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Background: Difficulty scoring systems are important for the safe, stepwise implementation of new procedures. We designed a retrospective observational study for building a difficulty score for robotic pancreatoduodenectomy. Methods: The difficulty score (PD-ROBOSCORE) aims at predicting severe postoperative complications after robotic pancreatoduodenectomy. The PD-ROBOSCORE was developed in a training cohort of 198 robotic pancreatoduodenectomies and was validated in an international multicenter cohort of 686 robotic pancreatoduodenectomies. Finally, all centers tested the model during the early learning curve (n = 300). Growing difficulty levels (low, intermediate, high) were defined using cut-off values set at the 33rd and 66th percentile (NCT04662346). Results: Factors included in the final multivariate model were a body mass index of ≥25 kg/m2 for males and ≥30 kg/m2 for females (odds ratio:2.39; P < .0001), borderline resectable tumor (odd ratio:1.98; P < .0001), uncinate process tumor (odds ratio:1.69; P < .0001), pancreatic duct size

Successful periodontal repair and regeneration requires the coordinated responses from soft and hard tissues as well as the soft tissue–to–bone interfaces. Inspired by the hierarchical structure of native periodontal tissues, tissue engineering technology provides unique opportunities to coordinate multiple cell types into scaffolds that mimic the natural periodontal structure in vitro. In this study, we designed and fabricated highly ordered multicompartmental scaffolds by melt electrowriting, an advanced 3-dimensional (3D) printing technique. This strategy attempted to mimic the characteristic periodontal microenvironment through multicompartmental constructs comprising 3 tissue-specific regions: 1) a bone compartment with dense mesh structure, 2) a ligament compartment mimicking the highly aligned periodontal ligaments (PDLs), and 3) a transition region that bridges the bone and ligament, a critical feature that differentiates this system from mono- or bicompartmental alternatives. The multicompartmental constructs successfully achieved coordinated proliferation and differentiation of multiple cell types in vitro within short time, including both ligamentous- and bone-derived cells. Long-term 3D coculture of primary human osteoblasts and PDL fibroblasts led to a mineral gradient from calcified to uncalcified regions with PDL-like insertions within the transition region, an effect that is challenging to achieve with mono- or bicompartmental platforms. This process effectively recapitulates the key feature of interfacial tissues in periodontium. Collectively, this tissue-engineered approach offers a fundament for engineering periodontal tissue constructs with characteristic 3D microenvironments similar to native tissues. This multicompartmental 3D printing approach is also highly compatible with the design of next-generation scaffolds, with both highly adjustable compartmentalization properties and patient-specific shapes, for multitissue engineering in complex periodontal defects.

ZusammenfassungDie COVID-19-Krise verdeutlichte die Schlüsselrolle des Öffentlichen Gesundheitsdienstes (ÖGD) mit den rund 375 kommunalen Gesundheitsämtern in der Pandemiebekämpfung. Dabei stellte neben fehlenden personellen Ressourcen auch die unzureichende digitale Reife vieler Gesundheitsämter eine Hürde für die effektive und skalierbare Infektionsmeldung und Kontaktnachverfolgung dar. In diesem Artikel stellen wir das Reifegradmodell (RGM) für die Digitalisierung von Gesundheitsämtern vor, dessen Erarbeitung im Zeitraum Januar 2021 bis Februar 2022 stattfand und durch das Bundesministerium für Gesundheit gefördert wurde. Es findet seit Anfang 2022 Anwendung mit dem Ziel der Stärkung der Digitalisierung des ÖGD. Das RGM zielt darauf ab, Gesundheitsämter Schritt für Schritt anzuleiten, ihre digitale Reife zu erhöhen, um so für zukünftige Herausforderungen gerüstet zu sein. Entwickelt und evaluiert wurde das RGM anhand qualitativer Interviews mit Mitarbeitenden der Gesundheitsämter und weiteren Expert*innen des ÖGD sowie in Workshops und im Rahmen einer quantitativen Umfrage. Das RGM erlaubt die Messung der digitalen Reife in 8 Dimensionen, welche jeweils in 2–5 Subdimensionen untergliedert sind. Innerhalb der Subdimensionen erfolgt eine Einstufung auf 5 verschiedenen Reifegradstufen. Derzeit dient das RGM neben der Erfassung der digitalen Reife der einzelnen Gesundheitsämter auch als Management-Tool für die Planung von Digitalisierungsprojekten. Ziel ist es, auf Basis des RGM eine zielgerichtete Kommunikation zwischen den Gesundheitsämtern zu fördern, um Best Practices für die einzelnen Dimensionen auszutauschen.

