Quality Assurance and Cost Reduction in Histopathology Laboratories Using Tissue Microarrays.

In the context of cost increases of both labor and consumables, cheaper and faster histopathology methods are needed. We implemented in our research laboratory the use of tissue microarrays (TMA) for the parallel processing and analysis of tissue samples. In this study, we used seven pre-processed, paraffinated biomimetic sectionable support matrices serving as "recipient" paraffin blocks to embed a total of 196 tissue cores from formalin-fixed paraffin-embedded tissue samples (serving as "donor" paraffin blocks) from seven different rabbit organs. These tissue samples were obtained using four different processing protocols: two 6 h protocols with xylene as the transition solvent, and two using butanol instead (one 10 h in duration and the other 72 h long). While the samples from protocols 1 and 2 (with xylene) quite regularly generated peeling of some of the cores from the slides (most likely because of substandard paraffin infiltration), butanol processing performed flawlessly for both processing protocols. Our proposed technique of using TMAs in the research laboratory brings with it a significant reduction in time and consumable costs (up to 77 and 64%, respectively), but also new challenges for all the upstream processes.

Effects of Plant-Based Antioxidants in Animal Diets and Meat Products: A Review.

The perceived level of risk associated with a food product can influence purchase and consumption decisions. Thus, current trends in food safety address an issue of general interest-the identification of healthy and economical alternatives to synthetic antioxidants that may have harmful effects on human health. Still, the processors' target is to increase the shelf life of food products using preserving substances. Natural antioxidants can be extracted and used in the food industry from different plants, such as blueberry, broccoli, chokeberry, cinnamon, ginger, olives, oregano, etc. The identification of the main natural antioxidant types that have been used in the food industry is very important in order to provide a comprehensive analysis of the researched topic. In this regard, the aim of this paper was to illustrate the positive aspects of using natural antioxidants with preservative roles in meat products, while, at the same time, highlighting the potential risks induced by these compounds. All of those aspects are correlated with the impact of sensorial attributes and the improvement of the nutritional value of meat products.

Identification of Uric Acid Crystals Accumulation in Human and Animal Tissues Using Combined Morphological and Raman Spectroscopy Analysis.

Gout is a metabolic condition, common to animals and humans, issuing from the excessive accumulation of end products of proteins degradation. In this study, histopathological and cytological examinations, combined with Raman spectroscopy, have been performed to investigate tissue samples from reptiles, chickens, and humans, presenting lesions produced by uric acid accumulation. As a result of classic processing and staining techniques commonly used in the anatomopathological diagnosis, uric acid crystals lose their structural characteristics, thus making difficult a precise diagnostic. Therefore, complementary diagnostic methods, such as Raman spectroscopy, are needed. This study compares from several perspectives the above mentioned diagnostic methods, concluding that Raman spectroscopy provides highlights in the diagnosis of gout in humans and animals, also adding useful information to differential diagnosis of lesions.

Spectral forecast: A general purpose prediction model as an alternative to classical neural networks.

Here, we describe a general-purpose prediction model. Our approach requires three matrices of equal size and uses two equations to determine the behavior against two possible outcomes. We use an example based on photon-pixel coupling data to show that in humans, this solution can indicate the predisposition to disease. An implementation of this model is made available in the supplementary material.

Tissue microarrays - brief history, techniques and clinical future.

INTRODUCTION AND AIM: There is a growing need for better, cheaper and faster histopathological diagnostic. The authors reviewed the main steps of the efforts towards the improvement of the pre-analytical phase of tissue processing for histological examination. RESULTS: Since their introduction decades ago tissue microarrays (TMAs) proved their value by increasing efficiency, standardization and accuracy of many histological techniques, such as histochemistry, histoenzymology, immunohistochemistry, in situ hybridization, etc. By allowing the simultaneous analysis and comparison of multiple different tissues on a single histology slide (up to 1000 individual samples), TMAs are also having a significant economic advantage (consumables and labor). From its first description until recent years, the TMA techniques have evolved steadily but slowly despite many attempts to adapt it for clinical diagnostics. In this paper, we are reviewing the main techniques of obtaining TMA blocks from the beginning to the present day, as well as recent developments that are expanding their scope into high accuracy/efficiency clinical diagnostics. CONCLUSIONS: Considering recent developments, we believe that the prospect of high-throughput histology might be achievable in the not-so-distant future.

Detection of Circulating Tumor Cells Using Microfluidics.

