Mehmet Dincer Bilgin

SINGLET OXYGEN GENERATION BY PHOTODYNAMIC AGENTS

The singlet oxygen quantum yield of photodynamic agents was measured at 546nm, 630 nm and on the far-red absorption peak. The technique employed is available in most laboratories, in which the photosensitization of lysozyme is used as an internal actinometer. Measurements in a pH 7.4 phosphate buffer plus 1% Triton X-100 (PB/X100) are scaled to 0.52 for methylene blue in the phosphate buffer. The average quantum yields are: hematoporphyrin IX (0.73), protoporphyrin IX (0.56), zinc protoporphyrin IX (0.91), mesotetra-(4-sulfonato-phenyl)porphine (0.61), PhotofrinR (0.89), benzoporphyrin derivative monoacid ring-A (0.84), chlorin e6 in PB (0.64), pheophorbide a (0.69), and aluminum phthalocyanine tetrasulfonate (0.38). Protection factors were measured for added azide ion, 1,4-diazabicyclo[2,2,2]-octane, and superoxide dismutase. Spectral evidence is presented for chlorin e6 interactions with PB/TX100 and for binding to lysozyme.

Singlet Oxygen Generation by Metallotexaphyrins

Abstract. Metallotexaphyrins have clinical applications as photo‐sensitizers of photodynamic therapy (PDT). The singlet oxygen quantum yield (φ>Δ) was determined for a series of metallotexaphyrin derivatives (Lu [III], Y [III], Cd [II], In [III] and Gd [in]) under conditions where the agents are believed to exist in monomeric form. The results show φΔ of metallotexaphyrins vary with the medium and the metal cation. Measurements on the Lu (III) texaphyrin led to φΔ= 0.38 in unbuffered 5% Tween 20 and φΔ= 0.58 in pH 7.4 phosphate buffer plus 1% Triton X‐100 (±10%). The in vitro photodynamic efficiency calculated from φΔ is compared to in vivo PDT efficacy in an animal tumor model.

The effect of diabetes mellitus on rat skeletal extensor digitorum longus muscle tissue: An FTIR study

Diabetes mellitus (DM) is a chronic disorder of carbohydrate, fat and protein metabolism, which is characterized by a defective insulin secretory response. Skeletal muscle takes role in determination of carbohydrate and lipid metabolism, therefore; it is one of the target tissues of diabetes. Herein this study, application of Fourier Transform Infrared (FTIR) spec- troscopy in diabetic skeletal Extensor Digitorum Longus (EDL) muscle tissues will be presented which highlight the promise of this technique in medical research. Type I DM was induced in rats by injection of streptozotocin (STZ) which is one of the most popular experimental models. In diabetes, a significant increase was observed in lipid order together with an increase in hydration of phospholipid molecules in membrane structure. There was a decrease in lipid and nucleic acid content in diabetic EDL muscles. A dramatic increase in the bandwidth of amide II band (1540 cm −1 ) and shifting of the position of this band to

Photosensitization of Red Blood Cell Hemolysis by Lutetium Texaphyrin

Lutetium (III) texaphyrin photosensitizes postirradiation or “delayed” photohemolysis (DPH) of human and bovine red blood cells at 730 nm by a Type‐2 pathway mediated by singlet molecular oxygen. The DPH rate increases with increasing incubation temperature and with the second power of the incident fluence. The experimental DPH curves are in good agreement with a multihit kinetics model based on target theory.

Electrophysiological and theoretical analysis of melatonin in peripheral nerve crush injury

Electrophysiological and theoretical studies have been performed to investigate peripheral nerve injuries and nerve regeneration. The aim of this work is to evaluate the effect of melatonin functionally and electrophysiologically on peripheral nerve crush injury. Adult male Wistar rats (n=32, 200+/-50 g) were randomly allocated into four groups. Sciatic crush was constituted on left sciatic nerves. Treatment groups received intraperitoneal melatonin at doses of 5 and 20mg/kg for 21 days. Functional nerve recovery was evaluated using sciatic functional index (SFI) every week during the experiment. In vivo electrophysiological measurements were performed at the end of the treatment. The electrophysiological data were also analyzed by wavelet analysis. Melatonin treatments increased the SFI values in the injured sciatic nerves. In vivo electrophysiological measurements showed that melatonin increased the conduction velocities and also decreased the latency values. The wavelet analysis showed that melatonin treatment reduced the densities of high frequency components of compound muscle action potential (CMAP). These results suggest that melatonin application is a promising strategy for the treatment of peripheral nerve crush injuries. Furthermore, analysis of EMG data with wavelet methods seems to give more reliable results to evaluate the nerve recovery.