The magnetically localized and wash-free fluorescence immunoassay (MLFIA) is a no-wash assay for directly measuring biomolecule (antigen and antibody) concentration, without mixing nor washing steps, from a body fluid (serum and plasma).

Helical aromatic oligoamide foldamers provide negative cavities that strongly bind dicationic guests, giving complexes as stable pseudofoldaxanes with few precedents.

The synthesis and characterization of an original π-extended cationic azahelicene is reported. The phenanthrene-fused aza[7]helicene derivative encompasses a total of ten aromatic fused rings leading to a dissymmetric yet helically folded structure, as revealed by NMR and X-ray diffraction analyses. The polyaromatic and cationic nature of the new azahelicenium makes it soluble in both organic and aqueous media, which allowed photophysical studies in solvents of different polarities. The extended chromophoric species demonstrates a broad absorption over the whole visible range and orange-red fluorescence emission. Chiral resolution of the racemate was performed subsequently, affording two optically pure and configurationally stable azahelicenium enantiomers. Multi-band circular dichroism and long-wavelength circularly polarized emission were observed, associated with remarkable absorption and luminescence dissymmetry factors, both in organic and aqueous media.

Fulfilling stabilizing hydrogen bonds in a synthetic helix–turn–helix structure may concomitantly generate conformational frustration.

We employ several galaxy formation models, in particular, L-GALAXIES, IllustrisTNG, and EAGLE, as well as observational samples from SDSS and DESI, to investigate galactic conformity, the observed large-scale correlation between the star-formation properties of central (primary) galaxies and those of their neighbours. To analyse the models and observations uniformly, we introduce CENSAT, a new algorithm to define whether a galaxy is a central or a satellite system based on an isolation criterion. We find that the conformity signal is present, up to at least 5 Mpc from the centres of low- and intermediate-mass centrals in the latest version of L-GALAXIES (Ayromlou et al. 2021), IllustrisTNG, and EAGLE, as well as in SDSS and DESI observational samples. In comparison, the conformity signal is substantially weaker in an older version of L-GALAXIES (Henriques et al. 2020). One of the main differences between this older model and the other models is its neglect of ram-pressure stripping of the gas reservoirs of galaxies except within the boundaries of massive cluster haloes. Our observational comparisons demonstrate that this difference significantly affects the observed large-scale conformity signal. Furthermore, by examining the contribution of backsplash, fly-by, central, and satellite galaxies to the conformity signal, we show that much, but not all, of it arises from primary galaxies near massive systems. Remaining tensions between the models and observations may be solved by modifying the physical prescriptions for how feedback processes affect the distribution and kinematics of gas and the environment around galaxies out to scales of several Megaparsecs., Comment: Submitted to MNRAS

Purpose A preoperative estimate of the risk of malignancy for intraductal papillary mucinous neoplasms (IPMN) is important. The present study carries out an external validation of the Shin score in a European multicenter cohort. Methods An observational multicenter European study from 2010 to 2015. All consecutive patients undergoing surgery for IPMN at 35 hospitals with histological-confirmed IPMN were included. Results A total of 567 patients were included. The score was significantly associated with the presence of malignancy (p < 0.001). In all, 64% of the patients with benign IPMN had a Shin score < 3 and 57% of those with a diagnosis of malignancy had a score >= 3. The relative risk (RR) with a Shin score of 3 was 1.37 (95% CI: 1.07-1.77), with a sensitivity of 57.1% and specificity of 64.4%. Conclusion Patients with a Shin score

Zusammenfassung. In diesem Artikel wird die onkologische Pflegeberatung in der Radioonkologie und Neuroonkologie anhand des kooperativen Beratungsmodells von Hüper & Hellige (2015) erläutert. Mittels eines Fallberichts aus der Radioonkologie und Neuroonkologie wird die Sinnhaftigkeit für systematisierte Pflegeberatung verdeutlicht. Abschließend wird anhand der Skizzierung einer kollegialen Beratung der Stellenwert der Pflegeberatung für die interdisziplinäre Zusammenarbeit hervorgehoben.

