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Supplementary Methods, radiologist's reading of CT images, supplementary figure legends, supplementary references

microCT scans

SFigure 1: Histological examination of mouse lungs; SFigure 2: Tissue sections from human lung adenocarcinoma; SFigure 3: Cabozantinib (CBT) and vandetanib (VDT) resistant cells; SFigure 4: Comparison of in vitro inhibition of RET, RETV804L and RETV804M by ponatinib, cabozantinib, vandetanib, and lenvatinib; SFigure 5: Ponatinib treatment schedule and body weight measurements; SFigure 6: Lung section histology from Dox-induced C/KR mice and treated with vehicle or ponatinib for 1 months; SFigure 7: μCT examination of dox-induced C/KR mice with lung tumors before and after one month treatment with vehicle or ponatinib.

Seventy-five wild tilapia samples from six rivers (ten sites) in Guangxi province were collected and analyzed for 53 organochlorine compounds. DDTs, endosulfan, and PCBs were the most dominant compounds found in this study. Tiandong County (TD) and Guigang City (GG) sites were found to be heavily contaminated with high levels of endosulfan (385–925 ng/g lw) and/or DDTs (20.1–422 ng/g lw). The diagnostic ratios indicated that the residues of DDTs and endosulfan in wild tilapia are associated with historical applications as well as the recent introduction of technical DDTs and endosulfan at some sampling sites. The correlation between total length, body mass, and organochlorines (OCs) was higher than the correlation between age and lipid content. There was no significant correlation between organochlorine pesticides (OCPs) and lipid content. Therefore, for organisms, the feeding intensity (related to length and mass) of fish could better reflect degree of pollution than exposure time (age) of fish. The hazardous ratios for the 50th and 95th percentile data of OCPs and PCBs in fish were both below 1, suggesting that daily exposure to OCPs and PCBs yields a lifetime cancer risk lower than 1 in 10,000.

Equol, which produced from daidzein (one of the principal isoflavones), is recognized to be the most resultful in stimulating an estrogenic and antioxidant response. The daidzein transformation was studied during fermentation of five growth media inoculated with feces from a healthy human, and a daidzein conversion strain was isolated. To enrich the bacterial population involved in daidzein metabolism in a complex mixture, fecal samples were treated with antibiotics. The improved propidium monoazide combined with the quantitative polymerase chain reaction (PMAxx-qPCR) assay showed that the ampicillin treatment of samples did result in a reduction of the total visible bacteria counts by 52.2% compared to the treatment without antibiotics. On this basis, the newly isolated rod-shaped, Gram-positive anaerobic bacterium, named strain Y11 (MN560033), was able to metabolize daidzein to equol under anaerobic conditions, with a conversion ratio (equol ratio: the amount of equol produced/amount of supplemented daizein) of 0.56 over 120 h. The 16S rRNA partial sequence of the strain Y11 exhibited 99.8% identity to that of Slackia equolifaciens strain DZE (NR116295). This study will provide new insights into the biotransformation of equol from daidzein by intestinal microbiota from the strain-level and explore the possibility of probiotic interventions.

This study reports the development and optimization of a real-time loop-mediated isothermal amplification (qLAMP) method for rapid detection of Acetobacter aceti strain in red wine samples. Our results showed that the primers and probes designed for 16S rRNA were effective for A. aceti detection. The quantification limit of real-time polymerase chain reaction (qPCR) and qLAMP in pure culture was 2.05 × 101 colony forming units (CFU) mL−1. qLAMP had a sensitivity of 6.88 × 101 CFU mL−1 in artificially contaminated Changyu dry red wine (CDRW) and Changyu red wine (CRW), and 6.88 × 102 CFU mL−1 in artificially contaminated Greatwall dry red wine (GDRW), which was 10 times higher than that of qPCR. In conclusion, this newly developed qLAMP is a reliable, rapid and accurate method for the detection and quantification of A. aceti species in red wine samples. Furthermore, our work provides a standard reference method for the quantitative detection of A. aceti and other acetic acid bacteria during the fermentation and storage of red wine samples.

Renal cell carcinoma (RCC) is a common and devastating disease characterized by a hypoxic microenvironment, epithelial-mesenchymal transition and potent resistance to therapy evidencing the presence of cancer stem cells (CSCs). Various CSC markers have been studied in RCC, but overall there is limited data on their role and most markers studied have been relatively nonspecific. Doublecortin-like kinase 1 (DCLK1) is a validated CSC marker in the gastrointestinal tract and evidence for an equivalent role in other cancers is accumulating. We used bioinformatics, immunohistochemistry, flow cytometry, spheroid self-renewal and chemoresistance assays in combination with overexpression and siRNA-knockdown to study the stem cell-supportive role of DCLK1 alternative splice variants (DCLK1 ASVs) in RCC. To target tumor cells expressing DCLK1 ASVs directly, we developed a novel monoclonal antibody (CBT-15) and delivered it systemically to RCC tumor xenografts. DCLK1 ASVs were overexpressed, enriched together with CSC markers and predictive of overall and recurrence-free survival in RCC patients. In vitro, DCLK1 ASVs were able to directly stimulate essential molecular and functional characteristics of renal CSCs including expression of aldehyde dehydrogenase, self-renewal and resistance to FDA-approved receptor tyrosine kinase and mTOR inhibitors, while targeted downregulation of DCLK1 reversed these characteristics. Finally, targeting DCLK1 ASV-positive cells with the novel CBT-15 monoclonal antibody blocked RCC tumorigenesis in vivo. These findings establish DCLK1 as a CSC marker with implications for therapy, disease progression and survival in RCC and demonstrate the therapeutic value of DCLK1-targeted monoclonal antibodies against renal CSCs.

