SUMMARY Evaluation of the therapeutic potential of RNAi for HIV infection has been hampered by the challenges of siRNA delivery and lack of suitable animal models. Using a novel delivery method in humanized mice, we show that siRNA treatment can dramatically suppress HIV infection. A CD7-specific single-chain antibody was conjugated to oligo-9-arginine peptide (scFvCD7-9R) for T cell-specific siRNA delivery in NOD/SCIDIL2rγ−/− mice reconstituted with human lymphocytes (Hu-PBL) or CD34+ hematopoietic stem cells (Hu-HSC). In HIV-infected Hu-PBL mice, treatment with anti-CCR5 and antiviral siRNAs complexed to scFvCD7-9R controlled viral replication and prevented the disease-associated CD4 T cell loss. This treatment also suppressed endogenous virus and restored CD4 T cell counts in mice reconstituted with HIV+ PBMC. Moreover, scFvCD7-9R could deliver antiviral siRNAs to naïve T cells in Hu-HSC mice and effectively suppress viremia in infected mice. Thus, siRNA therapy for HIV infection appears to be feasible in a preclinical animal model.

The pursuit of ever more precise measures of time and frequency is likely to lead to the eventual redefinition of the second in terms of an optical atomic transition. To ensure continuity with the current definition, based on a microwave transition between hyperfine levels in ground-state 133 Cs, it is necessary to measure the absolute frequency of candidate standards, which is done by comparing against a primary cesium reference. A key verification of this process can be achieved by performing a loop closure-comparing frequency ratios derived from absolute frequency measurements against ratios determined from direct optical comparisons. We measure the 1 S 0 → 3 P 0 transition of 171 Yb by comparing the clock frequency to an international frequency standard with the aid of a maser ensemble serving as a flywheel oscillator. Our measurements consist of 79 separate runs spanning eight months, and we determine the absolute frequency to be 518 295 836 590 863.71(11) Hz, the uncertainty of which is equivalent to a fractional frequency of 2.1 × 10 −16 .This absolute frequency measurement, the most accurate reported for any transition, allows us to close the Cs-Yb-Sr-Cs frequency measurement loop at an uncertainty of <3×10 −16 , limited by the current realization of the SI second. We use these measurements to tighten the constraints on variation of the electron-to-proton mass ratio, µ = m e /m p . Incorporating our measurements with the entire record of Yb and Sr absolute frequency measurements, we infer a coupling coefficient to gravitational potential of k µ = (−1.9 ± 9.4) × 10 −7 and a drift with respect to time oḟ µ µ = (5.3 ± 6.5) × 10 −17 /yr. arXiv:1811.05885v1 [physics.atom-ph]

N-cadherin and HER2/neu were found to be co-expressed in invasive breast carcinomas. To test the contribution of N-cadherin and HER2 in mammary tumor metastasis, we targeted N-cadherin expression in the mammary epithelium of the MMTV-Neu mouse. In the context of ErbB2/Neu, N-cadherin stimulated carcinoma cell invasion, proliferation and metastasis. N-cadherin caused fibroblast growth factor receptor (FGFR) upmodulation, resulting in epithelial-to-mesenchymal transition (EMT) and stem/progenitor like properties, involving Snail and Slug upregulation, mammosphere formation and aldehyde dehydrogenase activity. N-cadherin potentiation of the FGFR stimulated extracellular signal regulated kinase (ERK) and protein kinase B (AKT) phosphorylation resulting in differential effects on metastasis. Although ERK inhibition suppressed cyclin D1 expression, cell proliferation and stem/progenitor cell properties, it did not affect invasion or EMT. Conversely, AKT inhibition suppressed invasion through Akt 2 attenuation, and EMT through Snail inhibition, but had no effect on cyclin D1 expression, cell proliferation or mammosphere formation. These findings suggest N-cadherin/FGFR has a pivotal role in promoting metastasis through differential regulation of ERK and AKT, and underscore the potential for targeting the FGFR in advanced ErbB2-amplified breast tumors.