A cytorhabdovirus, tentatively named "patchouli chlorosis-associated cytorhabdovirus" (PCaCV), was identified in a patchouli plant, using high-throughput sequencing, and its genome sequence was confirmed by Sanger sequencing. The PCaCV genome consists of 12,913 nucleotides and contains six open reading frames in the order 3'-N-P'-P-P3-M-(G)-L-5'. The glycoprotein gene was found to contain stop codons in the coding frame; hence, this gene is considered defective. PCaCV is most closely related to tomato yellow mottle-associated virus, sharing 61.1% nucleotide sequence identity in the complete genome and 73.9% amino acid sequence identity in the L protein. These data suggest that PCaCV should be considered a new member of the genus Cytorhabdovirus, and the binomial species name "Cytorhabdovirus patchoulii" is proposed.

As páginas de um jornal congregam diversos gêneros/registros linguísticos especializados, seja qual for o meio de acesso para a sua leitura. Nem sempre, porém, percebe-se distinção clara entre gêneros/registros jornalísticos, mesmo entre os produtores desses textos, o que leva a especular sobre a conformação e estabilidade linguística de determinados gêneros/registros. O presente estudo investiga essa questão, ao analisar um corpus formado por textos de duas edições da "Folha de S.Paulo", classificados em termos de gêneros/ registros por especialistas. Os resultados foram tabulados por grau de concordância entre classificadores. A maioria das classificações recaiu sobre as categorias reportagem e notícia. Em seguida, escores que refletem o consenso na determinação do gêneros/registro foram mapeados segundo as dimensões de variação do texto jornalístico de Kauffmann (2005), verificando o quão separados linguisticamente estão os gêneros/registros menos consensuais.

An Online tool for Fragment-based Molecule Parametrization (OFraMP) is described. OFraMP is a web application for assigning atomic interaction parameters to large molecules by matching sub-fragments within the target molecule to equivalent sub-fragments within the Automated Topology Builder (ATB, atb.uq.edu.au) database. OFraMP identifies and compares alternative molecular fragments from the ATB database, which contains over 890,000 pre-parameterized molecules, using a novel hierarchical matching procedure. Atoms are considered within the context of an extended local environment (buffer region) with the degree of similarity between an atom in the target molecule and that in the proposed match controlled by varying the size of the buffer region. Adjacent matching atoms are combined into progressively larger matched sub-structures. The user then selects the most appropriate match. OFraMP also allows users to manually alter interaction parameters and automates the submission of missing substructures to the ATB in order to generate parameters for atoms in environments not represented in the existing database. The utility of OFraMP is illustrated using the anti-cancer agent paclitaxel and a dendrimer used in organic semiconductor devices. Graphical abstract: OFraMP applied to paclitaxel (ATB ID 35922).[Figure not available: see fulltext.]

The directionally solidified eutectic alloy NiAl-(Cr,Mo) is a promising candidate for structural applications at high temperatures, due to its increased creep resistance compared to its single phase B2ordered NiAl counterpart. This system yields an eutectic trough connecting the invariant reactions of the ternary alloys NiAl-Cr and NiAl-Mo. During directional solidification (DS) along this trough the evolved microstructures of the two-phase eutectic is changing from fibrous to lamellar and back to fibrous morphology while increasing and decreasing the amounts of Mo and Cr, respectively. To investigate these effects in the morphology, the phase-field method has proven to be predestined in the last decades. However, as the modeling of quaternary systems is challenging for the simulation with a grand potential based phase-field model, the focus of this work is on the generation of a material model for one defined compound namely NiAl-31Cr-3Mo. The modeling is validated by investigating the microstructure evolution in two- and three-dimensional simulations of the DS process for two different growth velocities and by investigating their undercooling spacing relationships. The evolving microstructures obtained from three-dimensional large-scale simulations are presented and validated with corresponding micrographs from scanning electron microscopy (SEM) of directionally solidified samples with the same growth velocities. The simulation results show the theoretically expected behaviors and are in qualitative and quantitative accordance with DS experiments. The study of NiAl-31Cr-3Mo serves as the basis for a comprehensive data-driven analysis of microstructure properties and system quantities of the entire quaternary material NiAl-(Cr,Mo). With this, an accelerated design of advanced materials is promoted.

