Short-term single treatment of chemotherapy results in the enrichment of ovarian cancer stem cell-like cells leading to an increased tumor burden

Over 80% of women diagnosed with advanced-stage ovarian cancer die as a result of disease recurrence due to failure of chemotherapy treatment. In this study, using two distinct ovarian cancer cell lines (epithelial OVCA 433 and mesenchymal HEY) we demonstrate enrichment in a population of cells with high expression of CSC markers at the protein and mRNA levels in response to cisplatin, paclitaxel and the combination of both. We also demonstrate a significant enhancement in the sphere forming abilities of ovarian cancer cells in response to chemotherapy drugs. The results of these in vitro findings are supported by in vivo mouse xenograft models in which intraperitoneal transplantation of cisplatin or paclitaxel-treated residual HEY cells generated significantly higher tumor burden compared to control untreated cells. Both the treated and untreated cells infiltrated the organs of the abdominal cavity. In addition, immunohistochemical studies on mouse tumors injected with cisplatin or paclitaxel treated residual cells displayed higher staining for the proliferative antigen Ki67, oncogeneic CA125, epithelial E-cadherin as well as cancer stem cell markers such as Oct4 and CD117, compared to mice injected with control untreated cells. These results suggest that a short-term single treatment of chemotherapy leaves residual cells that are enriched in CSC-like traits, resulting in an increased metastatic potential. The novel findings in this study are important in understanding the early molecular mechanisms by which chemoresistance and subsequent relapse may be triggered after the first line of chemotherapy treatment.

A copia-like element in Pisum demonstrates the uses of dispersed repeated sequences in genetic analysis

A DNA sequence between two legumin genes in Pisum is a member of the copia-like class of retrotransposons and represents one member of a polymorphic and heterogeneous dispersed repeated sequence family in Pisum. This sequence can be exploited in genetic studies either by RFLP analysis where several markers can be scored together, or the segregation of individual elements can be followed after PCR amplification of specific members.

Conservation and divergence of four kiwifruit SVP-like MADS-box genes suggest distinct roles in kiwifruit bud dormancy and flowering

MADS-box genes similar to Arabidopsis SHORT VEGETATIVE PHASE (SVP) have been implicated in the regulation of flowering in annual species and bud dormancy in perennial species. Kiwifruit (Actinidia spp.) are woody perennial vines where bud dormancy and out-growth affect flower development. To determine the role of SVP-like genes in dormancy and flowering of kiwifruit, four MADS-box genes with homology to Arabidopsis SVP, designated SVP1, SVP2, SVP3, and SVP4, have been identified and analysed in kiwifruit and functionally characterized in Arabidopsis. Phylogenetic analysis indicate that these genes fall into different sub-clades within the SVP-like gene group, suggesting distinct functions. Expression was generally confined to vegetative tissues, and increased transcript accumulation in shoot buds over the winter period suggests a role for these genes in bud dormancy. Down-regulation before flower differentiation indicate possible roles as floral repressors. Over-expression and complementation studies in Arabidopsis resulted in a range of floral reversion phenotypes arising from interactions with Arabidopsis MADS-box proteins, but only SVP1 and SVP3 were able to complement the svp mutant. These results suggest that the kiwifruit SVP-like genes may have distinct roles during bud dormancy and flowering.

Chemotaxis and chemokinesis of human prostate tumor cell lines in response to human prostate stromal cell secretory proteins containing a nerve growth factor-like protein

The migration of three human prostate tumor epithelial cell lines (TSU-pr1, PC-3, DU-145) in response to secreted protein from a human prostate stromal cell line was investigated by using the modified blind-well Boyden chamber assay. Migrated cells were quantified by spectrophotometrically measuring the concentration of crystal violet stain extracted from their nuclei. Cell number was correlated linearly with the concentration of extracted crystal violet stain. All three tumor cell lines showed intrinsic migratory ability in the absence of chemoattractants, such that approximately 1-7% of plated cells migrated across the filter of the Boyden chambers during a 5-h incubation period. Prostate tumor cell migration was significantly enhanced (3-13-fold) in response to stromal cell secretory protein in a dose-dependent manner, whereas bovine serum albumin had no effect on stimulating tumor cell migration. Immunoprecipitation of the stromal cell secreted protein with a nerve growth factor antibody partially and significantly reduced its stimulatory activity for tumor cell migration. A Zigmond-Hirsch matrix assay of tumor cell migration in response to various concentration gradients of stromal cell secreted protein demonstrated both chemotaxis and chemokinesis by all three cell lines. These results are consistent with the stromal cell secretory protein stimulation of chemokinetic tumor cell migration through the capsule of the prostate. Outside of the prostate gland metastasis of tumor cells may occur by chemotaxis to preferential sites containing chemoattractants similar to or related to maintenance factors that can substitute for components of stromal cell secretory protein.

