MIR-433 Predicts poor response to chemotherapy in ovarian cancer patients by the induction of cellular senescence

Annually, ovarian cancer (OC) affects 240,000 women worldwide and is the most lethal gynaecological malignancy. Such mortality is predominantly associated with the development of an intrinsic and acquired resistance to chemotherapy, the lack of targeted therapies and the lack of biomarkers predicting response to standard treatment.

Our clinical data demonstrates that increased miR-433 expression in primary high grade serous OC (HGSOCs) is significantly associated with poor PFS (n=46, p=0.024). Interestingly, the IHC analysis of two miR-433 targets: MAD2 [1] and HDAC6 shows that low IHC levels of both proteins is also significantly associated with worse outcome (p=0.002 and 0.002 respectively; n=43). Additionally, the analysis of miR 433 in the publicly available TCGA dataset corroborates that high miR-433 is significantly correlated with worse OS for patients presenting with OC (n=558 and p=0.027). In vito, in a panel of OC cell lines, higher miR-433 and lower MAD2 and HDAC6 levels were associated with resistance to paclitaxel.

To further investigate the role of miR-433 in the cellular response to chemotherapy, we generated an OC cell line stably expressing miR-433 or miR-control. MTT viability assays and Western Blot analyses established that miR-433 cells were more resistant to paclitaxel treatment (50nM) compared to miR-controls. Importantly, we have shown for the first time that miR 433 induced senescence resulting in a chracteristic flattened morphology and down-regulation of phosphorylated Retinoblastoma (p Rb), a molecular marker of senescence. Surprisingly, miR 433 induced senescence was independent from two well recognised senescent drivers: namely p53/p21 and p16. To explore this further we performed an in silico analysis of seven microRNA platforms which indicated that miR 433 potentially targets Cyclin-dependent kinase CDK6, which promotes sustained phosphorylation of Rb and thus cell cycle progression. In vitro, the overexpression of pre-miR-433 resulted in diminished CDK6 expression demonstrating a novel interaction between miR-433 and CDK6.

In conclusion, this study demonstrates that high miR-433 expression predicts poor outcome in OC patients by putatively rendering OC cells resistant to paclitaxel treatment through the induction of cellular senescence identifying this microRNA as a potential marker of chemoresponse.

Elucidation of the RamA Regulon in Klebsiella pneumoniae Reveals a Role in LPS Regulation

Klebsiella pneumoniae is a significant human pathogen, in part due to high rates of multidrug resistance. RamA is an intrinsic regulator in K. pneumoniae established to be important for the bacterial response to antimicrobial challenge; however, little is known about its possible wider regulatory role in this organism during infection. In this work, we demonstrate that RamA is a global transcriptional regulator that significantly perturbs the transcriptional landscape of K. pneumoniae, resulting in altered microbe-drug or microbe-host response. This is largely due to the direct regulation of 68 genes associated with a myriad of cellular functions. Importantly, RamA directly binds and activates the lpxC, lpxL-2 and lpxO genes associated with lipid A biosynthesis, thus resulting in modifications within the lipid A moiety of the lipopolysaccharide. RamA-mediated alterations decrease susceptibility to colistin E, polymyxin B and human cationic antimicrobial peptide LL-37. Increased RamA levels reduce K. pneumoniae adhesion and uptake into macrophages, which is supported by in vivo infection studies, that demonstrate increased systemic dissemination of ramA overexpressing K. pneumoniae. These data establish that RamA-mediated regulation directly perturbs microbial surface properties, including lipid A biosynthesis, which facilitate evasion from the innate host response. This highlights RamA as a global regulator that confers pathoadaptive phenotypes with implications for our understanding of the pathogenesis of Enterobacter, Salmonella and Citrobacter spp. that express orthologous RamA proteins.

Bit erasure analysis of binary adders in quantum-dot cellular automata

As a post-CMOS technology, the incipient Quantum-dot Cellular Automata technology has various advantages. A key aspect which makes it highly desirable is low power dissipation. One method that is used to analyse power dissipation in QCA circuits is bit erasure analysis. This method has been applied to analyse previously proposed QCA binary adders. However, a number of improved QCA adders have been proposed more recently that have only been evaluated in terms of area and speed. As the three key performance metrics for QCA circuits are speed, area and power, in this paper, a bit erasure analysis of these adders will be presented to determine their power dissipation. The adders to be analysed are the Carry Flow Adder (CFA), Brent-Kung Adder (B-K), Ladner-Fischer Adder (L-F) and a more recently developed area-delay efficient adder. This research will allow for a more comprehensive comparison between the different QCA adder proposals. To the best of the authors' knowledge, this is the first time power dissipation analysis has been carried out on these adders.

A First Step Towards Cost Functions for Quantum-dot Cellular Automata Designs

Quantum-dot cellular automata (QCA) is potentially a very attractive alternative to CMOS for future digital designs. Circuit designs in QCA have been extensively studied. However, how to properly evaluate the QCA circuits has not been carefully considered. To date, metrics and area-delay cost functions directly mapped from CMOS technology have been used to compare QCA designs, which is inappropriate due to the differences between these two technologies. In this paper, several cost metrics specifically aimed at QCA circuits are studied. It is found that delay, the number of QCA logic gates, and the number and type of crossovers, are important metrics that should be considered when comparing QCA designs. A family of new cost functions for QCA circuits is proposed. As fundamental components in QCA computing arithmetic, QCA adders are reviewed and evaluated with the proposed cost functions. By taking the new cost metrics into account, previous best adders become unattractive and it has been shown that different optimization goals lead to different “best” adders.

