Novel cancer stem cell marker MVP enhances temozolomide-resistance in glioblastoma

The resistance of highly aggressive glioblastoma multiforme (GBM) to chemotherapy is a major clinical problem resulting in a poor prognosis. GBM contains a rare population of self-renewing cancer stem cells (CSCs) that proliferate, spurring the growth of new tumors, and evade chemotherapy. In cancer, major vault protein (MVP) is thought to contribute to drug resistance. However, the role of MVP as CSCs marker remains unknown and whether MVP could sensitize GBM cells to Temozolomide (TMZ) also is unclear. We found that sensitivity to TMZ was suppressed by significantly increasing the MVP expression in GBM cells with TMZ resistance. Also, MVP was associated with the expression of other multidrug-resistant proteins in tumorsphere of TMZ-resistant GBM cell, and was highly co-expressed with CSC markers in tumorsphere culture. On the other hands, knockdown of MVP resulted in reduced sphere formation and invasive capacity. Moreover, high expression of MVP was associated with tumor malignancy and survival rate in glioblastoma patients. Our study describes that MVP is a potentially novel maker for glioblastoma stem cells and may be useful as a target for preventing TMZ resistance in GBM patients.     

Fitting host-parasitoid models with CV2 > 1 using hierarchical generalized linear models.

The powerful general Pacala-Hassell host-parasitoid model for a patchy environment, which allows host density-dependent heterogeneity (HDD) to be distinguished from between-patch, host density-independent heterogeneity (HDI), is reformulated within the class of the generalized linear model (GLM) family. This improves accessibility through the provision of general software within well-known statistical systems, and allows a rich variety of models to be formulated. Covariates such as age class, host density and abiotic factors may be included easily. For the case where there is no HDI, the formulation is a simple GLM. When there is HDI in addition to HDD, the formulation is a hierarchical generalized linear model. Two forms of HDI model are considered, both with between-patch variability: one has binomial variation within patches and one has extra-binomial, overdispersed variation within patches. Examples are given demonstrating parameter estimation with standard errors, and hypothesis testing. For one example given, the extra-binomial component of the HDI heterogeneity in parasitism is itself shown to be strongly density dependent.     

Phospholipase D2 induces stress fiber formation through mediating nucleotide exchange for RhoA

Phospholipase D (PLD) is involved in diverse cellular processes including cell movement, adhesion, and vesicle trafficking through cytoskeletal rearrangements. However, the mechanism by which PLD induces cytoskeletal reorganization is still not fully understood. Here, we describe a new link to cytoskeletal changes that is mediated by PLD2 through direct nucleotide exchange on RhoA. We found that PLD2 induces RhoA activation independent of its lipase activity. PLD2 directly interacted with RhoA, and the PX domain of PLD2 specifically recognized nucleotide-free RhoA. Finally, we found that the PX domain of PLD2 has guanine nucleotide-exchange factor (GEF) activity for RhoA in vitro. In addition, we verified that overexpression of the PLD2-PX domain induces RhoA activation, thereby provoking stress fiber formation. Together, our findings suggest that PLD2 functions as an upstream regulator of RhoA, which enables us to understand how PLD2 regulates cytoskeletal reorganization in a lipase activity-independent manner.     

Separation and simultaneous detection of anticancer drugs in a microfluidic device with an amperometric biosensor

A simple and highly sensitive method for simultaneous detection of anticancer drugs is developed by integrating the preconcentration and separation steps in a microfluidic device with an amperometric biosensor. An amperometric detection with dsDNA and cardiolipin modified screen printed electrodes are used for the detection of anticancer drugs at the end of separation channel. The preconcentration capacity is enhanced thoroughly using field amplified sample stacking and field amplified sample injection techniques. The experimental parameters affecting the analytical performances, such as pH, temperature, buffer concentration, water plug length, and detection potential are optimized. A reproducible response is observed during multiple injections of samples with a RSD <5%. The calibration plots are linear with the correlation coefficient between 0.9913 and 0.9982 over the range of 2-60 pM. The detection limits of four drugs are determined to be between 1.2 (± 0.05) and 5.5 (± 0.3) fM. The applicability of the device to the direct analysis of anticancer drugs is successfully demonstrated in a real spiked urine sample. Device was also examined for interference effect of common chemicals present in real samples.     

