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We provide a general overview of spatial distribution and the economic situation of aquaculture enterprises in Czechia (the Czech Republic) in terms of revenues, profitability, work productivity, indebtedness and grants. We analysed data available in the State Veterinary Administration and information system of the State Agricultural Intervention Fund. The most significant fish producers (enterprises) are located in the regions of South Bohemia. Aquaculture sector profitability and enterprises profits have been moderately rising since 2010. The profitability in 2015 is roughly the same as that in 2005. The amount of provided grants showed a higher value than the generated average annual profits. The enterprises were not able to generate sufficient resources to finance investments without grants resources. Enterprises that realised financing for larger investments from bank credits showed relatively high levels of indebtedness (micro-enterprises at 95.5%). On the contrary, work productivity was the highest for micro-enterprises. Micro-enterprises had the largest share of revenues in relation to the number of employees. Each group of enterprises is considerably specific and needs an individual approach for the subsequent evaluation.

The seasonality of fish marketing is a very important character for aquaculture production. We analysed data regarding the situation at the Czech market in 2015 and 2016 available in the information system of the State Agricultural Intervention Fund. The most significant volumes of freshwater fish were traded by traditional pond aquaculture companies (TPA) and approx. 1/3 of the all‑year production was sold at the end of the year (December). Another significant time when the supply of live fish from the TPA on the domestic market slightly increased was in March and then in April, i.e. before Easter, which represented approximately one fifth of all aquaculture production. The weakest periods in terms of marketing fish of TPA were the beginning of the year, i.e. during the first two months (January and February) and the period from May to September. On the contrary, the situation was different in intensive fish aquaculture companies (IA) which had the most significant volumes traded between April and September and then in November and December. However, the sold volume of fish from IA was not strictly concentrated in a single period.

PRIMA Trial Investigators and the PATH Study Group., Intravenous immunoglobulin (IVIG) is a potential therapy for chronic inflammatory demyelinating polyneuropathy (CIDP). To investigate the efficacy and safety of the IVIG IgPro10 (Privigen) for treatment of CIDP, results from Privigen Impact on Mobility and Autonomy (PRIMA), a prospective, open‐label, single‐arm study of IVIG in immunoglobulin (Ig)‐naïve or IVIG pre‐treated subjects (NCT01184846, n = 28) and Polyneuropathy And Treatment with Hizentra (PATH), a double‐blind, randomized study including an open‐label, single‐arm IVIG phase in IVIG pre‐treated subjects (NCT01545076, IVIG restabilization phase n = 207) were analyzed separately and together (n = 235). Efficacy assessments included change in adjusted inflammatory neuropathy cause and treatment (INCAT) score, grip strength and Medical Research Council (MRC) sum score. Adverse drug reactions (ADRs) and ADRs/infusion were recorded. Adjusted INCAT response rate was 60.7% in all PRIMA subjects at Week 25 (76.9% in IVIG pre‐treated subjects) and 72.9% in PATH. In the pooled cohort (n = 235), INCAT response rate was 71.5%; median time to INCAT improvement was 4.3 weeks. No clear demographic differences were noticed between early (responding before Week 7, n = 148) and late responders (n = 21). In the pooled cohort, median change from baseline to last observation was −1.0 (interquartile range −2.0; 0.0) point for INCAT score; +8.0 (0.0; 20.0) kPa for maximum grip strength; +3.0 (1.0; 7.0) points for MRC sum score. In the pooled cohort, 271 ADRs were reported in 105 subjects (44.7%), a rate of 0.144 ADRs per infusion. This analysis confirms the efficacy and safety of IgPro10, a recently FDA‐approved IVIG for CIDP, in a population of mainly pre‐treated subjects with CIDP [Correction added on 14 March 2019 after first online publication: the INCAT response rate has been corrected.].

International audience; Although invasive non-native species have been studied extensively, their monitoring and management are often inadequate (Pergl et al. 2020). Moreover, the great harm invasive non-native species cause tends to be

The Polyneuropathy And Treatment with Hizentra (PATH) study required subjects with chronic inflammatory demyelinating polyneuropathy (CIDP) to show dependency on immunoglobulin G (IgG) and then be restabilized on IgG before being randomized to placebo or one of two doses of subcutaneous immunoglobulin (SCIG). Nineteen of the 51 subjects (37%) randomized to placebo did not relapse over the next 24 weeks. This article explores the reasons for this effect. A post‐hoc analysis of the PATH placebo group was undertaken. A literature search identified other placebo‐controlled CIDP trials for review and comparison. In PATH, subjects randomized to placebo who did not relapse were significantly older, had more severe disease, and took longer to deteriorate in the IgG dependency period compared with those who relapsed. Published trials in CIDP, whose primary endpoint was stability or deterioration, had a mean non‐deterioration (placebo effect) of 43%, while trials with a primary endpoint of improvement had a placebo response of only 11%. Placebo is an important variable in the design of CIDP trials. Trials designed to show clinical improvement will have a significantly lower effect of this phenomenon than those designed to show stability or deterioration.

Continuous enzyme kinetic assays are often used in high-throughput applications, as they allow rapid acquisition of large amounts of kinetic data and increased confidence compared to discontinuous assays. However, data analysis is often rate-limiting in high-throughput enzyme assays, as manual inspection and selection of a linear range from individual kinetic traces is cumbersome and prone to user error and bias. Currently available software programs are specialized and designed for the analysis of complex enzymatic models. Despite the widespread use of initial rate determination for processing kinetic data sets, no simple and automated program existed for rapid analysis of initial rates from continuous enzyme kinetic traces. An Interactive Continuous Enzyme Kinetics Analysis Tool (ICEKAT) was developed for semi-automated calculation of initial rates from continuous enzyme kinetic traces with particular application to the evaluation of Michaelis-Menten and EC50/IC50 kinetic parameters, as well as the results of high-throughput screening assays. ICEKAT allows users to interactively fit kinetic traces using convenient browser-based selection tools, ameliorating tedious steps involved in defining ranges to fit in general purpose programs like Microsoft Excel and Graphpad Prism, while still maintaining simplicity in determining initial rates. As a test case, we quickly analyzed over 500 continuous enzyme kinetic traces resulting from experimental data on the response of the protein lysine deacetylase SIRT1 to small-molecule activators. ICEKAT allows simultaneous visualization of individual initial rate fits and the resulting Michaelis-Menten or EC50/IC50 kinetic model fits, as well as hits from high-throughput screening assays. In addition to serving as a convenient program for practicing enzymologists, ICEKAT is also a useful teaching aid to visually demonstrate in real-time how incorrect initial rate fits can affect calculated Michaelis-Menten or EC50/IC50 kinetic parameters. For the convenience of the research community, we have made ICEKAT freely available online at https://icekat.herokuapp.com/icekat .

The cyclinD:CDK4/6:Rb axis is dysregulated in a variety of human cancers. Targeting this pathway has proven to be a successful therapeutic approach in ER+ breast cancer. In this study, in vitro and in vivo preclinical breast cancer models were used to investigate the expanded use of the CDK4/6 inhibitor, abemaciclib. Using a panel of 44 breast cancer cell lines, differential sensitivity to abemaciclib was observed and was seen predominately in the luminal ER+/HER2− and ER+/HER2+ subtypes. However, a subset of triple-negative breast cancer (TNBC) cell lines with intact Rb signaling were also found to be responsive. Equivalent levels of tumor growth inhibition were observed in ER+/HER2−, ER+/HER2+ as well as biomarker selected TNBC xenografts in response to abemaciclib. In addition, abemaciclib combined with hormonal blockade and/or HER2-targeted therapy induced significantly improved antitumor activity. CDK4/6 inhibition with abemaciclib combined with antimitotic agents, both in vitro and in vivo, did not antagonize the effect of either agent. Finally, we identified a set of Rb/E2F-regulated genes that consistently track with growth inhibitory response and constitute potential pharmacodynamic biomarkers of response to abemaciclib. Taken together, these data represent a comprehensive analysis of the preclinical activity of abemaciclib, used alone or in combination, in human breast cancer models. The subtypes most likely to respond to abemaciclib-based therapies can be identified by measurement of a specific set of biomarkers associated with increased dependency on cyclinD:CDK4/6:Rb signaling. These data support the clinical development of abemaciclib as monotherapy or as a combination partner in selected ER+/HER2−, HER2+/ER+, and TNBCs. Mol Cancer Ther; 17(5); 897–907. ©2018 AACR.