We present an experimental study on non-equilibrium dynamics of a spinor condensate after it is quenched across a superfluid to Mott insulator (MI) phase transition in cubic lattices. Intricate dynamics consisting of spin-mixing oscillations at multiple frequencies are observed in time evolutions of the spinor condensate localized in deep lattices after the quantum quench. Similar spin dynamics also appear after spinor gases in the MI phase are suddenly moved away from their ground states via quenching magnetic fields. We confirm these observed spectra of spin-mixing dynamics can be utilized to reveal atom number distributions of an inhomogeneous system, and to study transitions from two-body to many-body dynamics. Our data also imply the non-equilibrium dynamics depend weakly on the quench speed but strongly on the lattice potential. This enables precise measurements of the spin-dependent interaction, a key parameter determining the spinor physics.

Cronobacter sakazakii (C. sakazakii) in viable but nonculturable (VBNC) state might evade traditional culture detection, and pose a serious threat to food safety. The objective of this study was to establish an improved propidium monoazide (PMAxx), combined with a single intact cell droplet digital PCR (SIC ddPCR) method for rapid and absolute quantification of VBNC C. sakazakii and apply it to the detection of several infant foods. In this study, we optimized the detection conditions for C. sakazakii, and determined the optimal PMAxx final concentration (8 μM), light exposure times (10 min), annealing temperature (58 °C) and elongation time (1 min). In addition, PMAxx-SIC ddPCR demonstrated a higher detection sensitivity and accuracy relative to real-time PCR (qPCR), with 23 CFU/mL in pure culture. Furthermore, PMAxx-SIC ddPCR was used to detect VBNC C. sakazakii in the actual infant food samples. A total of 240 samples were analyzed, out of which 6 samples turned out to be positive by the traditional culture method, whereas 11 samples were found as positive by PMAxx-SIC ddPCR. These results demonstrate that the presence of VBNC may cause the conventional culture-based method to underestimate the size of microbial population.

RET fusions have been found in lung adenocarcinoma, of which KIF5B–RET is the most prevalent. We established inducible KIF5B-RET transgenic mice and KIF5B–RET-dependent cell lines for preclinical modeling of KIF5B–RET-associated lung adenocarcinoma. Doxycycline-induced CCSP-rtTA/tetO-KIF5B-RET transgenic mice developed invasive lung adenocarcinoma with desmoplastic reaction. Tumors regressed upon suppression of KIF5B–RET expression. By culturing KIF5B–RET-dependent BaF3 (B/KR) cells with increasing concentrations of cabozantinib or vandetanib, we identified cabozantinib-resistant RETV804L mutation and vandetanib-resistant-RETG810A mutation. Among cabozantinib, lenvatinib, ponatinib, and vandetanib, ponatinib was identified as the most potent inhibitor against KIF5B–RET and its drug-resistant mutants. Interestingly, the vandetanib-resistant KIF5B-RETG810A mutant displayed gain-of-sensitivity (GOS) to ponatinib and lenvatinib. Treatment of doxycycline-induced CCSP-rtTA/tetO-KIF5B-RET bitransgenic mice with ponatinib effectively induced tumor regression. These results indicate that KIF5B-RET–associated lung tumors are addicted to the fusion oncogene and ponatinib is the most effective inhibitor for targeting KIF5B–RET in lung adenocarcinoma. Moreover, this study finds a novel vandetanib-resistant RETG810A mutation and identifies lenvatinib and ponatinib as the secondary drugs to overcome this vandetanib resistance mechanism. Mol Cancer Ther; 15(10); 2521–9. ©2016 AACR.

