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Glioblastoma is a highly aggressive brain tumor for which there is no cure. The metabolic enzyme 6-Phosphofructo-2-Kinase/Fructose-2,6-Biphosphatase 4 (PFKFB4) is essential for glioblastoma stem-like cell (GSC) survival but its mode of action is unclear. Understanding the role of PFKFB4 in tumor cell survival could allow it to be leveraged in a cancer therapy. Here, we show the importance of PFKFB4 for glioblastoma growth in vivo in an orthotopic patient derived mouse model. In an evaluation of patient tumor samples of different cancer entities, PFKFB4 protein was found to be overexpressed in prostate, lung, colon, mammary and squamous cell carcinoma, with expression level correlating with tumor grade. Gene expression profiling in PFKFB4-silenced GSCs revealed a downregulation of hypoxia related genes and Western blot analysis confirmed a dramatic reduction of HIF (hypoxia inducible factor) protein levels. Through mass spectrometric analysis of immunoprecipitated PFKFB4, we identified the ubiquitin E3 ligase, F-box only protein 28 (FBXO28), as a new interaction partner of PFKFB4. We show that PFKFB4 regulates the ubiquitylation and subsequent proteasomal degradation of HIF-1α, which is mediated by the ubiquitin ligase activity of FBXO28. This newly discovered function of PFKFB4, coupled with its cancer specificity, provides a new strategy for inhibiting HIF-1α in cancer cells. ispartof: ONCOGENESIS vol:11 issue:1 ispartof: location:United States status: published

Glioblastoma frequently exhibits therapy-associated subtype transitions to mesenchymal phenotypes with adverse prognosis. Here, we perform multi-omic profiling of 60 glioblastoma primary tumours and use orthogonal analysis of chromatin and RNA-derived gene regulatory networks to identify 38 subtype master regulators, whose cell population-specific activities we further map in published single-cell RNA sequencing data. These analyses identify the oligodendrocyte precursor marker and chromatin modifier SOX10 as a master regulator in RTK I-subtype tumours. In vitro functional studies demonstrate that SOX10 loss causes a subtype switch analogous to the proneural–mesenchymal transition observed in patients at the transcriptomic, epigenetic and phenotypic levels. SOX10 repression in an in vivo syngeneic graft glioblastoma mouse model results in increased tumour invasion, immune cell infiltration and significantly reduced survival, reminiscent of progressive human glioblastoma. These results identify SOX10 as a bona fide master regulator of the RTK I subtype, with both tumour cell-intrinsic and microenvironmental effects., Glioblastoma is divided into four subtypes based on molecular profiling at the methylome and transcriptome level. Here the authors perform an integrative analysis of these subtypes resulting in the identification of SOX10 whose loss induces a mesenchymal phenotype and promotes tumour progression.

Background Glioblastoma is the most common primary malignancy of the central nervous system with a dismal prognosis. Genomic signatures classify isocitrate dehydrogenase 1 (IDH)-wildtype glioblastoma into three subtypes: proneural, mesenchymal, and classical. Dasatinib, an inhibitor of proto-oncogene kinase Src (SRC), is one of many therapeutics which, despite promising preclinical results, have failed to improve overall survival in glioblastoma patients in clinical trials. We examined whether glioblastoma subtypes differ in their response to dasatinib and could hence be evaluated for patient enrichment strategies in clinical trials. Methods We carried out in silico analyses on glioblastoma gene expression (TCGA) and single-cell RNA-Seq data. In addition, in vitro experiments using glioblastoma stem-like cells (GSCs) derived from primary patient tumors were performed, with complementary gene expression profiling and immunohistochemistry analysis of tumor samples. Results Patients with the mesenchymal subtype of glioblastoma showed higher SRC pathway activation based on gene expression profiling. Accordingly, mesenchymal GSCs were more sensitive to SRC inhibition by dasatinib compared to proneural and classical GSCs. Notably, SRC phosphorylation status did not predict response to dasatinib treatment. Furthermore, serpin peptidase inhibitor clade H member 1 (SERPINH1), a collagen-related heat-shock protein associated with cancer progression, was shown to correlate with dasatinib response and with the mesenchymal subtype. Conclusion This work highlights further molecular-based patient selection strategies in clinical trials and suggests the mesenchymal subtype as well as SERPINH1 to be associated with response to dasatinib. Our findings indicate that stratification based on gene expression subtyping should be considered in future dasatinib trials.

The evolutionary processes that drive universal therapeutic resistance in adult patients with diffuse glioma remain unclear1,2. Here we analysed temporally separated DNA-sequencing data and matched clinical annotation from 222 adult patients with glioma. By analysing mutations and copy numbers across the three major subtypes of diffuse glioma, we found that driver genes detected at the initial stage of disease were retained at recurrence, whereas there was little evidence of recurrence-specific gene alterations. Treatment with alkylating agents resulted in a hypermutator phenotype at different rates across the glioma subtypes, and hypermutation was not associated with differences in overall survival. Acquired aneuploidy was frequently detected in recurrent gliomas and was characterized by IDH mutation but without co-deletion of chromosome arms 1p/19q, and further converged with acquired alterations in the cell cycle and poor outcomes. The clonal architecture of each tumour remained similar over time, but the presence of subclonal selection was associated with decreased survival. Finally, there were no differences in the levels of immunoediting between initial and recurrent gliomas. Collectively, our results suggest that the strongest selective pressures occur during early glioma development and that current therapies shape this evolution in a largely stochastic manner.