The pet trade has been responsible for many introductions of non-native species. Freshwater ornamental plants and animals originating from the pet trade are stocked to garden ponds. The present survey focused on awareness and responsible behaviour related to biological invasion risks of companies that designed, built, and stocked garden ponds. A representative number (n = 124) of companies (commercial garden pond architects and builders) in the Czech Republic were surveyed regarding the offer of non-native, invasive and threatened native species. The survey was conducted over the entire warm period (from 1 May 2015 to 31 September 2015) while using personal visits, correspondence and interviews to list the species on offer. ‘Traditional’ ornamental species were offered by 39.1% of surveyed vendors, non-traditional ‘marginal’ species by 5.6%, common native species by 6.5% and threatened native species by 2.4%. The findings of this study support the hypothesis that ‘garden pond’ vendors offer non-native species with a risk of biological invasion; moreover, threatened native animals were also on sale. It is probable that a similar situation exists in other countries. It is important for the conservation of native aquatic biota to raise awareness of the need to reduce risk through responsible behaviour of those involved in the pet trade. It is also essential to prohibit stocking of potential invaders and to enforce the illegal capture and sale of native species.

PATH study group., In patients with chronic inflammatory demyelinating polyneuropathy (CIDP), intravenous immunoglobulin (IVIG) is recommended to be periodically reduced to assess the need for ongoing therapy. However, little is known about the effectiveness of restabilization with IVIG in patients who worsen after IVIG withdrawal. In the Polyneuropathy And Treatment with Hizentra (PATH) study, the pre‐randomization period included sudden stopping of IVIG followed by 12 weeks of observation. Those deteriorating were then restabilized with IVIG. Of 245 subjects who stopped IVIG, 28 did not show signs of clinical deterioration within 12 weeks. Two hundred and seven received IVIG restabilization with an induction dose of 2 g/kg bodyweight (bw) IgPro10 (Privigen, CSL Behring, King of Prussia, Pennsylvania) and maintenance doses of 1 g/kg bw every 3 weeks for up to 13 weeks. Signs of clinical improvement were seen in almost all (n = 188; 91%) subjects. During IVIG restabilization, 35 subjects either did not show CIDP stability (n = 21, analyzed as n = 22 as an additional subject was randomized in error) or withdrew for other reasons (n = 14). Of the 22 subjects who did not achieve clinical stability, follow‐up information in 16 subjects after an additional 4 weeks was obtained. Nine subjects were reported to have improved, leaving a maximum of 27 subjects (13%) who either showed no signs of clinical improvement during the restabilization phase and 4 weeks post‐study or withdrew for other reasons. In conclusion, sudden IVIG withdrawal was effective in detecting ongoing immunoglobulin G dependency with a small risk for subjects not returning to their baseline 17 weeks after withdrawal., Editorial support was provided by Meridian HealthComms Ltd, funded by CSL Behring.

The South Caucasus (SC) region is recognized for its high biological diversity and various endemic animal taxa. The area has experienced many fish introductions over the years, but the overall information about non-native fishes in the three SC countries, Armenia, Azerbaijan, and Georgia did not exist. Although these three countries belong to the Kura River drainage, Caspian Sea basin (only the western half of Georgia drains into the Black Sea), the legislative framework for each country regarding introduction of non-native fish species and their treatment is different and poorly developed. The goal of the present study was to make an initial inventory of non-native fish species in the three SC countries, and summarize the existing knowledge as a basis for future risk assessment models and formulation of regional management policies. Here, we present a unified list of 27 non-native species recorded in the wild in Armenia, Azerbaijan, and Georgia. Among these 27 species, eight were translocated from the Black Sea basin to the Caspian Sea basin. Out of these 27 non-native fishes, 15 species have become established (three of them being considered invasive) and six fish species could not survive in the wild.

Background: Altered PI3K/mTOR signaling, through activating mutations in PIK3CA and/or PTEN loss, has been implicated in resistance to both hormonal and HER2-directed therapeutics for breast cancer. Recent data from the BOLERO-1 phase III clinical trials demonstrate an improvement in progression free survival (PFS) of 7.2 months in patients with HER2+/estrogen receptor negative (HER2+/ER-) advanced breast cancer when treated with trastuzumab and paclitaxel plus the mTORC1 inhibitor, everolimus (Afinitor ®). However, improvement in PFS was not observed in patients with HER2+/ER+ positive disease. These data suggest that hormone receptor levels may influence the response of HER2-amplifed cells to both HER2 and PI3K/mTOR directed therapies in the absence of hormonally directed therapies. In this study, we investigated the role of ER in HER2+ breast cancer by screening a large panel of HER2+/ER- and HER2+/ER+ breast cancer cell lines for responses to single agent and combined treatment with PI3K, mTOR, HER2 and ER-directed therapeutics. Materials and Methods: The anti-proliferative activity of BKM120 (pan-PI3K inhibitor), BYL719 (p110α-specific) and everolimus were assessed as single agents or in combination with trastuzumab and/or tamoxifen in a panel of six HER2+/ER+ versus six HER2+/ER- breast cancer cell lines in two-dimensional culture. Drug response IC50s were generated from actual cell counts as measured by Z2-particle counting. Biomarker analyses were conducted using baseline mRNA microarray (Agilent) and reverse phase protein array (RPPA) profiling of each of the cell lines to determine associations with response or resistance data. Results: RPPA analysis confirmed the presence of higher levels of ER protein in the cell lines designated as ER+ and significantly higher levels of PTEN protein were detected in those cell lines. Interestingly, each of the PI3K/mTOR pathway inhibitors tended to have increased single agent activity in the ER+ relative to the ER- lines. Combined activity with trastuzumab and either BKM120 or BYL719 was observed in 6 of the 12 cell lines tested and occurred independent of ER status. Similarly, combined activity of everolimus and trastuzumab was also observed in both HER2+/ER+ and HER2+/ER- cell lines. In 3/6 HER2+/ER+ cell lines the addition of tamoxifen provided no benefit to the combination of trastuzumab and PI3K/mTOR pathway inhibitor, whereas in two cell lines mild antagonism was observed with the triple combination. Finally, one cell line did show significant potentiation from the addition of endocrine therapy on top of HER2/PI3K/mTOR targeting. Discussion: These data confirm levels of estrogen receptor are likely playing a role in response to single agent PI3K/mTOR pathway inhibition and highlight the potential utility of combining endocrine therapy with HER2/PI3K/mTOR-directed therapeutics in a sub-group of HER2-amplified breast cancers. Further in depth biomarker analyses may reveal additional molecular alterations responsible for this differential sensitivity to the double and triple combinations and is underway. Citation Format: O'Brien NA, Nichols CM, Thomas J, Conklin D, Kalous O, Linnartz R, Di Tomasso E, Hurvitz SA, Hirawat S, Slamon DJ. Single agent and combined targeting of PI3K, mTOR, HER2 and ER signaling in a panel of HER2+/ER+ versus HER2+/ER- breast cancer cell lines. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-14-25.