// Janani Panneerselvam 1, 4 , Akhil Srivastava 1, 4 , Ranganayaki Muralidharan 1, 4 , Qi Wang 1, 4 , Wei Zheng 1 , Lichao Zhao 1, 4 , Alshine Chen 3, 4 , Yan D. Zhao 3, 4 , Anupama Munshi 2, 4 , Rajagopal Ramesh 1, 4, 5 1 Department of Pathology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA 2 Department of Radiation Oncology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA 3 Department of Biostatistics and Epidemiology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA 4 Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA 5 Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA Correspondence to: Rajagopal Ramesh, email: rajagopal-ramesh@ouhsc.edu Keywords: IL-24, lung cancer, HMGA1, miR222-3p, metastasis Received: April 21, 2016 Accepted: August 24, 2016 Published: September 02, 2016 ABSTRACT Interleukin (IL)-24, a novel tumor suppressor/cytokine exhibits antitumor activity against a broad-spectrum of human cancer cells. In a recent study, we showed that IL-24 inhibited AKT in lung cancer cells. However, the molecular mechanism of AKT inhibition by IL-24 remains elusive. The high mobility group (HMG) A1 a member of the non-histone chromosomal proteins and commonly referred to as architectural transcription factor, regulates transcription of various genes involved in cell growth and survival. Overexpression of HMGA1 has been shown to be associated with tumor progression and metastasis in several cancers, including human lung cancer. A recent study demonstrated that HMGA1 activates AKT function by reducing the activity of the protein phosphatase, phosphatase 2A subunit B (PPP2R2A) via the oncogenic micro (mi) RNA-222. Based on this report we hypothesized that IL-24-mediated AKT inhibition involved the HMGA1/miR-222 axis. To test our hypothesis, in the present study we used a H1299 lung cancer cell line that expressed exogenous human IL-24 when induced with doxycycline (DOX). Induction of IL-24 expression in the tumor cells markedly reduced HMGA1 mRNA and protein levels. Using a mechanistic approach, we found that IL-24 reduced miR-222-3p and -5p levels, as determined by qRT-PCR. Associated with HMGA1 and miR-222 inhibition was a marked increase in PPP2R2A, with a concomitant decrease in phosphorylated AKT T308/S473 expression. SiRNA-mediated knockdown of HMGA1 in combination with IL-24 significantly reduced AKT T308/S473 protein expression and greatly reduced cell migration and invasion compared with individual treatments. Further combination of IL-24 and a miR-222-3p inhibitor significantly increased PPP2R2A expression. Our results demonstrate for the first time that IL-24 inhibits AKT via regulating the HMGA1/miR-222 signaling node in human lung cancer cells and acts as an effective tumor suppressor. Thus, a therapy combining IL-24 with HMGA1 siRNA or miR-222-3p inhibitor should present effective treatment of lung cancer.

BACKGROUND Adequacy assessment for fine-needle aspiration procedures is a standard of care in large medical centers. Although the benefits of this approach include higher adequacy rates with fewer passes, it costs cytopathologist time and affects other clinical responsibilities. The objective of the current study was to evaluate the use of mobile video streaming (FaceTime) technology with the help of smartphone adapters attached to microscopes for remote adequacy assessment of cytologic samples. METHODS The study consisted of 2 phases: Phase 1 was a retrospective assessment of 25 samples by a primary pathologist with simultaneous streaming to a second pathologist using a smartphone (iPhone/iPad) FaceTime connection. Data on the adequacy of each sample and preliminary diagnoses were recorded. In phase 2, live cases were assessed prospectively by an onsite primary pathologist and by a remote pathologist using an iPhone/iPad-FaceTime connection. The testing phase involved prospective assessment of additional samples with a resident or cytotechnologist as the slide driver. RESULTS In phase 1, retrospective evaluation of 25 samples yielded considerable agreement (22 of 25 samples; 88%) between onsite and remote adequacy assessments. Three samples (12%) yielded results that did not agree, including 2 samples that were read as adequate in the onsite evaluation that were assessed as indeterminate using FaceTime. In phase 2 and in the testing phase, 14 samples exhibited considerable agreement on both adequacy and preliminary diagnosis (6 samples in phase 2 and 8 samples in the testing phase) and are currently available for reporting. Problems encountered include software version standardization, camera alignment, and (rarely) comprehension of the audio stream. CONCLUSIONS The current data indicate that iPhone/iPad FaceTime technology can be used to perform remote adequacy assessments of fine-needle aspirations and can help save valuable time for pathologists. Cancer Cytopathol 2016;124:213–220. © 2015 American Cancer Society.

The dynamic vibration absorbers have been applied to attenuate the rotor unbalance and torsional vibrations. The major purpose of this paper is to research the elimination of the fluid-induced vibration in the rotor/seal system using the absorber. The simplified rotor model with the absorber is established, and the Muszynska fluid force model is employed for the nonlinear seal force. The numerical method is used for the solutions of the nonlinear differential equations. The nonlinear responses of the rotor/seal system without and with the absorber are obtained, and then the rotating speed ranges by which the fluid-induced instability can be eliminated completely and partially are presented, respectively. The absorber parameters ranges by which the instability vibration can be eliminated completely and partially are obtained. The results show that the natural frequency vibration due to the fluid-induced instability in the rotor/seal system can be eliminated efficiently using the absorber. The appropriate natural frequency and damping ratio of the absorber can extend the complete elimination region of the instability vibration and postpone the occurrence of the instability vibration.