A coiled-coil-repeat protein 'Ccrp' in Bdellovibrio bacteriovorus prevents cellular indentation, but is not essential for vibroid cell morphology

Bdellovibrio bacteriovorus are small, vibroid, predatory bacteria that grow within the periplasmic space of a host Gram-negative bacterium. The intermediate-filament (IF)-like protein crescentin is a member of a broad class of IF-like, coiled-coil-repeat-proteins (CCRPs), discovered in Caulobacter crescentus, where it contributes to the vibroid cell shape. The B. bacteriovorus genome has a single ccrp gene encoding a protein with an unusually long, stutter-free, coiled-coil prediction; the inactivation of this did not alter the vibriod cell shape, but caused cell deformations, visualized as chiselled insets or dents, near the cell poles and a general 'creased' appearance, under the negative staining preparation used for electron microscopy, but not in unstained, frozen, hydrated cells. Bdellovibrio bacteriovorus expressing 'teal' fluorescent protein (mTFP), as a C-terminal tag on the wild-type Ccrp protein, did not deform under negative staining, suggesting that the function was not impaired. Localization of fluorescent Ccrp-mTFP showed some bias to the cell poles, independent of the cytoskeleton, as demonstrated by the addition of the MreB-specific inhibitor A22. We suggest that the Ccrp protein in B. bacteriovorus contributes as an underlying scaffold, similar to that described for the CCRP protein FilP in Streptomyces coelicolor, preventing cellular indentation, but not contributing to the vibroid shape of the B. bacteriovorus cells.

Distributed Massive MIMO in Cellular Networks: Impact of Imperfect Hardware and Number of Oscillators

Distributed massive multiple-input multiple-output (MIMO) combines the array gain of coherent MIMO processing with the proximity gains of distributed antenna setups. In this paper, we analyze how transceiver hardware impairments affect the downlink with maximum ratio transmission. We derive closed-form spectral efficiencies expressions and study their asymptotic behavior as the number of the antennas increases. We prove a scaling law on the hardware quality, which reveals that massive MIMO is resilient to additive distortions, while multiplicative phase noise is a limiting factor. It is also better to have separate oscillators at each antenna than one per BS.

Cellular signalling effects in high precision radiotherapy

Radiotherapy is commonly planned on the basis of physical dose received by the tumour and surrounding normal tissue, with margins added to address the possibility of geometric miss. However, recent experimental evidence suggests that intercellular signalling results in a given cell's survival also depending on the dose received by neighbouring cells. A model of radiation-induced cell killing and signalling was used to analyse how this effect depends on dose and margin choices. Effective Uniform Doses were calculated for model tumours in both idealised cases with no delivery uncertainty and more realistic cases incorporating geometric uncertainty. In highly conformal irradiation, a lack of signalling from outside the target leads to reduced target cell killing, equivalent to under-dosing by up to 10% compared to large uniform fields. This effect is significantly reduced when higher doses per fraction are considered, both increasing the level of cell killing and reducing margin sensitivity. These effects may limit the achievable biological precision of techniques such as stereotactic radiotherapy even in the absence of geometric uncertainties, although it is predicted that larger fraction sizes reduce the relative contribution of cell signalling driven effects. These observations may contribute to understanding the efficacy of hypo-fractionated radiotherapy.

Epigenetic Changes in Endothelial Progenitors as a Possible Cellular Basis for Glycemic Memory in Diabetic Vascular Complications

The vascular complications of diabetes significantly impact the quality of life and mortality in diabetic patients. Extensive evidence from various human clinical trials has clearly established that a period of poor glycemic control early in the disease process carries negative consequences, such as an increase in the development and progression of vascular complications that becomes evident many years later. Importantly, intensive glycemic control established later in the disease process cannot reverse or slow down the onset or progression of diabetic vasculopathy. This has been named the glycemic memory phenomenon. Scientists have successfully modelled glycemic memory using various in vitro and in vivo systems. This review emphasizes that oxidative stress and accumulation of advanced glycation end products are key factors driving glycemic memory in endothelial cells. Furthermore, various epigenetic marks have been proposed to closely associate with vascular glycemic memory. In addition, we comment on the importance of endothelial progenitors and their role as endogenous vasoreparative cells that are negatively impacted by the diabetic milieu and may constitute a "carrier" of glycemic memory. Considering the potential of endothelial progenitor-based cytotherapies, future studies on their glycemic memory are warranted to develop epigenetics-based therapeutics targeting diabetic vascular complications.

Cost-Efficient Decimal Adder Design in Quantum-dot Cellular Automata

Applications that cannot tolerate the loss of accuracy that results from binary arithmetic demand hardware decimal arithmetic designs. Binary arithmetic in Quantum-dot cellular automata (QCA) technology has been extensively investigated in recent years. However, only limited attention has been paid to QCA decimal arithmetic. In this paper, two cost-efficient binary-coded decimal (BCD) adders are presented. One is based on the carry flow adder (CFA) using a conventional correction method. The other uses the carry look ahead (CLA) algorithm which is the first QCA CLA decimal adder proposed to date. Compared with previous designs, both decimal adders achieve better performance in terms of latency and overall cost. The proposed CFA-based BCD adder has the smallest area with the least number of cells. The proposed CLA-based BCD adder is the fastest with an increase in speed of over 60% when compared with the previous fastest decimal QCA adder. It also has the lowest overall cost with a reduction of over 90% when compared with the previous most cost-efficient design.