A Study on Automated Process for Extracting White Blood Cellular Data from Microscopic Digital Injured Skeletal Muscle Images

Skeletal muscle injury is one of the common injuries caused by high-intensity sports activities, military related works, and natural disasters. In order to discover better therapies, it is important to study muscle regeneration process. Muscle regeneration process tracking is the act of monitoring injured tissue section over time, noting white blood cell behavior and cell-fiber relations. A large number of microscopic images are taken for tracking muscle regeneration process over multiple time instances. Currently, manual approach is widely used to analyze a microscopic image of muscle cross section, which is time consuming, tedious and buggy. Automation of this research methodology is essential to process a big amount of data. The objective of this thesis is to develop a framework to track the regeneration process automatically. The framework includes dynamic thresholding, morphological processing, and feature extraction.Based on the clinical assumptions, the threshold is calculated using standard deviation and mean of probable single cells. After thresholding, six parameters are calculated including average size, cell count, cell area density, cell count on the basis of size, the number of cytoplasmic and membrane cells as well as the average distance between cellular objects. All these parameters are estimated over the time, which helped to note the pattern of change in leukocytes (White blood cells) presence. Based on these results, a clear pattern of leukocyte movement is observed. Our future work includes deriving the mathematical equations using regression model on the data set of increased time points

Effect of a Large Dose of Di (2-ethylhexyl) phthalate (DEHP) on Hepatic Peroxisome in Cynomolgus Monkeys (Macaca Fascicularis)

To elucidate the effect of a large dose of di (2-ethylhexyl) phthalate (DEHP), a plasticizer and peroxisome proliferator-activated receptor-α (PPARα) agonist, on hepatic peroxisomes, we orally administered 1,000 mg/kg/day, once daily, to 3 male and 4 female cynomolgus monkeys for 28 days consecutively. Light-microscopic and electron microscopic examinations of the liver were carried out in conjunction with measurement of the hepatic fatty acid β-oxidation system (FAOS), carnitine acetyltransferase (CAT) and carnitine palmitoyltransferase (CPT) activities, which are peroxisomal and/or mitochondrial enzyme activities. Electron microscopically, enlargement of the mitochondria was observed with lamellar orientation of the cristae along the major axis. Although the number of peroxisomes showed a tendency to increase when compared with those in a biopsied specimen before treatment, no abnormality in morphology was observed. A slight increase in CPT activity was noted at termination. No changes were noted in hepatic FAOS or CAT activity. In conclusion, although repeated oral treatment of cynomolgus monkeys with a large dose of DEHP induced a subtle increase in the numbers of peroxisomes with slight enlargements of the mitochondria, this low-sensitivity response to peroxisome proliferators in cynomolgus monkeys was considered to be closer to the response in humans than that in rodents

A fast and robust hepatocyte quantification algorithm including vein processing

<p>Abstract</p> <p>Background</p> <p>Quantification of different types of cells is often needed for analysis of histological images. In our project, we compute the relative number of proliferating hepatocytes for the evaluation of the regeneration process after partial hepatectomy in normal rat livers.</p> <p>Results</p> <p>Our presented automatic approach for hepatocyte (HC) quantification is suitable for the analysis of an entire digitized histological section given in form of a series of images. It is the main part of an automatic hepatocyte quantification tool that allows for the computation of the ratio between the number of proliferating HC-nuclei and the total number of all HC-nuclei for a series of images in one processing run. The processing pipeline allows us to obtain desired and valuable results for a wide range of images with different properties without additional parameter adjustment. Comparing the obtained segmentation results with a manually retrieved segmentation mask which is considered to be the ground truth, we achieve results with sensitivity above 90% and false positive fraction below 15%.</p> <p>Conclusions</p> <p>The proposed automatic procedure gives results with high sensitivity and low false positive fraction and can be applied to process entire stained sections.</p

Liver Biopsy

Liver biopsy is recommended as the gold standard method to determine diagnosis, fibrosis staging, prognosis and therapeutic indications in patients with chronic liver disease. However, liver biopsy is an invasive procedure with a risk of complications which can be serious. This book provides the management of the complications in liver biopsy. Additionally, this book provides also the references for the new technology of liver biopsy including the non-invasive elastography, imaging methods and blood panels which could be the alternatives to liver biopsy. The non-invasive methods, especially the elastography, which is the new procedure in hot topics, which were frequently reported in these years. In this book, the professionals of elastography show the mechanism, availability and how to use this technology in a clinical field of elastography. The comprehension of elastography could be a great help for better dealing and for understanding of liver biopsy

Cytotoxicity and toxicokinetics of new psychoactive substances

To expand the knowledge about toxicokinetic and cytotoxic properties of new psychoactive substances (NPS), different in vitro and in vivo tests were (further) developed and applied on structurally different compounds. First, in vitro and in vivo models were used to determine the metabolic stability (in vitro half-life) and the metabolic fate to elucidate targets for urine-based screenings. Investigations on isozymes, which contribute to their metabolism, and on the plasma protein binding were conducted to identify potential interactions with other NPS or drugs (of abuse). Finally, an existing cytotoxicity high-content screening assay (HCSA) using HepG2 cells was optimized to specify the cytotoxic potential of NPS. Synthetic cannabinoids showed a more extensive metabolism compared to synthetic opioids, which was in accordance with the number of involved isozymes. Since plasma protein binding values exceeded 70% in all cases, relevant drug-drug interactions cannot be excluded. Based on the optimized HCSA, a cytotoxic potential was assigned for seven NPS. Therefore, for two NPS a previously reported in vivo hepatotoxicity can be confirmed in vitro. Furthermore, metabolism-based effects on their cytotoxicity were observed for two NPS.Zur Erforschung der toxikokinetischen und cytotoxischen Eigenschaften von neuen psychoaktiven Substanzen (NPS) wurden verschiedene in vitro und in vivo Tests (weiter-)entwickelt und an strukturell unterschiedlichen Verbindungen angewendet. Mittels Stoffwechselversuchen wurden unter anderem analytische Zielmoleküle für Urinuntersuchungen identifiziert. Um mögliche Interaktionen mit anderen NPS oder Drogen zu untersuchen, wurden die am Metabolismus beteiligte Isoenzyme sowie die Plasmaproteinbindung bestimmt. Im letzten Schritt ging es um die Optimierung eines bestehenden Hochdurchsatzverfahrens zur Bestimmung der zytotoxischen Eigenschaften von NPS an HepG2 Zellen. Synthetische Cannabinoide zeigten einen ausgeprägteren Metabolismus als synthetische Opioide, welches sich auch in der Anzahl der daran beteiligten Isoenzyme widerspiegelte. Die Plasmaproteinbindung der untersuchten NPS lag über 70%, weshalb Wechselwirkungen mit anderen Drogen nicht ausgeschlossen werden können. Mittels des optimierten Zytotoxizitäts-Assays konnten sieben NPS als potenziell zytotoxisch eingestuften werden. In zwei Fällen konnte somit eine zuvor beschriebene in vivo Hepatotoxizität in vitro bestätigt werden. Außerdem wurden Metabolismus-basierte Effekte an der Zytotoxizität zweier NPS beobachtet