Aquaponics is the integrated production of aquatic organisms and plants. The majority (>50 %) of nutrients sustaining optimal plant growth in aquaponics is derived from waste originating from the feeding of aquatic organisms kept in aquaculture. Currently, aquaponics is more popular as a hobby rather than for commercial-scale production. We used long-term data verification to present a qualitative in-depth study of the regional development of aquaponics. We used a questionnaire and in situ interviews to record farm information. In addition, we conducted an Internet search to explore the aquaponics sector and examine the trend in online interest. In contrast to the non-production part of the industry representing 71 % of the Internet search, commercial aquaponic farms only accounted for 2.7 %. In total, annual aquaponic production in Czechia peaked at 3 000 kg of fish and 276 kg of plants, produced mainly by one large-scale aquaponic farm that has been in long-term operation. Using Google Trends, we found a positive trend in online interest. However, a developed online “industry” with a positive image creates an unrealistic awareness of aquaponics. On the basis of this exemplary case, a similar trend can be expected in other countries.

Ornamental aquaculture is known to be one of the main sources of non-native species and Indonesia has been identified as one of the leading suppliers of these organisms worldwide. Released or escaped ornamental aquatic animals can establish new populations and become invasive. On the other hand, some invasive species can be also endangered in their native range, which is called the “Biodiversity Conservation Paradox”. This is true for Arapaima gigas, one of the popular ornamental creatures and the largest bony fish of all, which is threatened in its native range in parts of Amazonia and which has been found to occur in various localities in Java and Sumatra in Indonesia. Based on climate matching we found the vast majority of Indonesian territory to be suitable for this species establishment. Keeping in mind the size and predatory behaviour of A. gigas, we discussed possible consequences of its spread and impacts on native biota in Indonesia.

The information regarding the fish species occurrence in rivers and lakes depends on the quantity and quality of ichthyofaunal surveys. Non-native buffalo fishes, Ictiobus cyprinellus and I. niger (Catostomidae) were introduced to pond aquaculture in the 20th century but since that time they have never been recorded from the natural environment in Europe. Forensic analysis was performed using online data from Czech anglers websites to test whether these fishes were ever caught. In total 13 records were obtained but only 3 were verified in terms of species identification, locality and date. The found records are scattered throughout the area of Czechia and they are without an obvious distribution pattern. The analysis of online fishing web sites can be considered a suitable source of additional information on the occurrence of fishes.

Mika Saarela työryhmän jäsenenä. Background Approximately two-thirds of patients with chronic inflammatory demyelinating polyneuropathy (CIDP) need long-term intravenous immunoglobulin. Subcutaneous immunoglobulin (SCIg) is an alternative option for immunoglobulin delivery, but has not previously been investigated in a large trial of CIDP. The PATH study compared relapse rates in patients given SCIg versus placebo. Methods Between March 12, 2012, and Sept 20, 2016, we studied patients from 69 neuromuscular centres in North America, Europe, Israel, Australia, and Japan. Adults with definite or probable CIDP who responded to intravenous immunoglobulin treatment were eligible. We randomly allocated participants to 0.2 g/kg or 0.4 g/kg of a 20% SCIg solution (IgPro20) weekly versus placebo (2% human albumin solution) for maintenance treatment for 24 weeks. We did randomisation in a 1: 1:1 ratio with an interactive voice and web response system with a block size of six, stratified by region (Japan or non-Japan). The primary outcome was the proportion of patients with a CIDP relapse or who were withdrawn for any other reason during 24 weeks of treatment. Patients, caregivers, and study personnel, including those assessing outcomes, were masked to treatment assignment. Analyses were done in the intention-to-treat and per-protocol sets. This trial is registered with ClinicalTrials. gov, number NCT01545076. Findings In this randomised, double-blind, placebo-controlled trial, we randomly allocated 172 patients: 57 (33%) to the placebo group, 57 (33%) to the low-dose group, and 58 (34%) to the high-dose group. In the intention-to-treat set, 36 (63% [95% CI 50-74]) patients on placebo, 22 (39% [27-52]) on low-dose SCIg, and 19 (33% [22-46]) on high-dose SCIg had a relapse or were withdrawn from the study for other reasons (p=0.0007). Absolute risk reductions were 25% (95% CI 6-41) for low-dose versus placebo (p=0.007), 30% (12-46) for high-dose versus placebo (p=0.001), and 6% (-11 to 23) for high-dose versus low-dose (p=0.32). Causally related adverse events occurred in 47 (27%) patients (ten [18%] in the placebo group, 17 [30%] in the low-dose group, and 20 [34%] in the high-dose group). Six (3%) patients had 11 serious adverse events: one (2%) patient in the placebo group, three (5%) in the low-dose group, and two (3%) in the high-dose group; only one (an acute allergic skin reaction in the low-dose group) was assessed to be causally related. Interpretation This study, which is to our knowledge, the largest trial of CIDP to date and the first to study two administrations of immunoglobulins and two doses, showed that both doses of SCIg IgPro20 were efficacious and well tolerated, suggesting that SCIg can be used as a maintenance treatment for CIDP.

We thank Ravi Uniyal and colleagues for their comments on our results from the PATH trial1 on subcutaneous immunoglobulin (SCIg) for maintenance treatment in chronic inflammatory demyelinating polyneuropathy (CIDP). They express concern about the relapse rates in the treatment groups and hypothesise the cause being the pharmacokinetics of SCIg.

Everolimus (RAD001, Afinitor(®)) is an oral, selective mTOR inhibitor recently approved by the US-FDA in combination with exemestane for treatment of hormone receptor positive advanced breast cancer. To date, no molecular predictors of response to everolimus in breast cancer have been identified. We hypothesized predictive markers could be identified using preclinical models. Using a molecularly characterized panel of human breast cancer and immortalized breast epithelial cell lines, we determined sensitivity to everolimus alone or in combination with ER- or HER2- targeted therapy. Gene expression microarrays and comparative genomic hybridization were performed on the cell lines to identify predictors of response to everolimus. Among 13 everolimus-sensitive cell lines, 10/13(77 %) were luminal, while in 26 resistant cell lines, 16/26(62 %) were non-luminal, and 10/26(38 %) were luminal. Only 3/24 non-luminal lines were sensitive, two of which were HER2+. Everolimus enhanced the anti-proliferative effect of both tamoxifen (TAM) and fulvestrant (FUL) in ER+ breast cancer cell lines, as well as trastuzumab in HER2+ cell lines. Everolimus + FUL but not everolimus + TAM reversed acquired resistance to TAM. Everolimus inhibited mTOR in tested cell lines by decreasing S6 phosphorylation, mediating its anti-proliferative effect by G0/G1 cell cycle arrest and induction of apoptosis. Chromosomal amplifications of AURKA (p value = 0.04) and HER2 (p value = 0.03) were each associated with increased sensitivity to everolimus. Transcript expression microarrays identified GSK3A, PIK3R3, KLF8, and MAPK10 among the genes overexpressed in sensitive luminal lines, while PGP, RPL38, GPT, and GFAP were among the genes overexpressed in resistant luminal cell lines. These preclinical in vitro data provide further support for continued clinical development of everolimus in luminal (ER+ or HER2+) breast cancer in combination with targeted therapies. We identified several potential molecular markers associated with response to everolimus that will require validation in clinical material.