Metastasis is the main cause of death in cancer patients worldwide. During metastasis, cancer cells detach from the primary tumor and invade distant tissue. The cells that undergo this process are called circulating tumor cells (CTCs). Studies show that the number of CTCs in the peripheral blood can predict progression-free survival and overall survival and can be informative concerning the efficacy of treatment. Research is now concentrated on developing devices that can detect CTCs in the blood of cancer patients with improved sensitivity and specificity that can lead to improved clinical evaluation. This review focuses on devices that detect and capture CTCs using different cell properties (surface markers, size, deformability, electrical properties, etc.). We also discuss the process of tumor cell dissemination, the biology of CTCs, epithelial-mesenchymal transition (EMT), and several challenges and clinical applications of CTC detection.

Pharmacotoxicological screening on new derivatives of beta-phenylethylamine, potential agonists of beta3-adrenergic receptors.

BACKGROUND AND AIMS: Beta3-adrenergic receptors (beta3-ARs) have been initially characterized in 1989. Afterwards, their tissue distribution was established: white and brown adipose tissue, central nervous system, myocardium (atrial and ventricular), blood vessels, smooth gastrointestinal muscles (stomach, small intestine, colon), gallbladder, urinary bladder, prostate, skeletal muscles. Non-clinical trials have demonstrated the major implication of beta3-ARs in glucose metabolism, implicitly, in insulin release, and also in obesity. Therefore, new compounds were synthesized starting from beta-phenylethylamine nucleus and substituted in various positions, for possible antidiabetic and÷or antiobesity action. MATERIALS AND METHODS: In the present research, the antidiabetic action of newly synthesized compounds was investigated on an experimental model of alloxan-induced diabetes, administered in dose of 130 mg÷kg body weight (bw), intraperitoneally (i.p.). After 14 days of treatment, glycemia and enzymes involved in homeostasis of glucose metabolism, glucose-6-phosphate dehydrogenase (G6PD), glucose-6-phosphatase (G6Pase) and hexokinase were determined. Animals were then euthanized and histopathology examinations were performed on harvested liver, kidney, spleen and brain in order to document pathological changes induced by alloxan-induced diabetes and÷or by tested compounds. RESULTS AND CONCLUSIONS: Glycemia in animals treated with the tested compounds decreased statistically significant for groups C2 and C3 (-42.13% and -37.2%, respectively), compared to diabetic control group. C2 was also the compound to favorably modify the dynamics of determined enzymes, together with the display of very good safety profile supported by minor, non-significant, histopathological changes.

A 3D map of the islet routes throughout the healthy human pancreas.

Islets of Langerhans are fundamental in understanding diabetes. A healthy human pancreas from a donor has been used to asses various islet parameters and their three-dimensional distribution. Here we show that islets are spread gradually from the head up to the tail section of the pancreas in the form of contracted or dilated islet routes. We also report a particular anatomical structure, namely the cluster of islets. Our observations revealed a total of 11 islet clusters which comprise of small islets that surround large blood vessels. Additional observations in the peripancreatic adipose tissue have shown lymphoid-like nodes and blood vessels captured in a local inflammatory process. Our observations are based on regional slice maps of the pancreas, comprising of 5,423 islets. We also devised an index of sphericity which briefly indicates various islet shapes that are dominant throughout the pancreas.

Effect of diet and omega-3 fatty acids in NAFLD.

Nonalcoholic fatty liver disease (NAFLD) ranges from simple hepatic steatosis to steatohepatitis (NASH) and cirrhosis. The aim of this study is to test beneficial effects of omega-3 fatty acids (DHA 130 mg, EPA 25 mg) treatment in NAFLD, in a mouse model of non-alcoholic fatty liver disease. As pretreatment, 50 mice were fed for one month with a high-fat diet to induce NAFLD. Then, the mice were divided in different groups according to diet (normo- or hypercaloric), with and without treatment with omega-3 fatty acids, for another month, forming the post-treatment group. The liver and blood samples were collected for biochemical and histopathological analysis. Biochemical parameters including: glycemia, total cholesterol, triglycerides, uric acid, albumin, total plasma antioxidant capacity (TEAC) was measured in serum. Glutathione (GSH), total thiols and malonyldialdehyde (MDA) were determined in mouse liver homogenates. Mice from post-treatment group, on hypercaloric diet with or without omega-3 fatty acids treatment, had medium hepatopathy (granular and vacuolar degeneration of the hepatocytes) and hypertriglyceridemia. Omega-3 fatty acid treatment lowered the rise of triglycerides (p<0.03), glycemia (p<0.01) and cholesterol (p<0.02) in serum and MDA level of the liver (p<0.05). Mice from post-treatment group, on normocaloric diet with or without omega-3 fatty acid had different histopathological and biochemical results. Those with normocaloric/normolipidic diet and omega-3 fatty acids treatment had reversed liver histopathological results from NASH to normal aspect and had the best metabolic parameters results. In conclusion, omega-3 fatty acids treatment associated with a normocaloric/normolipidic diet has hepatoprotective action in nonalcoholic fatty liver disease.