We introduce phi-1, a new large language model for code, with significantly smaller size than competing models: phi-1 is a Transformer-based model with 1.3B parameters, trained for 4 days on 8 A100s, using a selection of ``textbook quality" data from the web (6B tokens) and synthetically generated textbooks and exercises with GPT-3.5 (1B tokens). Despite this small scale, phi-1 attains pass@1 accuracy 50.6% on HumanEval and 55.5% on MBPP. It also displays surprising emergent properties compared to phi-1-base, our model before our finetuning stage on a dataset of coding exercises, and phi-1-small, a smaller model with 350M parameters trained with the same pipeline as phi-1 that still achieves 45% on HumanEval., 26 pages

Despite the large-scale digitalization and automation of production lines, human operators still play a vital role in the shopfloor. Operators provide flexibility and agile responsiveness with their actions, as well as capability to react autonomously based on their technical knowledge and experience. Bridging the gap between modern digital systems of information creation and management and humans in the shopfloor is a current challenge for both the academia and the technology integrators. One emerging way for delivering instructions to the operators is Augmented Reality (AR). AR allows the visualization of the instructions in the operator's field of view, using digital designs, animations and text instructions. As this technology gains increasingly more ground in the shopfloor, there is a need for agile generation of content, automatizing most of the instructions' generation process. Towards that end, this paper presents an authoring tool for generating digital instructions for manual operations. The authoring tool allows the identification of the (dis-)assembly sequence of a product and also to include intermediate manual operations, such as surface treatment, and visualizations. In order to determine which assembly tasks and instructions are the most commonly needed, contextual inquiries were conducted in collaboration with the industry. These findings, together with the existing literature on manual processes on the shopfloor, served as a starting point for a taxonomy of assembly task types that are also presented in this paper. The taxonomy serves as a guide to define the processes on which digital instructions and visualizations can be provided. The result can be delivered as digital on-screen instructions or as an Augmented Reality application that may target mobile devices and headsets. The proposed approach is validated in an industrial use case of a compressor assembly.

Background Triangle pancreatoduodenectomy adds to the conventional procedure the en bloc removal of the retroperitoneal lympho-neural tissue included in the triangular area bounded by the common hepatic artery (CHA), the superior mesenteric artery (SMA), and the superior mesenteric vein/portal vein. We herein aim to show the feasibility of “cold” triangle robotic pancreaticoduodenectomy (C-Tr-RPD) for pancreatic cancer (PDAC). Methods Cold dissection corresponds to sharp arterial divestment performed using only the tips of robotic scissors. After division of the gastroduodenal artery, triangle dissection begins by lateral-to-medial divestment of the CHA and anterior-to-posterior clearance of the right side of the celiac trunk. Next, after a wide Kocher maneuver, the origin of the SMA, and the celiac trunk are identified. After mobilization of the first jejunal loop and attached mesentery, the SMA is identified at the level of the first jejunal vein and is divested along the right margin working in a distal-to-proximal direction. Vein resection and reconstruction can be performed as required. C-Tr-RPD was considered feasible if triangle dissection was successfully completed without conversion to open surgery or need to use energy devices. Postoperative complications and pathology results are presented in detail. Results One hundred twenty-seven consecutive C-Tr-RPDs were successfully performed. There were three conversions to open surgery (2.3%), because of pneumoperitoneum intolerance (n = 2) and difficult digestive reconstruction. Thirty-four patients (26.7%) required associated vascular procedures. No pseudoaneurysm of the gastroduodenal artery was observed. Twenty-eight patients (22.0%) developed severe postoperative complications (≥ grade III). Overall 90-day mortality was 7.1%, declining to 2.3% after completion of the learning curve. The median number of examined lymph nodes was 42 (33–51). The rate of R1 resection (7 margins Conclusion C-Tr-RPD is feasible, carries a risk of surgical complications commensurate to the magnitude of the procedure, and improves staging of PDAC. Graphical abstract

