Molecular manipulation of keratin 8/18 intermediate filaments: modulators of FAS-mediated death signaling in human ovarian granulosa tumor cells

Background: Granulosa cell tumors (GCT) are a rare ovarian neoplasm but prognosis is poor following recurrence. Keratin intermediate filaments expressed in these tumors are a diagnostic marker, yet paradoxically, may also constitute a target for therapeutic intervention. In the current study, we evaluated keratin 8/18 (K8/18) filament expression as a mechanism of resistance to apoptosis in GCT, specifically focusing on regulation of the cell surface death receptor, Fas (FAS). Methods: The GCT cell line, KGN, was transiently transfected with siRNA to KRT8 and KRT18 to reduce K8/18 filament expression. Expression of K8/18, FAS, and apoptotic proteins (PARP, cleaved PARP) were evaluated by fluorescence microscopy, flow cytometric analysis, and immunoblotting, respectively. The incidence of FAS-mediated apoptosis in KGN cells was measured by caspase 3/7 activity. All experiments were performed independently three to six times, using a fresh aliquot of KGN cells for each experiment. Quantitative data were analyzed by one- or two-way analysis of variance (ANOVA), followed by a Tukey’s post-test for multiple comparisons; differences among means were considered statistically significant at P \u3c 0.05. Results: Control cultures of KGN cells exhibited abundant K8/18 filament expression (~90 % of cells), and minimal expression of FAS (\u3c25 % of cells). These cells were resistant to FAS-activating antibody (FasAb)-induced apoptosis, as determined by detection of cleaved PARP and measurement of caspase 3/7 activity. Conversely, siRNA-mediated knock-down of K8/18 filament expression enhanced FAS expression (\u3e 70 % of cells) and facilitated FasAb-induced apoptosis, evident by increased caspase 3/7 activity (P \u3c 0.05). Additional experiments revealed that inhibition of protein synthesis, but not MEK1/2 or PI3K signaling, also prompted FasAb-induced apoptosis. Conclusions: The results demonstrated that K8/18 filaments provide resistance to apoptosis in GCT by impairing FAS expression. The abundance of keratin filaments in these cells and their role in apoptotic resistance provides a greater mechanistic understanding of ovarian tumorgenicity, specifically GCT, as well as a clinically-relevant target for potential therapeutic intervention

Joint measurement of multiple noncommuting parameters

Although quantum metrology allows us to make precision measurements beyond the standard quantum limit, it mostly works on the measurement of only one observable due to the Heisenberg uncertainty relation on the measurement precision of noncommuting observables for one system. In this paper, we study the schemes of joint measurement of multiple observables which do not commute with each other using the quantum entanglement between two systems. We focus on analyzing the performance of a SU(1,1) nonlinear interferometer on fulfilling the task of joint measurement. The results show that the information encoded in multiple noncommuting observables on an optical field can be simultaneously measured with a signal-to-noise ratio higher than the standard quantum limit, and the ultimate limit of each observable is still the Heisenberg limit. Moreover, we find a resource conservation rule for the joint measurement

An efficient optimization approach for designing machine learning models based on genetic algorithm

Machine learning (ML) methods have shown powerful performance in different application. Nonetheless, designing ML models remains a challenge and requires further research as most procedures adopt a trial and error strategy. In this study, we present a methodology to optimize the architecture and the feature configurations of ML models considering a supervised learning process. The proposed approach employs genetic algorithm (GA)-based integer-valued optimization for two ML models, namely deep neural networks (DNN) and adaptive neuro-fuzzy inference system (ANFIS). The selected variables in the DNN optimization problems are the number of hidden layers, their number of neurons and their activation function, while the type and the number of membership functions are the design variables in the ANFIS optimization problem. The mean squared error (MSE) between the predictions and the target outputs is minimized as the optimization fitness function. The proposed scheme is validated through a case study of computational material design. We apply the method to predict the fracture energy of polymer/nanoparticles composites (PNCs) with a database gathered from the literature. The optimized DNN model shows superior prediction accuracy compared to the classical one-hidden layer network. Also, it outperforms ANFIS with significantly lower number of generations in GA. The proposed method can be easily extended to optimize similar architecture properties of ML models in various complex systems

