A novel Monoclonal Antibody against Notch1 Targets Leukemia-associated Mutant Notch1 and Depletes Therapy Resistant Cancer Stem Cells in Solid Tumors

Higher Notch signaling is known to be associated with hematological and solid cancers. We developed a potential immunotherapeutic monoclonal antibody (MAb) specific for the Negative Regulatory Region of Notch1 (NRR). The MAb604.107 exhibited higher affinity for the ``Gain-offunction'' mutants of Notch1 NRR associated with T Acute lymphoblastic Leukemia (T-ALL). Modeling of the mutant NRR with 12 amino-acid insertion demonstrated ``opening'' resulting in exposure of the S2-cleavage site leading to activated Notch1 signaling. The MAb, at low concentrations (1-2 mu g/ml), inhibited elevated ligand-independent Notch1 signaling of NRR mutants, augmented effect of Thapsigargin, an inhibitor of mutant Notch1, but had no effect on the wild-type Notch1. The antibody decreased proliferation of the primary T-ALL cells and depleted leukemia initiating CD34/CD44 high population. At relatively high concentrations, (10-20 mu g/ml), the MAb affected Notch1 signaling in the breast and colon cancer cell lines. The Notch-high cells sorted from solid-tumor cell lines exhibited characteristics of cancer stem cells, which were inhibited by the MAb. The antibody also increased the sensitivity to Doxorubucinirubicin. Further, the MAb impeded the growth of xenografts from breast and colon cancer cells potentiated regression of the tumors along with Doxorubucin. Thus, this antibody is potential immunotherapeutic tool for different cancers.

BLOC-2 targets recycling endosomal tubules to melanosomes for cargo delivery

Hermansky-Pudlak syndrome (HPS) is a group of disorders characterized by the malformation of lysosome-related organelles, such as pigment cell melanosomes. Three of nine characterized HPS subtypes result from mutations in subunits of BLOC-2, a protein complex with no known molecular function. In this paper, we exploit melanocytes from mouse HPS models to place BLOC-2 within a cargo transport pathway from recycling endosomal domains to maturing melanosomes. In BLOC-2-deficient melanocytes, the melanosomal protein TYRP1 was largely depleted from pigment granules and underwent accelerated recycling from endosomes to the plasma membrane and to the Golgi. By live-cell imaging, recycling endosomal tubules of wild-type melanocytes made frequent and prolonged contacts with maturing melanosomes; in contrast, tubules from BLOC-2-deficient cells were shorter in length and made fewer, more transient contacts with melanosomes. These results support a model in which BLOC-2 functions to direct recycling endosomal tubular transport intermediates to maturing melanosomes and thereby promote cargo delivery and optimal pigmentation.

Identification of microRNAs and their targets in Finger millet by high throughput sequencing

MicroRNAs are short non-coding RNAs which play an important role in regulating gene expression by mRNA cleavage or by translational repression. The majority of identified miRNAs were evolutionarily conserved; however, others expressed in a species-specific manner. Finger millet is an important cereal crop; nonetheless, no practical information is available on microRNAs to date. In this study, we have identified 95 conserved microRNAs belonging to 39 families and 3 novel microRNAs by high throughput sequencing. For the identified conserved and novel miRNAs a total of 507 targets were predicted. 11 miRNAs were validated and tissue specificity was determined by stem loop RT-qPCR, Northern blot. GO analyses revealed targets of miRNA were involved in wide range of regulatory functions. This study implies large number of known and novel miRNAs found in Finger millet which may play important role in growth and development. (C) 2015 Elsevier B.V. All rights reserved.

Recognizing drug targets using evolutionary information: implications for repurposing FDA-approved drugs against Mycobacterium tuberculosis H37Rv

Drug repurposing to explore target space has been gaining pace over the past decade with the upsurge in the use of systematic approaches for computational drug discovery. Such a cost and time-saving approach gains immense importance for pathogens of special interest, such as Mycobacterium tuberculosis H37Rv. We report a comprehensive approach to repurpose drugs, based on the exploration of evolutionary relationships inferred from the comparative sequence and structural analyses between targets of FDA-approved drugs and the proteins of M. tuberculosis. This approach has facilitated the identification of several polypharmacological drugs that could potentially target unexploited M. tuberculosis proteins. A total of 130 FDA-approved drugs, originally intended against other diseases, could be repurposed against 78 potential targets in M. tuberculosis. Additionally, we have also made an attempt to augment the chemical space by recognizing compounds structurally similar to FDA-approved drugs. For three of the attractive cases we have investigated the probable binding modes of the drugs in their corresponding M. tuberculosis targets by means of structural modelling. Such prospective targets and small molecules could be prioritized for experimental endeavours, and could significantly influence drug-discovery and drug-development programmes for tuberculosis.