HSP90 enables the activation of many client proteins of which the most clinically validated is HER2. NVP-AUY922, a potent HSP90 inhibitor, is currently in phase II clinical trials. To explore its potential clinical use in HER2-amplified breast and gastric cancers, we evaluated the effect of AUY922 alone and in combination with trastuzumab in both trastuzumab-sensitive and -resistant models. A panel of 16 human gastric and 45 breast cancer cell lines, including 16 HER2-amplified (3 and 13, respectively) cells, was treated with AUY922 over various concentrations. In both breast and gastric cancer, we used cell lines and xenograft models with conditioned trastuzumab-resistance to investigate the efficacy of AUY922 alongside trastuzumab. Effects of this combination on downstream markers were analyzed via Western blot analysis. AUY922 exhibited potent antiproliferative activity in the low nanomolar range (

Freshwater fishes of the genus Carassius , widespread throughout Europe and Asia, are important aquaculture fishes and include the world's most important pet fish, the goldfish. The high morphologic similarity between the species, however, has up to now prevented reliable conclusions on their taxonomy, biogeography and introduction history. A phylogeny of the fish genus Carassius based on the cytochrome b sequence of 404 specimens collected from aquaculture and open water localities across Eurasia identifies most of the presently recognised species as monophyletic lineages, but also that at least one lineage exists that does not correspond to any described species. Within Europe, feral populations of Carassius auratus occur mainly in the Mediterranean area and Great Britain, while Carassius gibelio is found in most of non-Mediterranean Europe and some localities in Italy. Carassius langsdorfii has very scattered points of occurrence in at least six European countries. C. auratus and C. langsdorfii are not native to Europe. The populations of C. gibelio in eastern Central Europe and parts of Eastern Europe are considered as resulting from a natural postglacial range expansion, while the rest of Europe was colonised due to anthropogenic impact. The presence of diploid (2n = 100) as well as triploid (3n = 150) specimens in the three most widespread species indicates that ploidy level is not a character to identify the species of Carassius . A remarkably low genetic divergence in C. gibelio can be the result of clone selection in the gynogenetic populations. In general, our data present the first comprehensive overview about the genus Carassius in Europe based on genetic data.

Background: Aberrant activity of the PI3K/mTOR pathway has been implicated in resistance to trastuzumab and anti-hormonal therapy for HER2-amplified and ER-positive breast cancer, respectively. Previous studies in our laboratory and others have shown that increased PI3K/mTOR signaling, either through PTEN loss or activating PIK3CA mutations, can confer resistance to trastuzumab therapy. In this study, we assessed the potential of targeting the PI3K/mTOR pathway in overcoming both de novo and acquired trastuzumab resistance. Materials and Methods: The in vitro activity of the pan-PI3K inhibitor BKM120, the mTORC1 inhibitor RAD001 and the dual PI3K/mTORC1/2 inhibitor BEZ235 were evaluated in a panel of 49 human breast cancer and immortalized cell lines. The in vivo activity of these molecules was assessed in six cell line xenografts models representing, ER+/HER2− (KPL-1, ZR75-1), ER+/HER2+ (UACC812, MDA361), ER−/HER2+ (SUM190), PIK3CA mutant (SUM190, MDA361), PTEN-null (ZR75-1) and trastuzumab resistant (BT-TR) breast cancer. Finally, the effect of PI3K/mTOR inhibition on feedback activation of PI3K signaling and compensatory pathways was measured by Western blot, immunohistochemistry (IHC) and reverse phase protein analysis (RPPA) of control and treated cell lysates/tumors. Results: Using a sensitivity cut-off of an IC50 of < 1 µmol/L, 16 of the 18 HER2-amplified breast cancer cell lines and 8 of 10 cell lines with activating mutations in PIK3CA were sensitive to the pan-PI3K inhibitor BKM120. BEZ235 showed the most potent efficacy across the panel with IC50s < 100 nmol/L for each of the 49 cell lines tested. The HER2-amplified/PIK3CA mutant cell lines were also unexpectedly sensitive to the mTORC1 inhibitor RAD001, this was despite the silencing of mTORC1 signaling being followed by a feedback increase in phospho-AKT signaling. Furthermore, each of these molecules showed remarkable in vivo activity across the panel of xenografts models. BKM120, RAD001 and BEZ235 induced both tumor stabilization and regression independent of PTEN, PI3K, ER and HER2 status. Pharmacodynamic analysis of tumor tissue revealed that BEZ235 and RAD001 both inhibited mTORC1 signaling as indicated by a reduction in the levels of phosphorylated ribosomal protein S6 (pS6). However, in contrast to BKM120 and BEZ235, RAD001 did not induce a reduction in the levels of pAKT (S473 or TH308) yet showed comparable in vivo efficacy to each of these molecules. Finally, combined targeting of HER2 and PI3K/mTOR in vitro increased the anti-proliferative activity of the molecules and led to an increased induction of apoptosis. The efficacy of these molecules (alone or in combination with trastuzumab) was assessed in a model of in vivo trastuzumab resistance generated through long-term treatment of the trastuzumab sensitive BT474 cells (BT-TR). All PI3Ki induced complete inhibition of tumor proliferation in monotherapy, while the combination of trastuzumab and each of these molecules induced tumor regression in the trastuzumab resistant tumors. Discussion: These pre-clinical data indicate that targeting the PI3K/mTOR pathway either alone or in combination with trastuzumab is effective strategy for overcoming trastuzumab resistance. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P4-08-01.

The human EGF (HER) family of receptors has been pursued as therapeutic targets in breast cancer and other malignancies. Trastuzumab and lapatinib are standard treatments for HER2-amplified breast cancer, but a significant number of patients do not respond or develop resistance to these drugs. Here we evaluate the in vitro activity of dacomitinib (PF-00299804), an irreversible small molecule pan-HER inhibitor, in a large panel of human breast cancer cell lines with variable expression of the HER family receptors and ligands, and with variable sensitivity to trastuzumab and lapatinib. Forty-seven human breast cancer and immortalized breast epithelial lines representing the known molecular subgroups of breast cancer were treated with dacomitinib to determine IC50 values. HER2-amplified lines were far more likely to respond to dacomitinib than nonamplified lines (RR, 3.39; P < 0.0001). Furthermore, HER2 mRNA and protein expression were quantitatively associated with response. Dacomitinib reduced the phosphorylation of HER2, EGFR, HER4, AKT, and ERK in the majority of sensitive lines. Dacomitinib exerted its antiproliferative effect through a combined G0–G1 arrest and an induction of apoptosis. Dacomitinib inhibited growth in several HER2-amplified lines with de novo and acquired resistance to trastuzumab. Dacomitinib maintained a high activity in lines with acquired resistance to lapatinib. This study identifies HER2-amplified breast cancer lines as most sensitive to the antiproliferative effect of dacomitinib and provides a strong rationale for its clinical testing in HER2-amplified breast cancers resistant to trastuzumab and lapatinib. Mol Cancer Ther; 11(9); 1978–87. ©2012 AACR.

Distal parts of the fish intestine were analyzed for presence of bifidobacteria and lactobacilli using selective agars. Seventy seven samples from Cyprinus carpio, Oncorhynchus mykiss, Carassius auratus, Tinca tinca, Perca fluviatilis, Rutilus rutilus, Scardinius erythrophthalmus, Oreochromis niloticus, and Squalius cephalus were collected randomly throughout years 2008 and 2009. Bifidobacteria were detected in 5 samples from 4 fish species at counts 2.18–4.29 log CFU/g, lactobacilli were present in 6 fish species at counts 1.21–3.65 log CFU/g. Seven bifidobacterial isolates were identified to the species level using biochemical tests and by sequencing of 16S rRNA gene. Three strains belonged to species B. longum, two isolates were identified as B. dentium, one strain as B. asteroides and one isolate was not determined to the species level by employed methods. As identified bifidobacterial species are considered to be of human, animal or honeybee origin, they probably derived as contamination from sewage or other sources. After further more detail testing, the possible use of isolated bifidobacteria as probiotics is promising since they were able to pass through the digestive tract successfully.