BACKGROUND Glioblastoma is the most common primary malignancy of the central nervous system with a dismal prognosis, even with surgical and chemoradiotherapy. Expression profiling studies classify IDH-wildtype Glioblastoma into three subtypes: Proneural (PN), mesenchymal (MES) and classical (CL). A promising target to inhibit in Glioblastoma is the non-receptor tyrosine kinase and proto-oncogene SRC. After robust pre-clinical results, SRC inhibitors like dasatinib did not improve survival of Glioblastoma patients after recurrence in clinical trials. MATERIAL AND METHODS Consolidating efforts to personalize cancer therapy, we use in silico analyses backed by in vitro and in vivo experiments on Glioblastoma stem-like cells (GSCs) derived from primary patient tumors to present a novel stratification strategy for dasatinib therapy in glioblastoma. To further tackle dasatinib resistance in GSCs, a pooled shRNA library against 5000 genes was combined with dasatinib to identify genes whose knockdown sensitizes GSCs to dasatinib. This was integrated with proteomics and phosphoproteomics data of dasatinib inhibited GSCs. RESULTS We found MES tumors with high expression of SERPINH1 to be sensitive to dasatinib inhibition, compared to the CL and PN subtypes. Interestingly, SRC phosphorylation status did not predict the efficacy of dasatinib inhibition. Computational analyses integrating data from the loss-of-function dropout viability screen and proteomics/phosphoproteomics using a novel modification of the SamNet algorithm identified Wee1, a tyrosine kinase involved in cell-cycle signaling, as a potential combination inhibition target with dasatinib. Further validation experiments showed a robust synergistic effect through combination of dasatinib and the wee1 inhibitor, MK-1775 in PN GSCs. CONCLUSION This study highlights strategies to optimize dasatinib treatment in different glioblastoma subtypes. While the stratification of patients harboring mesenchymal glioblastoma with SERPINH1 overexpression could provide an option in this particular subtype, combining dasatinib or other SRC inhibitors with Wee1 inhibitors could present an additional possibility for treating resistant proneural tumors

Significance How species differ from each other is a key question in biology. However, genetic mapping between species often fails because of sterile hybrid crosses. Here, we have developed a technique called in vitro recombination to circumvent breeding. We induced genetic reshuffling through mitotic recombination with the drug ML216 and mapped trait variations in a dish. Starting with hybrid embryonic stem cells between the Mus musculus laboratory mouse and Mus spretus spanning ∼1.5 million years of divergence, we show that it is possible to map the gene responsible for differential resistance to the drug tioguanine in as few as 21 days. Our technique opens up experimental avenues in genetic mapping of various traits and diseases across mouse species., Discovering the genetic changes underlying species differences is a central goal in evolutionary genetics. However, hybrid crosses between species in mammals often suffer from hybrid sterility, greatly complicating genetic mapping of trait variation across species. Here, we describe a simple, robust, and transgene-free technique to generate “in vitro crosses” in hybrid mouse embryonic stem (ES) cells by inducing random mitotic cross-overs with the drug ML216, which inhibits the DNA helicase Bloom syndrome (BLM). Starting with an interspecific F1 hybrid ES cell line between the Mus musculus laboratory mouse and Mus spretus (∼1.5 million years of divergence), we mapped the genetic basis of drug resistance to the antimetabolite tioguanine to a single region containing hypoxanthine–guanine phosphoribosyltransferase (Hprt) in as few as 21 d through “flow mapping” by coupling in vitro crosses with fluorescence-activated cell sorting (FACS). We also show how our platform can enable direct study of developmental variation by rederiving embryos with contribution from the recombinant ES cell lines. We demonstrate how in vitro crosses can overcome major bottlenecks in mouse complex trait genetics and address fundamental questions in evolutionary biology that are otherwise intractable through traditional breeding due to high cost, small litter sizes, and/or hybrid sterility. In doing so, we describe an experimental platform toward studying evolutionary systems biology in mouse and potentially in human and other mammals, including cross-species hybrids.

Discovering the genetic changes underlying species differences is a central goal in evolutionary genetics. However, hybrid crosses between species in mammals often suffer from hybrid sterility, greatly complicating genetic dissection of trait variation. Here we describe a simple, robust and transgene-free technique to make “in vitrocrosses” in hybrid mouse embryonic stem cells by inducing random mitotic crossovers with the drug ML216, which inhibitsBloom syndrome(BLM). Starting with an interspecific hybrid (betweenMus musculusandMus spretus) embryonic stem cell line spanning 1.5 million years of divergence, we demonstrate the feasibility of mapping enzymatic differences across species within weeks and the possibility of re-deriving whole mice. Our work shows howin vitrocrosses can overcome major bottlenecks like hybrid sterility in traditional mouse breeding to address fundamental questions in evolutionary biology.Impact StatementBy mixing hybrid mouse genomes in stem cells via mitotic recombination, genetic mapping and hybrid mosaic mice can be achieved in weeks, even across species barriers.

