Determination of the Maturation Status of Dendritic Cells by Applying Pattern Recognition to High-Resolution Images.

The maturation or activation status of dendritic cells (DCs) directly correlates with their behavior and immunofunction. A common means to determine the maturity of dendritic cells is from high-resolution images acquired via scanning electron microscopy (SEM) or atomic force microscopy (AFM). While direct and visual, the determination has been made by directly looking at the images by researchers. This work reports a machine learning approach using pattern recognition in conjunction with cellular biophysical knowledge of dendritic cells to determine the maturation status of dendritic cells automatically. The determination from AFM images reaches 100% accuracy. The results from SEM images reaches 94.9%. The results demonstrate the accuracy of using machine learning for accelerating data analysis, extracting information, and drawing conclusions from high-resolution cellular images, paving the way for future applications requiring high-throughput and automation, such as cellular sorting and selection based on morphology, quantification of cellular structure, and DC-based immunotherapy.

Applying Pattern Recognition to High-Resolution Images to Determine Cellular Signaling Status.

Two frequently used tools to acquire high- resolution images of cells are scanning electron microscopy (SEM) and atomic force microscopy (AFM). The former provides a nanometer resolution view of cellular features rapidly and with high throughput, while the latter enables visualizing hydrated and living cells. In current practice, these images are viewed by eye to determine cellular status, e.g., activated versus resting. Automatic and quantitative data analysis is lacking. This paper develops an algorithm of pattern recognition that works very effectively for AFM and SEM images. Using rat basophilic leukemia cells, our approach creates a support vector machine to automatically classify resting and activated cells. Ten-fold cross-validation with cells that are known to be activated or resting gives a good estimate of the generalized classification results. The pattern recognition of AFM images achieves 100% accuracy, while SEM reaches 95.4% for our images as well as images published in prior literature. This outcome suggests that our methodology could become an important and frequently used tool for researchers utilizing AFM and SEM for structural characterization as well as determining cellular signaling status and function.

Electroporation-induced cell lysis in SWLA-2 hybridomas.

This paper describes experimental results involving the percentage cell lysis in SWLA-2 murine hybridomas produced by square wave electric field pulses of 100, 200, and 300 V across a 1 mm gap width in a standard cuvette. Pulse lengths were of 0.2 and 0.6 ms duration; 1, 2, or 3 pulses were applied with 100 ms time interval between pulses. Cells were cultured and separate samples examined at 48 hours to determine cell mortality. Nearly 90% cell mortality was produced by applying 3 pulses at of 0.6 ms duration at 300 V.

Detection of Vesicular Stomatitis Virus using a Capacitive Immunosensor.

This paper describes the detection of Vesicular Stomatitis Virus using a novel capacitive immunosensor technology, whereby the respective antibodies for the antigens were used as a means for chemical detection on separate sensors. Devices were fabricated using standard etching and metal plating techniques, followed by immobilization of antibodies. Detection of antigen was performed by measuring voltage change due to changes in capacitance as antigen bound to the antibody surface. Capacitance changes were detected upon binding of specific antigen to the surface. This rugged, prototype device detected VSV down to the 2 pg/ml range.

Cell Lysis in SWLA-2 Hybridomas due to 1 kHz AC Electric Fields.

This paper describes results involving the percentage cell lysis of SWLA-2 murine hybridomas produced by AC electric field pulses at 1 kHz with pulse widths ranging from 1 ms to 1 second. Cells that had been exposed to the electric fields were cultured and replicate samples were examined at 48 hours to determine the number of viable cells.

An examination of the effect of an AC pulsed electric field on cell mortality in SWLA-2 hybridomas.

This work describes the initial experimental setup and results involving the percentage cell lysis in SWLA-2 murine hybridomas produced by AC electric field pulses of varying amplitudes and pulse widths. Cells were cultured and separate samples examined at 24 hours. The frequency, pulse width and peak-to-peak voltage were varied. AC electric fields producing at least 1 V across the cell membrane appear to be more effective in producing cell lysis than similar fields producing lower membrane voltages. Additionally, higher frequencies, in the 10 kHz range, appear to be more effective than lower frequencies at membrane voltages above 1 V in producing cell lysis.

An examination of the effect of decaying exponential pulse electric fields on cell mortality in murine spleenocytes, hybridomas, and human natural killer cells.

This work describes the percentage cell lysis produced by exponentially decaying electric field pulses of varying amplitudes and time constants. Three different cell types were examined: murine spleenocytes, hybridomas, and human natural killer. Cells were cultured and separate samples examined at 24 hours and 48 hours. Two sets of experiments were performed for each cell type. At 0.3 kV, the spleenocytes exhibited a mortality of roughly 50% twenty-four hours after exposure to the pulse; while at forty-eight hours the spleenocyte cell count had reduced to roughly 25% viable cells. All other cell types showed mortality consistently in excess of 80% at field pulse strengths of about 0.3 V/m.

Engineering managed care automation via the treatment planning process: a bottom-up approach.

This paper describes implementing technology used for developing a personalized treatment plan using a specific treatment planning process and methodology. This methodology involves defining categories of treatment, goals that a program can assist patients to achieve, client objectives that evidence movement towards achieving specific goals, and interventions that can be performed by the program to accomplish these objectives. Each goal is issued a start and end (review) date. At the review date, whether a patient accomplished the goal or not will be assessed by a clinician using acceptable testing methods. Staff intervene according to the objectives for which the intervention has been designated and chart against these objectives by comments and assigning a progress indicator. Using this methodology which includes these two disjoint measures, care facilities can utilize technology to employ a paperless system that will standardize the process for developing a person-centered plan, identify program and staff deficiencies, evaluate the effectiveness of changes made to the program with respect to staff and patients, and capture information necessary to produce detailed billing. Software that implements this method has successfully been employed at the Rose Hill Center for Psychiatric Treatment & Rehabilitation in less than six months. Facilities using this software can transfer patients between programs and keep a longitudinal patient record. Funding has been granted to use this software to study best practices in the treatment of schizophrenia and schizoaffective disorder.

Modeling HIV-1 dynamics and the effects of decreasing activated infected T-cell count by filtration.

Over the past 10 years mathematical models have been developed using differential equations for the progression of HIV-1 to AIDS in an infected patient. Additional terms and formulations are presented over time, as the disease is better understood. Experimentation has been used to obtain many of the modeling parameters. These previous works focus primarily on modeling the progression of the disease, some with intervention such as HAART drugs. Attempts have been made to filter HIV-1 and HIV-1-infected T-cells from the blood. A model is developed that characterizes the rate of filtrations impact on disease progression. Such studies could be also used to investigate the impact of an implantable HIV-1 filter. Proliferation rates of actively and latently infected T-cells are significant and have been incorporated into the models developed in this paper.

Using a system-on-a-chip implantable device to filter circulating infected cells in blood or lymph.

This paper describes a system on a chip (SoC) that makes use of nanoscale cellular adhesion mechanisms in an integrated electronic microsystem to filter infected cells from blood or lymph. An example of a human immunodeficiency virus-specific SoC is explored in depth. Such systems work in vivo, and blood and lymph are filtered on a continuous basis. With the intelligence on the chip, captured cells can be identified and lyzed, expelled, or otherwise acted upon. These types of systems transfer the burden of research from traditional chemotherapy to bioengineering and system design.