Cancer therapy combining the modalities of hyperthermia and chemotherapy: In vitro cellular response after rapid heat accumulation in the cancer cell

Hyperthermia is usually used at a sub-lethal level in cancer treatment to potentiate the effects of chemotherapy. The purpose of this study is to investigate the role of heating rate in achieving synergistic cell killing by chemotherapy and hyperthermia. For this purpose, in vitro cell culture experiments with a uterine cancer cell line (MES-SA) and its multidrug resistant (MDR) variant MES-SA/Dx5 were conducted. The cytotoxicity, mode of cell death, induction of thermal tolerance and P-gp mediated MDR following the two different modes of heating were studied. Doxorubicin (DOX) was used as the chemotherapy drug. Indocyanine green (ICG), which absorbs near infrared light at 808nm (ideal for tissue penetration), was chosen for achieving rapid rate hyperthermia. A slow rate hyperthermia was provided by a cell culture incubator. The results show that the potentiating effect of hyperthermia to chemotherapy can be maximized by increasing the rate of heating as evident by the results from the cytotoxicity assay. When delivered at the same thermal dose, a rapid increase in temperature from 37°C to 43°C caused more cell membrane damage than gradually heating the cells from 37°C to 43°C and thus allowed for more intracellular accumulation of the chemotherapeutic agents. Different modes of cell death are observed by the two hyperthermia delivery methods. The rapid rate laser-ICG hyperthermia @ 43°C caused cell necrosis whereas the slow rate incubator hyperthermia @ 43°C induced very mild apoptosis. At 43°C a positive correlation between thermal tolerance and the length of hyperthermia exposure is identified. This study shows that by increasing the rate of heating, less thermal dose is needed in order to overcome P-gp mediated MDR.

Thermal imaging as a biometrics approach to facial signature authentication

This dissertation develops an image processing framework with unique feature extraction and similarity measurements for human face recognition in the thermal mid-wave infrared portion of the electromagnetic spectrum. The goals of this research is to design specialized algorithms that would extract facial vasculature information, create a thermal facial signature and identify the individual. The objective is to use such findings in support of a biometrics system for human identification with a high degree of accuracy and a high degree of reliability. This last assertion is due to the minimal to no risk for potential alteration of the intrinsic physiological characteristics seen through thermal infrared imaging. The proposed thermal facial signature recognition is fully integrated and consolidates the main and critical steps of feature extraction, registration, matching through similarity measures, and validation through testing our algorithm on a database, referred to as C-X1, provided by the Computer Vision Research Laboratory at the University of Notre Dame. Feature extraction was accomplished by first registering the infrared images to a reference image using the functional MRI of the Brain’s (FMRIB’s) Linear Image Registration Tool (FLIRT) modified to suit thermal infrared images. This was followed by segmentation of the facial region using an advanced localized contouring algorithm applied on anisotropically diffused thermal images. Thermal feature extraction from facial images was attained by performing morphological operations such as opening and top-hat segmentation to yield thermal signatures for each subject. Four thermal images taken over a period of six months were used to generate thermal signatures and a thermal template for each subject, the thermal template contains only the most prevalent and consistent features. Finally a similarity measure technique was used to match signatures to templates and the Principal Component Analysis (PCA) was used to validate the results of the matching process. Thirteen subjects were used for testing the developed technique on an in-house thermal imaging system. The matching using an Euclidean-based similarity measure showed 88% accuracy in the case of skeletonized signatures and templates, we obtained 90% accuracy for anisotropically diffused signatures and templates. We also employed the Manhattan-based similarity measure and obtained an accuracy of 90.39% for skeletonized and diffused templates and signatures. It was found that an average 18.9% improvement in the similarity measure was obtained when using diffused templates. The Euclidean- and Manhattan-based similarity measure was also applied to skeletonized signatures and templates of 25 subjects in the C-X1 database. The highly accurate results obtained in the matching process along with the generalized design process clearly demonstrate the ability of the thermal infrared system to be used on other thermal imaging based systems and related databases. A novel user-initialization registration of thermal facial images has been successfully implemented. Furthermore, the novel approach at developing a thermal signature template using four images taken at various times ensured that unforeseen changes in the vasculature did not affect the biometric matching process as it relied on consistent thermal features.

A novel method for rapid and selective extraction of male DNA from rape kits using alkaline lysis and pressure cycling technology (PCT)

There is an increasing demand for DNA analysis because of the sensitivity of the method and the ability to uniquely identify and distinguish individuals with a high degree of certainty. But this demand has led to huge backlogs in evidence lockers since the current DNA extraction protocols require long processing time. The DNA analysis procedure becomes more complicated when analyzing sexual assault casework samples where the evidence contains more than one contributor. Additional processing to separate different cell types in order to simplify the final data interpretation further contributes to the existing cumbersome protocols. The goal of the present project is to develop a rapid and efficient extraction method that permits selective digestion of mixtures. ^ Selective recovery of male DNA was achieved with as little as 15 minutes lysis time upon exposure to high pressure under alkaline conditions. Pressure cycling technology (PCT) is carried out in a barocycler that has a small footprint and is semi-automated. Typically less than 10% male DNA is recovered using the standard extraction protocol for rape kits, almost seven times more male DNA was recovered from swabs using this novel method. Various parameters including instrument setting and buffer composition were optimized to achieve selective recovery of sperm DNA. Some developmental validation studies were also done to determine the efficiency of this method in processing samples exposed to various conditions that can affect the quality of the extraction and the final DNA profile. ^ Easy to use interface, minimal manual interference and the ability to achieve high yields with simple reagents in a relatively short time make this an ideal method for potential application in analyzing sexual assault samples.^

A Novel Method for Rapid and Selective Extraction of Male DNA from Rape Kits using Alkaline Lysis and Pressure Cycling Technology (PCT)

There is an increasing demand for DNA analysis because of the sensitivity of the method and the ability to uniquely identify and distinguish individuals with a high degree of certainty. But this demand has led to huge backlogs in evidence lockers since the current DNA extraction protocols require long processing time. The DNA analysis procedure becomes more complicated when analyzing sexual assault casework samples where the evidence contains more than one contributor. Additional processing to separate different cell types in order to simplify the final data interpretation further contributes to the existing cumbersome protocols. The goal of the present project is to develop a rapid and efficient extraction method that permits selective digestion of mixtures. Selective recovery of male DNA was achieved with as little as 15 minutes lysis time upon exposure to high pressure under alkaline conditions. Pressure cycling technology (PCT) is carried out in a barocycler that has a small footprint and is semi-automated. Typically less than 10% male DNA is recovered using the standard extraction protocol for rape kits, almost seven times more male DNA was recovered from swabs using this novel method. Various parameters including instrument setting and buffer composition were optimized to achieve selective recovery of sperm DNA. Some developmental validation studies were also done to determine the efficiency of this method in processing samples exposed to various conditions that can affect the quality of the extraction and the final DNA profile. Easy to use interface, minimal manual interference and the ability to achieve high yields with simple reagents in a relatively short time make this an ideal method for potential application in analyzing sexual assault samples.