Bimolecular interaction of Insulin-Like Growth Factor (IGF) binding protein-2 with αvβ3 negatively modulates IGF-I-Mediated migration and tumor growth

Both the integrin and insulin-like growth factor binding protein (IGFBP) families independently play important roles in modulating tumor cell growth and progression. We present evidence for a specific cell surface localization and a bimolecular interaction between the αvβ3 integrin and IGFBP-2. The interaction, which could be specifically perturbed using vitronectin and αvβ3 blocking antibodies, was shown to modulate IGF-mediated cellular migration responses. Moreover, this interaction was observed in vivo and correlated with reduced tumor size of the human breast cancer cells, MCF-7β3, which overexpressed the αvβ3 integrin. Collectively, these results indicate that αvβ3 and IGFBP-2 act cooperatively in a negative regulatory manner to reduce tumor growth and the migratory potential of breast cancer cells.

Art of sleeping - "Like a Thief"

Research background Art of Sleeping’s “Like a Thief” is an EP of popular music produced at Airlock studios and released in 2012 by the record label Dew Process. “Like a Thief” contributes to studies in the field of music production, as well as furthering research into the role of of national radio play in the development of emerging musicians and artists. Using Australian bands such as Boy and Bear as a reference point, the EP showcases the vocal range of Caleb Hodges and investigates possibilities in extending the modes of musicality possible within the recording studio through use of multiple layering of processed keyboards and electric guitar. Scholars such as Ben Eltham and journalists such as Everett True have written about the importance of the support of Australia’s national youth broadcaster Triple J in the early stages of a band’s development. The approach of the production of the EP was conceived with this agenda in mind, and asked, what can we learn about the relationships between popular music production, distribution and consumption, and the role that national public radio has come to play in the Australian context? Research contribution This project has identified that certain avenues of the music industry continue to aligned with Triple J airplay, that these avenues play an important role in the level of success that artists are likely to achieve, and that targeted understanding of music production techniques and aesthetics may contribute to success in this regard. Research significance The EP’s two singles were both in the top 100 most frequently played tracks on Triple J in 2012, and the band were able to leverage this exposure in securing a record deal with Dew Process, along with being selected as part of the competitive peer review process for festival performances such as Splendour in the Grass. The work received positive peer reviews, with indieshuffle writing: “Brisbane's Art of Sleeping are a truly talented band. … the five-piece gained plenty of local attention earlier this year with their stunning single "Empty Hands.” This was the first single off their new EP, Like a Thief, which was produced by the renowned Yanto Browning (The Medics, The Jungle Giants) … The EP includes five tracks of incredible power and sentiment … Title track "Like a Thief” is … [a] standout, again a vast and expansive song which showcases the band's completeness”

Cisplatin treatment of primary and metastatic epithelial ovarian carcinomas generates residual cells with mesenchymal stem cell-like profile

Epithelial mesenchymal transition (EMT) and cancer stem cells (CSC) have been associated with resistance to chemotherapy. Eighty percent of ovarian cancer patients initially respond to platinum-based combination therapy but most return with recurrence and ultimate demise. To better understand such chemoresistance we have assessed the potential role of EMT in tumor cells collected from advanced-stage ovarian cancer patients and the ovarian cancer cell line OVCA 433 in response to cisplatin in vitro. We demonstrate that cisplatin-induced transition from epithelial to mesenchymal morphology in residual cancer cells correlated with reduced E-cadherin, and increased N-cadherin and vimentin expression. The mRNA expression of Snail, Slug, Twist, and MMP-2 were significantly enhanced in response to cisplatin and correlated with increased migration. This coincided with increased cell surface expression of CSC-like markers such as CD44, α2 integrin subunit, CD117, CD133, EpCAM, and the expression of stem cell factors Nanog and Oct-4. EMT and CSC-like changes in response to cisplatin correlated with enhanced activation of extracellular signal-regulated kinase (ERK)1/2. The selective MEK inhibitor U0126 inhibited ERK2 activation and partially suppressed cisplatin-induced EMT and CSC markers. In vivo xenotransplantation of cisplatin-treated OVCA 433 cells in zebrafish embryos demonstrated significantly enhanced migration of cells compared to control untreated cells. U0126 inhibited cisplatin-induced migration of cells in vivo, suggesting that ERK2 signaling is critical to cisplatin-induced EMT and CSC phenotypes, and that targeting ERK2 in the presence of cisplatin may reduce the burden of residual tumor, the ultimate cause of recurrence in ovarian cancer patients.