Although the survival of patients diagnosed with breast cancer has greatly improved over the last decades, the metastatic stage of the disease still remains incurable. Blood circulating tumor cells (CTCs) are assumed to be the source for metastatic onset and the enumeration of CTCs has already proven its prognostic value for survival in metastatic breast cancer (mBC) patients. However, the biological profiles of CTCs are so far not considered in therapy selection, but hold the potential to display the disease stage in a minimally invasive procedure. Here we serially collect liquid biopsies of mBC patients throughout their course of therapy with the CDK4/6 inhibitor palbociclib to monitor treatment response and drug resistance development. To this end, we combine transcriptomic, genomic and proteomic analyses of primary CTCs on bulk but also on the single cell level. Furthermore, we generated patient-specific, in vitro expanded CTC cell lines to intensively study mechanisms of drug resistance and identify new therapeutic targets by drug sensitivity assays. By analyzing mBC CTCs of serial liquid biopsies we monitored cell cycle arrest upon palbociclib treatment and drug resistance development. Drug sensitivity and successive drug resistance can be observed in primary and in vitro expanded, patient-specific CTCs. As revealed by RNA-seq of bulk and single CTCs, acquired resistance to palbociclib in mBC CTCs is associated with altered transcriptional programs including RB transcriptional corepressor 1 (RB1) function and can be linked to a loss-of-function mutation in the RB1 gene. Interestingly, apart from altered RB1 function, we can identify a global transcriptional transition from palbociclib sensitivity to resistance. Based on the transcriptomic information, pathways that are upregulated in resistant CTCs were determined and the patient-derived CTC cell lines were used to validate and specifically target tumor cells that are resistant to CDK4/6 inhibitors. Using drug screening assays on the CTC-derived cell lines, we identified potential drugs that in combination with palbociclib could circumvent the acquired resistance and help to prolong patient survival. Our study indicates that information about the biological state of serially collected CTCs can help to earlier detect the onset of resistance and should be included in clinical disease management. This will help to spare the patient severe side effects from ineffective drug treatment. Citation Format: Andreas Trumpp, Lisa Becker, Michael Fletcher, Roberto Würth, Anand Thippeswamy, Simon Haas, Lars Velten, Laura Michel, Frederik Marme, Sarah Jauch, Mirjam Becker, Lars Steinmetz, Andreas Schneeweiss, Marc Zapatka, Peter Lichter, Martin Sprick. Exploring CDK4/6 inhibitor therapy response and drug resistance development at the single cell level in metastatic breast cancer CTCs [abstract]. In: Proceedings of the 2019 San Antonio Breast Cancer Symposium; 2019 Dec 10-14; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2020;80(4 Suppl):Abstract nr PD2-01.

We studied how intratumoral genetic heterogeneity shapes tumor growth and therapy response for isocitrate dehydrogenase (IDH)-wild-type glioblastoma, a rapidly regrowing tumor. We inferred the evolutionary trajectories of matched pairs of primary and relapsed tumors based on deep whole-genome-sequencing data. This analysis suggests both a distant origin of de novo glioblastoma, up to 7 years before diagnosis, and a common path of early tumorigenesis, with one or more of chromosome 7 gain, 9p loss, or 10 loss, at tumor initiation. TERT promoter mutations often occurred later as a prerequisite for rapid growth. In contrast to this common early path, relapsed tumors acquired no stereotypical pattern of mutations and typically regrew from oligoclonal origins, suggesting sparse selective pressure by therapeutic measures.

Summary The fibroblast growth factor receptor 2 (FGFR2) locus is the ‘top hit’ in genome-wide association studies for breast cancer. Here, we examine the effect of FGFR2 signalling on transcriptional networks in breast cancer and propose a mechanism for FGFR2 risk single-nucleotide polymorphism function., The fibroblast growth factor receptor 2 (FGFR2) locus is consistently the top hit in genome-wide association studies for oestrogen receptor-positive (ER+) breast cancer. Yet, its mode of action continues to be controversial. Here, we employ a systems biology approach to demonstrate that signalling via FGFR2 counteracts cell activation by oestrogen. In the presence of oestrogen, the oestrogen receptor (ESR1) regulon (set of ESR1 target genes) is in an active state. However, signalling by FGFR2 is able to reverse the activity of the ESR1 regulon. This effect is seen in multiple distinct FGFR2 signalling model systems, across multiple cells lines and is dependent on the presence of FGFR2. Increased oestrogen exposure has long been associated with an increased risk of breast cancer. We therefore hypothesized that risk variants should reduce FGFR2 expression and subsequent signalling. Indeed, transient transfection experiments assaying the three independent variants of the FGFR2 risk locus (rs2981578, rs35054928 and rs45631563) in their normal chromosomal context show that these single-nucleotide polymorphisms (SNPs) map to transcriptional silencer elements and that, compared with wild type, the risk alleles augment silencer activity. The presence of risk variants results in lower FGFR2 expression and increased oestrogen responsiveness. We thus propose a molecular mechanism by which FGFR2 can confer increased breast cancer risk that is consistent with oestrogen exposure as a major driver of breast cancer risk. Our findings may have implications for the clinical use of FGFR2 inhibitors.

