ramifications are improved systemic absorption and higher therapeutic efficacy. Herein, we review the progress and advances related to nanoparticle based inhalable formulations for systemic delivery of therapeutics, and also discusses the associated challenges. Methods: We performed detailed searches in PubMed and compiled the literature on inhalable nanoparticles for systemic delivery of therapeutic agents. Results: To date, multiple inhalable nanocarriers have been explored for systemic delivery of therapeutics; and can be broadly classified into three categories, viz. lipid based nanoparticles, polymeric nanoparticles and porous nanoparticle aggregate particles. Conclusion: In spite of the promising data, there are still multiple challenges, including poor understanding of nanotoxicology of therapeutic nanoparticles. Overcoming these challenges can lead to successful clinical translation of inhalable nanoparticles for systemic drug delivery, leading to the development of more effective and patient compliant therapies.

Background and Objective: Chlorella vulgaris is a multi-cellular edible algal species with abundant proteins. Extraction of high value protein fractions for pharmaceutical and nutritional applications can significantly increase the commercial value of microalga biomasses. There is no known report on the anticancer peptides derived from the Chlorella vulgaris abundant protein.Materials and Methods: This study examined the antimicrobial and anticancer effects of peptides from a hydrolyzed Chlorella vulgaris protein with 62 kDa molecular weight. Protein hydrolysis was done by pepsin as a gastrointestinal protease, and was monitored through protein content measurement, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and high performance liquidchromatography measurements. Inhibitory effect of the produced peptides on Escherichia coli cells and breast cancer cell lines was assayed.Results and Conclusion: Hydrolyzed peptides induced a decrease of about 34.1% in the growth of Escherichia coli, and the peptides of 3 to 5 kDa molecular weight had strong impact on the viability of breast cancer cells with IC50 value of 50 μg μl-1. The peptide fractions demonstrating antimicrobial and anti-cancer activities have the potential for use as functional food ingredients for health benefits. These results demonstrate that inexpensive algae proteinscould be a new alternative to produce anticancer peptides.Conflict of interest: The authors declare that there is no conflict of interest.

Isoprenoids represent one of the largest classes of phytochemicals. The structural diversity of these compounds, as well as their remarkable biological activities, makes them suitable candidates for the development of novel therapeutic agents. Several isoprenoids have demonstrated promising potential in the modulation of angiogenesis processes, and therefore provide an appealing alternative and/or addition to the available pharmacotherapies. These compounds could be used per se or combined with standard therapies, which can potentially reduce the undesired secondary effects. Compounds like the sesquiterpenoid artemisinin, and its derivatives, or the diterpenoid triptolide have been successfully tested in a broad range of models (in vitro and in vivo). Moreover, sesquiterpenoids seem to be a promising resource of natural angiogenic modulators, as it can be attested by the significant number of recent publications in this subject. On the other hand, other isoprenoids, such as the triterpenoid ursolic acid, are still under-explored and further studies are needed to understand their role within angiogenic process. Further insights into isoprenoids mode of action in angiogenesis will hopefully pave the way towards their successful clinical use.

Objective. We have previously shown that adenoviral-mediated melanoma differentiation-associated gene-7 (Ad.mda-7) therapy induces apoptosis in ovarian cancer cells. However, the apoptosis induction was low and directly correlated with infectivity of Ad.mda-7. The objective of this study was to derive ovarian cancer targeted infectivity-enhanced adenoviral vectors encoding mda-7 and evaluate their enhancement in therapeutic efficacy for ovarian carcinoma.Methods. Infectivity-enhanced adenoviral vectors encoding mda-7 Ad.RGD.mda-7 and Ad.RGD.pK7.mda-7 were derived by incorporation of RGD and or RGD and Pk7 motifs in the fiber knobs by genetic modification. Viruses were validated by PCR for presence of mda-7 and by Western blot for expression of MDA-7 protein. To test the enhancement of therapeutic efficacy of these viruses, a panel of human ovarian carcinoma cells, OV-4, HEY, SKOV3, SKOV3.ipl, were infected by either Ad.mda-7 or Ad.RGD.mda-7 and Ad.RGD.pK7.mda-7 or their respective control viruses and the cell killing was evaluated by crystal violet staining in vitro. Further, therapeutic efficacy was evaluated in vivo using human ovarian cancer xenograft mouse models.Results. Both Ad.RGD.pK7.mda-7 and Ad.RGD.mda-7 showed significant increase in cell killing in vitro compared to unmodified Ad.mda-7 with Ad.RGD.pK7.mda-7 showing highest cell killing. Further, Ad.RGD.pK7.mda-7 showed a significant increase in survival of mice bearing human ovarian cancer xenografts compared to Ad.mda-7 and other control groups.Conclusion. Infectivity-enhanced Ad.RGD.rnda-7 and Ad.RGD.pK7.mda-7 viruses significantly enhanced ovarian cancer tumor cell killing in vitro. Significant prolongation of survival by Ad.RGD.pK7.mda-7 in murine ovarian cancer models demonstrates the high clinical translational potential of these viruses for ovarian cancer therapy. (C) 2005 Elsevier Inc. All rights reserved.