Reversible transdifferentiation of blood vascular endothelial cells to a lymphatic-like phenotype in vitro

Blood vascular cells and lymphatic endothelial cells (BECs and LECs, respectively) form two separate vascular systems and are functionally distinct cell types or lineages with characteristic gene expression profiles. Interconversion between these cell types has not been reported. Here, we show that in conventional in vitro angiogenesis assays, human BECs of fetal or adult origin show altered gene expression that is indicative of transition to a lymphatic-like phenotype. This change occurs in BECs undergoing tubulogenesis in fibrin, collagen or Matrigel assays, but is independent of tube formation per se, because it is not inhibited by a metalloproteinase inhibitor that blocks tubulogenesis. It is also reversible, since cells removed from 3D tubules revert to a BEC expression profile upon monolayer culture. Induction of the lymphatic-like phenotype is partially inhibited by co-culture of HUVECs with perivascular cells. These data reveal an unexpected plasticity in endothelial phenotype, which is regulated by contact with the ECM environment and/or cues from supporting cells.

Feather-like He plasma plumes in surrounding N2 gas

Effects of surrounding gases on the propagation of room-temperature atmospheric-pressure plasma jets are reported. A highly unusual feather-like plasma plume is observed only when N2 is used as surrounding gas. The He concentration on the axis at the starting point of the feather-like plume is ∼0.85 of the maximum value and is independent on the He flow rates. High-speed optical imaging reveals that dim diffuse plasmas emerge just behind the bright head of the plasma bullet at the starting point of the feather-like plume. These results help tailoring surface exposure in emerging applications of plasma jets in medicine and nanotechnology.

Carbon nanorods and graphene-like nanosheets by hot filament CVD : growth mechanisms and electron field emission

Carbon nanorods and graphene-like nanosheets are catalytically synthesized in a hot filament chemical vapor deposition system with and without plasma enhancement, with gold used as a catalyst. The morphological and structural properties of the carbon nanorods and nanosheets are investigated by field-emission scanning electron microscopy, transmission electron microscopy and micro-Raman spectroscopy. It is found that carbon nanorods are formed when a CH4 + H2 + N2 plasma is present while carbon nanosheets are formed in a methane environment without a plasma. The formation of carbon nanorods and carbon nanosheets are analyzed. The results suggest that the formation of carbon nanorods is primarily a precipitation process while the formation of carbon nanosheets is a complex process involving surface-catalysis, surface diffusion and precipitation influenced by the Gibbs–Thomson effect. The electron field emission properties of the carbon nanorods and graphene-like nanosheets are measured under high-vacuum; it is found that the carbon nanosheets have a lower field emission turn-on than the carbon nanorods. These results are important to improve the understanding of formation mechanisms of carbon nanomaterials and contribute to eventual applications of these structures in nanodevices.

Core-leaf onion-like carbon/MnO2 hybrid nano-urchins for rechargeable lithium-ion batteries

A hybrid nano-urchin structure consisting of spherical onion-like carbon and MnO2 nanosheets is synthesized by a facile and environmentally-friendly hydrothermal method. Lithium-ion batteries incorporating the hybrid nano-urchin anode exhibit reversible lithium storage with superior specific capacity, enhanced rate capability, stable cycling performance, and nearly 100% Coulombic efficiency. These results demonstrate the effectiveness of designing hybrid nano-architectures with uniform and isotropic structure, high loading of electrochemically-active materials, and good conductivity for the dramatic improvement of lithium storage.

Inhibition of the JAK2/STAT3 pathway in ovarian cancer results in the loss of cancer stem cell-like characteristics and a reduced tumor burden