ObjectivesBarbecuing and smoked meat continues to grow in popularity for food service and consumers at home. However, little research has examined the eating quality differences of point (pectoralis superficialis) and flat (pectoralis profundi) muscles across USDA quality grade. The objective of this study was to investigate differences in smoked beef brisket palatability from three USDA quality grades.Materials and MethodsBeef briskets from the USDA Prime, Average Choice, and Select quality grades (n = 54; 18 per treatment) were collected at a commercial abattoir in Omaha, NE. Briskets were trimmed to 6 mm of external fat, seasoned with a blend of 1:1 coarse kosher salt/coarse black pepper by hand (0.05% of the brisket raw weight), and were held at 2–4° for 12 h prior to cooking. Briskets were cooked in an electric pellet smoker utilizing Gold Blend Hardwood Pellets (red oak, hickory, and maple wood) for ∼4 h to an internal temperature of 63°C; wrapped in aluminum foil, placed back in the smoker for ∼4 h, and cooked to 93°C, then held in an insulated cooler until slicing. Approximately 90 min prior to serving, briskets were separated in point and flat portions, and then sliced (6 mm × 50 mm x cooked depth) perpendicular to the muscle fiber for consumer evaluation and held in warmers at (∼50°C) until serving. Each consumer (n = 360) received six test samples representing all quality grade × muscle combinations to evaluate tenderness, juiciness, flavor liking, overall liking, as well as the acceptability of these traits. Additionally, willingness to pay (WTP) was collected on an individual sample basis.ResultsAn interaction between quality grade and muscle was observed (P ≤ 0.03) for all palatability traits, proportion of acceptable samples, and WTP. Consumers could not distinguish between quality grades of the point portions for tenderness, juiciness, flavor and overall liking (P > 0.05). Point samples, regardless of quality grade were scored greater than Prime flat samples, which were intermediate (P < 0.05). Consumers similarly (P > 0.05) scored Choice and Select flat samples lower for all palatability traits compared to all other treatment combinations. In alignment with palatability traits, consumers were willing to pay the most for point portions, regardless of quality grade (P < 0.05). Consumers WTP of the Prime flat portion was intermediate, and consumers were willing to pay the least for Choice and Select flat portions (P < 0.05).Consumer acceptability followed similar trends as palatability scores. However, a greater proportion of consumers classified Choice and Select point samples as acceptable than that of Prime point samples in all categories of acceptability (P < 0.05). Consumers struggled to distinguish differences in acceptability for Choice and Select flat portions (P > 0.05) in all factors except juiciness acceptability.ConclusionQuality grade had no effect on the eating quality of the point portions of smoked briskets, and point portions received superior palatability scores to flat portions. Briskets from the Prime flat portions had greater eating quality than Choice and Select briskets from the flat portion, and consumers were willing to pay more for what they perceived as superior eating quality. This data suggests that unless consumers prefer the flat portion of the brisket there is no benefit to paying the premium for a prime brisket from a palatability standpoint.

SUMMARY The clinical features and family histories of 20 adults with dyskinetic cerebral palsy from 20 families were studied. The majority of the patients showed progressive neurological deterioration in adult life. In only three did the condition stabilise by 10 years of âge and in seven there was deterioration after the âge of 30. Two patients developed a secondary cervical spondylotic myelopathy. Four patients had affected relatives and there were similar proportions of affected parents and siblings. The family data suggest genetic heterogeneity with autosomal recessive and dominant variants. The existence of an X-linked form cannot be excluded, and the demonstration of an increased paternal âge effect among single cases suggests that some of these may arise because of fresh dominant genetic mutation. RESUME IMC dyskinetique: une Aetude clinique et genetique Les caracteristiques cliniques et l'anamnese familiale de 20 IMC adultes dyskinetiques provenant de 20 families ont eteetudees. Une deterioration neurologique progressive durant la vie adulte fut constated chez la majority des patients. La stabilisation apres l'âge de 10 ans ne fut observed que trois fois, la deterioration apres l'âge de 30 ans sept fois. Une myelopathie par spondylose cervicale secondaire apparait chez deux patients. On notait des cas familiaux chez quatre patients, avec une proportion identique chez les parents et dans la fratrie. Les donnees familiales suggerent une heterogeneite genetique avec une variation autosomale recessive et dominante. L'existence d'une forme liee a l'X ne peut etre exclue et la mise en evidence d'un effet d'âge paternel accru parmi les cas isoles suggere un role partiel de mutations genetiques dominantes recentes. ZUSAMMENFASSUNG Dyskinetische Cerebralparese: Eine klinische und genetische Studie Es wurden die klinischen Merkmale und Familienanamnesen von 20 Erwachsenen mit dyskinetischer Cerebralparese aus 20 Familien untersucht. Die Mehrzahl der Patienten zeigte eine progressive neurologische Verschlechterung im Erwachsenenalter. Nur bei drei Patienten stabilisierte sich der Zustand mit 10 Jahren und bei sieben verschlechterte er sich nach dem 30ten Lebensjahr. Zwei Patienten entwickelten eine sekundare cervikale Spondylo-Myelopathie. Vier Patienten hatten ebenfalls betroffene Verwandte und es fanden sich ahnliche Verhaltnisse bei betroffenen Eltern und Geschwistern. Die Familiendaten lassen eine Heterogenic mit autosomal rezessiven oder dominaten Varianten vermuten. Eine X-gebundene Form kann nicht ausgeschlossen werden und der Nachweis, das in einigen Fallen ein erhohtes Alter der Eltern eine Rolle spielt, laBt vermuten, das einige Falle durch neu entstandene dominante genetische Mutationen entstanden sind. RESUMEN Paralisis cerebral disqinetica: estudio clfnico y genetico Se estudiaron los caracteres clinicos y las historias familiares de 20 adultos con paralisis cerebral disquinetica procedentes de 20 familias. La mayoria de los pacientes mostraron una deteriorizacion neurologica progresiva al llegar a adultos. Solamente en tres la situacion se estabilizo a los 10 afios de edad y en siete hubo una deteriorizacion despues de los 30. Dos pacientes desarrollaron secundariamente una mielopatia espondilotica cervical. Cuatro pacientes tenfan familiares afectados y habfa una proporcione similar de padres y hermanos afectados. Los datos familiares sugerian una heterogeneidad genetica, con variantes autosomicas recesivas y dominantes. La existencia de una forma ligada al cromosoma X no puede ser excluida y la demostracion de una edad paterna aumentada en los casos aislados, sugiere que algunos de ellos pueden deberse a una mutacion dominante reciente.