Activation of the innate immune system by different homotopes may direct the subsequent specific immune response, and as a consequence may have significant implications on vaccination. In this study, effects of i.v. administered lipopolysaccharide (LPS), and lipoteichoic acid (LTA), respectively, and s.c. administered Keyhole Limpet Hemocyanin (KLH) on total and isotype-specific (IgM and IgG) primary and secondary antibody responses of chickens to human serum albumin (HuSA) were determined. Similarly, effects of pre-treatment of the birds with the homotopes on primary antibody responses to rabbit gamma globulin (RGG) were measured. In addition, antigen-specific (HuSA and RGG) and mitogenic (concanavalin A, Con A) responses of peripheral blood leucocytes in vitro were also measured. Our findings confirm earlier observations that both LTA as well as LPS have immunomodulatory features in chickens, albeit in an opposite direction. LTA enhanced primary and to a minor degree secondary antigen-specific antibody titers, but LPS was found to suppress these responses. In RGG-primed birds, LPS enhanced a non-antigen-specific recall response to HuSA. In addition, KLH acted in an immunomodulatory role too, enhancing primary and secondary antibody responses, but suppressing non-specific cellular immunity in a non-antigen-specific fashion. Possible mechanisms underlying our observations, and the role of LTA, LPS, and KLH in polarization of the specific antibody response in chickens are discussed.

After a decade of clinical trials, gene therapy seems to have found its place between excessive ambitions and feasible aims, with encouraging results obtained in recent years. Intracellular delivery of genetic material is the key step in gene therapy. Optimization of delivery vectors is of major importance for turning gene therapy into a successful therapeutic method. Nonviral gene delivery relies mainly on the complexes formed from cationic liposomes (or cationic polymers) and DNA, i.e., lipoplexes (or polyplexes). Many lipoplex formulations have been studied, but in vivo activity is generally low compared to that of viral systems. This review gives a concise overview of studies on the application of cationic liposomes in vivo in animal models of diseases and in clinical studies. The transfection efficiency, the pharmacokinetic and pharmacodynamic properties of the lipid-DNA complexes, and potentially relevant applications for cationic liposomes are discussed. Furthermore, the toxicity of, and the induction of an inflammatory response in association with the administration of lipoplexes are described. Increasing understanding of lipoplex behavior and gene transfer capacities in vivo offers new possibilities to enhance their efficiency and paves the path to more extensive clinical applications in the future.

After a decade of clinical trials, gene therapy seems to have found its place between excessive ambitions and feasible aims, with encouraging results obtained in recent years. Intracellular delivery of genetic material is the key step in gene therapy. Optimization of delivery vectors is of major importance for turning gene therapy into a successful therapeutic method. Nonviral gene delivery relies mainly on the complexes formed from cationic liposomes (or cationic polymers) and DNA, i.e., lipoplexes (or polyplexes). Many lipoplex formulations have been studied, but in vivo activity is generally low compared to that of viral systems. This review gives a concise overview of studies on the application of cationic liposomes in vivo in animal models of diseases and in clinical studies. The transfection efficiency, the pharmacokinetic and pharmacodynamic properties of the lipid-DNA complexes, and potentially relevant applications for cationic liposomes are discussed. Furthermore, the toxicity of, and the induction of an inflammatory response in association with the administration of lipoplexes are described. Increasing understanding of lipoplex behavior and gene transfer capacities in vivo offers new possibilities to enhance their efficiency and paves the path to more extensive clinical applications in the future.