Background: Atypical EGFR mutations occur in 10%-30% of non-small-cell lung cancer (NSCLC) patients with EGFR mutations and their sensitivity to classical epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKI) is highly heterogeneous. Patients harboring one group of uncommon, recurrent EGFR mutations (G719X, S768I, L861Q) respond to EGFR-TKI. Exon 20 insertions are mostly insensitive to EGFR-TKI but display sensitivity to exon 20 inhibitors. Clinical outcome data of patients with very rare point and compound mutations upon systemic treatments are still sparse to date. Patients and methods: In this retrospective, multicenter study of the national Network Genomic Medicine (nNGM) in Germany, 856 NSCLC cases with atypical EGFR mutations including co-occurring mutations were reported from 12 centers. Clinical follow-up data after treatment with different EGFR-TKIs, chemotherapy and immune checkpoint inhibitors were available from 260 patients. Response to treatment was analyzed in three major groups: (i) uncommon mutations (G719X, S7681, L861Q and combinations), (ii) exon 20 insertions and (iii) very rare EGFR mutations (very rare single point mutations, compound mutations, exon 18 deletions, exon 19 insertions). Results: Our study comprises the largest thus far reported real-world cohort of very rare EGFR single point and compound mutations treated with different systemic treatments. We validated higher efficacy of EGFR-TKI in comparison to chemotherapy in group 1 (uncommon), while most exon 20 insertions (group 2) were not EGFR-TKI responsive. In addition, we found TKI sensitivity of very rare point mutations (group 3) and of complex EGFR mutations containing exon 19 deletions or L858R mutations independent of the combination partner. Notably, treatment responses in group 3 (very rare) were highly heterogeneous. Co-occurring TP53 mutations exerted a non-significant trend for a detrimental effect on outcome in EGFR-TKI-treated patients in groups 2 and 3 but not in group 1. Conclusions: Based on our findings, we propose a novel nNGM classification of atypical EGFR mutations.

BACKGROUND: Robot-assisted distal pancreatectomy (RDP) is increasingly used as an alternative to laparoscopic distal pancreatectomy (LDP) in patients with resectable pancreatic cancer but comparative multicenter studies confirming the safety and efficacy of RDP are lacking. METHODS: An international, multicenter, retrospective, cohort study, including consecutive patients undergoing RDP and LDP for resectable pancreatic cancer in 33 experienced centers from 11 countries (2010-2019). The primary outcome was R0-resection. Secondary outcomes included lymph node yield, major complications, conversion rate, and overall survival. RESULTS: In total, 542 patients after minimally invasive distal pancreatectomy were included: 103 RDP (19%) and 439 LDP (81%). The R0-resection rate was comparable (75.7% RDP vs. 69.3% LDP, p = 0.404). RDP was associated with longer operative time (290 vs. 240 min, p < 0.001), more vascular resections (7.6% vs. 2.7%, p = 0.030), lower conversion rate (4.9% vs. 17.3%, p = 0.001), more major complications (26.2% vs. 16.3%, p = 0.019), improved lymph node yield (18 vs. 16, p = 0.021), and longer hospital stay (10 vs. 8 days, p = 0.001). The 90-day mortality (1.9% vs. 0.7%, p = 0.268) and overall survival (median 28 vs. 31 months, p = 0.599) did not differ significantly between RDP and LDP, respectively. CONCLUSIONS: In selected patients with resectable pancreatic cancer, RDP and LDP provide a comparable R0-resection rate and overall survival in experienced centers. Although the lymph node yield and conversion rate appeared favorable after RDP, LDP was associated with shorter operating time, less major complications, and shorter hospital stay. The specific benefits associated with each approach should be confirmed by multicenter, randomized trials.