The application of multivariate statistical projection based techniques has been recognized as one approach to contributing to an increased understanding of process behaviour. The key methodologies have included multi-way principal component analysis (PCA), multi-way partial least squares (PLS) and batch observation level analysis. Batch processes typically exhibit nonlinear, time variant behaviour and these characteristics challenge the aforementioned techniques. To address these challenges, dynamic PLS has been proposed to capture the process dynamics. Likewise approaches to removing the process nonlinearities have included the removal of the mean trajectory and the application of nonlinear PLS. An alternative approach is described whereby the batch trajectories are sub-divided into operating regions with a linear/linear dynamic model being fitted to each region. These individual models are spliced together to provide an overall nonlinear global model. Such a structure provides the potential for an alternative approach to batch process performance monitoring. In the paper a number of techniques are considered for developing the local model, including multi-way PLS and dynamic multi-way PLS. Utilising the most promising set of results from a simulation study of a batch process, the local model comprising individual linear dynamic PLS models was benchmarked against global nonlinear dynamic PLS using data from an industrial batch fermentation process. In conclusion the results for the local operating region techniques were comparable to the global model in terms of the residual sum of squares but for the global model structure was evident in the residuals. Consequently, the local modelling approach is statistically more robust. L'application de techniques basees sur la projection statistique multivariee est reconnue comme etant une approche qui contribue a une meilleure comprehension du comportement des procedes. Les methodologies cles incluent l'analyse des composantes principales (PCA) a plusieurs criteres de classification, les moindres carres partiels (PLS) a plusieurs criteres de classification et l'analyse des niveaux d'observation discontinus. Les procedes discontinus presentent typiquement un comportement non lineaire et variable dans le temps et ces caracteristiques mettent au defi les techniques mentionnees ci-dessus. Devant ces defis, la methode PLS dynamique est proposee pour saisir la dynamique des procedes. Des approches semblables pour supprimer la non linearite des procedes incluent le retrait de la trajectoire principale et l'application des PLS non lineaires. On decrit une autre approche ou les trajectoires discontinues sont subdivisees en regions operatoires avec un modele dynamique lineaire/lineaire adapte a chaque region. Ces modeles individuels sont raccordes pour obtenir un modele non lineaire global. Une telle structure presente un potentiel pour une approche differente du suivi des performances des procedes discontinus. Dans cet article, plusieurs techniques sont considerees pour la mise au point du modele local, incluant les PLS a plusieurs criteres de classification et les PLS a plusieurs criteres de classification dynamique. En utilisant la serie de resultats les plus prometteurs d'une etude de simulation d'un procede discontinu, le modele local comprenant les modeles de PLS dynamiques lineaires individuels a ete compare a la methode de PLS non lineaires dynamique globale utilisant des donnees d'un procede de fermentation discontinu industriel. En conclusion, les resultats pour les techniques des regions operatoires locales sont comparables au modele global en termes de somme des carres des residus mais pour le modele global, la presence d'une structure dans les residus est evidente. En consequence, l'approche de modelisation locale est statistiquement plus robuste.

This paper reports on a study of the statistical properties of single photons emitted by a nitrogen vacancy centre using a confocal microscope and Hanbury-Brown and Twiss interferometer. High signal-to-noise photon correlation histograms from a single NV centre have been recorded for various pump laser powers. The form of these signals can be described by modelling the nitrogen vacancy as a three-level system. The parameters that characterize the model have been measured as a function of pump laser power.

Sterile painted apple moths have been released in Auckland in an attempt to assist with the eradication of this species from New Zealand Quality as measured by wind tunnel flight performance of irradiated insects decreased from 66 of unirradiated males in 20032004 to approximately 43 in 20052006 and recapture rates were lower A decrease in quality measures (ie emergence rate wind tunnel flight performance and recapture rate) was related to an increase in the length of pre and posttreatment time that the insects were held in the irradiation container potentially caused by container conditions Altering the irradiation container conditions to improve aeration led to an increase in posttreatment quality The surveillance grid in place for monitoring caught the sterile moths Higher recapture rates have increased confidence in the effectiveness of the sterile insect technique against this insect which is continuing in southeastern Auckland

$\ensuremath{\alpha}$-particle decays from several excited states in ${}^{11}\mathrm{C}$, ${}^{13}\mathrm{C}$, and ${}^{15}\mathrm{N}$ have been observed in three-body final state reactions induced by heavy-ion beams at 6.5--9.0 MeV/nucleon from the FSU Tandem/LINAC. Where possible, branching fraction estimates are made. Improvements in the energy resolution of the detection system have allowed the observation of many previously unreported $\ensuremath{\alpha}$-particle decays, several new total energy width determinations, and a few new excited state energies.

A new method proposed for the elemental analysis of liquid organic samples has been used in this pilot investigation to determine the concentration of several non-volatile light elements contained in human amniotic fluid (AF) from a single measurement. The technique is based on Rutherford forward scattering (RFS) at 28° of a 16 MeV 7 Li beam from vacuum dried AF samples deposited on a formvar support. The risk of contamination of the AF sample due to its manipulation is minimum since it is not required to treat the sample chemically. Statistically significant yields were obtained for beam currents of 5 nA and irradiation periods of 30 min approximately. An energy calibration with the spectrum obtained from the irradiation of an AF target was carried out to take into account any effect due to the AF organic matrix. AF spectra were quantified by a chi square fitting of Gaussian curves to element signals. According to this study, the RFS method may be considered as a potential method for light trace element analysis of non-volatile fractions where standard techniques such as PIXE/PIGE and TXRF are difficult to apply.

