Double coupling Edman chemistry for high-sensitivity automated protein sequencing

Capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) is a promising new method for the analysis of protein sequencing products. It gives 10 zmol (1 zmol = 10(-21) mol) limits of detection (3 sigma) for fluorescein thiohydantoin (FTH) amino acids. We have developed a separation for the (FTH)-amino acid products generated from 18 of the 20 coded amino acids. The extremely low volume requirement associated with CE-LIF makes it incompatible with commercial sequencers. For this reason, we have also been developing a miniaturized sequencer that can be more easily coupled to our detection system. Both the CE-LIF system and the miniaturized sequencer are described.     

Characterization and screening for mutations of the growth arrest-specific 11 (GAS11) and C16orf3 genes at 16q24.3 in breast cancer

BACKGROUND: Both fibroblast-mediated cytokine gene therapy and bone marrow transplantation (BMT) have proven to be efficient protocols for the recovery of bone marrow depression. In this report, the effects of fibroblast-mediated interleukin (IL)-6 gene therapy, in combination with BMT, on the recovery of irradiation-induced bone marrow depression were investigated. METHODS: NIH3T3 fibroblast cells engineered to secrete IL-6 (NIH3T3-IL-6) or NIH3T3 cells transduced with the neomycin gene (NIH3T3-Neo), in combination with 10(7), 10(6), or 10(5) syngeneic bone marrow cells, were implanted into irradiated mice. RESULTS: The platelets and white blood cells in the peripheral blood of the irradiated mice increased greatly 12 days after implantation of NIH3T3-IL-6 cells and BMT, the white blood cell counts were restored to a normal level 32 days after the combined therapy, and the platelet number was obviously higher than that in mice implanted with NIH3T3-Neo and BMT. Twenty and 25 days after the combined therapy, the mice showed accelerated recovery of colony-forming unit (CFU)-granulocyte/macrophages and CFU-megakaryocytes when compared with the mice implanted with NIH3T3-Neo cells and BMT. Ten days after lethal irradiation with gamma rays, the spleens formed more CFU-spleen in mice implanted with NIH3T3-IL-6 cells and BMT than in mice injected with phosphate-buffered saline or NIH3T3-Neo cells. Combined therapy with NIH3T3-IL-6 cell implantation and BMT delayed the survival period of the hematopoietic-depressed mice significantly when compared with therapy with NIH3T3-Neo cell implantation and BMT. CONCLUSIONS: These data demonstrated that the combined therapy of fibroblast-mediated IL-6 gene therapy and BMT could significantly promote the recovery of irradiation-induced hematopoietic depression.     

High-resolution analysis of T-cell receptor beta-chain repertoires using DNA heteroduplex tracking: generally stable, clonal CD8+ expansions in all healthy young adults

Harringtonine (HT), an anticancer drug with high chemotherapeutic efficiency to human chronic granulocytic/myelomonocytic leukemia, has been reported to rapidly induce apoptosis in HL-60 cells in a wide scope/range of dosage by investigators from our lab and others. In the present studies, by using video enhancement contrast (VEC) microscopy, we dynamically analyzed changes in intracellular calcium distribution in a single HL-60 cell over the period from the initiation of apoptosis to the obvious appearance of chromatin condensation. The results from this paper demonstrated the striking distinction of intracellular calcium distribution at different time points after treatment with HT. Before treatment in normal HL-60 cells the highest [Ca2+]i accumulation was observed in the peri-nuclear area and the lowest was observed in the nucleus; after treatment with 1 microg/ml HT for 30 min intracellular calcium diffused all over the cell compartments, while intranuclear calcium increased comparatively and significantly. The phenomenon of intranuclear calcium accumulation was further confirmed by using laser scanning confocal microscopy (LSCM). In addition, co-localization of the highest calcium region with condensed chromatin in apoptotic HL-60 cells was also observed by LSCM. Our results suggest that two sequential alterations of intracellular calcium distribution occurred in apoptotic HL-60 cells induced by HT, i.e. (a) accumulation of calcium in the nucleus and (b) regionalization in a specific nuclear region.     

Synthesis and biological activities of phenyl piperazine-based peptidomimetic growth hormone secretagogues

A new class of potent, orally active phenyl piperazine-based GH secretagogues have been discovered from attempts to mimic the arrangement of the phenyl substituent in the spiroindanyl piperidine and spiroindoline sulfonamide privileged structures of 4 and 1, respectively. The best of these compounds, 18 (EC50 = 2.8 nM) is nearly as potent as MK-0677 for releasing GH from rat pituitary cells.     

A visual study of the breakdown of emulsions in porous coalescers

A two-dimensional photoetched glass flow cell was used to observe the process by which an emulsion was broken down by a granular porous coalescer. The effects of wettability, emulsion and collector zeta potentials, and emulsion droplet size on the coalescence phenomenon were studied. The possible use of a graded medium or a coalescer with mixed wettability to enhnce the coalescence process was also examined. Based on these visual studies, a set of guidelines was developed to aid in the selection of a porous coalescer for a given separation duty.     