Background Current treatment of ovarian cancer patients with chemotherapy leaves behind a residual tumor which results in recurrent ovarian cancer within a short time frame. We have previously demonstrated that a single short-term treatment of ovarian cancer cells with chemotherapy in vitro resulted in a cancer stem cell (CSC)-like enriched residual population which generated significantly greater tumor burden compared to the tumor burden generated by control untreated cells. In this report we looked at the mechanisms of the enrichment of CSC-like residual cells in response to paclitaxel treatment. Methods The mechanism of survival of paclitaxel-treated residual cells at a growth inhibitory concentration of 50% (GI50) was determined on isolated tumor cells from the ascites of recurrent ovarian cancer patients and HEY ovarian cancer cell line by in vitro assays and in a mouse xenograft model. Results Treatment of isolated tumor cells from the ascites of ovarian cancer patients and HEY ovarian cancer cell line with paclitaxel resulted in a CSC-like residual population which coincided with the activation of Janus activated kinase 2 (JAK2) and signal transducer and activation of transcription 3 (STAT3) pathway in paclitaxel surviving cells. Both paclitaxel-induced JAK2/STAT3 activation and CSC-like characteristics were inhibited by a low dose JAK2-specific small molecule inhibitor CYT387 (1 μM) in vitro. Subsequent, in vivo transplantation of paclitaxel and CYT387-treated HEY cells in mice resulted in a significantly reduced tumor burden compared to that seen with paclitaxel only-treated transplanted cells. In vitro analysis of tumor xenografts at protein and mRNA levels demonstrated a loss of CSC-like markers and CA125 expression in paclitaxel and CYT387-treated cell-derived xenografts, compared to paclitaxel only-treated cell-derived xenografts. These results were consistent with significantly reduced activation of JAK2 and STAT3 in paclitaxel and CYT387-treated cell-derived xenografts compared to paclitaxel only-treated cell derived xenografts. Conclusions This proof of principle study demonstrates that inhibition of the JAK2/STAT3 pathway by the addition of CYT387 suppresses the ‘stemness’ profile in chemotherapy-treated residual cells in vitro, which is replicated in vivo, leading to a reduced tumor burden. These findings have important implications for ovarian cancer patients who are treated with taxane and/or platinum-based therapies. Keywords: Ovarian carcinoma, Cancer stem cell, Metastasis, Ascites, Chemoresistance, Recurrence, JAK2/STAT3 pathway

Unconventional Ni–P alloy-catalyzed CVD of carbon coil-like micro- and nano-structures

Conventional catalyzed thermal CVD of carbon microcoils commonly suffers from poor control of the coil shape and morphology and rarely reaches the nanoscale size range. This article reports on an unconventional Ni-P alloy-catalyzed, high-throughput, highly reproducible CVD of ultra-long carbon coil-like micro- and nano-structures using acetylene precursor at relatively low process temperatures. Helical carbon microcoils with consistently uniform, circular cross-sections and a high degree of crystallinity have been synthesized at 750 °C. A further reduction of the temperature to 650 °C led to the growth of ultra-long (up to several mm) wave-like carbon nanofibers made of two nanowires with the diameters in the 100-200 nm range. The results of the XRD and Raman analysis reveal that the nanofibers feature only a slightly more disordered structure compared to the microcoils. Our results suggest that morphology and structure of the carbon coil-like micro- and nano-structures can be tailored by the appropriate alloying of the catalyst and the choice of the CVD process parameters.

Growth kinetics of carbon nanowall-like structures in low-temperature plasmas

The results of a hybrid numerical simulation of the growth kinetics of carbon nanowall-like nanostructures in the plasma and neutral gas synthesis processes are presented. The low-temperature plasma-based process was found to have a significant advantage over the purely neutral flux deposition in providing the uniform size distribution of the nanostructures. It is shown that the nanowall width uniformity is the best (square deviations not exceeding 1.05) in high-density plasmas of 3.0× 1018 m-3, worsens in lower-density plasmas (up to 1.5 in 1.0× 1017 m-3 plasmas), and is the worst (up to 1.9) in the neutral gas-based process. This effect has been attributed to the focusing of ion fluxes by irregular electric field in the vicinity of plasma-grown nanostructures on substrate biased with -20 V potential, and differences in the two-dimensional adatom diffusion fluxes in the plasma and neutral gas-based processes. The results of our numerical simulations are consistent with the available experimental reports on the effect of the plasma process parameters on the sizes and shapes of relevant nanostructures.

E and H regimes of plasma enhanced chemical vapor deposition of diamond-like carbon film in low frequency inductively coupled plasma reactor

This paper reports on the efficient deposition of hydrogenated diamond-like carbon (DLC) film in a plasma reactor that features both the capacitively and inductively coupled operation regimes. The hydrogenated DLC films have been prepared on silicon wafers using a low-frequency (500 kHz) inductively coupled plasma (ICP) chemical vapor deposition (CVD) system. At low RF powers, the system operates as an asymmetric capacitively coupled plasma source, and the film deposition process is undertaken in the electrostatic (E) discharge regime. Above the mode transition threshold, the high-density inductively coupled plasma is produced in the electromagnetic (H) discharge regime. It has been shown that the deposition rate and hardness of the DLC film are much higher in the H-mode deposition regime. For a 2.66-Pa H-mode CH4 + Ar gas mixture discharge, the deposited DLC film exhibits a mechanical hardness of 18 GPa, Young's modulus of 170 GPa, and compressive stress of 1.3 GPa.