Enhancing chemosensitivity to gemcitabine via RNA interference targeting the catalytic subunits of protein kinase CK2 in human pancreatic cancer cells

<p>Abstract</p> <p>Background</p> <p>Pancreatic cancer is a complex genetic disorder that is characterized by rapid progression, invasiveness, resistance to treatment and high molecular heterogeneity. Various agents have been used in clinical trials showing only modest improvements with respect to gemcitabine-based chemotherapy, which continues to be the standard first-line treatment for this disease. However, owing to the overwhelming molecular alterations that have been reported in pancreatic cancer, there is increasing focus on targeting molecular pathways and networks, rather than individual genes or gene-products with a combination of novel chemotherapeutic agents.</p> <p>Methods</p> <p>Cells were transfected with small interfering RNAs (siRNAs) targeting the individual CK2 subunits. The CK2 protein expression levels were determined and the effect of its down-regulation on chemosensitization of pancreatic cancer cells was investigated.</p> <p>Results</p> <p>The present study examined the impact on cell death following depletion of the individual protein kinase CK2 catalytic subunits alone or in combination with gemcitabine and the molecular mechanisms by which this effect is achieved. Depletion of the CK2α or -α' subunits in combination with gemcitabine resulted in marked apoptotic and necrotic cell death in PANC-1 cells. We show that the mechanism of cell death is associated with deregulation of distinct survival signaling pathways. Cellular depletion of CK2α leads to phosphorylation and activation of MKK4/JNK while down-regulation of CK2α' exerts major effects on the PI3K/AKT pathway.</p> <p>Conclusions</p> <p>Results reported here show that the two catalytic subunits of CK2 contribute differently to enhance gemcitabine-induced cell death, the reduced level of CK2α' being the most effective and that simultaneous reduction in the expression of CK2 and other survival factors might be an effective therapeutic strategy for enhancing the sensitivity of human pancreatic cancer towards chemotherapeutic agents.</p

Using Argo data to investigate the Meridional Overturning Circulation in the North Atlantic

Author Posting. © The Author(s), 2009. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Deep Sea Research Part I: Oceanographic Research Papers 57 (2010): 29-36, doi:10.1016/j.dsr.2009.10.003.Using a variety of oceanographic data, including direct volume transports in the Florida 19 Strait, and Argo float profiles and drift velocities at 24 and 36N in the North Atlantic, inverse calculations are presented in which the net meridional transport, 20 down to a depth of approximately 1600 m, is estimated at both latitudes for a five year period 2003-2007. The upper ocean is divided into 7 layers using neutral density, and mass conservation constraints have been applied to a closed box bounded by these latitudes, including the Florida Strait. Ekman layer transports have been included in the top-most layer, and the inverse calculation has solved for changes from the initial reference velocities, Ekman and Florida Strait transports, given a priori estimates on the accuracy of each of these quantities. Solutions with and without transformations due to Mediterranean Water (MW) formation are made. Our results indicate that 1) time-averaged transport estimates derived from Argo have significant less eddy noise than individual hydrographic sections, 2) Argo drift velocities provide information to the inverse solution for the ocean interior, and 3) comparison of the total integrated interior mass transports in the thermocline waters for the period 2003-2007 with the previous estimates based on trans-ocean hydrographic sections shows that the Meridional Overturning Circulation has not significantly changed since 1957.TJ would like to acknowledge support from NSF Grant OCE-0241354 and NOAA/CICOR grant NA17RJ1223

Immune-based therapies for hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the third most frequent cause of cancer-related death. The immune-rich contexture of the HCC microenvironment makes this tumour an appealing target for immune-based therapies. Here, we discuss how the functional characteristics of the liver microenvironment can potentially be harnessed for the treatment of HCC. We will review the evidence supporting a therapeutic role for vaccines, cell-based therapies and immune-checkpoint inhibitors and discuss the potential for patient stratification in an attempt to overcome the series of failures that has characterised drug development in this disease area

Gender Dimorphism in Skeletal Muscle Leptin Receptors, Serum Leptin and Insulin Sensitivity