Synthesis of Hollow Nanotubes of Zn2SiO4 or SiO2: Mechanistic Understanding and Uranium Adsorption Behavior

We report a facile synthesis of Zn2SiO4 nanotubes using a two-step process consisting of a wet-chemical synthesis of core-shell ZnO@SiO2 nanorods followed by thermal annealing. While annealing in air leads to the formation of hollow Zn2SiO4, annealing under reducing atmosphere leads to the formation of SiO2 nanotubes. We rationalize the formation of the silicate phase at temperatures much lower than the temperatures reported in the literature based on the porous nature of the silica shell on the ZnO nanorods. We present results from in situ transmission electron microscopy experiments to clearly show void nucleation at the interface between ZnO and the silica shell and the growth of the silicate phase by the Kirkendall effect. The porous nature of the silica shell is also responsible for the etching of the ZnO leading to the formation of silica nanotubes under reducing conditions. Both the hollow silica and silicate nanotubes exhibit good uranium sorption at different ranges of pH making them possible candidates for nuclear waste management.

Composite Guidance for Impact Angle Control Against Higher Speed Targets

In the literature, the impact angle control problem has been addressed mostly against lower speed or stationary targets. However, in the current defense scenario, targets of much higher speeds than interceptors are a reality. Moreover, approaching a higher speed target from a specified angle is important for effective seeker acquisition and enhanced warhead effectiveness. This paper proposes a composite proportional navigation guidance law using a combination of the standard proportional navigation and the recently proposed retroproportional navigation guidance laws for intercepting higher speed nonmaneuvering targets at specified impact angles in three-dimensional engagements. An analysis of the set of achievable impact angles by the composite proportional navigation guidance law is presented. It is shown that there exists an impulse bias that, when added to the composite proportional navigation guidance command, expands this set further by reversing the direction of the line-of-sight angular rotation vector. A bound on the magnitude of the bias is also derived. Finally, an implementation of this impulse bias, in the form of a series of pulses, is proposed and analyzed. Simulation results are also presented to support the analysis.

Composite Guidance for Impact Angle Control Against Higher Speed Targets

In the literature, the impact angle control problem has been addressed mostly against lower speed or stationary targets. However, in the current defense scenario, targets of much higher speeds than interceptors are a reality. Moreover, approaching a higher speed target from a specified angle is important for effective seeker acquisition and enhanced warhead effectiveness. This paper proposes a composite proportional navigation guidance law using a combination of the standard proportional navigation and the recently proposed retroproportional navigation guidance laws for intercepting higher speed nonmaneuvering targets at specified impact angles in three-dimensional engagements. An analysis of the set of achievable impact angles by the composite proportional navigation guidance law is presented. It is shown that there exists an impulse bias that, when added to the composite proportional navigation guidance command, expands this set further by reversing the direction of the line-of-sight angular rotation vector. A bound on the magnitude of the bias is also derived. Finally, an implementation of this impulse bias, in the form of a series of pulses, is proposed and analyzed. Simulation results are also presented to support the analysis.

Formation of nanoparticles from ultrafast laser condensates of metal targets

Optimised ultrafast laser ablation can result in almost complete ionisation of the target material and the formation of a high velocity plasma jet. Collisions with the ambient gas behind the shock front cools the material resulting in the formation of mainly spherical, single crystal nanoscale particles in the condensate. This work characterises the nanoscale structures produced by the ultrafast laser interactions in He atmospheres at STP with Ni and Al. High resolution transmission electron microscopy was employed to study the microstructure of the condensates and to classify the production of particles forms as a function of the illumination conditions.