Renal cell carcinoma (RCC) is a common and devastating disease characterized by a hypoxic microenvironment, epithelial-mesenchymal transition and potent resistance to therapy evidencing the presence of cancer stem cells (CSCs). Various CSC markers have been studied in RCC, but overall there is limited data on their role and most markers studied have been relatively nonspecific. Doublecortin-like kinase 1 (DCLK1) is a validated CSC marker in the gastrointestinal tract and evidence for an equivalent role in other cancers is accumulating. We used bioinformatics, immunohistochemistry, flow cytometry, spheroid self-renewal and chemoresistance assays in combination with overexpression and siRNA-knockdown to study the stem cell-supportive role of DCLK1 alternative splice variants (DCLK1 ASVs) in RCC. To target tumor cells expressing DCLK1 ASVs directly, we developed a novel monoclonal antibody (CBT-15) and delivered it systemically to RCC tumor xenografts. DCLK1 ASVs were overexpressed, enriched together with CSC markers and predictive of overall and recurrence-free survival in RCC patients. In vitro, DCLK1 ASVs were able to directly stimulate essential molecular and functional characteristics of renal CSCs including expression of aldehyde dehydrogenase, self-renewal and resistance to FDA-approved receptor tyrosine kinase and mTOR inhibitors, while targeted downregulation of DCLK1 reversed these characteristics. Finally, targeting DCLK1 ASV-positive cells with the novel CBT-15 monoclonal antibody blocked RCC tumorigenesis in vivo. These findings establish DCLK1 as a CSC marker with implications for therapy, disease progression and survival in RCC and demonstrate the therapeutic value of DCLK1-targeted monoclonal antibodies against renal CSCs.

BACKGROUND Recent proposals to lengthen the interval in cervical cancer screening highlight the importance of the accurate interpretation of screening tests. Tumor debris present in Papanicolaou (Pap) tests from women with invasive cancer is known to hamper interpretation. The current study evaluated limiting factors in Pap tests from women with invasive cervical cancer. METHODS A total of 3003 women with the spectrum of cervical lesions who had ThinPrep (Hologic Inc, Marlborough, Mass) Pap and human papillomavirus (HPV) genotyping tests performed were grouped by their most severe histologic diagnosis. Cytologic and HPV results were analyzed by cross-sectional analysis. RESULTS The unsatisfactory rate of cytology specimens from patients with cancer (3.1%) was significantly higher than those from patients with cervical intraepithelial neoplasia of type 3 or less (0.8%) (P 30 years can help to identify high-risk women with unsatisfactory Pap tests. Cancer (Cancer Cytopathol) 2014;122:694–701. © 2014 American Cancer Society.

The antioxidant activity of Sweet Potato Extracts (SPE) can be enhanced by the presence of these other active antioxidants such as Tea Polyphones (TP) and Pueraria Flavonoid (PF). Since many of these natural antioxidants are consumed together in foods, the potential for synergistic interactions is high in the human diet. The aim of this study was to determine what concentrations and combinations of antioxidants among SPE, TP and PF are capable of producing synergistic antioxidant effects, based on potato-based food products. Solutions of the antioxidant activity of SPE, TP and PF, alone and in different combinations were measured using the stable free radical 2, 2-Diphenyl-1-Picrylhydrazyl (DPPH) and Ferric Reducing Anti-oxidant Power (FRAP) method. A comparison of the antioxidant activity of the combinations of antioxidants to the arithmetic sum of the antioxidant activity of the individual antioxidants was used to calculate the Synergistic Effects (SEs) between the antioxidants. The results showed that all concentrations of TP and PF combination with SPE (1 and 1.5%) could produce significant SEs (p

We observe evidence of first-order superfluid-to-Mott-insulator quantum phase transitions in a lattice-confined antiferromagnetic spinor Bose-Einstein condensate. The observed signatures include the hysteresis effect, significant heatings across the phase transitions, and changes in spin populations due to the formation of spin singlets in the Mott-insulator phase. The nature of the phase transitions is found to strongly depend on the ratio of the quadratic Zeeman energy to the spin-dependent interaction. Our observations are qualitatively understood by the mean field theory and suggest tuning the quadratic Zeeman energy is a new approach to realize superfluid-to-Mott-insulator phase transitions.

BACKGROUND: To evaluate the efficacy and the limitation of fine-needle aspiration (FNA) biopsy in thyroid bed lesions, a retrospective review was performed of the medical records of thyroid cancer patients who underwent ultrasound-guided FNA biopsy of the thyroid bed at The University of Texas MD Anderson Cancer Center over a 5-year period. METHODS: Data were reviewed on 220 FNA biopsies taken from thyroid bed lesions in 195 patients who had undergone thyroidectomy for thyroid carcinoma. Thyroid bed FNA results were compared with clinical follow-up, including neck dissection results. RESULTS: Recurrent carcinoma was diagnosed by FNA biopsy in 139 of 220 (63%) cases. Neck dissections were performed for 112 sites identified by FNA biopsies, and recurrent carcinoma was confirmed in 110 sites. The concordance between positive and/or suspicious FNA diagnosis and positive neck dissection results was 98% (118 of 120 cases). A false-positive FNA occurred in one patient with follicular thyroid carcinoma. The other discrepancy was attributed to failure to remove the lesion by neck dissection. The diagnostic accuracy of thyroid bed FNA was 100% in papillary and medullary thyroid carcinoma and 93% in follicular thyroid carcinoma. Suspicious and rare false-negative FNA results were attributed to low cellularity and lack of characteristic cytomorphologic features of thyroid carcinoma. CONCLUSIONS: Ultrasound-guided thyroid bed FNA biopsy is accurate and efficient in triaging patients who require post-thyroidectomy follow-up for recurrent thyroid carcinoma. Caution should be taken in the interpretation of FNA specimens that have low cellularity and lack characteristic cytologic features of thyroid carcinoma. Cancer (Cancer Cytopathol) 2013. © 2012 American Cancer Society.

