Developmental gene expression in the eyes of the pygmy squid Xipholeptos notoides.

The eyes of squids, octopuses, and cuttlefish are a textbook example for evolutionary convergence, due to their striking similarity to those of vertebrates. For this reason, studies on cephalopod photoreception and vision are of importance for a broader audience. Previous studies showed that genes such as pax6, or certain opsin-encoding genes, are evolutionarily highly conserved and play similar roles during ontogenesis in remotely related bilaterians. In this study, genes that encode photosensitive proteins and Reflectins are identified and characterized. The expression patterns of rhodopsin, xenopsin, retinochrome, and two reflectin genes have been visualized in developing embryos of the pygmy squid Xipholeptos notoides by in situ hybridization experiments. Rhodopsin is not only expressed in the retina of X. notoides but also in the olfactory organ and the dorsal parolfactory vesicles, the latter a cephalopod apomorphy. Both reflectin genes are expressed in the eyes and in the olfactory organ. These findings corroborate previous studies that found opsin genes in the transcriptomes of the eyes and several extraocular tissues of various cephalopods. Expression of rhodopsin, xenopsin, retinochrome, and the two reflectin genes in the olfactory organ is a finding that has not been described so far. In other organisms, it has been shown that Retinochrome and Rhodopsin proteins are obligatorily associated with each other as both molecules rely on each other for Retinal isomerisation. In addition, we demonstrate that retinochrome is expressed in the retina of X. notoides and in the olfactory organ. This study shows numerous new expression patterns for Opsin-encoding genes in organs that have not been associated with photoreception before, suggesting that either Opsins may not only be involved in photoreception or organs such as the olfactory organ are involved in photoreception.

Autophagy inhibition overcomes sorafenib resistance in S45F-mutated desmoid tumors.

BACKGROUND: Desmoid tumors (DTs) are rare and understudied fibroblastic lesions that are frequently recurrent and locally invasive. DT patients often experience chronic pain, organ dysfunction, decrease in quality of life, and even death. METHODS: Sorafenib has emerged as a promising therapeutic strategy, which has led to the first randomized phase 3 clinical trial devoted to DTs. Concurrently, we conducted a comprehensive analysis of sorafenib efficacy in a large panel of desmoid cell strains to probe for response mechanism. RESULTS: We found distinctive groups of higher- and lower-responder cells. Clustering the lower-responder group, we observed that CTNNB1 mutation was determinant of outcome. Our results revealed that a lower dose of sorafenib was able to inhibit cell viability, migration, and invasion of wild-type and T41A-mutated DTs. Apoptosis induction was observed in those cells after treatment with sorafenib. On the other hand, the lower dose of sorafenib was not able to inhibit cell viability, migration, or invasion or to induce apoptosis in the S45F-mutated DTs. The investigation of autophagy showed the dependency of S45F-mutated DTs on this pathway as a part of cell survival mechanism. Significantly, when autophagy was inhibited genetically or pharmacologically in the S45F mutant cell strains, sensitivity to sorafenib was restored. CONCLUSIONS: Our findings suggest that the response to sorafenib differs when comparing S45F-mutated DTs and T41A-mutated or wild-type DTs. Furthermore, the combination of hydroxychloroquine and sorafenib enhances the antiproliferative and proapoptotic effects in S45F-mutated DT cells, suggesting that profiling ß-catenin status could guide clinical management of desmoid patients who are considering sorafenib treatment.

miR-133a function in the pathogenesis of dedifferentiated liposarcoma.