The fibroblast growth factor receptor 2 (FGFR2) locus has been consistently identified as a breast cancer risk locus in independent genome-wide association studies. However, the molecular mechanisms underlying FGFR2-mediated risk are still unknown. Using model systems we show that FGFR2-regulated genes are preferentially linked to breast cancer risk loci in expression quantitative trait loci analysis, supporting the concept that risk genes cluster in pathways. Using a network derived from 2,000 transcriptional profiles we identify SPDEF, ERα, FOXA1, GATA3 and PTTG1 as master regulators of fibroblast growth factor receptor 2 signalling, and show that ERα occupancy responds to fibroblast growth factor receptor 2 signalling. Our results indicate that ERα, FOXA1 and GATA3 contribute to the regulation of breast cancer susceptibility genes, which is consistent with the effects of anti-oestrogen treatment in breast cancer prevention, and suggest that fibroblast growth factor receptor 2 signalling has an important role in mediating breast cancer risk., FGFR2 gene variation is associated with breast cancer risk but the molecular mechanism is unknown. Fletcher et al. provide a link between FGFR2 signalling and breast cancer susceptibility by demonstrating that FGFR2 signalling activates the ERa transcriptional network, which drives transcription of risk genes.

Recent publications have proposed that adult glioblastoma (GBM) can be classified into four methylation subtypes; however, the underlying biological differences between these subtypes remains unclear. Our project has produced a rich set of genomic data (WGBS, strand-specific RNAseq, WES, histone mark ChIPseq) for 48 adult GBMs that is now being analysed to further characterise these four subtypes. We have established a transcriptome assembly pipeline using StringTie, followed by differential gene expression analysis with limma to identify novel subtype-specific transcripts. The pipeline finds 14209 novel transcripts in 6631 genes, 90% of which have not been previously annotated. The majority of these transcripts appear to be non-coding, with a subset predicted as containing protein domains. The subtype-specific novel transcripts identified by limma are supported by subtype-specific enrichment of histone modifications. A selection of candidate novel transcripts will be further characterised in silico and, potentially, in cell line models.

Hepatitis C virus (HCV) is an enveloped, positive-strand RNA virus of the family Flaviviridae that primarily infects hepatocytes, causing acute and chronic liver disease. HCV is also associated with a variety of extrahepatic symptoms including central nervous system (CNS) abnormalities, cognitive dysfunction, fatigue and depression. These symptoms do not correlate with the severity of liver disease and are independent of hepatic encephalopathy. HCV RNA has been associated with CNS tissue, and reports of viral sequence diversity between brain and liver tissue suggest independent viral evolution in the CNS and liver. This review will explore the data supporting HCV infection of the CNS and how this fits into our current understanding of HCV pathogenesis.

Parkinson’s disease (PD) is a neurodegenerative disorder that results in the loss of nigrostriatal dopamine neurons. The etiology of this cell loss is unknown, but it involves abnormalities in mitochondrial function. In this study, we have demonstrated that the administration of a novel noncoding p137 RNA, derived from the human cytomegaloviral β2.7 transcript, can prevent and rescue dopaminergic cell death in vitro and in animal models of PD by protecting mitochondrial Complex I activity. Furthermore, as this p137 RNA is fused to a rabies virus glycoprotein peptide that facilitates delivery of RNA across the blood–brain barrier, such protection can be achieved through a peripheral intravenous administration of this agent after the initiation of a dopaminergic lesion. This approach has major implications for the potential treatment of PD, especially given that this novel agent could have the same protective effect on all diseased neurons affected as part of this disease process, not just the dopaminergic nigrostriatal pathway.

The differential cross section for the primordial reaction [sup 8]Li([ital p],[ital n])[sup 8]Be(g.s.) has been determined at [ital E][sub c.m.]=1.5 MeV in the c.m. angular range 38[degree] to 90[degree] by use of the radioactive nuclear beam facility at the University of Notre Dame which utilizes a superconducting solenoid lens system designed at the University of Michigan, and a [sup 8]Be detection technique developed at the Florida State University. A [sup 8]Li beam of approximately 10[sup 7] pps is used to infer a total cross section of 21[plus minus]2 mb, with a 20% absolute cross section uncertainty. This total cross section is a factor of two smaller than a conventional Hauser-Feshbach calculation and it is also much smaller than that of some other [sup 8]Li burning reactions.

Generalised, multifocal or segmental idiopathic torsion dystonia (ITD), is caused by an autosomal dominant gene with reduced penetrance in about 85% of cases. Of 104 patients with these types of ITD, 17 (16.4%) gave a history which suggested that dystonic movements had been precipitated or exacerbated by trauma. Eight of these 17 patients had affected relatives. If precipitated, dystonia appeared first in the injured part of the body within days or up to 12 months after the trauma and later became more widespread. Peripheral injuries may influence basal ganglia function and provoke the onset of dystonic movements in individuals who are ITD gene carriers.

A genetic study of idiopathic focal dystonias was undertaken by examining 153 first-degree relatives of 40 index patients with torticollis (14 patients), other focal cranial dystonias (16 patients), and writer's cramp (10 patients). Nine relatives with dystonia were identified in 6 families; 8 of these had symptoms such as clumsiness or tremor, but none were aware of any dystonia. A further 4 relatives, now decreased, were affected by history. Overall, 25% of index patients had relatives with dystonia. The results of segregation analysis suggested the presence of an autosomal dominant gene or genes with reduced penetrance as a common cause for focal dystonia. Segregation ratios were not significantly different from those ratios observed in generalized or segmental dystonia in the United Kingdom, and it is possible that a single autosomal dominant gene mutation is responsible for inherited dystonia in the majority of patients irrespective of distribution or severity.

For most patients presenting with a spinal cord syndrome MR scanning has become the key investigation in establishing the diagnosis. However, myelopathy with normal spinal imaging remains a common clinical conundrum. In this review we discuss the diagnoses to consider for the neurologist presented with a patient with "MR normal myelopathy". We will illustrate this scenario with a series of short cases and consider the further investigation of "MRI normal" myelopathy.