The limited capacity to divide is one of the major differences between normal somatic cells and cancerous cells. This finite life span' of somatic cells is closely linked to loss of telomeric DNA at telomeres, the 'chromosome caps' consisting of repeated (TTAGGG) sequences. In more than 85% of advanced cancers, this telomeric attrition is compensated by telomerase, 'the immortality enzyme', implying that telomerase inhibition may restore mortality in tumor cells. This review discusses the progress in research on the structure and function of telomeres and the telomerase holoenzyme. In addition, new developments in telomere/telomerase targeting compounds such as antisense oligonucleotides and G-quadritplex stabilizing substances, but also new telomerase expression-related strategies such as telomerase promoter-driven suicide gene therapy and telomerase immunotherapy will be presented. It will be discussed how these data call be implemented in telomerase-directed therapies.

The limited capacity to divide is one of the major differences between normal somatic cells and cancerous cells. This finite life span' of somatic cells is closely linked to loss of telomeric DNA at telomeres, the 'chromosome caps' consisting of repeated (TTAGGG) sequences. In more than 85% of advanced cancers, this telomeric attrition is compensated by telomerase, 'the immortality enzyme', implying that telomerase inhibition may restore mortality in tumor cells. This review discusses the progress in research on the structure and function of telomeres and the telomerase holoenzyme. In addition, new developments in telomere/telomerase targeting compounds such as antisense oligonucleotides and G-quadritplex stabilizing substances, but also new telomerase expression-related strategies such as telomerase promoter-driven suicide gene therapy and telomerase immunotherapy will be presented. It will be discussed how these data call be implemented in telomerase-directed therapies.

The intervertebral disc (ND) is the most avascular and acellular tissue in the body and therefore prone to degeneration. During IVD degeneration, the balance between anabolic and catabolic processes in the disc is deregulated, amongst others leading to alteration of extracellular matrix production, abnormal enzyme activities and production of pro-inflammatory substances like cytokines. The established treatment strategy for ND degeneration consists of physiotherapy, pain medication by drug therapy and if necessary surgery. This approach, however, has shown limited success. Alternative strategies to increase and prolong the effects of bioactive agents and to reverse the process of IVD degeneration include the use of delivery systems for drugs, proteins, cells and genes. In view of the specific anatomy and physiology of the ND and depending on the strategy of the therapy, different delivery systems have been developed which are reviewed in this article. (C) 2014 Elsevier B.V. All rights reserved.

Telomerase, a ribonucleoprotein expressed in 85% of advanced cancers but not in most somatic cells, compensates for telomeric DNA erosion and as such stabilizes cell immortality. Telomerase inhibition might restore mortality in tumor cells. Recent progress is illustrated in studies on telomerase and telomere targeting with differentiation induction, reverse transcriptase inhibitors, promoter down regulation, antisense inhibition, and blockage of telomere/telomerase interactions. Also, new developments are described indicating that anti-telomerase treatment can induce apoptosis in tumor cells and fan chemosensitize drug-resistant cell lines. Implications of these findings for anti-telomerase-based therapeutic applications, in particular in combination therapies, are discussed.

Telomerase, a ribonucleoprotein expressed in 85% of advanced cancers but not in most somatic cells, compensates for telomeric DNA erosion and as such stabilizes cell immortality. Telomerase inhibition might restore mortality in tumor cells. Recent progress is illustrated in studies on telomerase and telomere targeting with differentiation induction, reverse transcriptase inhibitors, promoter down regulation, antisense inhibition, and blockage of telomere/telomerase interactions. Also, new developments are described indicating that anti-telomerase treatment can induce apoptosis in tumor cells and fan chemosensitize drug-resistant cell lines. Implications of these findings for anti-telomerase-based therapeutic applications, in particular in combination therapies, are discussed.