Influence of planets on debris disks in star clusters I: the 50 AU Jupiter

Although debris disks may be common in exoplanet systems, only a few systems are known in which debris disks and planets coexist. Planets and the surrounding stellar population can have a significant impact on debris disk evolution. Here we study the dynamical evolution of debris structures around stars embedded in star clusters, aiming to determine how the presence of a planet affects the evolution of such structures. We combine NBODY6++GPU and REBOUND to carry out N-body simulations of planetary systems in star clusters (N=8000; Rh=0.78 pc) for a period of 100 Myr, in which 100 solar-type stars are assigned 200 test particles. Simulations are carried out with and without a Jupiter-mass planet at 50 au. We find that the planet destabilizes test particles and speeds up their evolution. The planet expels most particles in nearby and resonant orbits. Remaining test particles tend to retain small inclinations when the planet is present, and fewer test particles obtain retrograde orbits. Most escaping test particles with speeds smaller than the star cluster's escape speed originate from cold regions of the planetary system or from regions near the planet. We identify three regions within planetary systems in star clusters: (i) the private region of the planet, where few debris particles remain (40 - 60 au), (ii) the reach of the planet, in which particles are affected by the planet (0 - 400 au), and (iii) the territory of the planetary system, most particles outside which will eventually escape (0 - 700 au).Comment: 17 pages, 20 figures, and 3 tables. Accepted for publication in MNRA

Decoupling Cross-Quadrature Correlations using Passive Operations

Quadrature correlations between subsystems of a Gaussian quantum state are fully characterised by its covariance matrix. For example, the covariance matrix determines the amount of entanglement or decoherence of the state. Here, we establish when it is possible to remove correlations between conjugate quadratures using only passive operations. Such correlations are usually undesired and arise due to experimental cross-quadrature contamination. Using the Autonne--Takagi factorisation, we present necessary and sufficient conditions to determine when such removal is possible. Our proof is constructive, and whenever it is possible we obtain an explicit expression for the required passive operation

Optical-fiber source of polarization-entangled photon pairs in the 1550nm telecom band

We present a fiber based source of polarization-entangled photon pairs that is well suited for quantum communication applications in the 1550nm band of standard fiber-optic telecommunications. Polarization entanglement is created by pumping a nonlinear-fiber Sagnac interferometer with two time-delayed orthogonally-polarized pump pulses and subsequently removing the time distinguishability by passing the parametrically scattered signal-idler photon pairs through a piece of birefringent fiber. Coincidence detection of the signal-idler photons yields biphoton interference with visibility greater than 90%, while no interference is observed in direct detection of either the signal or the idler photons. All four Bell states can be prepared with our setup and we demonstrate violations of CHSH form of Bell's inequalities by up to 10 standard deviations of measurement uncertainty.Comment: 12 pages, 4 figures, to be submitted to Phys. Rev. Lett. See also paper QTuB4 in QELS'03 Technical Digest (OSA, Washington, D.C., 2003). This is a more complete versio

CRISPR/Cas Derivatives as Novel Gene Modulating Tools:Possibilities and In Vivo Applications

The field of genome editing started with the discovery of meganucleases (e.g., the LAGLIDADG family of homing endonucleases) in yeast. After the discovery of transcription activator-like effector nucleases and zinc finger nucleases, the recently discovered clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated proteins (Cas) system has opened a new window of applications in the field of gene editing. Here, we review different Cas proteins and their corresponding features including advantages and disadvantages, and we provide an overview of the different endonuclease-deficient Cas protein (dCas) derivatives. These dCas derivatives consist of an endonuclease-deficient Cas9 which can be fused to different effector domains to perform distinct in vitro applications such as tracking, transcriptional activation and repression, as well as base editing. Finally, we review the in vivo applications of these dCas derivatives and discuss their potential to perform gene activation and repression in vivo, as well as their potential future use in human therapy

Analysis of Clock-Regulated Genes in Neurospora Reveals Widespread Posttranscriptional Control of Metabolic Potential

Neurospora crassa has been for decades a principal model for filamentous fungal genetics and physiology as well as for understanding the mechanism of circadian clocks. Eukaryotic fungal and animal clocks comprise transcription-translation-based feedback loops that control rhythmic transcription of a substantial fraction of these transcriptomes, yielding the changes in protein abundance that mediate circadian regulation of physiology and metabolism: Understanding circadian control of gene expression is key to understanding eukaryotic, including fungal, physiology. Indeed, the isolation of clock-controlled genes (ccgs) was pioneered in Neurospora where circadian output begins with binding of the core circadian transcription factor WCC to a subset of ccg promoters, including those of many transcription factors. High temporal resolution (2-h) sampling over 48 h using RNA sequencing (RNA-Seq) identified circadianly expressed genes in Neurospora, revealing that from ∼10% to as much 40% of the transcriptome can be expressed under circadian control. Functional classifications of these genes revealed strong enrichment in pathways involving metabolism, protein synthesis, and stress responses; in broad terms, daytime metabolic potential favors catabolism, energy production, and precursor assembly, whereas night activities favor biosynthesis of cellular components and growth. Discriminative regular expression motif elicitation (DREME) identified key promoter motifs highly correlated with the temporal regulation of ccgs. Correlations between ccg abundance from RNA-Seq, the degree of ccg-promoter activation as reported by ccg-promoter-luciferase fusions, and binding of WCC as measured by ChIP-Seq, are not strong. Therefore, although circadian activation is critical to ccg rhythmicity, posttranscriptional regulation plays a major role in determining rhythmicity at the mRNA level