Shear-resistant adhesion and extravasation of disseminated cancer cells at the target organ is a crucial step in hematogenous metastasis. We found that the vascular adhesion molecule E-selectin preferentially promoted the shear-resistant adhesion and transendothelial migration of the estrogen receptor (ER)–/CD44+ hormone-independent breast cancer cells, but not of the ER+/CD44-/low hormone-dependent breast cancer cells. Coincidentally, CD44+ breast cancer cells were abundant in metastatic lung and brain lesions in ER– breast cancer, suggesting that E-selectin supports hematogenous metastasis of ER–/CD44+ breast cancer. In an attempt to prevent hematogenous metastasis through the inhibition of a shear-resistant adhesion of CD44+ cancer cells to E-selectin-expressing blood vessels on the premetastatic niche, an E-selectin targeted aptamer (ESTA) was developed. We demonstrated that a single intravenous injection of ESTA reduced metastases to a baseline level in both syngeneic and xenogeneic forced breast cancer metastasis models without relocating the site of metastasis. The effect of ESTA was absent in E-selectin knockout mice, suggesting that E-selectin is a molecular target of ESTA. Our data highlight the potential application of an E-selectin antagonist for the prevention of hematogenous metastasis of ER–/CD44+ breast cancer.

Doublecortin-like kinase 1 (DCLK1) is a putative pancreatic stem cell marker and is upregulated in pancreatic cancer, colorectal cancer, and many other solid tumors. It marks tumor stem cells in mouse models of intestinal neoplasia. Here we sought to determine whether DCLK1 protein can be detected in the bloodstream and if its levels in archived serum samples could be quantitatively assessed in pancreatic cancer patients. DCLK1 specific ELISA, western blotting, and immunohistochemical analyses were used to determine expression levels in the serum and staining intensity in archived tumor tissues of pancreatic ductal adenocarcinoma (PDAC) patients and in pancreatic cancer mouse models. DCLK1 levels in the serum were elevated in early stages of PDAC (stages I and II) compared to healthy volunteers (normal controls). No differences were observed between stages III/IV and normal controls. In resected surgical tissues, DCLK1 expression intensity in the stromal cells was significantly higher than that observed in tumor epithelial cells. Circulating tumor cells were isolated from KPCY mice and approximately 52% of these cells were positive for Dclk1 staining. Dclk1 levels in the serum of KPC mice were also elevated. We have previously demonstrated that DCLK1 plays a potential role in regulating epithelial mesenchymal transition (EMT). Given the increasingly recognized role of EMT derived stem cells in cancer progression and metastasis, we hypothesize that DCLK1 may contribute to the metastatic process. Taken together, our results suggest that DCLK1 serum levels and DCLK1 positive circulating tumor cells should be further assessed for their potential diagnostic and prognostic significance.

We experimentally demonstrate that spin dynamics and the phase diagram of spinor condensates can be conveniently tuned by a two-dimensional optical lattice. Spin population oscillations and a lattice-tuned separatrix in phase space are observed in every lattice where a substantial superfluid fraction exists. In a sufficiently deep lattice, we observe a phase transition from a longitudinal polar phase to a broken-axisymmetry phase in steady states of lattice-confined spinor condensates. The steady states are found to depend sigmoidally on the lattice depth and exponentially on the magnetic field. We also introduce a phenomenological model that semi-quantitatively describes our data without adjustable parameters.

// Nathaniel Weygant 1 , Dongfeng Qu 1,2,3 , Randal May 1,2 , Ryan M. Tierney 1 , William L. Berry 4 , Lichao Zhao 5 , Shweta Agarwal 5 , Parthasarathy Chandrakesan 1 , Harisha R. Chinthalapally 1 , Nicholas T. Murphy 1 , James D. Li 1 , Sripathi M. Sureban 1,2,3 , Michael J. Schlosser 6 , James J. Tomasek 4 and Courtney W. Houchen 1,2,3,6 1 Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma, OK 2 Department of Veterans Affairs Medical Center, Oklahoma, OK 3 Peggy and Charles Stephenson Oklahoma Cancer Center, Oklahoma, OK 4 Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma, OK 5 Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma, OK 6 COARE Biotechnology, Oklahoma, OK Correspondence: Courtney W. Houchen, email: // Keywords : DCLK1, Renal Carcinoma, Tumor Stem Cell, Focal Adhesion, EMT Received : October 31, 2014 Accepted : December 10, 2014 Published : December 11, 2014 Abstract Renal clear cell carcinoma (RCC) is the most common type of kidney cancer and the 8 th most common cancer overall in the US. RCC survival rates drop precipitously with regional and distant spread and recent studies have demonstrated that RCC presents an epithelial-mesenchymal transition (EMT) phenotype linked to increased recurrence and decreased survival. EMT is a key characteristic of tumor stem cells (TSCs) along with chemo-resistance and radio-resistance, which are also phenotypic of RCC. Targeting these factors is key to increasing the survival of RCC patients. Doublecortin-like kinase 1 (DCLK1) marks TSCs in pancreatic and colorectal cancer and regulates EMT and stemness. Analysis of the Cancer Genome Atlas’ RCC dataset revealed that DCLK1 is overexpressed and dysregulated on the mRNA and epigenetic level in more than 93% of RCC tumors relative to adjacent normal tissue. Immunohistochemistry using α-DCLK1 antibody confirmed overexpression and demonstrated a major increase in immunoreactivity in stage II-III tumors compared to normal kidney and stage I tumors. Small-interfering RNA (siRNA) mediated knockdown of DCLK1 resulted in decreased expression of EMT and pluripotency factors and significantly reduced invasion, migration, focal adhesion, drug-resistance, and clonogenic capacity. These findings suggest that DCLK1 is a novel, overexpressed factor in RCC progression that may be targeted to suppress EMT, metastasis, and stemness in early-stage and advanced RCC to increase patient survival. Moreover, the possibility that DCLK1 may mark a population of tumor stem-like cells in RCC should be further investigated in light of these findings.

