Preincubation of cut tobacco leaf explants promotes Agrobacterium-mediated transformation by increasing vir gene induction

The effects of preincubation of cut tobacco leaf explants on Agrobacterium transformation efficiency and induction of Agrobacterium virE-lacZ fusion were evaluated. Transformation efficiency was evaluated by histochemical and fluorometric analysis of beta-glucuronidase in leaf rings transformed with Agrobacterium tumefaciens strain LBA4404(pKIWI105). The transformation efficiency increased by 2-fold, 5-fold, and 4.3-fold upon preincubation for 24, 48, and 72 h, respectively. Preincubation for 24, 48, and 72 h increased the ability of tobacco leaf segments to induce Agrobacterium virE by 2.3-fold, 3.5-fold and 4.5-fold, respectively. The requirement of preincubation for increased transformation efficiency was obviated by the addition of 100 mu M acetosyringone to the freshly cut leaf rings cocultivated with Agrobacterium. The production of vii gene inducers by the leaf rings during the preincubation period is an important factor that contributes to increased transformation efficiency of Agrobacterium upon preincubation. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.

California bearing ratio behavior of cement-stabilized fly ash-soil mixes

Thermal power stations using pulverized coal as fuel generate large quantities of fly ash as a byproduct, which has created environmental and disposal problems. Using fly ash for gainful applications will solve these problems. Among the various possible uses for fly ash, the most massive and effective utilization is in geotechnical engineering applications like backfill material, construction of embankments, as a subbase material, etc. A proper understanding of fly ash-soil mixes is likely to provide viable solutions for its large-scale utilization. Earlier studies initiated in the laboratory have resulted in a good understanding of the California Bearing Ratio (CBR) behavior of fly ash-soil mixes. Subsequently, in order to increase the CBR value, cement has been tried as an additive to fly ash-soil mixes. This paper reports the results.

Water content - void ratio swell-shrink paths of compacted expansive soils

This paper addresses the behaviour of compacted expansive soils under swell-shrink cycles. Laboratory cyclic swell-shrink tests were conducted on compacted specimens of two expansive soils at surcharge pressures of 6.25, 50.00, and 100.00 kPa. The void ratio and water content of the specimens at several intermediate stages during swelling until the end of swelling and during shrinkage until the end of shrinkage were determined to trace the water content versus void ratio paths with an increasing number of swell-shrink cycles. The test results showed that the swell-shrink path was reversible once the soil reached an equilibrium stage where the vertical deformations during swelling and shrinkage were the same. This usually occurred after about four swell-shrink cycles. The swelling and shrinkage path of each specimen subjected to full swelling - full shrinkage cycles showed an S-shaped curve (two curvilinear portions and a linear portion). However, the swelling and shrinkage path occurred as a part of the S-shaped curve, when the specimen was subjected to full swelling - partial shrinkage cycles. More than 80% of the total volumetric change and more than 50% of the total vertical deformation occurred in the central linear portion of the S-shaped curve. The volumetric change was essentially parallel to the saturation line within a degree of saturation range of 50-80% for the equilibrium cycle. The primary value of the swell-shrink path is to provide information regarding the void ratio change that would occur for a given change in water content for any possible swell-shrink pattern. It is suggested that these swell-shrink paths can be established with a limited number of tests in the laboratory.

Expediency of Penetration Ratio and Evaluation of Mean Throughput for Safety and Commercial Applications in VANETs

Vehicular ad hoc network (VANET) applications are principally categorized into safety and commercial applications. Efficient traffic management for routing an emergency vehicle is of paramount importance in safety applications of VANETs. In the first case, a typical example of a high dense urban scenario is considered to demonstrate the role of penetration ratio for achieving reduced travel time between source and destination points. The major requirement for testing these VANET applications is a realistic simulation approach which would justify the results prior to actual deployment. A Traffic Simulator coupled with a Network Simulator using a feedback loop feature is apt for realistic simulation of VANETs. Thus, in this paper, we develop the safety application using traffic control interface (TraCI), which couples SUMO (traffic simulator) and NS2 (network simulator). Likewise, the mean throughput is one of the necessary performance measures for commercial applications of VANETs. In the next case, commercial applications have been considered wherein the data is transferred amongst vehicles (V2V) and between roadside infrastructure and vehicles (I2V), for which the throughput is assessed.