BACKGROUND: Sarcomas are malignant heterogeneous tumors of mesenchymal derivation. Dedifferentiated liposarcoma (DDLPS) is aggressive with recurrence in 80% and metastasis in 20% of patients. We previously found that miR-133a was significantly underexpressed in liposarcoma tissues. As this miRNA has recently been shown to be a tumor suppressor in many cancers, the objective of this study was to characterize the biological and molecular consequences of miR-133a underexpression in DDLPS. METHODS: Real-time PCR was used to evaluate expression levels of miR-133a in human DDLPS tissue, normal fat tissue, and human DDLPS cell lines. DDLPS cells were stably transduced with miR-133a vector to assess the effects in vitro on proliferation, cell cycle, cell death, migration, and metabolism. A Seahorse Bioanalyzer system was also used to assess metabolism in vivo by measuring glycolysis and oxidative phosphorylation (OXPHOS) in subcutaneous xenograft tumors from immunocompromised mice. RESULTS: miR-133a expression was significantly decreased in human DDLPS tissue and cell lines. Enforced expression of miR-133a decreased cell proliferation, impacted cell cycle progression kinetics, decreased glycolysis, and increased OXPHOS. There was no significant effect on cell death or migration. Using an in vivo xenograft mouse study, we showed that tumors with increased miR-133a expression had no difference in tumor growth compared to control, but did exhibit an increase in OXPHOS metabolic respiration. CONCLUSIONS: Based on our collective findings, we propose that in DDPLS, loss of miR-133a induces a metabolic shift due to a reduction in oxidative metabolism favoring a Warburg effect in DDLPS tumors, but this regulation on metabolism was not sufficient to affect DDPLS.

Photoluminescence quenching in compressed MgO nanoparticle systems.

Efficient use of highly dispersed metal oxides for lighting, energy conversion and catalysis requires knowledge about the impact of density and microstructure of the powders on the optical nanoparticle properties. For MgO nanocube powders we present a combined photoluminescence (PL) and electron paramagnetic resonance (EPR) approach which enables for samples of different aggregation states the quantification of the fractional powder volume that becomes illuminated with UV and visible light during the PL measurements. Using O2 as a PL emission quencher and - after light induced exciton separation and oxygen adsorption - as an EPR active adsorbate we observed clear aggregation dependent trends in PL emission quenching that originate from particle-particle contacts. Upon interaction of low coordinated surface elements with the surfaces of adjacent MgO nanocubes, which occurs even at powder consolidation levels that escape sorption analysis, the radiative decay of excited surface states becomes quenched down to 15% of the original intensity. Our results underline the critical role of microstructure and the aggregation state of a nanoparticle ensemble with respect to spectroscopic properties and related adsorption induced changes.

[Day care clinics central and/or distributed. A standard element of child and adolescent psychiatric care]. / Tageskliniken zentral und/oder disloziert. Ein Standardelement der kinder- und jugendpsychiatrischen Versorgung.

BACKGROUND: Day-care clinics as specialized care units of child and adolescent psychiatric care in Austria represent an important component for person-orientated treatment offers. In addition to numerous advantages, they also put higher demands on the young patients and their relatives. METHODS: The Austrian structure for day-care settings is recorded in the Austrian structure plan for health. Therefore, particular attention is paid to structural quality criteria, which are based on the Austrian concept of performance-oriented hospital financing. RESULTS: A high demand and need for readiness for transdisciplinary, multimodal treatment concepts and forms is discussed. It becomes ovious that there is a need for large spatial resource requirements. The establishment of several day clinic groups at one location and spatial requirements cannot be found in the concept of performance-oriented hospital financing. CONCLUSION: To ensure and evaluate the quality of treatment, recommendations are made on a patient-related and team- or organization-related level. The workgroup "day-care-clinic" of the Austrian Society for Child and Adolescent Psychiatry, Psychosomatics and Psychotherapy recommends a treatment structure characterized by transdisciplinarity and participation. Further, it regionalized and local access and the integration of the day clinic into the community is necessary. Networking with other care structures and using social-psychiatric networks is essential. Regional characteristics should be taken into account and specialized, topic-specific day clinic groups should be increasingly included in further planning and concepts.

Preclinical efficacy of the Wnt/ß-catenin pathway inhibitor BC2059 for the treatment of desmoid tumors.