We experimentally study two quantum phase transitions in a sodium spinor condensate immersed in a microwave dressing field. We also demonstrate that many previously unexplored regions in the phase diagram of spinor condensates can be investigated by adiabatically tuning the microwave field across one of the two quantum phase transitions. This method overcomes two major experimental challenges associated with some widely used methods, and is applicable to other atomic species. Agreements between our data and the mean-field theory for spinor Bose gases are also discussed.

We experimentally study spin dynamics in a sodium antiferromagnetic spinor condensate as a result of spin-dependent interactions $c$ and microwave dressing field interactions characterized by the net quadratic Zeeman effect ${q}_{\mathrm{net}}$. In contrast to magnetic fields, microwave dressing fields enable us to access both negative and positive values of ${q}_{\mathrm{net}}$. We find an experimental signature to determine the sign of ${q}_{\mathrm{net}}$ and observe harmonic spin population oscillations at every ${q}_{\mathrm{net}}$ except near each separatrix in phase space where spin oscillation period diverges. No spin domains and spatial modes are observed in our system. Our data in the negative ${q}_{\mathrm{net}}$ region exactly resembles what is predicted to occur in a ferromagnetic spinor condensate in the positive ${q}_{\mathrm{net}}$ region. This observation agrees with an important prediction derived from the mean-field theory: spin dynamics in spin-1 condensates substantially depends on the sign of ${q}_{\mathrm{net}}/c$. This work uses only one atomic species to reveal mean-field spin dynamics, especially the remarkably different relationship between each separatrix and the magnetization, of spin-1 antiferromagnetic and ferromagnetic spinor condensates.

We present a simple and optimal experimental scheme for an all-optical production of a sodium spinor Bose-Einstein condensate (BEC). With this scheme, we demonstrate that the number of atoms in a pure BEC can be greatly boosted by a factor of 5 in a simple setup that includes a single-beam optical trap or a crossed optical trap. This optimal scheme avoids technical challenges associated with some all-optical BEC methods, and can be applied to other optically trappable atomic species. In addition, we find a good agreement between our theoretical model and data. The upper limit for the efficiency of evaporative cooling in all-optical BEC approaches is also discussed., 5 pages, 5 figures

Background: High mobility group A1 (HMGA1), a member of the non-histone chromosomal proteins and commonly referred to as architectural transcription factor, regulates transcription of various genes involved in cell growth and survival. Overexpression of HMGA1 has been shown to be associated with tumor progression and metastasis in several cancers, including human lung cancer. A recent study demonstrated that HMGA1 activates AKT function by reducing the activity of the protein phosphatase, phosphatase 2A subunitB (PPPR2A) via the oncogenic micro (mi) RNA-222. We demonstrated that interleukin (IL)-24, a novel tumor suppressor/cytokine, inhibited AKT in lung cancer cells. However, the molecular mechanism of AKT inhibition by IL-24 remains elusive. Aim: To determine the molecular mechanism of IL-24-mediated AKT inhibition involved the HMGA1/miR-222 axis. Methods: Human H1299 lung tumor cell line was stably transfected with a tetracycline-inducible plasmid vector carrying the IL-24. After the induced expression of IL-24 protein, expression levels of HMGA1 and its downstream molecular mechanisms were analyzed at the RNA and protein levels in lung cancer cell lines. The inhibitory effect of IL-24 on HMG A1/miR222 /AKT axis in the lung cancer cells is determined by RT-qPCR, western blot, reporter assay, and immunocytochemistry. Mechanistic approaches on overexpression and knockdown of HMGA1 and or miR-222 were utilized and the consequences of its inhibition/overexpression were analyzed on HMGA1/miR222 /AKT signaling axis and in vitro migration and invasion. Results: Upon induction of IL-24 expression in the H1299 lung tumor cells, we observed a marked reduction in HMGA1protein and mRNA levels. Using a mechanistic approach, we found that IL-24 reduced miR-222-3p and -5p levels, as determined by qRT-PCR. Associated with HMGA1 and miR-222 inhibition was a marked increase in PPP2R2A, with a concomitant decrease in phosphorylated AKTT308/S473 expression. SiRNA-mediated knockdown of HMGA1in combination with IL-24 significantly reduced AKT T308/S473 protein expression and greatly reduced cell migration and invasion compared with individual treatments. Further combination of IL-24 and a miR-222-3p inhibitor significantly increased PPP2R2A expression. Conclusion: Our results demonstrate for the first time that IL-24 inhibits AKT via regulating the HMGA1/miR-222 signaling node in human lung cancer cells and acts as an effective tumor suppressor. HMGA1 should present a novel target for the effective treatment of lung cancer. Citation Format: Janani Panneerselvam, Akhil Srivastava, Ranganayaki Muralidharan, Qi Wang, Wei Zheng, Lichao Zhao, Allshine Chen, Yan Zhao, Anupama Munshi, Rajagopal Ramesh. IL-24 modulates the high mobility group (HMG) A1/miR222 /AKT signaling in lung cancer cells. [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 3738.