Background: Genetic and epigenetic alterations in the PI3K/mTOR and cyclin D:CDK-4/6:Rb signaling axes occur frequently in breast cancer and have been attributed to resistance to both ER- and HER2-directed therapeutics. Pharmacologically targeting CDK-4/6 in combination with hormonal blockade provides clinical benefit in patients with advanced ER+ breast cancer. In this study, we evaluated the activity of the pan-class I PI3K inhibitor, copanlisib (BAY-80-6946), with potent alpha and delta activity as a single agent or in combination with CDK-4/6 inhibition and hormonal blockade in a panel of breast cancer cell lines. Methods: The growth inhibitory activity of copanlisib was evaluated against a large panel of 48 breast cancer cell lines molecularly characterized by genomic, transcriptomic and proteomic profiling. IC50 values were determined from direct cell counts using a Z1-particle counter. The activity of copanlisib in combination with hormone blockade and CDK-4/6 inhibition, by palbociclib, was assessed in two cell line xenograft models of ER+ breast cancer; MCF7(PIK3CA-E545K) and ZR751(PIK3CA WT). For xenograft studies, tumor bearing mice were treated once weekly (BID) by intravenous injection with clinically achievable doses of copanlisib (10 mg/kg) as single agent or in combination with tamoxifen or fulvestrant with or without 75 mg/kg daily palbociclib for 21 days. Results: A broad range of IC50 values (0.491-895 nM), with a high degree of separation between sensitive and resistant histologically defined subgroups were determined for copanlisib, indicating the potential for a wide therapeutic window. Luminal subtype, the presence of activating mutations in PIK3CA, high levels of ER, HER2, HER3 and EGFR protein enriched for sensitivity to copanlisib. Activating mutations of KRAS and BRAF were associated with resistance to copanlisib. Single agent copanlisib induced significant tumor growth inhibition (TGI) relative to vehicle control in each of the xenograft models. Modest increases in anti-tumor activity were achieved when copanlisib was combined with hormonal blockade by either tamoxifen or fulvestrant. However, robust tumor regressions were observed with the triple combinations of copanlisib-palbociclib-tamoxifen and copanlisib-palbociclib-fulvestrant. Furthermore, these triple combinations achieved a statistically significant improvement in anti-tumor activity over the standard of care combination of palbociclib plus fulvestrant. Each of the single agent and treatment combinations tested were well tolerated in animals. Discussion: These preclinical data illustrate the potent and selective activity of the pan class I PI3K inhibitor copanlisib in luminal breast cancers and support the clinical investigation of copanlisib in combination with CDK-4/6 inhibition and hormonal blockade in ER+ breast cancer. Citation Format: O'Brien NA, Conklin D, Luo T, Ayala R, Issakhanian S, Kalous O, Von Euw E, Politz O, Wilhelm S, Childs BH, Hurvitz SA, Slamon DJ. The PI3K-inhibitor, copanlisib, has selective activity in luminal breast cancer cell lines and shows robust combined activity with hormonal blockade and CDK-4/6 inhibition in ER+ breast cancer cell line xenografts [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P3-04-15.

Background: Although trastuzumab and lapatinib provide clinical benefit for women with HER2−positive breast cancer, both de novo and acquired resistance to these agents exist. There is increasing evidence to suggest that aberrant activity of the PI3K/AKT/mTOR signaling pathway is one of the key mechanisms responsible for resistance. Thus, pharmacologically targeting the PI3K pathway is a rational approach for overcoming resistance to HER2−targeted therapy. However, specifically targeting individual elements of the PI3K pathway may result in feedback activation or in activation of compensatory pathways, whereas targeting the pathway at multiple points may be more effective in permanently shutting down proliferation and inducing cell death. Materials and Methods: In this study we evaluated the activity of three small molecule tyrosine kinase inhibitors that target specific critical portions of the PI3K pathway; BKM120 (pan-PI3K), RAD001 (mTORC1 specific) and BEZ235 (dual PI3K & mTOR) both individually and in combination with trastuzumab in a panel of HER2−amplified breast cancer cell lines that have previously been characterized for trastuzumab response/resistance. Results: In the trastuzumab sensitive BT-474 and SKBR3 cell lines, 72 hours of trastuzumab treatment efficiently inhibited cell proliferation and deactivated AKT (S473 & Th308), PS6K and ERK signaling. In contrast, in the trastuzumab-resistant MDA453 and SUM225 cell lines, as well as the BT-474 cells conditioned to acquire trastuzumab resistance (BT-TR), trastuzumab treatment did not inhibit pAKT, pS6K and pERK levels. RAD001 had significant antiproliferative activity in both trastuzumab sensitive and resistant cell lines and resulted in rapid down regulation of pPS6K (mTOR) activity (15 min). However, this deactivation was followed by a reactivation of AKT and ERK signaling (24 to 72 hours). BKM120 also demonstrated anti-proliferative activity independent of trastuzumab sensitivity. Specifically targeting PI3K with BKM120 directly decreased pAKT/pS6K/pERK signaling and feedback activation was less pronounced than that observed with RAD001. Of the three molecules tested, BEZ235 was the most efficacious in the cell line panel (all IC50gs < 20 nM). BEZ235 rapidly (15 min) decreased pAKT/pS6K/pERK signaling in both trastuzumab sensitive and resistant cells lines and this inhibition was maintained at 72 hours. Thus, BEZ235 is effective in shutting down PI3K feedback activation and efficiently shuts down PI3K signaling in trastuzumab resistant HER2−amplified breast cancer cell lines. The combined treatment of trastuzumab and each of the PI3K/mTOR targeting agents resulted in greater inhibition of proliferation and induction of cell death in the HER2−amplified cell lines than each agent used alone. No increase in activity was observed in the HER2−normal control cell lines, MCF-7 and KPL-1. Discussion: Together, these data indicate that targeting the PI3K/AKT/mTOR pathway either alone or in combination with trastuzumab is effective in overcoming trastuzumab resistance. These findings are currently being validated in in vivo models of trastuzumab resistance. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P1-12-03.

The freshwater characiform fish Prochilodus lineatus is a detritivorous species that has its native distribution area in South America but has been imported to China for aquaculture purposes. This is the first time that it is being reported in Vietnam, both from aquaculture and captured from a river channel. According to local authorities, the species is becoming increasingly important in local aquaculture and its spread can be expected. Keeping in mind the high biomass production in the rivers of its native distribution area, a successful establishment of P. lineatus into Vietnamese fresh waters may exert serious impacts on the local ecosystems.

Spinal cord injury (SCI) is a major cause of persistent neuropathic pain of central origin. Recent evidence suggests neuropathic pain in clinically complete SCI patients correlates with limited sensory function below the lesion (sensory discomplete). On this basis we examined if the onset of mechanical hyperalgesia was different in rodents after a severe incomplete clip-compression SCI versus a complete spinal cord transection at thoracic segment T13. Above-level withdrawal behaviors evoked by forepaw stimulation provided evidence of mechanical hyperalgesia after incomplete but not complete SCI, whereas below-level responses evoked by hindpaw stimulation revealed hypersensitivity after both injuries. The latency of the above-level response was 4–5 wks but was longer after a moderate clip-compression injury. Mechanical hyperalgesia was fully reversed by three analgesic drugs used in treating neuropathic SCI pain, but their duration of action differed significantly, showing a rank order of amitriptyline (24–48 h) ≫ morphine (6 h) > gabapentin (2 h). Evidence of central sensitization in cervical spinal cord segments that receive sensory projections from the forelimbs was provided by immunohistochemistry for Zif268, a functional marker of neuroplasticity. Zif268-immunoreactive neurons in laminae I/II increased in response to repetitive noxious forepaw stimulation in the incomplete SCI group, and this response was reduced in the complete transection and sham-operated groups. These data are consistent with the hypothesis that neuropathic pain of cord origin is more likely to develop after SCI when there is an incomplete loss of axons traversing the lesion.

Summary The objective of this study was to develop a fast, reliable and cost effective photographic documentation tool to complement identification methods on preserved material. We tested the applicability of a flatbed desktop scanner as an easy means to provide a voucher in form of a whole body image for further morphological analysis. Similar species of European perch and zander were included in the test trial, accompanied by molecular genetic analysis as reference identification. The outcome of the study reveals the general utility of the method.