To determine if there is a gender dimorphism in the expression of leptin receptors (OB-R170, OB-R128 and OB-R98) and the protein suppressor of cytokine signaling 3 (SOCS3) in human skeletal muscle, the protein expression of OB-R, perilipin A, SOCS3 and alpha-tubulin was assessed by Western blot in muscle biopsies obtained from the m. vastus lateralis in thirty-four men (age = 27.1±6.8 yr) and thirty-three women (age = 26.7±6.7 yr). Basal serum insulin concentration and HOMA were similar in both genders. Serum leptin concentration was 3.4 times higher in women compared to men (P<0.05) and this difference remained significant after accounting for the differences in percentage of body fat or soluble leptin receptor. OB-R protein was 41% (OB-R170, P<0.05) and 163% (OB-R128, P<0.05) greater in women than men. There was no relationship between OB-R expression and the serum concentrations of leptin or 17β-estradiol. In men, muscle OB-R128 protein was inversely related to serum free testosterone. In women, OB-R98 and OB-R128 were inversely related to total serum testosterone concentration, and OB-R128 to serum free testosterone concentration. SOCS3 protein expression was similar in men and women and was not related to OB-R. In women, there was an inverse relationship between the logarithm of free testosterone and SCOS3 protein content in skeletal muscle (r = −0.46, P<0.05). In summary, there is a gender dimorphism in skeletal muscle leptin receptors expression, which can be partly explained by the influence of testosterone. SOCS3 expression in skeletal muscle is not up-regulated in women, despite very high serum leptin concentrations compared to men. The circulating form of the leptin receptor can not be used as a surrogate measure of the amount of leptin receptors expressed in skeletal muscles

Ethnic Concentration, Cultural Identity and Immigrant Self-Employment in Switzerland

Immigrant self-employment rates vary considerably across regions in Switzerland. Business ownership provides an alternative to wage labour, where immigrants have to face structural barriers such as the limited knowledge of the local language, or difficulties in fruitfully making use of their own human capital. Despite their historically high unemployment rates with respect to natives, immigrants in Switzerland are less entrepreneurial. It is therefore important to uncover factors that may facilitate the transition from the status of immigrant to the one of economic agent. Among others factors, concentration in ethnic enclaves, as well as accumulated labour market experience and time elapsed since immigration, have been associated to higher business ownership rates. In this paper, we use a cross-section of 2,490 Swiss municipalities in order to investigate the role played by the ethnic concentration of immigrants, as well as cultural factors, in determining self-employment rates

Optimization of Pathogenicity Tests for Selection of Native Isolates of Entomopathogenic Fungi Isolated from Citrusgrowing Areas of México on Adults ofDiaphorina citriKuwayama (Hemiptera: Liviidae)

Huanglonbing (HLB), es considerado una de las más letales enfermedades de los cítricos alrededor del mundo, y ha alcanzado las principales áreas de producción de limón Mexicano (Citrus latifolia Tanaka) en la costa del pacifico de México. Los productores han iniciado el uso de insecticidas para controlar las poblaciones del psílido asiático de los cítricos, Diaphorina citri Kuwayama (Hemiptera: Liviidae), el vector del patógeno ‘Candidatus Liberibacter asiaticus’ asociado con el HLB. Actualmente los costos de los insecticidas y los efectos secundarios de su uso son las principales preocupaciones, ya que podrían perjudicar la estrategia de manejo contra el vector; y por lo tanto, alternativas ecológicas y económicamente viables a los insecticidas convencionales serian necesarias a corto plazo. Por tanto, el objetivo de este estudio fue evaluar la patogenicidad de 27 aislados nativos y 3 cepas de hongos entomopatógenos para determinar su potencial como agentes de control biológico sobre Diaphorina citri usando 2 diferentes métodos de bioensayo. Los bioensayos fueron realizados bajo condiciones de laboratorio (26 ± 2 °C, 60 ± 5% H.R y 16:8 h L:O) mediante la exposición de insectos adultos a una concentración de 1 × 108 conidios por mililitro utilizando 2 diferentes métodos de aplicación, es decir, por asperjado de esporas en las plántulas de cítricos y por asperjado directo a los psílidos adultos. Los resultados mostraron que para el asperjado directo a los adultos los aislados HIB-24 (B. bassiana) y HIB-32 (I. fumosorosea) mostraron el mayor porcentaje de mortalidad (60.66%). Respecto al asperjado de plántulas el aislado HIB-19 (I. fumosorosea) mostró el mayor porcentaje de mortalidad (62.02%). Los resultados de este estudio demuestran el potencial para el uso de hongos entomopatógenos en el manejo de D. citri en México. ABSTRACT Huanglongbing (HLB), considered one of the most lethal diseases of citrus worldwide, has reached the main areas of Mexican lime (Citrus latifolia Tanaka) fruit production on the Pacific coast of México. Growers have initiated intensive use of insecticides in order to control populations of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), the vector of the pathogen, ‘Candidatus Liberibacter asiaticus’ associated with huanglongbing. Presently, costs of insecticides and the side effects of their use are major concerns, because they could impair the management strategy against the vector; and thus, ecologically and economically viable alternatives to conventional insecticides are required in the short term. Therefore the goal of this study was to evaluate the pathogenicity of 27 native isolates and 3 strains of entomopathogenic fungi and determine their potential as biological control agents of D. citri by using 2 different bioassay methods. Bioassays were performed under laboratory conditions (26 ± 2 °C, 60 ± 5% RH and 16:8 h L:D) by exposing adult insects to a concentration of 1 × 108 conidia per milliliter using 2 different application methods, i.e., spraying the spores onto the citrus seedlings and spraying the spores directly onto the adult psyllids. The results showed that by direct spraying the adults, HIB-24 (B. bassiana) and HIB-32 (I. fumosorosea) isolates showed the highest mortality (60.66%). Regarding spraying of the seedlings, HIB-19 (I. fumosorosea) showed the highest percentage of mortality (62.02%). The results from this study demonstrate potential for using entomopathogenic fungi in the management of D. citri in México