Development and characterization of microsatellite markers for genetic analysis of the Korean black scraper, Thamnaconus modestus

Black scraper is a commercially important oceanodromous fish in Korea. The commercial catch of this fish has decreased continuously since 1990. However, its genetic characteristics have never been studied. For population genetic analysis to assist effective aquaculture and stock management efforts, we isolated and characterized 19 microsatellite loci using an microsatellite-enrichment method based on magnetic/biotin capture of microsatellite sequences from a size-selected genomic library. To characterize each locus, 30 individuals from a natural T. modestus population in southern Korea were genotyped. All loci except two, KTm213 and KTm284, were polymorphic, with an average of 15.18 alleles per locus (range, 2–41). The mean observed and expected heterozygosities were 0.67 (range, 0.33–0.97) and 0.79 (range, 0.40–1.00), respectively. A significant deviation from Hardy-Weinberg equilibrium was observed at six loci (KTm142, KTm145, KTm158, KTm24, KTm221, and KTm250). This high variability indicates that these microsatellites may be useful for high-resolution studies of population genetics.     

A new vicious cycle involving glutamate excitotoxicity, oxidative stress and mitochondrial dynamics

Glutamate excitotoxicity leads to fragmented mitochondria in neurodegenerative diseases, mediated by nitric oxide and S-nitrosylation of dynamin-related protein 1, a mitochondrial outer membrane fission protein. Optic atrophy gene 1 (OPA1) is an inner membrane protein important for mitochondrial fusion. Autosomal dominant optic atrophy (ADOA), caused by mutations in OPA1, is a neurodegenerative disease affecting mainly retinal ganglion cells (RGCs). Here, we showed that OPA1 deficiency in an ADOA model influences N-methyl-D-aspartate (NMDA) receptor expression, which is involved in glutamate excitotoxicity and oxidative stress. Opa1^enu/+ mice show a slow progressive loss of RGCs, activation of astroglia and microglia, and pronounced mitochondrial fission in optic nerve heads as found by electron tomography. Expression of NMDA receptors (NR1, 2A, and 2B) in the retina of Opa1^enu/+ mice was significantly increased as determined by western blot and immunohistochemistry. Superoxide dismutase 2 (SOD2) expression was significantly decreased, the apoptotic pathway was activated as Bax was increased, and phosphorylated Bad and BcL-xL were decreased. Our results conclusively demonstrate that not only glutamate excitotoxicity and/or oxidative stress alters mitochondrial fission/fusion, but that an imbalance in mitochondrial fission/fusion in turn leads to NMDA receptor upregulation and oxidative stress. Therefore, we propose a new vicious cycle involved in neurodegeneration that includes glutamate excitotoxicity, oxidative stress, and mitochondrial dynamics.     

Elevated CO2 does not affect stem CO2 efflux nor stem respiration in a dry Eucalyptus woodland,but it shifts the vertical gradient in xylem [CO2]

To quantify stem respiration (R_S) under elevated CO₂ (eCO₂), stem CO₂ efflux (E_A) and CO₂ flux through the xylem (F_T) should be accounted for, because part of respired CO₂ is transported upwards with the sap solution. However, previous studies have used E_A as a proxy of R_S, which could lead to equivocal conclusions. Here, to test the effect of eCO₂ on R_S, both E_A and F_T were measured in a free‐air CO₂ enrichment experiment located in a mature Eucalyptus native forest. Drought stress substantially reduced E_A and R_S, which were unaffected by eCO₂, likely as a consequence of its neutral effect on stem growth in this phosphorus‐limited site. However, xylem CO₂ concentration measured near the stem base was higher under eCO₂, and decreased along the stem resulting in a negative contribution of F_T to R_S, whereas the contribution of F_T to R_S under ambient CO₂ was positive. Negative F_T indicates net efflux of CO₂ respired below the monitored stem segment, likely coming from the roots. Our results highlight the role of nutrient availability on the dependency of R_S on eCO₂ and suggest stimulated root respiration under eCO₂ that may shift vertical gradients in xylem [CO₂] confounding the interpretation of E_A measurements.