STEM+: A Fair and Efficient Algorithm for Scheduling on the DSCH in UMTS Networks

In Universal Mobile Telecommunication Systems (UMTS), the Downlink Shared Channel (DSCH) can be used for providing streaming services. The traffic model for streaming services is different from the commonly used continuously- backlogged model. Each connection specifies a required service rate over an interval of time, k, called the "control horizon". In this paper, our objective is to determine how k DSCH frames should be shared among a set of I connections. We need a scheduler that is efficient and fair and introduce the notion of discrepancy to balance the conflicting requirements of aggregate throughput and fairness. Our motive is to schedule the mobiles in such a way that the schedule minimizes the discrepancy over the k frames. We propose an optimal and computationally efficient algorithm, called STEM+. The proof of the optimality of STEM+, when applied to the UMTS rate sets is the major contribution of this paper. We also show that STEM+ performs better in terms of both fairness and aggregate throughput compared to other scheduling algorithms. Thus, STEM+ achieves both fairness and efficiency and is therefore an appealing algorithm for scheduling streaming connections.

Effect of N/Ga flux ratio on transport behavior of Pt/GaN Schottky diodes

GaN films were grown on c-plane sapphire by plasma-assisted molecular beam epitaxy (PAMBE). The effect of N/Ga flux ratio on structural, morphological, and optical properties was studied. The dislocation density found to increase with increasing the N/Ga ratio. The surface morphology of the films as seen by scanning electron microscopy shows pits on the surface and found that the pit density on the surface increases with N/Ga ratio. The room temperature photoluminescence study reveals the shift in band-edge emission toward the lower energy with increase in N/Ga ratio. This is believed to arise from the reduction in compressive stress in the films as is evidenced by room temperature Raman study. The transport studied on the Pt/GaN Schottky diodes showed a significant increase in leakage current with an increase in N/Ga ratio and was found to be caused by the increase in pit density as well as increase in dislocation density in the GaN films. (C) 2011 American Institute of Physics. [doi:10.1063/1.3634116]

The determination of stem cell fate by 3D scaffold structures through the control of cell shape

Stem cell response to a library of scaffolds with varied 3D structures was investigated. Microarray screening revealed that each type of scaffold structure induced a unique gene expression signature in primary human bone marrow stromal cells (hBMSCs). Hierarchical cluster analysis showed that treatments sorted by scaffold structure and not by polymer chemistry suggesting that scaffold structure was more influential than scaffold composition. Further, the effects of scaffold structure on hBMSC function were mediated by cell shape. Of all the scaffolds tested, only scaffolds with a nanofibrous morphology were able to drive the hBMSCs down an osteogenic lineage in the absence of osteogenic supplements. Nanofiber scaffolds forced the hBMSCs to assume an elongated, highly branched morphology. This same morphology was seen in osteogenic controls where hBMSCs were cultured on flat polymer films in the presence of osteogenic supplements (OS). In contrast, hBMSCs cultured on flat polymer films in the absence of OS assumed a more rounded and less-branched morphology. These results indicate that cells are more sensitive to scaffold structure than previously appreciated and suggest that scaffold efficacy can be optimized by tailoring the scaffold structure to force cells into morphologies that direct them to differentiate down the desired lineage. Published by Elsevier Ltd.

C:N ratio of Sediments in a sewage fed Urban Lake

C:N ratio of lake sediments provide valuable information about the source and proportions of terrestrial, phytogenic and phycogenic carbon and nitrogen. This study has been carried out in Varthur lake which is receiving sewage since many decades apart from large scale land cover changes. C:N profile of the surficial sediment layer collected in the rainy and the dry seasons revealed higher C:N values[43] due to the accumulation of autochthonous organic material mostly at the deeper portions of the lake. This also highlights N limitation in the sludge either due to uptake by micro and macro-biota or rapid volatilization, denitrification and possible leaching in water. Organic Carbon was lower towards the inlets and higher near the deeper zones. This pattern of Organic C deposition was aided by gusty winds and high flow conditions together with impacts by the land use land cover changes in the watershed. Spatial variability of C:N in surficial sediments is significant compared to its seasonal variability. This communication provides an insight to the pattern in which nutrients are distributed in the sludge/sediment and its variation across seasons and space impacted by the biotic process accompanied by the hydrodynamic changes in the lake.

A Monoclonal Antibody against Human Notch1 Ligand-Binding Domain Depletes Subpopulation of Putative Breast Cancer Stem-like Cells