An Evolutionarily Conserved Function of Polycomb Silences the MHC Class I Antigen Presentation Pathway and Enables Immune Evasion in Cancer.

Loss of MHC class I (MHC-I) antigen presentation in cancer cells can elicit immunotherapy resistance. A genome-wide CRISPR/Cas9 screen identified an evolutionarily conserved function of polycomb repressive complex 2 (PRC2) that mediates coordinated transcriptional silencing of the MHC-I antigen processing pathway (MHC-I APP), promoting evasion of T cell-mediated immunity. MHC-I APP gene promoters in MHC-I low cancers harbor bivalent activating H3K4me3 and repressive H3K27me3 histone modifications, silencing basal MHC-I expression and restricting cytokine-induced upregulation. Bivalent chromatin at MHC-I APP genes is a normal developmental process active in embryonic stem cells and maintained during neural progenitor differentiation. This physiological MHC-I silencing highlights a conserved mechanism by which cancers arising from these primitive tissues exploit PRC2 activity to enable immune evasion.Cancer Research UK Clinician Scientist Fellowship C53779/A20097 (M.L.B), Leukaemia Foundation Australia Senior Fellowship and Howard Hughes Medical Institute International Research Scholarship 55008729 (M.A.D), Peter and Julie Alston Centenary fellowship (K.D.S.), Wellcome Trust Principal Research Fellowship 101835/Z/13/Z (P.J.L), Peter MacCallum Postgraduate Scholarship (C.E.S), NHMRC Postgraduate Scholarship (K.L.C.), Maddie Riewoldt's Vision 064728 (Y-C.C), Victorian Cancer Agency (E.Y.N.L), CSL Centenary fellowship (S-J.D), National Breast Cancer Foundation Fellowship ECF-17-005 (P.A.B.), Addenbrooke’s Charitable Trust and NIHR Cambridge BRC (M.L.B., P.J.L), NHMRC grant 1085015, 1106444 (M.A.D) and 1128984 (M.A.D, S-J.D)

The Small GTPase RhoA Localizes to the Nucleus and Is Activated by Net1 and DNA Damage Signals

Rho GTPases control many cellular processes, including cell survival, gene expression and migration. Rho proteins reside mainly in the cytosol and are targeted to the plasma membrane (PM) upon specific activation by guanine nucleotide exchange factors (GEFs). Accordingly, most GEFs are also cytosolic or associated with the PM. However, Net1, a RhoA-specific GEF predominantly localizes to the cell nucleus at steady-state. Nuclear localization for Net1 has been seen as a mechanism for sequestering the GEF away from RhoA, effectively rendering the protein inactive. However, considering the prominence of nuclear Net1 and the fact that a biological stimulus that promotes Net1 translocation out the nucleus to the cytosol has yet to be discovered, we hypothesized that Net1 might have a previously unidentified function in the nucleus of cells.Using an affinity precipitation method to pulldown the active form of Rho GEFs from different cellular fractions, we show here that nuclear Net1 does in fact exist in an active form, contrary to previous expectations. We further demonstrate that a fraction of RhoA resides in the nucleus, and can also be found in a GTP-bound active form and that Net1 plays a role in the activation of nuclear RhoA. In addition, we show that ionizing radiation (IR) specifically promotes the activation of the nuclear pool of RhoA in a Net1-dependent manner, while the cytoplasmic activity remains unchanged. Surprisingly, irradiating isolated nuclei alone also increases nuclear RhoA activity via Net1, suggesting that all the signals required for IR-induced nuclear RhoA signaling are contained within the nucleus.These results demonstrate the existence of a functional Net1/RhoA signaling pathway within the nucleus of the cell and implicate them in the DNA damage response