A conditioning lesion improves regeneration of central and peripheral axons of dorsal root ganglion (DRG) neurons after a subsequent injury by enhancing intrinsic growth capacity. This enhanced growth state is also observed in cultured DRG neurons, which support a more sparsely and rapidly elongating mode of growth after a prior conditioning lesion in vivo. Here we examined differences in the capacity or requirements of specific types of sensory neurons for regenerative growth, which has important consequences for development of strategies to improve recovery after injury. We showed that after partial or complete injury of the sciatic nerve in mice, an elongating mode of growth in vitro was activated only in DRG neurons that did not express calcitonin gene-related peptide (CGRP) or bind Bandeiraea simplicifolia I-isolectin B4 (IB4). We also directly examined the response of conditioned sensory neurons to nerve growth factor (NGF), which does not enhance growth in injured peripheral nerves in vivo. We showed that after partial injury, NGF stimulated a highly branched and linearly restricted rather than elongating mode of growth. After complete injury, the function of NGF was impaired, which immunohistochemical studies of DRG indicated was at least partly due to downregulation of the NGF receptor, tropomyosin-related kinase A (TrkA). These results suggest that, regardless of the type of conditioning lesion, each type of DRG neuron has a distinct intrinsic capacity or requirement for the activation of rapidly elongating growth, which does not appear to be influenced by NGF.

Objective: Chronic lung disease caused by volutrauma is one of the most important consequences of preterm delivery. In this pilot study a new method is presented that consists of flushing part of the dead space with fresh gas in order to reduce high tidal volumes, the chief cause of volutrauma. The aim of the study was to evaluate if the new method could reduce ventilatory effort in preterm infants by diminishing dead space. Design and setting: In split-flow ventilation, gas required for dead-space washout is split off from the regular ventilation circuit. The split flow bypasses the apparatus dead space and fills it retrogradely with fresh breathing gas, mainly in the pause between exhalation and inspiration. The mean per-minute ventilation and ventilation index after 12 h of conventional ventilation were compared with corresponding mean values after 12 h of split-flow ventilation in 17 preterm infants weighing

Superoxide radicals (O2-) are generated during reoxygenation following asphyxia, possibly more when higher concentrations of O2 are used during resuscitation. Superoxide dismutase (SOD) is an antioxidant enzyme, which scavenges O2-. We tested the hypothesis that a single intravenous dose of recombinant human Cu,Zn SOD (rhSOD) could influence microcirculation and biochemical markers of asphyxia in piglets reoxygenated with 21% or 100% O2 after combined cerebral hypoxemia-ischemia-hypercapnia.Anesthetized newborn piglets were randomized to asphyxia (n = 40) or control (n = 3). Asphyxia was induced by ventilation with 8% O2, adding CO2, and temporary occlusion of both common carotid arteries. After 20 min, 16 piglets received rhSOD 5 mg/kg intravenously and reoxygenation with 21% O2 (rhSOD, 21%; n = 8) or 100% O2 (rhSOD, 100%; n = 8), and 24 piglets received saline and reoxygenation with 21% O2 (21%, n = 13) or 100% O2 (100%, n = 11). The cortical microcirculation was assessed by laser Doppler flowmetry, and glutamate in the striatum and hypoxanthine in the cortex were measured by in vivo microdialysis.rhSOD peaked in plasma after 5 min. No rhSOD was detected in brain tissue. There were no significant differences between rhSOD and non-rhSOD groups in any measured variable.

The selective cyclin dependent kinases 4 and 6 (CDK-4/6) inhibitor, palbociclib, has recently been approved in combination with endocrine-based therapies for the treatment of advanced ER+/HER2- breast cancer (BC). Despite the improvements in progression-free and overall survivals, a significant number of patients on CDK-4/6 therapy will go on to develop progressive disease. As such, it is critical to identify the mechanisms by which tumor cells evade CDK-4/6 targeted therapy and to develop therapeutic strategies to overcome resistance. In this study, we evaluated the efficacy of a p110α-selective PI3K inhibitor, alpelisib (BYL719) and the mTORC1 specific inhibitor, everolimus (RAD001), in ER+/HER2- xenograft tumors conditioned in vitro and in vivo to acquire resistance to combined palbociclib + endocrine-based therapy. The EFM19 (ER+/HER2-, PIK3CA mt), MDA134 (ER+/HER2-, PIK3CA wt), and MDA361 (ER+/HER2+, PIK3CA mt) BC cell lines were conditioned to acquire palbociclib resistance through long-term culture in the presence of increasing concentrations of drug. Baseline total/phosphoprotein levels of over 280 cancer-associated analytes were measured in the parental and resistant cell lines by Reverse Phase Protein Array (RPPA). In vivo resistant models were developed from xenografts of the MCF7 and HCC1500 ER+ BC cell lines, treated until progression with palbociclib (75 mg/kg) + fulvestrant (5 mg/mse). EFM19-PR cells maintained palbociclib resistance in vivo and were included in the xenograft studies. Resistance to palbociclib was confirmed in vitro by a shift in growth inhibitory IC50 from an average of less 50nM to over 1 µM. Significantly reduced Rb and ER protein levels were detected in each of these palbociclib resistant cell lines by RPPA. Cross-resistance to an alternative CDK-4/6 inhibitor (ribociclib) was found in in vitro studies. In addition, switching treatment of the xenografts progressing on palbociclib + fulvestrant to ribociclib (75mg/kg) + fulvestrant did not impact tumor progression. However, when progressing tumors were switched to BYL719 (30mg/kg) + fulvestrant or RAD001 (10mg/kg) + fulvestrant, sustained tumor regression was observed for over 45 days of treatment in every model tested. The triplet combination of ribociclib + fulvestrant + BYL719 or RAD001 provided marginal additional benefit over the doublet combination of BYL719 or RAD001 + fulvestrant. RPPA analysis of xenograft tissue collected from these studies will help to identify additional molecular alterations involved in resistance to CDK-4/6 targeted therapy. These preclinical data indicate that acquired resistance to CDK-4/6 inhibition occurs through a loss of dependence on Rb-signaling. However, targeting an alternative pathway like PI3K/mTOR with molecules such as BYL719 or RAD001 may be a viable strategy for further clinical investigation. Citation Format: Neil A. O'Brien, Dylan Conklin, Tong Luo, Raul Ayala, Shawnt Issakhanian, Ondrej Kalous, Erika Von Euw, Sara A. Hurvitz, Emmanuelle DiTomaso, Faye Su, Ronald Linnartz, Stefan Scherer, Samit Hirawat, Dennis J. Slamon. Anti-tumor activity of the PI3K/mTOR pathway inhibitors alpelisib (BYL719) and everolimus (RAD001) in xenograft models of acquired resistance to CDK-4/6 targeted therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4150. doi:10.1158/1538-7445.AM2017-4150

In this study, records on the occurrence of the Japanese ginbuna Carassius langsdorfii from northern Germany, north-western Italy and southern Bosnia and Herzegovina are presented. The new findings, in addition to former studies reported in the Czech Republic and Greece, show that C. langsdorfii is much more widespread in Europe than was previously believed.

Approximately 25,000 ovarian cancers are diagnosed in the U.S. annually, and 75% are in the advanced stage and largely incurable. There is critical need for early detection tools and novel treatments. Proteasomal ubiquitin receptor ADRM1 is a protein that is encoded by the ADRM1 gene. Recently, we showed that among 20q13-amplified genes in ovarian cancer, ADRM1 overexpression was the most highly correlated with amplification and was significantly upregulated with respect to stage, recurrence, and metastasis. Its overexpression correlated significantly with shorter time to recurrence and overall survival. Array-CGH and microarray expression of ovarian cancer cell lines provided evidence consistent with primary tumor data that ADRM1 is a 20q13 amplification target. Herein, we confirm the ADRM1 amplicon in a second ovarian cancer cohort and define a minimally amplified region of 262 KB encompassing seven genes. Additionally, using RNAi knock-down of ADRM1 in naturally amplified cell line OAW42 and overexpression of ADRM1 via transfection in ES2, we show that (1) ADRM1 overexpression increases proliferation, migration, and growth in soft agar, and (2) knock-down of ADRM1 results in apoptosis. Proteomic analysis of cells with ADRM1 knock-down reveals dysregulation of proteins including CDK-activating kinase assembly factor MAT1. Taken together, the results indicate that amplified ADRM1 is involved in cell proliferation, migration and survival in ovarian cancer cells, supporting a role as an oncogene and novel therapeutic target for ovarian cancer.