The active form of vitamin D3, 1,25-dihydroxyvitamin D3, is a major regulator of bone and calcium homeostasis. In addition, this hormone also inhibits the proliferation and stimulates the differentiation of normal as well as malignant cells. Supraphysiological doses of 1,25-dihydroxyvitamin D3 are required to reduce cancer cell proliferation. However, these doses will lead in vivo to calcemic side effects such as hypercalcemia and hypercalciuria. During the last 25 years, many structural analogs of 1,25-dihydroxyvitamin D3 have been synthesized by the introduction of chemical modifications in the A-ring, central CD-ring region or side chain of 1,25-dihydroxyvitamin D3 in the hope to find molecules with a clear dissociation between the beneficial antiproliferative effects and adverse calcemic side effects. One example of such an analog with a good dissociation ratio is calcipotriol (Daivonex®), which is clinically used to treat the hyperproliferative skin disease psoriasis. Other vitamin D analogs were clinically approved for the treatment of osteoporosis or secondary hyperparathyroidism. No vitamin D analog is currently used in the clinic for the treatment of cancer although several analogs have been shown to be potent drugs in animal models of cancer. Transcriptomics studies as well as in vitro cell biological experiments unraveled basic mechanisms involved in the antineoplastic effects of vitamin D and its analogs. 1,25-dihydroxyvitamin D3 and analogs act in a cell type- and tissue-specific manner. Moreover, a blockade in the transition of the G0/1 toward S phase of the cell cycle, induction of apoptosis, inhibition of migration and invasion of tumor cells together with effects on angiogenesis and inflammation have been implicated in the pleiotropic effects of 1,25-dihydroxyvitamin D3 and its analogs. In this review we will give an overview of the action of vitamin D analogs in tumor cells and look forward how these compounds could be introduced in the clinical practice. ispartof: Frontiers in physiology vol:5 pages:122- ispartof: location:Switzerland status: published

We present an integrated approach that predicts and validates novel anti-cancer drug targets. We first built a classifier that integrates a variety of genomic and systematic datasets to prioritize drug targets specific for breast, pancreatic and ovarian cancer. We then devised strategies to inhibit these anti-cancer drug targets and selected a set of targets that are amenable to inhibition by small molecules, antibodies and synthetic peptides. We validated the predicted drug targets by showing strong anti-proliferative effects of both synthetic peptide and small molecule inhibitors against our predicted targets. Electronic supplementary material The online version of this article (doi:10.1186/s13073-014-0057-7) contains supplementary material, which is available to authorized users.

Metal-based compounds represent promising anticancer therapeutic agents. In this study, the mechanism of action of a novel metal-based drug, a palladium(II) (Pd) complex ([PdCl(terpy)](sac)2H(2)O, terpy=2,2':6',2 ''-terpyridine and sac=saccharinate), was elucidated. The tested compound induced cytotoxicity in nine different human cancer cell lines that originated from various organs, suggesting a broad spectrum of activity. The IC50 values were significantly higher for noncancerous cells when compared with cancer cells. We found that cells treated with the Pd(II) complex exhibited increased caspase 3/7 activities and condensed/fragmented nuclei, as demonstrated by nuclear staining and DNA laddering. Morphological features, such as cellular shrinkage and blebbing, were also observed, indicating that apoptosis was the primary mechanism of cell death. Pd(II) treatment induced DNA double-stranded breaks both in vitro and in vivo, potentially accounting for the source of stress in these cells. Although caspase 3/7 activities were elevated after Pd(II) treatment, silencing or using inhibitors of caspase 3 did not block apoptosis. Other molecules that could potentially play a role in Pd(II)-induced apoptosis, such as p53 and Bax, were also tested using silencing technology. However, none of these proteins were essential for cell death, indicating either that these molecules do not participate in Pd(II)-induced apoptosis or that other pathways were activated in their absence. Hence, this new molecule might represent a promising anticancer agent that exhibits cytotoxicity in p53-mutant, Bax-mutant, and/or caspase 3-mutant cancer cells. (C) 2013 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

One of the barriers to the development of protein therapeutics is effective delivery to mammalian cells. The proteins must maintain a careful balance of polar moieties to enable administration and distribution and hydrophobic character to minimize cell toxicity. Numerous strategies have been applied to this end, from appending additional cationic peptides to supercharging the protein itself, sometimes with limited success. Here we present a strategy that combines these methods, by equipping a protein with supercharged elastin-like polypeptide (ELP) tags. We monitored cellular uptake and cell viability for GFP reporter proteins outfitted with a range of ELP tags and demonstrated enhanced uptake that correlates with the number of positive charges, while maintaining remarkably low cytotoxicity and resistance to degradation in the cell. GFP uptake proceeded mainly through caveolae-mediated endocytosis and we observed GFP emission inside the cells over extended time (up to 48 h). Low toxicity combined with high molecular weights of the tag opens the way to simultaneously optimize cell uptake and pharmacokinetic parameters. Thus, cationic supercharged ELP tags show great potential to improve the therapeutic profile of protein drugs leading to more efficient and safer biotherapeutics. (C) 2013 Elsevier Ltd. All rights reserved.