Photo Sequencing

A group of people taking pictures of a dynamic event with their mobile phones is a popular sight. The set of still images obtained this way is rich in dynamic content but lacks accurate temporal information. We propose a method for photo-sequencing—temporally ordering a set of still images taken asynchronously by a set of uncalibrated cameras. Photo-sequencing is an essential tool in analyzing (or visualizing) a dynamic scene captured by still images. The first step of the method detects sets of corresponding static and dynamic feature points across images. The static features are used to determine the epipolar geometry between pairs of images, and each dynamic feature votes for the temporal order of the images in which it appears. The partial orders provided by the dynamic features are not necessarily consistent, and we use rank aggregation to combine them into a globally consistent temporal order of images. We demonstrate successful photo-sequencing on several challenging collections of images taken using a number of mobile phones.     

Water quality prediction capabilities of WASP model for a tropical lake system

Water quality modelling facilitates our better understanding of the processes taking place in a lake system, and conservation plans to address them. The water quality analysis simulation programme (WASP) was used in this study to predict daily variations in water quality parameters, namely dissolved oxygen, nitrate, phosphate and chlorophyll‐a and biochemical oxygen demand concentrations in a tropical lake system. The lake was divided into eight segments with the respective morphological, environmental and flow details being model inputs. The monthly concentration of each water quality parameter also comprised model input. The model output was daily spatiotemporal variation in these parameters over a period of 476 days. This study also indicated that the occurrence of precipitation plays a major role in defining the water quality of a tropical lake. The heavy precipitation after a long gap, especially during the summer season, results in a large quantity of organic matter entering the lake through drains, thereby increasing the organic matter and phosphate in the water body, and subsequently resulting in high chlorophyll‐a concentrations in the lake. A reduced chlorophyll‐a concentration was observed during the heavy rains. The water quality fluctuations are more pronounced with precipitation, especially where polluted drains enter the lake. An improved water quality can be observed downstream, including increased dissolved oxygen and nitrate concentrations. Improved water quality was observed during the postmonsoon period, with increased salinity and dissolved oxygen concentrations, a finding that confirms generalized and specific conclusions can be achieved with the use of the WASP model.     

A scalable data structure for real-time estimation of resource availability in build-to-order environments

This paper defines a highly scalable interval index structure called the Temporal Bin tree (TB-tree) that can be embedded in any resource planning application whose algorithms require efficiently estimating either the time that a resource will be available to process a specific task of known length or the net availability of a resource during a specified period of time. It is specifically engineered to meet the real-time response and space efficiency requirements of large-scale resource planning applications that are required for mass customization. Basically, the TB-tree is a binary tree structure that represents availability of a resource across a planning horizon. Representing intervals of availability hierarchically using a tree structure increases the efficiency of search for resource availability when the discretization of time is fine-grained or the planning horizon is long. The tree forms a backbone structure that does not require disruptive rebalancing during update operations, which would mitigate the ability of the tree to respond to queries in real time. Its specific implementation allows for random access at any level of the tree to further improve scalability. An application of planning to real-time promising of order due dates for custom built products provides the context for empirical evaluation. Results of analytical evaluations and simulation experiments clearly demonstrate the scalability of the TB-tree relative to existing index structures in terms of both time and space.     

Family psychosocial screening: should we focus on high-risk settings?

The objective of this study was to address the question of whether or not psychosocial screening should be focused on "high-risk" populations. A cross-sectional survey of mothers of young children was conducted in various clinics: 758 in teaching clinics, 444 in private practices, and 202 at a military clinic. The self-administered questionnaire covered demographic factors, problems in mothers family of origin, maternal depression, and substance abuse. Mothers in the teaching clinics were younger and had less education and lower incomes than mothers in private practices, with intermediate levels in the military clinic. However, a substantial proportion of mothers seen in all sites reported psychosocial problems. Approximately 20% of mothers in all sites reported a family history of alcoholism. Positive screens for maternal depression ranged from about 15% to 35%. Binge drinking was reported by 10% to 20% at different sites. Psychosocial problems were common even among families seen in "low-risk" settings. Focusing screening only on high-risk clinics would miss many families with psychosocial problems.     

Fuzzy Fruit Fly Optimized Node Quality-Based Clustering Algorithm for Network Load Balancing

Recently, the fundamental problem with Hybrid Mobile Ad-hoc Networks (H-MANETs) is to find a suitable and secure way of balancing the load through Internet gateways. Moreover, the selection of the gateway and overload of the network results in packet loss and Delay (DL). For optimal performance, it is important to load balance between different gateways. As a result, a stable load balancing procedure is implemented, which selects gateways based on Fuzzy Logic (FL) and increases the efficiency of the network. In this case, since gateways are selected based on the number of nodes, the Energy Consumption (EC) was high. This paper presents a novel Node Quality-based Clustering Algorithm (NQCA) based on Fuzzy-Genetic for Cluster Head and Gateway Selection (FGCHGS). This algorithm combines NQCA with the Improved Weighted Clustering Algorithm (IWCA). The NQCA algorithm divides the network into clusters based upon node priority, transmission range, and neighbour fidelity. In addition, the simulation results tend to evaluate the performance effectiveness of the FFFCHGS algorithm in terms of EC, packet loss rate (PLR), etc.