Two patients fulfilling suggested clinical diagnostic criteria for corticobasal degeneration (CBD) are presented, who were found at postmortem to have alternative pathological diagnoses not suspected during life, namely, Alzheimer's disease and Pick's disease, respectively. The nosological position of these cases is considered in light of a literature review of previous reports of clinically diagnosed corticobasal degeneration with atypical (not corticobasal degeneration) pathology. Since such phenocopies may be common, we suggest that all clinically diagnosed cases of corticobasal degeneration should initially be labelled as “corticobasal degeneration syndrome” (CBDS) to emphasize that this is a diagnosis based on clinical phenotype, with the term corticobasal degeneration being reserved for the specific neuropathological phenotype, which itself may have a variety of clinical presentations.

A project based on low mass multielemental analysis of liquid samples by using forward elastic scattering is under progress. Elastic cross section measurements for the reactions 13 MeV 6,7Li +A and 24 MeV 16O +A, A being any nucleus lighter than Na, are necessary due to the existence of non‐elastic processes. The non‐Rutherford character of some of the reactions quoted above has been established from experiments carried out at 12.45°, 16.45°, 20.45° and 28.0° where A = Be, C, O and Si. The accuracy and precision of the measured cross sections have been tested. In the future, accurate cross section determinations will be required for lighter target elements such as B, Li and H. The dilution of a liquid sample improves the spectrum resolution. Efforts are currently concentrated on determining if the relative yields corresponding to various elements present in a given sample remain constant as dilution changes.

As culturally unique individuals, our culture impacts and even dictates our responses to normal events of daily living. We are frequently unaware of the way we value time, perceive spatial boundaries, and communicate with others. Health care professionals must be culturally sensitive to all individuals. Culturally diverse nursing care acknowledges the individual's cultural uniqueness and incorporates a variety of approaches to provide culturally sensitive care. This paper will discuss culturally diverse health care with a client experiencing grief and loss.

Free rotational aerophones such as the bullroarer, which consists of a wooden slat whirled around on the end of a string, and which emits a loud pulsating roar, have been used in many ancient and traditional societies for ceremonial purposes. This article presents an experimental and theoretical investigation of this instrument. The aerodynamics of rotational behavior is elucidated, and relates slat rotation frequency to slat width and velocity through the air. Analysis shows that sound production is due to generation of an oscillating-rotating dipole across the slat, the role of the vortices shed by the slat being relatively minor. Apparent discrepancies between the behavior of a bullroarer slat and a slat mounted on an axle in a wind tunnel are shown to be due to viscous friction in the bearings of the wind-tunnel experiment.

Six years after the publication of the first breast cancer GWAS, skepticism is expressed about the usefulness of the results. The GWAS loci identified to date explain only a minority of the estimated genetic variance, and the risks conferred by even the strongest individual variants are small. Classical biochemical pathway approaches to elucidating mechanism have had modest success. I will argue that an attempt to examine the combined effects by common and rare variants and their interactions through analysis based on gene regulatory networks may yield new and useful information. Although still speculative, I will present initial data that provide support for this approach. Citation Format: Bruce A. J. Ponder, Kerstin Meyer, Michael Fletcher, Florian Markowetz, Ines de Santiago, Mauro Castro. Gene regulatory network approaches to interpreting breast cancer GWAS results. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Susceptibility and Cancer Susceptibility Syndromes; Jan 29-Feb 1, 2014; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(23 Suppl):Abstract nr IA01. doi:10.1158/1538-7445.CANSUSC14-IA01

For most insects, size determines the call frequency. This paper describes the acoustics of a small brachypterous cricket (Rufocephalus sp.; body length 9.6 mm) producing a call with a carrier frequency of approximately 3.2 kHz from a subterranean burrow. Crickets such as Gryllus campestris are approximately twice this length and produce a call frequency close to 5 kHz. The burrow of Rufocephalus opens via a small hole with a diameter of 3.2 mm. The neck of the hole at approximately 1.4 mm depth opens to a vertical two-part burrow with an upper vase-shaped chamber 16.1 mm in height with a diameter of 9.4 mm. This top chamber connects via a 6.4mm high (diameter 5.2 mm) neck to a more irregular chamber approximately 18 mm high with a width of approximately 11 mm. The walls of the top chamber neck and of the upper part of the lower chamber are smooth and appear to be sealed with saliva. The song has a mean centre frequency of 3.2 kHz and is made up of variable-length trills of pulses of mean duration 15.8 ms. Many song pulses had smooth envelopes and their frequency did not vary by more than +/-40Hz from the centre frequency, with a relative bandwidth Q-3dB of over 50. Other pulses showed considerable amplitude and frequency modulation within the pulse. When driven by external sound, burrows resonated at a mean frequency of 3.5 kHz with a mean quality factor Q of 7.4. Natural-size model burrows resonated at similar frequencies with similar Q values. One cricket, which had previously called from its own burrow at 2.95 kHz, sang at 3.27 kHz from a burrow that resonated at the same frequency. Life-size model burrows driven by external sound resonated at similar frequencies to the actual burrows; models three times life size resonated at one-third of this frequency. In all models, the sound pressure was more-or-less constant throughout the top chamber but fell rapidly in the neck of the burrow; the phase of the sound was effectively constant in the top chamber and neck and fell through approximately 180 degrees in passing from the neck into the lower chamber. A numerical model of the sound flow from region to region gave essentially similar results. A resonant electrical model fed from a high-impedance source with discrete tone bursts at different frequencies showed similar amplitude and frequency modulation to the various types of song pulses that were observed. It is suggested that the high purity of the songs results from close entrainment of the sound-producing mechanism of the insect's wings to the sharply resonant burrow.