Overexpression of Notch receptors and ligands has been associated with various cancers and developmental disorders, making Notch a potential therapeutic target. Here, we report characterization of Notch1 monoclonal antibodies (mAb) with therapeutic potential. The mAbs generated against epidermal growth factor (EGF) repeats 11 to 15 inhibited binding of Jagged1 and Delta-like4 and consequently, signaling in a dose-dependent manner, the antibodies against EGF repeats 11 to 12 being more effective than those against repeats 13 to 15. These data emphasize the role of EGF repeats 11 to 12 in ligand binding. One of the mAbs, 602.101, which specifically recognizes Notch1, inhibited ligand-dependent expression of downstream target genes of Notch such as HES-1, HES-5, and HEY-L in the breast cancer cell line MDA-MB-231. The mAb also decreased cell proliferation and induced apoptotic cell death. Furthermore, exposure to this antibody reduced CD44(Hi)/CD24(Low) subpopulation in MDA-MB-231 cells, suggesting a decrease in the cancer stem-like cell subpopulation. This was confirmed by showing that exposure to the antibody decreased the primary, secondary, and tertiary mammosphere formation efficiency of the cells. Interestingly, effect of the antibody on the putative stem-like cells appeared to be irreversible, because the mammosphere-forming efficiency could not be salvaged even after antibody removal during the secondary sphere formation. The antibody also modulated expression of genes associated with stemness and epithelial-mesenchymal transition. Thus, targeting individual Notch receptors by specific mAbs is a potential therapeutic strategy to reduce the potential breast cancer stem-like cell subpopulation. Mol Cancer Ther; 11(1); 77-86. (C) 2011 AACR.

Composition of Extrafloral Nectar Influences Interactions between the Myrmecophyte Humboldtia brunonis and its Ant Associates

Ant-plant interactions often are mediated by extrafloral nectar (EFN) composition that may influence plant visitation by ants. Over a 300 km range in the Indian Western Ghats, we investigated the correlation between the EFN composition of the myrmecophytic ant-plant Humboldtia brunonis (Fabaceae) and the number and species of ants visiting EFN. EFN composition varied among H. brunonis populations and between plant organs (floral bud vs. young leaf EFN). In general, EFN was rich in sugars with small quantities of amino acids, especially essential amino acids, and had moderate invertase activity. In experiments at the study sites with sugar and amino acid solutions and with leaf or floral bud EFN mimics, dominant EFN-feeding ants differentiated between solutions as well as between mimics. The castration parasite Crematogaster dohrni (northern study site) was the least selective and did not exhibit any clear feeding preferences, while the largely trophobiont-tending non-protective Myrmicaria brunnea (middle study site) preferred higher sucrose concentrations and certain essential/non-essential amino acid mixtures. The mutualistic Technomyrmex albipes (southern study site) preferred sucrose over glucose or fructose solutions and consumed the leaf EFN mimic to a greater extent than the floral bud EFN mimic. This young leaf EFN mimic had low sugar concentrations, the lowest viscosity and sugar: amino acid ratio, was rich in essential amino acids, and appeared ideally suited to the digestive physiology of T. albipes. This preference for young leaf EFN may explain the greater protection afforded to young leaves than to floral buds by T. albipes, and may also help to resolve ant-pollinator conflicts. The differential response of dominant ants to sugar, amino acids, or solution viscosity suggests that plants can fine-tune their interactions with local ants via EFN composition. Thus, EFN can mediate local partner-choice mechanisms in ant-plant interactions.

Introduction of SV40ER and hTERT into mammospheres generates breast cancer cells with stem cell properties

Emerging evidence suggests that cancers arise in stem/progenitor cells. Yet, the requirements for transformation of these primitive cells remains poorly understood. In this study, we have exploited the `mammosphere' system that selects for primitive mammary stem/progenitor cells to explore their potential and requirements for transformation. Introduction of Simian Virus 40 Early Region and hTERT into mammosphere-derived cells led to the generation of NBLE, an immortalized mammary epithelial cell line. The NBLEs largely comprised of bi-potent progenitors with long-term self-renewal and multi-lineage differentiation potential. Clonal and karyotype analyses revealed the existence of heterogeneous population within NBLEs with varied proliferation, differentiation and sphere-forming potential. Significantly, injection of NBLEs into immunocompromised mice resulted in the generation of invasive ductal adenocarcinomas. Further, these cells harbored a sub-population of CD44(+)/CD24(-) fraction that alone had sphere- and tumor-initiating potential and resembled the breast cancer stem cell gene signature. Interestingly, prolonged in vitro culturing led to their further enrichment. The NBLE cells also showed increased expression of stemness and epithelial to mesenchymal transition markers, deregulated self-renewal pathways, activated DNA-damage response and cancer-associated chromosomal aberrations-all of which are likely to have contributed to their tumorigenic transformation. Thus, unlike previous in vitro transformation studies that used adherent, more differentiated human mammary epithelial cells our study demonstrates that the mammosphere-derived, less-differentiated cells undergo tumorigenic conversion with only two genetic elements, without requiring oncogenic Ras. Moreover, the striking phenotypic and molecular resemblance of the NBLE-generated tumors with naturally arising breast adenocarcinomas supports the notion of a primitive breast cell as the origin for this subtype of breast cancer. Finally, the NBLEs represent a heterogeneous population of cells with striking plasticity, capable of differentiation, self-renewal and tumorigenicity, thus offering a unique model system to study the molecular mechanisms involved with these processes. Oncogene (2012) 31, 1896-1909; doi:10.1038/onc.2011.378; published online 29 August 2011