No abstract

Dengue is a common arthropod-borne flaviviral infection in the tropics, for which there is no vaccine or specific antiviral drug. The infection is often associated with serious complications such as dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS), in which both viral and host factors have been implicated. RNA interference (RNAi) is a potent antiviral strategy and a potential therapeutic option for dengue if a feasible strategy can be developed for delivery of small interfering RNA (siRNA) to dendritic cells (DCs) and macrophages, the major in vivo targets of the virus and also the source of proinflammatory cytokines. Here we show that a dendritic cell-targeting 12-mer peptide ( Dengue is a mosquito-borne flavivirus infection that has emerged as a serious public health problem worldwide. Four serotypes of dengue virus (DEN-1 to DEN-4) are capable of causing human disease varying in severity from acute selflimiting febrile illness to life-threatening dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). The plasma leakage, hemorrhagic manifestations, and shock that characterize DHF/DSS are considered to have an immunological basis, as they are more common during secondary infection with a heterologous dengue virus strain (15,28,33). However, severe clinical manifestations can also occur during primary dengue infection, pointing to a contributory role of viral virulence factors. The WHO estimates that more than 20,000 people worldwide, mainly children, die each year from serious complications of dengue. No specific antiviral therapies are currently available for treating the infection, and efforts to develop a safe prophylactic vaccine have been hindered by the complex role of the immune system in disease pathogenesis (39,52,57). Thus, novel treatment strategies that block viral replication and/or to attenuate the exaggerated cytokine response associated with DHF/DSS complications are urgently needed.Potent and specific gene silencing mediated by RNA interference (RNAi) has generated a great deal of interest in development of RNAi as a therapeutic strategy against viral infections (50,54). Many studies have demonstrated the effectiveness of the RNAi approach to suppress flavivirus infection, including dengue virus replication in experimental cell lines (3,23,26,42,60). In addition, the versatility of RNAi could also be exploited to block important host mediators that contribute to dengue pathogenesis. However, the existence of * Corresponding author. Present address:

N-cadherin is a cell-cell adhesion molecule that plays a role in breast cancer metastasis. Here, we show that in vivo expression of N-cadherin in the PyMT mouse model, which enhances mammary tumor metastasis, results in selective inhibition of Akt3 expression and phosphorylation. Similarly, exogenous expression of N-cadherin in PyMT or MCF-7 mammary tumor cells enhanced cell motility and caused a dramatic reduction in Akt3 expression and phosphorylation. Moreover, knockdown of Akt3 in PyMT tumor cells increased cell motility and disrupted mammary morphogenesis, but had no effect on cell proliferation. Conversely, overexpression of wild-type Akt3 in PyMT-N-cadherin cells inhibited cell motility promoted by N-cadherin. Taken altogether, these findings demonstrate that N-cadherin suppresses Akt3 to promote cell motility and highlight the intricate regulation of Akt isoforms by N-cadherin during metastasis.

Global positioning system (GPS) carrier-phase time transfer, as a widely accepted highprecision time transfer method, frequently shows a data-batch boundary discontinuity of up to 1 ns, because of the inconsistency of the phase ambiguities between two consecutive data batches. To eliminate the data-batch boundary discontinuity, several techniques have been proposed in recent years. The question is how much the solutions of these techniques differ from each other and how well the solutions are faithful to clocks. To answer these questions, this paper chooses two techniques to study: revised RINEX-shift (RRS) technique [1, 2], and phase integer common-view (Phase-CV) technique [3]. This paper shows that the time deviation of the difference between the two techniques is below 100 ps, for an averaging time of less than 10 d. Especially, for an averaging time of less than 1 d, the time deviation is less than 30 ps. We also find that both RRS and Phase-CV match TWSTFT (two-way satellite time and frequency transfer) and TWOTFT (two-way optical-fiber time and frequency transfer) quite well. Especially, the difference between RRS/Phase-CV and TWOTFT is less than ±0.25 ns for more than 20 d, for a baseline of 268 km. These results show that both RRS and Phase-CV agree very well, and they are both faithful to clocks. However, this is based on the assumption that there is no obvious change in the GPS receiver reference time. When there is a sudden change in the reference time, Phase-CV cannot follow the time change. In contrast, RRS still follows the time change and represents the clock well.

Tumor cells must overcome apoptosis to survive throughout metastatic dissemination and distal organ colonization. Here we show in the Polyoma Middle T mammary tumor model that N-cadherin expression causes Slug upregulation, which in turn promotes carcinoma cell survival. Slug was dramatically upregulated in metastases relative to primary tumors. Consistent with a role in metastasis, Slug knockdown in carcinoma cells suppressed lung colonization by decreasing cell survival at metastatic sites, but had no effect on tumor cell invasion or extravasation. In support of this idea, Slug inhibition by shRNA, sensitized tumor cells to apoptosis by DNA damage, resulting in caspase-3 and PARP cleavage. The pro-survival effect of Slug was found to be caused by direct repression of the pro-apoptotic gene, Puma, by Slug. Consistent with a pivotal role for a Slug-Puma axis in metastasis, inhibition of Puma by RNA interference in Slug-knockdown cells rescued lung colonization, whereas Puma overexpression in control tumor cells suppressed lung metastasis. The survival function of the Slug-Puma axis was confirmed in human breast cancer cells, where Slug knockdown increased Puma expression and inhibited lung colonization. This study demonstrates a pivotal role for Slug in carcinoma cell survival, implying that disruption of the Slug-Puma axis may impinge on the survival of metastatic cells.