The design and development of water dispersible, pH responsive peptide mimic shell cross-linked magnetic nanocarriers (PMNCs) using a facile soft-chemical approach is reported. These nanocarriers have an average size about 10 nm, are resistant to protein adsorption in physiological medium, and transform from a negatively charged to a positively charged form in the acidic environment. The terminal amino acid on the shell of the magnetic nanocarriers allows us to create functionalized exteriors with high densities of organic moieties (both amine and carboxyl) for conjugation of drug molecules. The drug-loading efficiency of the nanocarriers is investigated using doxorubicin hydrochloride (DOX) as a model drug to evaluate their potential as a carrier system. Results show high loading affinity of nanocarriers for anticancer drug, their sustained release profile, magnetic-field-induced heating, and substantial cellular internalization. Moreover, the enhanced toxicity to tumor cells by DOX-loaded PMNCs (DOX-PMNCs) under an AC magntic field suggest their potential for combination therapy involving hyperthermia and chemotherapy.

International audience; Cardiovascular disease (CVD) mortality rates are lower in Asian countries where dietary patterns are very different from Western diet. A number of studies have linked these lower rates to the inclusion of soy products as a staple food in those countries. Soy is the richest dietary source of isoflavones, a type of phytoestrogen associated with many potentially beneficial effects. Isoflavone-containing soy protein consumption has been linked to reduced levels of LDL cholesterol in hypercholesterolemic patients. This effect is increased with the concomitant administration of isoflavones, and seems to be also complemented by the isoflavone capacity to restore the endothelial function in patients with weak and moderated endothelial dysfunction. The effects are variable depending on individuals' metabolism and in particular to their ability to convert daidzein to equol that seems to be restricted to approximately 1/3 of the population. Equol production has been indeed linked to a decreased arterial stiffness and antiatherosclerotic effects via NO production. Because the relevance of isoflavones consumption on the modulation of cardiovascular risk still remains unclear, this paper aims to review the existing knowledge on the biological activity of the isoflavones on the human cardiovascular system from an epidemiological, clinical and -omics point of view.

The second messenger diacylglycerol (DAG) plays a central role in the signal transduction of G-protein coupled receptors and receptor tyrosine kinases by binding to C1 domain of effector proteins. C1 domain was first identified in protein kinase C (PKC) which comprises a family of ten isoforms that play roles in diverse cellular processes such as proliferation, apoptosis and differentiation. Aberrant signaling through PKC isoforms and other C1 domain-containing proteins has been implicated in several pathological disorders. Drug discovery concerning C1 domains has exploited both natural products and rationally designed compounds. Currently, molecules from several classes of C1 domain-binding compounds are in clinical trials; however, still more have the potential to enter the drug development pipeline. This review gives a summary of the recent developments in C1 domain-binding compounds.

In the absence of any ideal gene delivery carrier despite the recent explosion of novel carrier systems, the current trend is to explore the complementary synergy promised by a combination of delivery systems such as liposomes, which are the most widely researched versatile non-viral carriers, and tissue-engineered scaffolds with macrostructures of defined architecture comprised of natural or synthetic macromolecules. Here, we discuss the recent advances in liposomal gene delivery and the possible benefits of a combined liposome–scaffold approach, such as long-term expression, enhanced stability, reduction in toxicity and ability to produce spatio-temporal expression patterns. This approach is generating significant impact in the field as a result of its potential for extended localised gene delivery for applications in a variety of clinical conditions. Deposited by bulk import