HuR, an mRNA binding protein regulates the stability of many oncoproteins associated with cell survival, proliferation, migration and angiogenesis. HuR overexpression is a marker for poor prognosis in patients diagnosed with cancer of lung, ovary, breast and colon. We hypothesized that the silencing of HuR using small interfering RNA (siRNA) could be a promising approach for lung cancer therapy. To test our hypothesis, we developed a tumor-targeted nanoparticle (NP) system that is targeted to transferrin receptor (TfR) for delivering HuRsiRNA (HuR-TfNP) in human lung cancer cells. Human lung cancer cells (A549, HCC827) and normal lung fibroblast (MRC-9) cell lines expressing varying levels of TfR were used in the present study. TfR expression was highest in A549, moderate in HCC827, and low to undetectable in MRC9 cells. In vitro studies demonstrated enhanced uptake of Tf-NP (51%) in TfR overexpressing A549 cells, compared to the non-targeted NP. Specificity studies using desferrioxamine (DFO; 100 µM), a stimulator of TfR, showed a two-fold increased uptake of Tf-NP whereas blocking TfR with exogenous transferrin (1 µg/well) reduced the uptake by 3 fold in A549 cells. Further, HuR-TfNP treatment reduced HuR expression and significantly suppressed cell proliferation at 24h and 48h compared to control siRNA containing NP (C-Tf-NP) in tumor cells but not in normal cells. Greatest inhibition was observed in A549 cells (23% and 30% inhibition at 24 and 48 h respectively) compared to 15% and 25% in HCC827. In MRC-9 cells, only 4% inhibition was observed. HuR-TfNP induced G1 cell-cycle arrest in tumor cells that correlated with marked reduction in Cyclin D1, and Cyclin E protein expression. Further, tumor cell migration and invasion was significantly inhibited in HuR-TfNP treated tumor cells compared to C-TfNP treatment (p

Despite the success of CD20 antibody rituximab in immunotherapy, acquired resistance is one of the prime obstacles for the successful treatment of B-cell malignancies. There is an urgent need to intensify efforts against resistance in cancer treatment. Growing evidence indicated that lysosomes may form an “Achilles heel” for cancer cells by sensitizing them to death pathways. Here, we uncover an important role of CD20 in initiation of ceramide/lysosomal membrane permeabilization (LMP)-mediated cell death, showing that colocalization of CD20-TNFR1 after type II CD20 antibody ligation can stimulate de novo ceramide synthesis by ceramide synthase and consequently induce remarkable lysosomal permeabilization (LMP) and lysosome-mediated cell death. Further studies show that the potent lysosome-mediated cell death induced by CD20 antibodies exhibits a profound killing effect against both rituximab-sensitive and -resistant (RR) lymphoma. Furthermore, engineering of rituximab by introducing a point mutation endows it with the ability to induce potent ceramide/LMP-mediated cell death in both RR lymphoma and primary B-cell malignancies from patients with rituximab-refractory, suggesting the potential clinical application to combat rituximab resistance.

Background— The mechanism(s) for acute changes in electrophysiological properties of the atria during rapid pacing induced atrial fibrillation (AF) is not completely understood. We sought to evaluate the contribution of the intrinsic cardiac autonomic nervous system in acute atrial electrical remodeling and AF induced by 6-hour rapid atrial pacing. Methods and Results— Continuous rapid pacing (1200 bpm, 2� threshold [TH]) was performed at the left atrial appendage. Group 1 (n=7) underwent 6-hour pacing immediately followed by ganglionated plexi (GP) ablation; group 2 (n=7) underwent GP ablation immediately followed by 6-hour pacing; and group 3 (n=4) underwent administration of autonomic blockers, atropine (1 mg/kg), and propranolol (0.6 mg/kg) immediately followed by 6-hour pacing. The effective refractory period (ERP) and window of vulnerability (WOV, in milliseconds), ie, the difference between the longest and the shortest coupling interval of the premature stimulus that induced AF, were measured at 2�TH and 10�TH at the left atrium, right atrium, and pulmonary veins every hour before and after GP ablation or autonomic blockade. In group 1, ERP was markedly shortened in the first 2 hours and then stabilized both at 2�TH and 10�TH; however, WOV was progressively widened throughout the 6-hour period. After GP ablation, ERP was significantly longer than before ablation and AF could not be induced (WOV=0) at either 2�TH or 10�TH. In groups 2 and 3, rapid atrial pacing failed to shorten the ERP. AF could not be induced in 6 of 7 dogs in group 2 and all 4 dogs in group 3 during the 6-hour pacing period. Conclusion— The intrinsic cardiac autonomic nervous system plays a crucial role in the acute stages of atrial electrical remodeling induced by rapid atrial pacing.