The cyclin D:CDK-4/6:Rb axis is dysregulated in a number of different cancers and is implicated in resistance to hormonal therapy in breast cancer. Pharmacologically targeting cyclin dependent kinase 4 and 6 (CDK4 and 6) has proven to be a successful therapeutic approach in ER+ breast cancer (BC). This study aimed to identify the molecular subtypes of BC that are sensitive to the novel CDK4 and 6 inhibitor, abemaciclib, and identify the best combination strategies for the clinical development. Growth inhibition activity of abemaciclib was assessed in a panel of 46 BC cell lines molecularly characterized by genomic, transcriptomic and proteomic profiling. IC50 values were determined from direct cell counts using a Z1-particle counter. In vivo activity of abemaciclib was assessed in cell line xenograft models of ER+ and HER2+/ER+ BCs. For ER+ BC, mice were treated daily with clinically achievable doses of abemaciclib (50-75 mg/kg) as a single agent or in combination with tamoxifen or fulvestrant. Combinations with trastuzumab, docetaxel and tamoxifen were assessed in the HER2+/ER+ xenografts. Sensitivity to abemaciclib was observed predominately in multiple luminal BC cell lines and a small subset of triple negative cell lines that had intact Rb signaling. Activating mutations in PIK3CA also marked for abemaciclib sensitivity. Abemaciclib potentiated the anti-proliferative effects of cytotoxic/anti-mitotic agents when given simultaneously or 48 hours prior to treatment in vitro. Significant tumor growth inhibition (TGI) was observed with single agent abemaciclib in the ER+ BC cell line xenografts. In ZR751 xenografts, the addition of either tamoxifen or fulvestrant to abemaciclib induced complete inhibition of tumor growth for the 12 weeks of treatment. In the MCF7 model, treatment was withdrawn after five weeks, which triggered tumor regrowth in each of the single agent arms. However, complete responses were maintained in the combination arms for a further six weeks post drug withdrawal. In HER2 amplified xenografts, abemaciclib single agent treatment induced significant TGI in trastuzumab sensitive and resistant xenografts, and combination with trastuzumab further increased this anti-tumor effect. The addition of tamoxifen to this combination induced a further increment in TGI. Consistent with the in vitro findings, the combination of abemaciclib and the anti-mitotic agent docetaxel was not antagonistic in vivo, and the addition of docetaxel to the triple combination of abemaciclib, trastuzumab, and tamoxifen induced the most efficacy of any of the treatment arms tested. Combinations were well tolerated in animals. These data highlight the potential of abemaciclib to have single agent activity in addition to combined activity with anti-mitotic or targeted therapies for breast cancer. Citation Format: Neil A. O’Brien, Dylan Conklin, Tong Luo, Ondrej Kalous, Erika von Euw, Sara A. Hurvitz, Richard P. Beckmann, Colleen Mockbee, Dennis J. Slamon. Preclinical activity of abemaciclib as a single agent or in combination with anti-mitotic or targeted therapies for breast cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 2828.

BACKGROUND: Aurora kinases (AK) A and B play essential roles in multiple stages of mitosis and are frequently overexpressed in a subset of human cancers, including ovarian cancer (OC). AMG 900, a potent and highly selective small molecule inhibitor of AKs, showed promising single-agent activity in heavily pretreated patients with advanced OC in a Phase 1b clinical trial. In this study, we report the preclinical effects of AMG 900 in a panel of well-characterized human cancer cell lines representing clinically-relevant OC subtypes. METHODS: The anti-proliferative effects of AMG 900 were evaluated using a 5-day cell count assay. Cell lines were classified as sensitive to AMG 900 when lethality was > 15% at 10 nM. Molecular markers were profiled including TP53 mutation status, AURKA, CCNE1, MYC copy number, and p53, p21 and cyclin E1 protein by reverse phase protein array. Flow cytometry and imaging methods were used to evaluate the mechanism of action of AMG 900 alone and in combination with chemotherapy. The combination of AMG 900 plus docetaxel was evaluated in an IGROV-1 ovarian endometrioid carcinoma xenograft model. RESULTS: One third of the cell lines (11 of 35) were classified as sensitive to AMG 900 and showed enrichment for TP53 mutations and serous OC subtype. However, 10 of 24 resistant cell lines harbored TP53 mutations, indicating that TP53 mutational status alone was not sufficient for predicting AMG 900 sensitivity. Inhibition of AK activity by AMG 900 in OC cells resulted in aborted cell division leading to polyploidy and cell death (suggestive of aurora-B dominant phenotype). Re-plating of remnant cells after AMG 900 treatment showed an attenuation of cell regrowth, where TP53mut IGROV-1 cells showed minimal regrowth compared to TP53wt OVCAR-5 cells. AMG 900 inhibited proliferation at low nanomolar concentrations in the majority of OC cell lines and enhanced the effects of paclitaxel, carboplatin, and doxorubicin in IGROV-1 cells. In tumor-bearing mice, administration of AMG 900 at 7.5 mg/kg (PO) for two days per week or docetaxel at 10 mg/kg (IP) weekly for four cycles significantly inhibited the growth of IGROV-1 tumor xenografts (P < .0001 vs. vehicle alone). Notably, co-administration of AMG 900 with docetaxel enhanced efficacy and induced a delay in tumor regrowth compared to docetaxel alone. Single-agent-treated mice showed minimal body weight loss (BWL), whereas combination-treated mice showed moderate BWL (average < 10%) that was largely reversible (2 of 12 animals removed due to toxicity). CONCLUSIONS: AMG 900 alone or in combination with chemotherapy such as paclitaxel may be a promising clinical strategy to treat patients with ovarian cancer. Citation Format: Ondrej Kalous, Dylan Conklin, Kanthinh Manivong, William Wayne, Kelly Hanestad, Jude Canon, Robert Loberg, Gregory Friberg, Erick Gamelin, Florian D. Vogl, Gloria Juan, Angela Coxon, Dennis Slamon, Richard Finn, Marc Payton. Preclinical characterization of AMG 900, a pan-aurora kinase inhibitor, alone and in combination with taxanes in ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3008.

An unusual case of a primary cutaneous Absidia corymbifera infection in a premature twin successfully treated with low doses of intravenous amphotericin B and topical natamycin is described. Epidemiological and therapeutical aspects of the case are discussed and in vitro antifungal susceptibility data are presented.

The paper deals with a time-domain (transient) model of a microturbine power unit for the design, optimization, and verification of control equipment. The model includes the starter motor, generator, gearbox, and single-shaft gas microturbine. In addition to the normal operational performance the model allows simulating various faults including failed ignition in the combustion chamber. Explicit formulation of the differential equations of the model allows computationally effective implementations in a hardware emulator.

Purpose: Fenretinide [N-(4-hydroxyphenyl)retinamide (4-HPR)] is a cytotoxic retinoid that suffers from a wide interpatient variation in bioavailability when delivered orally in a corn oil capsule. The poor bioavailability of the capsule formulation may have limited responses in clinical trials, and the large capsules are not suitable for young children. To support the hypothesis that a novel organized lipid matrix, LYM-X-SORB, can increase the oral bioavailability of fenretinide, fenretinide in LYM-X-SORB matrix and in a powderized LYM-X-SORB formulation was delivered to mice. Experimental Design: Fenretinide was delivered orally to mice as the contents of the corn oil capsule, in LYM-X-SORB matrix (4-HPR/LYM-X-SORB matrix) or in a LYM-X-SORB matrix powderized with sugar and flour (4-HPR/LYM-X-SORB oral powder). Levels of 4-HPR, and its principal metabolite, N-(4-methoxyphenyl)retinamide, were assayed in plasma and tissues. Results: In a dose-responsive manner, from 120 to 360 mg/kg/d, delivery to mice of 4-HPR in LYM-X-SORB matrix, or as 4-HPR/LYM-X-SORB oral powder, increased 4-HPR plasma levels up to 4-fold (P < 0.01) and increased tissue levels up to 7-fold (P < 0.01) compared with similar doses of 4-HPR delivered using capsule contents. Metabolite [N-(4-methoxyphenyl)retinamide] levels mirrored 4-HPR levels. Two human neuroblastoma murine xenograft models showed increased survival (P < 0.03), when treated with 4-HPR/LYM-X-SORB oral powder, confirming the bioactivity of the formulation. Conclusions: 4-HPR/LYM-X-SORB oral powder is a novel, oral drug delivery formulation, suitable for pediatric use, which warrants further development for the delivery of fenretinide in the treatment of cancer. A phase I clinical trial in pediatric neuroblastoma is in progress.