Mutation in the CTNNB1 gene, leading to a deregulation of the WTN/ß-catenin pathway, is a common feature of desmoid tumors (DTs). Many ß-catenin inhibitors have recently been tested in clinical studies; however, BC2059 (also referred as Tegavivint), a selective inhibitor of nuclear ß-catenin that works through binding TBL-1, is the only one being evaluated in a clinical study, specifically for treatment of desmoid tumor patients. Preclinical studies on BC2059 have shown activity in multiple myeloma, acute myeloid leukemia and osteosarcoma. Our preclinical studies provide data on the efficacy of BC2059 in desmoid cell lines, which could help provide insight regarding antitumor activity of this therapy in desmoid tumor patients. In vitro activity of BC2059 was evaluated using desmoid tumor cell lines. Ex vivo activity of BC2059 was assessed using an explant tissue culture model. Pharmacological inhibition of the nuclear ß-catenin activity using BC2059 markedly inhibited cell viability, migration and invasion of mutated DT cells, but with lower effect on wild-type DTs. The decrease in cell viability of mutated DT cells caused by BC2059 was due to apoptosis. Treatment with BC2059 led to a reduction of ß-catenin-associated TBL1 in all mutated DT cells, resulting in a reduction of nuclear ß-catenin. mRNA and protein levels of AXIN2, a ß-catenin target gene, were also found to be downregulated after BC2059 treatment. Taken together, our results demonstrate that nuclear ß-catenin inhibition using BC2059 may be a novel therapeutic strategy for desmoid tumor treatment, especially in patients with CTNNB1 mutation.

ß-catenin S45F mutation results in apoptotic resistance.

Wnt/ß-catenin signaling is one of the key cascades regulating embryogenesis and tissue homeostasis; it has also been intimately associated with carcinogenesis. This pathway is deregulated in several tumors, including colorectal cancer, breast cancer, and desmoid tumors. It has been shown that CTNNB1 exon 3 mutations are associated with an aggressive phenotype in several of these tumor types and may be associated with therapeutic tolerance. Desmoid tumors typically have a stable genome with ß-catenin mutations as a main feature, making these tumors an ideal model to study the changes associated with different types of ß-catenin mutations. Here, we show that the apoptosis mechanism is deregulated in ß-catenin S45F mutants, resulting in decreased induction of apoptosis in these cells. Our findings also demonstrate that RUNX3 plays a pivotal role in the inhibition of apoptosis found in the ß-catenin S45F mutants. Restoration of RUNX3 overcomes this inhibition in the S45F mutants, highlighting it as a potential therapeutic target for malignancies harboring this specific CTNNB1 mutation. While the regulatory effect of RUNX3 in ß-catenin is already known, our results suggest the possibility of a feedback loop involving these two genes, with the CTNNB1 S45F mutation downregulating expression of RUNX3, thus providing additional possible novel therapeutic targets for tumors having deregulated Wnt/ß-catenin signaling induced by this mutation.

Mocetinostat combined with gemcitabine for the treatment of leiomyosarcoma: Preclinical correlates.

Leiomyosarcoma (LMS) is a malignant soft tissue sarcoma (STS) with a dismal prognosis following metastatic disease. Chemotherapeutic intervention has demonstrated to have modest clinical efficacy with no curative potential in LMS patients. Previously, we demonstrated pan-HDAC inhibition to have a superior effect in various complex karyotypic sarcomas. In this study, our goal is to evaluate the therapeutic efficacy of mocetinostat alone and in combination with gemcitabine in LMS. Human leiomyosarcoma (LMS) cell lines were used for in vitro and in vivo studies. Compounds tested included the class I HDAC inhibitor, mocetinostat, and nucleoside analog, gemcitabine. MTS and clonogenic assays were used to evaluate the effect of mocetinostat on LMS cell growth. Cleaved caspase 3/7 analysis was used to determine the effects of mocetinostat on apoptosis. Compusyn software was used to determine in vitro synergy studies for the combination of mocetinostat plus gemcitabine. A LMS xenograft model in SCID mice was used to test the impact of mocetinostat alone, gemcitabine alone and the combination of mocetinostat plus gemcitabine. Mocetinostat abrogated LMS cell growth and clonogenic potential, and enhanced apoptosis in LMS cell lines. The combination of mocetinostat plus gemcitabine exhibited a synergistic effect in LMS cells in vitro. Similarly, mocetinostat combined with gemcitabine resulted in superior anti-LMS effects in vivo. Mocetinostat reduced the expression of gemcitabine-resistance markers RRM1, RRM2, and increased the expression of gemcitabine-sensitivity marker, hENT1, in LMS cells. LMS are aggressive, metastatic tumors with poor prognosis where effective therapeutic interventions are wanting. Our studies demonstrate the potential utility of mocetinostat combined with gemcitabine for the treatment of LMS.