The composition of the fluid within the oviduct is largely determined by the secretory and absorptive activities of the oviduct epithelium. The present study explored the effects of basolateral nucleotide stimulation on ion transport in the bovine oviduct using the chamber short-circuit current technique. Basolateral application of ATP induced a rapid transient increase in ion secretion by oviduct epithelial monolayers in a concentration-dependent manner. The ATP-induced short-circuit current (I SC ) response was preserved in the presence of amiloride, whereas it was reduced in the absence of extracellular chloride or in the presence of bumetanide. The channels underlying the chloride secretory response were identified as Ca 2! -activated Cl " channels and CFTR. The ATP-induced Cl " secretory response was largely preserved in the absence of extracellular Ca 2! but was significantly reduced in the presence of BAPTA-AM(1,2-bis(2-aminophenoxy)ethaneN,N,N 0 ,N 0 -tetraacetic acid-acetomethoxy ester), thapsigargin, or 2-APB (2-aminoethoxydiphenylborate), demonstrating an important role for intracellular Ca 2! signaling in mediating these effects. A nucleotide potency profile of ATP#UTP (uridine triphosphate) .ADP, sensitivity to suramin, and cross-desensitization by basolateral UTP suggests that ATP exerted its effects on chloride secretion through the purinergic receptor P2Y, G proteincoupled 2, and the presence of theP2RY2gene was confirmed by RT-PCR. These results provide strong evidence that purinergic signaling constitutes a key mechanism of regulating chloride secretion and thus fluid formation in the bovine oviduct. ATP, chloride, female reproductive tract, oviduct, purinoceptors, secretory epithelia, signal transducers, signal transduction

La(1-x)Sr(x)MnO(3), a ferromagnet with high magnetization and Curie temperature T(C) below 70 degrees C, enables its use for magnetic hyperthermia treatment of cancer with a possibility of in vivo temperature control. We found that La(0.73)Sr(0.27)MnO(3) particles of size range 20-100 nm showed saturation magnetization around 38 emu/g at 20 kOe and a T(C) value of 45 degrees C. Aqueous suspension of these nanoparticles was prepared using a polymer, acrypol 934, and the biocompatibility of the suspension was examined using HeLa cells. A good heating ability of the magnetic suspension was obtained in the presence of AC magnetic field, and it was found to increase with the amplitude of field. The suspension having concentration of 0.66 mg/mL (e.g., 0.66 mg of nanoparticles with acropyl per milliliter of culture media) was observed to be biocompatible even after 96 h of treatment, as estimated by sulforhodamine B and trypan blue dye exclusion assays. Further, the treatment with the aforementioned concentration did not alter the microtubule cytoskeleton or the nucleus of the cells. However, the bare particles (concentration of 0.66 mg of nanoparticles per milliliter of culture media, but without acropyl) decreased the viability of cell significantly. Our in vitro studies suggest that the suspension (concentration of 0.66 mg/mL) may further be analyzed for in vivo studies.

Prostate cancer is the most common cancer of men in the Western world, and novel approaches for prostate cancer risk reduction are needed. Plant-derived phenolic compounds attenuate prostate cancer growth in preclinical models by several mechanisms, which is in line with epidemiological findings suggesting that consumption of plant-based diets is associated with low risk of prostate cancer. The objective of this study was to assess the effects of a novel lignan-stilbenoid mixture in PC-3M-luc2 human prostate cancer cells in vitro and in orthotopic xenografts. Lignan and stilbenoid -rich extract was obtained from Scots pine (Pinus sylvestris) knots. Pine knot extract as well as stilbenoids (methyl pinosylvin and pinosylvin), and lignans (matairesinol and nortrachelogenin) present in pine knot extract showed antiproliferative and proapoptotic efficacy at ≥ 40 μM concentration in vitro. Furthermore, pine knot extract derived stilbenoids enhanced tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induced apoptosis already at ≥ 10 μM concentrations. In orthotopic PC-3M-luc2 xenograft bearing immunocompromized mice, three-week peroral exposure to pine knot extract (52 mg of lignans and stilbenoids per kg of body weight) was well tolerated and showed anti-tumorigenic efficacy, demonstrated by multivariate analysis combining essential markers of tumor growth (i.e. tumor volume, vascularization, and cell proliferation). Methyl pinosylvin, pinosylvin, matairesinol, nortrachelogenin, as well as resveratrol, a metabolite of pinosylvin, were detected in serum at total concentration of 7-73 μM, confirming the bioavailability of pine knot extract derived lignans and stilbenoids. In summary, our data indicates that pine knot extract is a novel and cost-effective source of resveratrol, methyl pinosylvin and other bioactive lignans and stilbenoids. Pine knot extract shows anticarcinogenic efficacy in preclinical prostate cancer model, and our in vitro data suggests that compounds derived from the extract may have potential as novel chemosensitizers to TRAIL. These findings promote further research on health-related applications of wood biochemicals.