BACKGROUND: Robot-assisted distal pancreatectomy (RDP) is increasingly used as an alternative to laparoscopic distal pancreatectomy (LDP) in patients with resectable pancreatic cancer but comparative multicenter studies confirming the safety and efficacy of RDP are lacking. METHODS: An international, multicenter, retrospective, cohort study, including consecutive patients undergoing RDP and LDP for resectable pancreatic cancer in 33 experienced centers from 11 countries (2010-2019). The primary outcome was R0-resection. Secondary outcomes included lymph node yield, major complications, conversion rate, and overall survival. RESULTS: In total, 542 patients after minimally invasive distal pancreatectomy were included: 103 RDP (19%) and 439 LDP (81%). The R0-resection rate was comparable (75.7% RDP vs. 69.3% LDP, p = 0.404). RDP was associated with longer operative time (290 vs. 240 min, p < 0.001), more vascular resections (7.6% vs. 2.7%, p = 0.030), lower conversion rate (4.9% vs. 17.3%, p = 0.001), more major complications (26.2% vs. 16.3%, p = 0.019), improved lymph node yield (18 vs. 16, p = 0.021), and longer hospital stay (10 vs. 8 days, p = 0.001). The 90-day mortality (1.9% vs. 0.7%, p = 0.268) and overall survival (median 28 vs. 31 months, p = 0.599) did not differ significantly between RDP and LDP, respectively. CONCLUSIONS: In selected patients with resectable pancreatic cancer, RDP and LDP provide a comparable R0-resection rate and overall survival in experienced centers. Although the lymph node yield and conversion rate appeared favorable after RDP, LDP was associated with shorter operating time, less major complications, and shorter hospital stay. The specific benefits associated with each approach should be confirmed by multicenter, randomized trials.

Background Prophylactic central neck dissection in clinically low-risk cT1bT2N0 papillary thyroid carcinoma is controversial, due to a large number of conflicting retrospective studies, some showing an advantage in terms of locoregional recurrence, others showing no advantage. These previous studies all show high rates of excellent response. We aim to demonstrate the non-inferiority of thyroidectomy alone as compared to total thyroidectomy with prophylactic central neck dissection in conjunction with adjuvant RAI 30 mCi with rTSH stimulation in terms of excellent response at 1 year. Trial design and methods Prospective randomized open multicenter phase III trial including patients with 11–40-mm papillary thyroid carcinoma (Bethesda VI) or suspicious cytology (Bethesda V) confirmed malignant on intra-operative frozen section analysis, with no suspicious lymph nodes on a specialized preoperative ultrasound examination. Patients will be randomized 1:1 into two groups: the reference group total thyroidectomy with bilateral prophylactic central neck dissection, and the comparator group total thyroidectomy alone. All patients will receive an ablative dose of 30mCi of radioactive iodine (RAI) within 4 months of surgery. The primary outcome is to compare the rate of excellent response at 1 year after surgery between the groups, as defined by an unstimulated serum thyroglobulin (Tg) level ≤ 0.2 ng/mL with no anti-Tg antibodies, an normal neck ultrasound and no ectopic uptake on the post-RAI scintiscan. Non-inferiority will be demonstrated if the rate of patients with excellent response at 1 year after randomization does not differ by more than 5%. Setting the significance level at 0.025 (one-sided) and a power of 80% requires a sample size of 598 patients (299 per group). Secondary outcomes are to compare Tg levels at 8 +/− 2 postoperative weeks, before RAI ablation, the rate of excellent response at 3 and 5 years, the rate of other responses at 1, 3, and 5 years (biochemical incomplete, indeterminate, and structurally incomplete responses), complications, quality of life, and cost-utility. Discussion (potential implications) If non-inferiority is demonstrated with this high-level evidence, prophylactic neck dissection will have been shown to not be necessary in clinically low-risk papillary thyroid carcinoma. Trial registration NCT 03570021. June 26,2018

PDF - 60K, Linear correlation observed between tumor growth inhibition (% T/C) or tumor regression and the fraction of time over the in vivo S473P-Akt IC80 in different cancer cell line-derived tumor xenografts implanted in nude mice (represented as dots) and nude rats (represented as triangles) following NVP-BYL719 treatment (R2=0.77, p