Hybrid functionals for solids with an optimized Hartree-Fock mixing parameter.

(Screened) hybrid functionals are being used more and more for solid-state calculations. Usually the fraction α of Hartree-Fock exchange is kept fixed during the calculation; however, there is no single (universal) value for α which systematically leads to satisfying accuracy. Instead, one could use a property of the system under consideration to determine α, and in this way the functional would be more flexible and potentially more accurate. Recently, it was proposed to use the static dielectric constant ε for the calculation of α (Shimazaki and Asai 2008 Chem. Phys. Lett. 466 91 and Marques et al 2011 Phys. Rev. B 83 035119). We explore this idea further and propose a scheme where the connection between ε and α is optimized based on experimental band gaps. ε, and thus α, is recalculated at each iteration of the self-consistent procedure. We present results for the bandgap and lattice constant of various semiconductors and insulators with this procedure. In addition, we show that this approach can also be combined with a non-self-consistent hybrid approximation to speed up the calculations considerably, while retaining an excellent accuracy in most cases.

Optical properties of nanocrystal interfaces in compressed MgO nanopowders.

The optical properties and charge trapping phenomena observed on oxide nanocrystal ensembles can be strongly influenced by the presence of nanocrystal interfaces. MgO powders represent a convenient system to study these effects due to the well-defined shape and controllable size distributions of MgO nanocrystals. The spectroscopic properties of nanocrystal interfaces are investigated by monitoring the dependence of absorption characteristics on the concentration of the interfaces in the nanopowders. The presence of interfaces is found to affect the absorption spectra of nanopowders more significantly than changing the size of the constituent nanocrystals and, thus, leading to the variation of the relative abundance of light-absorbing surface structures. We find a strong absorption band in the 4.0-5.5 eV energy range, which was previously attributed to surface features of individual nanocrystals, such as corners and edges. These findings are supported by complementary first-principles calculations. The possibility to directly address such interfaces by tuning the energy of excitation may provide new means for functionalization and chemical activation of nanostructures and can help improve performance and reliability for many nanopowder applications.

Single-chamber ventricular pacing increases markers of left ventricular dysfunction compared with dual-chamber pacing.

AIMS: Large randomized trials comparing DDD with VVI pacing have shown no differences in mortality, but conflicting evidence exists in regard to heart failure endpoints. Here we evaluated the effect of pacing mode on serum levels of brain natriuretic peptide (BNP) and amino-terminal-proBNP (NT-proBNP). Methods Forty-one patients (age 73 +/- 10 years) with dual-chamber pacemakers were included in a prospective, single-blind, randomized crossover study evaluating the impact of DDD(R)/VDD versus VVI(R) mode on objective and functional parameters. Data were collected after a 2-week run-in phase and after 2 weeks each of VVI(R) and DDD(R)/VDD pacing or vice versa. Results BNP and NT-proBNP levels during DDD(R)/VDD stimulation (151 +/- 131 and 547 +/- 598 pg/mL) showed no change compared with baseline (154 +/- 130 and 565 +/- 555 pg/mL), but a significant 2.4-fold increase was observed during VVI(R) mode [360 +/- 221 and 1298 +/- 1032 pg/mL; P < 0.001 compared with DDD(R)/VDD]. The assessment of functional class, the presence of pacemaker syndrome [49% in VVI(R) mode] and the patients' preferred pacing mode showed significant differences in favour of DDD(R)/VDD pacing. CONCLUSION: Patients can differentiate between DDD(R)/VDD and VVI(R) pacing, and prefer the former. Compared with DDD(R)/VDD pacing, VVI(R) stimulation induces a two- to three-fold increase in serum BNP and NT-proBNP levels.

Respiratory dependent compression of a venous bypass: therapy by stenting.

Although coronary artery bypass surgery has provided major advances in the treatment of coronary artery disease, narrowing of bypass vessels still constitutes a drawback of this therapy. Although this event is most frequently caused by intraluminal processes, obstruction from external structures is extremely rare. We report such a case in which external bypass compression was provoked by deep inspiration causing typical anginal symptoms. Percutaneous coronary intervention including stent placement provided bypass patency independent from the patient's respiratory phase. Disappearance of symptoms and absence of myocardial ischemia in perfusion scans confirmed successful treatment.