Multielemental analysis of evaporated liquid samples is possible by irradiating the samples with a 16 MeV 7Li beam and detecting the elastically scattered ions at 28°. The method is easily applied when Rutherford scattering dominates. To prepare the targets, the liquid sample is deposited on a formvar backing and dried with vacuum. Preliminary results indicate a possible relationship between sample concentration, uniformity and spectrum energy resolution. Details on the spectrum analysis will be given. The method is mainly useful for multielemental quantification in the low mass region from lithium to fluorine where standard techniques such as PIXE and TXRF are useless.

A family is described in which dopa-responsive dystonia affected six members and segregated in an autosomal dominant fashion. Patients either presented in childhood with dystonia of the legs, going to develop parkinsonism and pseudo-pyramidal deficits, or in adult life with parkinsonian tremor and rigidity, with pseudo-pyramidal signs. Remarkably, in the three cases with childhood onset the symptoms and signs of the condition were abolished 36 to 52 years later by small doses of levodopa. No long term side effects of levodopa have appeared after 15 years of treatment.

Idiopathic torsion dystonia (ITD) is a dominantly inherited disorder with variable penetrance and expressivity. Factors affecting the penetrance of the ITD gene have not yet been identified. The present study used four published series of cases to test specific hypotheses regarding factors that could affect the expression of ITD. Among the combined 253 families, transmission of ITD did not depend on either the sex of the affected offspring or that of the transmitting parent. Furthermore, neither the specific type of dystonia manifested, the site at which clinical signs of dystonia first appeared, nor age of onset differed significantly as a function of the gender of the transmitting parent. However, in familial cases of later onset (ageor = 20 years), nearly all involved a transmitting mother. There is evidence for consistency of age of onset within the subset of Jewish families. Although anticipation effects are apparent, sampling bias cannot be ruled out.

Three patients with childhood onset symptomatic dystonia responded to levodopa. None fulfilled criteria for a diagnosis of "dopa responsive dystonia" (Segawa's disease). One may have had athetoid cerebral palsy for almost 25 years. All obtained dramatic and sustained benefit from levodopa therapy. A therapeutic trial of levodopa is advised in all patients in whom dystonia has developed in childhood or early adult life, regardless of suspected aetiology or duration of symptoms.

Idiopathic torsion dystonia is most commonly caused by an autosomal dominant gene or genes with reduced penetrance. An idiopathic torsion dystonia locus has been mapped to chromosome 9q34 in one large non-Jewish and several Jewish kindreds in the USA. Linkage analysis was performed in 27 (26 British, one French) small families with idiopathic torsion dystonia, three of which were Ashkenazi Jewish, using the highly polymorphic loci argininosuccinate synthetase (ASS) and Abelson oncogene (ABL) which map to 9q34. The cumulative lod score for the more informative ASS locus at a recombination fraction of 0.001 was —6.72. A large component of this score was derived from three non-Jewish families, indistinguishable clinically from the others, in which individual lod scores excluded a disease locus tightly linked to ASS. Analysis of all the data using HOMOG showed significant heterogeneity, but evidence for linkage of an idiopathic torsion dystonia gene to 9q34 in a subset of families. The allelic association observed between ASS/ABL and idiopathic torsion dystonia in Ashkenazi families in the USA was also present in British Jewish kindreds. These data suggest genetic heterogeneity in idiopathic torsion dystonia but indicate the existence of a locus for idiopathic torsion dystonia at 9q34 in both Jewish and non-Jewish kindreds in the UK.

Parental age and birth order were studied in 251 patients with cerebral palsy. No parental age or birth order effects were observed in spastic quadriplegia or diplegia, but a paternal age effect was detected in those with athetoid/dystonic cerebral palsy and congenital hemiplegia. These observations indicate that some cases of athetoid/dystonic or hemiplegic cerebral palsy might arise by fresh dominant genetic mutation.

Age of onset and severity of idiopathic torsion dystonia (ITD) were studied in 100 British families containing 107 index cases and 79 secondary cases. Analysis of variance of these clinical features did not suggest that ITD is genetically heterogenous, and they were similar in Jewish and non-Jewish patients. Intrafamilial correlation for age of onset was low, particularly between parents and their offspring, suggesting that the ITD phenotype may be determined in part by nongenetic factors or an allelic modifying gene.

A study of 71 patients with idiopathic torsion dystonia (ITD) and 71 matched controls was performed to investigate the range of possible clinical expression of ITD and the role of environmental factors in the development of the disease. A family history of tremor and stuttering were the only factors significantly associated with ITD. No associated environmental factor was identified.

Fifty two cases of haemangioblastoma were reviewed for their clinical, genetic and prognostic features. Of 34 patients with apparently isolated cerebellar lesions, postoperative outcome was good in 79%. Six isolated spinal lesions presented more insidiously and morbidity was related to incomplete resection. Twelve (23%) of the patients definitely had von Hippel-Lindau disease (VHLD). The true proportion may be higher as this diagnosis was not definitely excluded in many of the remainder; only ten patients with seemingly isolated cerebellar tumours were appropriately investigated and two had evidence of VHLD. Four out of 26 cases (15%) with apparently completely resected, isolated, cerebellar lesions later developed recurrent tumours. Brainstem and supratentorial haemangioblastomas were rare and were always associated with VHLD. The cerebellar or spinal haemangioblastomas due to VHLD had no distinctive clinical features compared with isolated tumours and there was considerable overlap in age of onset between the two groups of cases. All patients with an apparently isolated CNS haemangioblastoma should be investigated for evidence of von Hippel-Lindau disease.