Somatic PIK3CA mutations are frequently found in solid tumors, raising the hypothesis that selective inhibition of PI3Kα may have robust efficacy in PIK3CA-mutant cancers while sparing patients the side-effects associated with broader inhibition of the class I phosphoinositide 3-kinase (PI3K) family. Here, we report the biologic properties of the 2-aminothiazole derivative NVP-BYL719, a selective inhibitor of PI3Kα and its most common oncogenic mutant forms. The compound selectivity combined with excellent drug-like properties translates to dose- and time-dependent inhibition of PI3Kα signaling in vivo, resulting in robust therapeutic efficacy and tolerability in PIK3CA-dependent tumors. Novel targeted therapeutics such as NVP-BYL719, designed to modulate aberrant functions elicited by cancer-specific genetic alterations upon which the disease depends, require well-defined patient stratification strategies in order to maximize their therapeutic impact and benefit for the patients. Here, we also describe the application of the Cancer Cell Line Encyclopedia as a preclinical platform to refine the patient stratification strategy for NVP-BYL719 and found that PIK3CA mutation was the foremost positive predictor of sensitivity while revealing additional positive and negative associations such as PIK3CA amplification and PTEN mutation, respectively. These patient selection determinants are being assayed in the ongoing NVP-BYL719 clinical trials. Mol Cancer Ther; 13(5); 1117–29. ©2014 AACR.

PDF file, 300KB, A. Upon infection with viral particles obtained from the pBabe-puro-myr-Akt retroviral vectors, pools were selected upon addition of puromycin and maintained in culture for several weeks. Extracts from pools were generated for the indicated time of culture and analyzed by Western-blot for the expression of the exogenously expressed HA-tagged form of the dominant active myr-Akt protein. B. 2.5x106 MCF7-BP (upper panel) or MCF7-myr-Akt (bottom panel) were seeded in 10 cm dishes and exposed for 1 h to the indicated compounds at the indicated concentrations (in nM). Cells were then lyzed and cell extracts were analyzed by Western-blot for pathway inhibition as revealed by S473P-Akt levels.

The pan-phosphoinositide 3-kinase (PI3K) inhibitor BKM120 was found, at high concentrations, to cause cell death in various cellular systems, irrespective of their level of PI3K addiction. Transcriptional and biochemical profiling studies were used to identify the origin of these unexpected and apparently PI3K-independent effects. At 5- to 10-fold, the concentration needed to half-maximally inhibit PI3K signaling. BKM120 treatment caused changes in expression of mitotic genes and the induction of a robust G2–M arrest. Tubulin polymerization assays and nuclear magnetic resonance-binding studies revealed that BKM120 inhibited microtubule dynamics upon direct binding to tubulin. To assess the contribution of this off-target activity vis-à-vis the antitumor activity of BKM120 in PI3K-dependent tumors, we used a mechanistic PI3K-α–dependent model. We observed that, in vivo, daily treatment of mice with doses of BKM120 up to 40 mg/kg led to tumor regressions with no increase in the mitotic index. Thus, strong antitumor activity can be achieved in PI3K-dependent models at exposures that are below those necessary to engage the off-target activity. In comparison, the clinical data indicate that it is unlikely that BKM120 will achieve exposures sufficient to significantly engage the off-target activity at tolerated doses and schedules. However, in preclinical settings, the consequences of the off-target activity start to manifest themselves at concentrations above 1 μmol/L in vitro and doses above 50 mg/kg in efficacy studies using subcutaneous tumor–bearing mice. Hence, careful concentration and dose range selection is required to ensure that any observation can be correctly attributed to BKM120 inhibition of PI3K. Mol Cancer Ther; 11(8); 1747–57. ©2012 AACR.

Correlation analysis matrix displaying features that correlate most strongly with sgRNA's having off target effects.

Activity of FGF401 in biochemical kinase assays

Activity of FGF401 in PDXs

Details on materials and methods

Additional method details for ATAC-Seq Data Analysis and ChIP-qPCR

PDF file, 53KB, A and B. 5x105 A2058 cells were seeded in 10 cm dishes and incubated for 24 h either with increasing amounts of BKM120 (A) or with 5 �M of either GDC-0941 (B, top panel) or BEZ235 (B, bottom panel). Cells were then fixed, prepared as described for quantification of the population in the different phases of the cell cycle by fluorescence-activated cell sorting. G1, S and G2/M distribution for control untreated cells are described in the mean text and in Figure 4A. The activities of BKM120 on the cell cycle were plotted along to the inhibitory effects on pAkt levels (A).