Ranan Gulhan Aktas

Correlation between the mechanical and histological properties of liver tissue.

In order to gain further insight into the mechanisms of tissue damage during the progression of liver diseases as well as the liver preservation for transplantation, an improved understanding of the relation between the mechanical and histological properties of liver is necessary. We suggest that this relation can only be established truly if the changes in the states of those properties are investigated dynamically as a function of post mortem time. In this regard, we first perform mechanical characterization experiments on three bovine livers to investigate the changes in gross mechanical properties (stiffness, viscosity, and fracture toughness) for the preservation periods of 5, 11, 17, 29, 41 and 53h after harvesting. Then, the histological examination is performed on the samples taken from the same livers to investigate the changes in apoptotic cell count, collagen accumulation, sinusoidal dilatation, and glycogen deposition as a function of the same preservation periods. Finally, the correlation between the mechanical and histological properties is investigated via the Spearmans Rank-Order Correlation method. The results of our study show that stiffness, viscosity, and fracture toughness of bovine liver increase as the preservation period is increased. These macroscopic changes are very strongly correlated with the increase in collagen accumulation and decrease in deposited glycogen level at the microscopic level. Also, we observe that the largest changes in mechanical and histological properties occur after the first 11-17h of preservation.

Ultrastructural immunolocalization of basic fibroblast growth factor in fibroblasts and extracellular matrix

Abstract Basic fibroblast growth factor (bFGF) is thought to play an important role in normal tissue repair and wound healing. It is a potent mitogenic and chemotactic factor for fibroblasts, regulating proliferation and extracellular matrix (ECM) production by these cells. In this study, we present morphologic evidence of the ultrastructural location of bFGF in fibroblasts and ECM using several antibodies, tissues, and species. Distinct labeling is seen in the nuclei of fibroblasts and some labeling in the cytosol. Immunolabeling of the cytosol excludes organelles involved in the usual secretory pathway, such as rough endoplasmic reticulum, Golgi apparatus, and secretory vacuoles. The same labeling is observed with either polyclonal or monoclonal antibodies. We suggest that bFGF functions as a nuclear protein in fibroblasts and is not secreted by a normal secretory pathway. Fibroblasts may export bFGF via unique cellular pathways that are clearly distinct from classic signal peptide mediated secretion. This may provide a source for ECM-resident bFGF. The same antibodies show different labeling intensity in the ECM. This protein, through integration into the ECM, may act as a local regulator and promote regeneration of these tissues after wounding. Direct evidence is the dramatic reduction of bFGF labeling in axotomized rat ECM collagen fibers versus control animals.

Electron microscopic immunolocalization of basic fibroblast growth factor in peripheral nerves

Abstract. In spite of ample information about the distribution and the effects of basic fibroblast growth factor (bFGF) in the central nervous system, few data are available concerning the localization of this protein in the peripheral nervous system. In view of the role of bFGF in the regulation of trophic and non-trophic functions, we focused on the presence and precise localization of this growth factor in normal peripheral nerves at the electron microscopic level. The study shows that bFGF is mainly located in the Schwann cells, especially in the nuclei. There is slight labeling in the myelin sheath and in the axon cytoplasm. The study provides morphologic evidence for an association between bFGF expression and Schwann cells. Such an association argues for a role of this peptide in the maintenance or regeneration of peripheral nerves.

Use of iodine-123 metaiodobenzylguanidine scintigraphy for the detection of amiodarone induced pulmonary toxicity in a rabbit model : a comparative study with technetium-99m diethyltriaminepenta acetic acid radioaerosol scintigraphy

The purpose of the study was; (i) to determine whether123I-MIBG scintigraphy is sensitive for detection of amiodarone induced pulmonary toxicity (AIPT) and (ii) to compare it with99mTc-DTPA radioaerosol. Twelve white New Zealand rabbit with initial mean body weight 4.24 ± 0.47 g were divided into two groups. AIPT group (n = 7) was administered amiodarone (20 mg/kg BW). The control group (n = 5) received the same amount of 0.9% saline. All animals underwent123I-MIBG and99mTc-DTPA radioaerosol scintigraphy at the end of the treatment period.123I-MIBG static thorax images were obtained during 10 minutes at 15 minutes and 3-hours after intravenous injection of the radiopharmaceutical. Lung to heart ratios (LHR) and lung to mediastinum ratios (LMR), and retention index (LRI) of123I-MIBG were determined. Two days after123I-MIBG scintigraphy,99mTc-DTPA radioaerosol scintigraphy was performed, and clearance from the lungs was measured for 10 min (1 min/frame) following termination of inhalation.123I-MIBG lung retention index (LRI) was significantly higher in the AIPT group than the control (61 ± 4.6 vs. 40 ± 4.5, p = 0.01). Early LHR and LMR were significantly lower in the AIPT group than in the control group (p = 0.04, p = 0.01, respectively), whereas those of late LHR and LMR were not significantly different. T1/2 values of DTPA clearance were significantly increased in AIPT group according to the control group (55 ± 7.2 vs. 86.6 ± 18.5, p = 0.02).123I-MIBG scintigraphy is a valuable tool for detecting AIPT in a rabbit model. Additionally,99mTc-DTPA radioaerosol scintigraphy is an excellent comprehensive investigational tool for detecting AIPT with the added advantage of lower cost.