Renal clear cell carcinoma (RCC) is the most common type of kidney cancer and the 8th most common cancer overall in the US. RCC survival rates drop precipitously with regional and distant spread and recent studies have demonstrated that RCC presents an epithelial-mesenchymal transition (EMT) phenotype linked to increased recurrence and decreased survival. EMT is a key characteristic of tumor stem cells (TSCs) along with chemo-resistance and radio-resistance, which are also phenotypic of RCC. Targeting these factors is key to increasing the survival of RCC patients. Doublecortin-like kinase 1 (DCLK1) marks TSCs in pancreatic and colorectal cancer and regulates EMT and stemness. Analysis of the Cancer Genome Atlas’ RCC dataset revealed that DCLK1 is overexpressed and dysregulated on the mRNA and epigenetic level in more than 93% of RCC tumors relative to adjacent normal tissue. Immunohistochemistry using α-DCLK1 antibody confirmed overexpression and demonstrated a major increase in immunoreactivity in stage II-III tumors compared to normal kidney and stage I tumors. Small-interfering RNA (siRNA) mediated knockdown of DCLK1 resulted in decreased expression of EMT and pluripotency factors and significantly reduced invasion, migration, focal adhesion, drug-resistance, and clonogenic capacity. These findings suggest that DCLK1 is a novel, overexpressed factor in RCC progression that may be targeted to suppress EMT, metastasis, and stemness in early-stage and advanced RCC to increase patient survival. Moreover, the possibility that DCLK1 may mark a population of tumor stem-like cells in RCC should be further investigated in light of these findings. Citation Format: Nathaniel Weygant, Dongfeng Qu, Randal May, Ryan M. Tierney, William L. Berry, Lichao Zhao, Shweta Agarwal, Parthasarathy Chandrakesan, Sripathi M. Sureban, Michael J. Schlosser, James J. Tomasek, Courtney W. Houchen. DCLK1 is a broadly dysregulated target against epithelial-mesenchymal transition, focal adhesion, and stemness in clear cell renal carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4374. doi:10.1158/1538-7445.AM2015-4374

Although trastuzumab has succeeded in breast cancer treatment, acquired resistance is one of the prime obstacles for breast cancer therapies. There is an urgent need to develop novel HER2 antibodies against trastuzumab resistance. Here, we first rational designed avidity-imporved trastuzumab and pertuzumab variants, and explored the correlation between the binding avidity improvement and their antitumor activities. After characterization of a pertuzumab variant L56TY with potent antitumor activities, a bispecific immunoglobulin G-like CrossMab (Tras-Permut CrossMab) was generated from trastuzumab and binding avidity-improved pertuzumab variant L56TY. Although, the antitumor efficacy of trastuzumab was not enhanced by improving its binding avidity, binding avidity improvement could significantly increase the anti-proliferative and antibody-dependent cellular cytotoxicity (ADCC) activities of pertuzumab. Further studies showed that Tras-Permut CrossMab exhibited exceptional high efficiency to inhibit the progression of trastuzumab-resistant breast cancer. Notably, we found that calreticulin (CRT) exposure induced by Tras-Permut CrossMab was essential for induction of tumor-specific T cell immunity against tumor recurrence. These data indicated that simultaneous blockade of HER2 protein by Tras-Permut CrossMab could trigger CRT exposure and subsequently induce potent tumor-specific T cell immunity, suggesting it could be a promising therapeutic strategy against trastuzumab resistance.

Sustaining the release of therapeutic nanoparticles in a cell-, tissue-, or disease-specific manner is a potentially powerful technology. A new drug carrier-dialdehyde starch nanoparticle (DASNP) that can sustain the loading and release of 5-fluorouracil (5-Fu) antitumor drug is reported in this study. IR spectrophotometer and 1H NMR confirmed the formation of aldehyde groups, and scan electron microscope determinations showed that the dialdehyde starch nanoparticles obtained had an average diameter of 90 nm. 5-Fu, the model drug, was conjugated into nanoparticles by aldehyde groups. These 5-Fu-binding nanoparticles significantly enhanced breast cancer cell (MCF-7) inhibition in vitro compared with free 5-Fu. After subcutaneous 0 injection in the breast tumor-loaded rats, 5-Fu-DASNP exhibited remarkable tumor-inhibitory efficacy determined by measuring tumor weight in vivo. The tumor inhibition of 5-Fu-DASNP was 61%±6%, whereas that of free 5-Fu was only 42%±4%. Bcl-2/Bax immunohistochemistry studies indicated that 5-Fu-DASNP remarkably induced tumor tissue necrosis. These results demonstrated that the DASNP prepared in this work is a potentially effective drug carrier.