Left lateralized cerebral glucose metabolism declines in amyloid-β positive persons with mild cognitive impairment

Background: Previous publications indicate that Alzheimer\u27s Disease (AD) related cortical atrophy may develop in asymmetric patterns, with accentuation of the left hemisphere. Since fluorodeoxyglucose positron emission tomography (FDG PET) measurements of the regional cerebral metabolic rate of glucose (rCMRgl) provide a sensitive and specific marker of neurodegenerative disease progression, we sought to investigate the longitudinal pattern of rCMRgl in amyloid-positive persons with mild cognitive impairment (MCI) and dementia, hypothesizing asymmetric declines of cerebral glucose metabolism. Methods: Using florbetapir PET and cerebrospinal fluid (CSF) measures to define amyloid-β (Aβ) positivity, 40 Aβ negative (Aβ-) cognitively unimpaired controls (CU; 76 ± 5y), 76 Aβ positive (Aβ+) persons with MCI (76 ± 7y) and 51 Aβ+persons with probable AD dementia (75 ± 7y) from the AD Neuroimaging Initiative (ADNI) were included in this study with baseline and 2-year follow-up FDG PET scans. The degree of lateralization of longitudinal rCMRgl declines in subjects with Aβ+MCI and AD in comparison with Aβ- CU were statistically quantified via bootstrapped lateralization indices [(LI); range−1 (right) to 1 (left)]. Results: Compared to Aβ- CU, Aβ+MCI patients showed marked left hemispheric lateralization (LI: 0.78). In contrast, modest right hemispheric lateralization (LI: −0.33) of rCMRgl declines was found in Aβ+persons with probable AD dementia. Additional comparisons of Aβ+groups (i.e. MCI and probable AD dementia) consequently indicated right hemispheric lateralization (LI: −0.79) of stronger rCMRgl declines in dementia stages of AD. For all comparisons, voxel-based analyses confirmed significant (pFWE\u3c0.05) declines of rCMRgl within AD-typical brain regions. Analyses of cognitive data yielded predominant decline of memory functions in both MCI and dementia stages of AD. Conclusions: These data indicate that in early stages, AD may be characterized by a more lateralized pattern of left hemispheric rCMRgl declines. However, metabolic differences between hemispheres appear to diminish with further progression of the disease

Left lateralized cerebral glucose metabolism declines in amyloid-β positive persons with mild cognitive impairment

Background: Previous publications indicate that Alzheimer\u27s Disease (AD) related cortical atrophy may develop in asymmetric patterns, with accentuation of the left hemisphere. Since fluorodeoxyglucose positron emission tomography (FDG PET) measurements of the regional cerebral metabolic rate of glucose (rCMRgl) provide a sensitive and specific marker of neurodegenerative disease progression, we sought to investigate the longitudinal pattern of rCMRgl in amyloid-positive persons with mild cognitive impairment (MCI) and dementia, hypothesizing asymmetric declines of cerebral glucose metabolism. Methods: Using florbetapir PET and cerebrospinal fluid (CSF) measures to define amyloid-β (Aβ) positivity, 40 Aβ negative (Aβ-) cognitively unimpaired controls (CU; 76 ± 5y), 76 Aβ positive (Aβ+) persons with MCI (76 ± 7y) and 51 Aβ+persons with probable AD dementia (75 ± 7y) from the AD Neuroimaging Initiative (ADNI) were included in this study with baseline and 2-year follow-up FDG PET scans. The degree of lateralization of longitudinal rCMRgl declines in subjects with Aβ+MCI and AD in comparison with Aβ- CU were statistically quantified via bootstrapped lateralization indices [(LI); range−1 (right) to 1 (left)]. Results: Compared to Aβ- CU, Aβ+MCI patients showed marked left hemispheric lateralization (LI: 0.78). In contrast, modest right hemispheric lateralization (LI: −0.33) of rCMRgl declines was found in Aβ+persons with probable AD dementia. Additional comparisons of Aβ+groups (i.e. MCI and probable AD dementia) consequently indicated right hemispheric lateralization (LI: −0.79) of stronger rCMRgl declines in dementia stages of AD. For all comparisons, voxel-based analyses confirmed significant (pFWE\u3c0.05) declines of rCMRgl within AD-typical brain regions. Analyses of cognitive data yielded predominant decline of memory functions in both MCI and dementia stages of AD. Conclusions: These data indicate that in early stages, AD may be characterized by a more lateralized pattern of left hemispheric rCMRgl declines. However, metabolic differences between hemispheres appear to diminish with further progression of the disease