Effect of solution and post-mortem time on mechanical and histological properties of liver during cold preservation.

BACKGROUND In liver transplantation, the donor and recipient are in different locations most of the time, and longer preservation periods are inevitable. Hence, the choice of the preservation solution and the duration of the preservation period are critical for the success of the transplant surgery. OBJECTIVE In this study, we examine the mechanical and histological properties of the bovine liver tissue stored in Lactated Ringers (control), HTK and UW solutions as a function of preservation period. METHODS The mechanical experiments are conducted with a shear rheometer on cylindrical tissue samples extracted from 3 bovine livers and the change in viscoelastic material properties of the bovine liver is characterized using the fractional derivative Kelvin-Voigt Model. Also, the histological examinations are performed on the same liver samples under a light microscope. RESULTS The results show that the preservation solution and period have a significant effect on the mechanical and histological properties of the liver tissue. The storage and loss shear moduli, the number of the apoptotic cells, the collagen accumulation, and the sinusoidal dilatation increase, and the glycogen deposition decreases as the preservation period is longer. CONCLUSIONS Based on the statistical analyses, we observe that the liver tissue is preserved well in all three solutions for up to 11 h. After then, UW solution provides a better preservation up to 29 h. However, for preservation periods longer than 29 h, HTK is a more effective preservation solution based on the least amount of change in mechanical properties. On the other hand, the highest correlation between the mechanical and histological properties is observed for the liver samples preserved in UW solution.

Ultrastructural immunolocalization of basic fibroblast growth factor in endothelial cells: morphologic evidence for unconventional secretion of a novel protein

Basic fibroblast growth factor (bFGF) is one of the most potent angiogenic factors. Unlike many other growth factors, bFGF lacks a classic peptide sequence for its secretion. Recent studies suggest that there is an unconventional secretory pathway for this growth factor. The aim of this study was to identify the specific location of bFGF in endothelial cells and to find morphologic evidences concerning its synthesis, storage and release from endothelial cells. The capillaries in hippocampus, adrenal gland, kidney, peripheral nerves as well as the vessels in connective tissues were analysed by using immunogold labeling techniques at electron microscope level. Results show that endogenous bFGF is mainly located in the nuclei of endothelial cells. Slight immunoreactivity is found in the cytoplasm. Immunolabeling is notably absent in pinocytotic vesicles, Golgi complexes, endoplasmic reticulum, nuclear membrane and intercellular junctions. These results provide morphologic evidence suggesting that endothelial cells might export bFGF via unique cellular pathways that are clearly distinct from classical signal peptide mediated secretion and/or release of this protein could be directly through mechanically induced disruptions of these cells. The current study support the recent hypothesis related with unconventional secretory pathway for bFGF as some other “cargo” proteins.

Blister Artifact Formation Due to Organic Solvent Effects on Spurr's Epoxy Resin

Wrinkles and air bubble artifacts may occur when preparing slides of semithin sections (0.5 microm) from blocks embedded in different resins. More than aesthetically annoying, wrinkles and air bubble artifacts may prohibit study of small structures. Present observations suggest that organic solvent based mounting media may interact with the resin of the section. This sometimes causes wrinkles and air bubble artifacts in the sections that degrade the quality of light microscope images. We compared the quality of semithin sections of several tissues in different resins using various types of mounting media. We observed that sections from Spurrs resin have many more artifacts. In particular, small, 2-10 microm round or oblong blister-like artifacts often plague our Spurrs resin sections. We demonstrate that Spurrs resin sections react with toluene and xylene in organic solvent based mounting media forming blisters, while sections from Araldite and L. R. White do not. We suggest combinations of embedding and mounting media for successful preparation of semithin sections for light microscopy without wrinkles, blisters, or air bubble artifacts.