We tested if reoxygenation with 100% O2 was superior to 21% O2 after combined cerebral hypoxemia-ischemia-hypercapnia (HIH) in newborn piglets. Twenty-eight piglets were randomized to reoxygenation with 100 or 21% O2 following asphyxia. Asphyxia was induced by ventilation with 8% O2, adding CO2, and temporary occlusion of both common carotid arteries. After 20 min, reoxygenation-reperfusion was started with 21% O2 (HIH 21% group, n = 13) or 100% O2 (HIH 100% group, n = 11) for 30 min followed by 21% O2. All piglets were observed for 2 h. We measured mean arterial blood pressure (MABP), changes in microcirculation in the cerebral cortex (laser Doppler), and extracellular concentrations of hypoxanthine in the cortex and amino acids in the striatum (microdialysis). We found significantly higher MABP and better restoration of microcirculation after reoxygenation with 100% compared with 21% O2, but no differences in biochemical markers were found between the groups. This indicates that the brain tolerated reoxygenation with 21% as well as with 100% O2 in the present model of experimental asphyxia in spite of the differences in MABP and cerebral microcirculation.

To determine whether the administration of lisofylline (1-[5R-hydroxyhexyl]-3,7-dimethylxanthine) would decrease mortality in patients with acute lung injury (ALI) or acute respiratory distress syndrome (ARDS).A prospective, randomized, double-blind, placebo-controlled, multicenter study.Intensive care units at 21 hospitals at the ten centers constituting the ARDS Clinical Trials Network.A total of 235 patients who met eligibility criteria were enrolled in the study (116 into the lisofylline group, 119 into the placebo group).Patients were randomized to receive either lisofylline or placebo. The dose of lisofylline was 3 mg/kg with a maximum dose of 300 mg intravenously every 6 hrs. The intravenous solution of study drug was administered over 10 mins every 6 hrs. Dosing was continued for 20 days or until the patient achieved 48 hrs of unassisted breathing.The trial was stopped by the Data Safety Monitoring Board for futility at the first scheduled interim analysis. The patient groups had similar characteristics at enrollment. No significant safety concerns were associated with lisofylline therapy. There was no significant difference between groups in the number of patients who had died at 28 days (31.9% lisofylline vs. 24.7% placebo, p = .215). There was no significant difference between the lisofylline and placebo groups in terms of resolution of organ failures, ventilator-free days, infection-related deaths, or development of serious infection during the 28-day study period. The median number of organ failure-free days for the five nonpulmonary organ failures examined (cardiovascular, central nervous system, coagulation, hepatic, and renal) was not different between the lisofylline and placebo groups. Although lisofylline has been reported to decrease circulating free fatty acid levels, we did not find any such treatment effect compared with placebo.In this study, there was no evidence that lisofylline had beneficial effects in the treatment of established ALI/ARDS.

This paper describe an application of a neural network approach to SM (standard model) and MSSM (minimal supersymetry standard model) Higgs search in the associated production ttH with H ? bb. This decay channel is considered as a discovery channel for Higgs scenarios for Higgs boson masses in the range 80 - 130 GeV. Neural network model with a special type of data flow is used to separate ttjj background from H ? bb events. Used neural network combine together a classical neural network approach and linear decision tree separation process. Parameters of these neural networks are randomly generated and population of predefined size of those networks is learned to get initial generation for the following genetic algorithm optimization process. A genetic algorithm principles are used to tune parameters of further neural network individuals derived from previous neural networks by GA operations of crossover and mutation. The goal of this GA process is optimization of the final neural network performance.Our results show that NN approach is applicable to the problem of Higgs boson detection. Neural network filters can be used to emphasize difference of Mbb distribution for events accepted by filter (with better signal/background rate) and Mbb distribution for original events (with original stgna/background rate) under condition that there is no loss of significance. This improvement of the shape of Mbb distribution can be used as a criterion of existence of Higgs boson decay in considered discovery channel.

We studied drug-induced cardiotoxic effects in 22 pregnant women having tocolysis with intravenous fenoterol and verapamil. Because CK-MB is released from the uterus and placenta, we used the determination of cardiac troponin T (cTnT) as it is one of the most sensitive and specific indicators of myocardial necrosis. Cardiac troponin T levels were within physiological range (0.08 +/- 0.01 microgram/l) in all healthy pregnant women tested between 32 and 36 weeks of gestation (control group). In the pregnant women having tocolysis cTnT levels started to increase slightly during the first day of treatment (0.10 +/- 0.03 microgram/l) and were significantly higher (p0.05) during the third day (0.35 +/- 0.14 microgram/l) of tocolytic therapy. The cTnT levels in cord blood (0.13 +/- 0.03 microgram/l) did not correspond with maternal cTnT concentrations.

Background: Heat Shock Protein 90 (HSP90) enables the activation of a large number of client proteins of which the most clinically validated is HER2. NVP-AUY922 is a a highly potent, non-geldamycin HSP90 inhibitor that has shown preliminary activity in HER2 positive metastatic breast cancer. To explore its potential clinical utility specifically in HER2 amplified gastric and breast cancers, we evaluated the effect of AUY922 alone and in combination with trastuzumab in both trastuzumab-sensitive and resistant in vitro and in vivo models. Methods: A panel of 16 human gastric and 45 breast cancer cell lines, with 3 and 13 HER2-amplified lines respectively, were exposed in vitro to AUY922 over various concentrations to generate dose response curves. In both breast and gastric cancer, we used cell lines and xenograft models with conditioned trastuzumab-resistance to investigate the efficacy of AUY922 alongside trastuzumab. In all in vivo experiments, trastuzumab was dosed at 10 mg/kg (IP; twice/week) and AUY922 at 50 mg/kg, (IP; 5 days on, 2 days off). Effects of this combination on downstream markers (HER2, AKT, ERK) were analyzed via western blot. Results: AUY922 was found to have potent anti-proliferative activity in the low nanomolar range (< 40 nM) for 59 of the 61 gastric and breast cancer cell lines tested. Additionally, in both gastric and breast cancer, HER2-amplified cells expressed greater sensitivity to the compound when compared to the HER2-negative cells. In conditioned trastuzumab-resistant cell line models, AUY922 continued to have potent activity and exhibited a synergistic effect with trastuzumab. The in vitro combination also induced greater decreases in HER2, induced a G0/G1 cell cycle arrest, and increased the rates of apoptosis. In a conditioned trastuzumab-resistant gastric cancer in vivo model (N87-Res), the combination of AUY922 and trastuzumab showed greater anti-tumor efficacy than either drug alone. Conclusions: These data suggest that AUY922 in combination with trastuzumab has unique efficacy in trastuzumab-resistant models. The combination of HSP90 inhibitors and trastuzumab represents a novel approach to the treatment of HER2 amplified cancers. Clinical trials combining AUY922 and trastuzumab in HER2 amplified breast and gastric cancers are currently ongoing. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3668. doi:1538-7445.AM2012-3668

14688 Background: Elevated levels of Src kinase have been implicated in the malignant potential of colorectal cancer. Dasatinib is an orally active, multi-targeted tyrosine kinase inhibitor of ABL ...