Protoporphyrin IX-loaded magnetoliposomes as a potential drug delivery system for photodynamic therapy: Fabrication, characterization and in vitro study

BACKGROUND Protoporphyrin IX (PpIX) is a well-known photosensitizer that has great potential for use in photodynamic therapy (PDT). However, aggregation behavior of PpIX in neutral water makes it inappropriate for physiological studies. PpIX-loaded magnetoliposomes (MLs) were fabricated to increase PpIX biocompatibility. PpIX-loaded ML physical properties were characterized, and PpIX-loaded ML drug release behavior was investigated under the influence of an external magnetic field and heat. Toxicity and photodynamic effects of the complex were also examined using in vitro experiments with MCF-7 human breast cancer cells. METHODS The magnetoliposomes were prepared with DPPC, DSPE-PEG2000 lipids and Fe3O4 nanoparticles. The toxicity and in vitro photodynamic effects of the PpIX-loaded MLs at various concentrations were studied using the MCF-7 cell line. RESULTS The produced PpIX-loaded MLs exhibited an average hydrodynamic diameter of 221nm; however, TEM measurements indicated that the diameter of the PpIX-loaded MLs varied between 166 and 720nm. The iron content of the MLs affected cell viability less than the content of the iron free liposomes. Cell viability was reduced to 66% when the concentration of the PpIX-loaded MLs was 350nM, but when white light was applied for 5min, all of the cells that were exposed to concentrations of 250nM and higher PpIX died within 24h. CONCLUSION The results of this study demonstrated the effective application of PpIX-loaded MLs for in vitro photodynamic therapy at nanomolar concentrations. The results also indicated that an LED light source provided sufficient energy to stimulate the PpIX molecules.

Restoring effect of selenium on the molecular content, structure and fluidity of diabetic rat kidney brush border cell membrane

Diabetic kidney disease (DKD) is a dominant factor standing for kidney impairments during diabetes. In this study, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy was used to disclose the diabetes-induced structural changes in the kidney and evaluate the effects of selenium on diabetes. The increase in the area of the olefinic band indicated increased amount of lipid peroxidation end products in diabetic kidney brush border cell membrane. Moreover, saturated lipid content of this cell membrane considerably diminished. DKD was found to disrupt lipid order and cause a decrease in membrane dynamics. However, the administration of selenium at low and medium doses was shown to improve these conditions by changing the lipid contents toward control values, restoring the ordered structure of the lipids and membrane dynamics. Curve-fitting and artificial neural network (ANN) analyses of secondary structures of proteins demonstrated a relative increase in α-helix and reduction in the β-sheet during diabetes in comparison to the control group, which were ameliorated following selenium treatment at low and medium doses. These findings were further confirmed by applying hierarchical cluster analysis (HCA) and principal component analysis (PCA). A clear separation of the experimental groups was obtained with high heterogeneity in the lipid and protein regions. These chemometric analyses showed that the low and medium doses of selenium-treated diabetic groups are successfully segregated from the diabetic group and clustered closer to the control. The study suggests that medium and, more predominantly, low-dose selenium treatment can be efficient in eliminating diabetes-induced structural alterations.

Early Alterations in Bone Characteristics of Type I Diabetic Rat Femur: A Fourier Transform Infrared (FT-IR) Imaging Study.

Alterations in microstructure and mineral features can affect the mechanical and chemical properties of bones and their capacity to resist mechanical forces. Controversial results on diabetic bone mineral content have been reported and little is known about the structural alterations in collagen, maturation of apatite crystals, and carbonate content in diabetic bone. This current study is the first to report the mineral and organic properties of cortical, trabecular, and growth plate regions of diabetic rat femurs using Fourier transform infrared (FT-IR) microspectroscopy and the Vickers microhardness test. Femurs of type I diabetic rats were embedded into polymethylmethacrylate blocks, which were used for FT-IR imaging and microhardness studies. A lower mineral content and microhardness, a higher carbonate content especially labile type carbonate content, and an increase in size and maturation of hydroxyapatite crystals were observed in diabetic femurs, which indicate that diabetes has detrimental effects on bone just like osteoporosis. There was a decrease in the level of collagen maturity in diabetic femurs, implying a decrease in bone collagen quality that may contribute to the decrease in tensile strength and bone fragility. Taken together, the findings revealed alterations in structure and composition of mineral and matrix components, and an altered quality and mechanical strength of rat femurs in an early stage of type I diabetes. The results contribute to the knowledge of structure–function relationship of mineral and matrix components in diabetic bone disorder and can further be used for diagnostic or therapeutic purposes.

Effect of Pulsed Electromagnetic Field on MMP-9 and TIMP-1 Levels in Chondrosarcoma Cells Stimulated with IL-1β

Chondrosarcoma, the second most common type of bone malignancy, is characterized by distant metastasis and local invasion. Previous studies have shown that treatment by pulsed electromagnetic field (PEMF) has beneficial effects on various cancer cells. In this study, we investigated the effects of PEMF applied for 3 and 7 days on the matrix metalloproteinase (MMP) levels in chondrosarcoma SW1353 cells stimulated with two different doses of IL-1β. SW1353 cells were treated with (0.5 and 5 ng/ml) IL-1β and PEMF exposure was applied either 3 or 7 days. MMP-9 and TIMP-1 levels were measured in conditioned media by enzyme-linked immunosorbent assay. The results were relative to protein levels. Statistical analyses were performed using one-way analysis of variance (ANOVA). P<0.05 was considered significant. PEMF treatment significantly decreased MMP-9 protein levels in human chondrosarcoma cells stimulated with 0.5 ng/ml IL-1β at day 7, whereas it did not show any effect on cells stimulated with 5 ng/ml IL-1β. There was no significant change in TIMP-1 protein levels either by IL-1β stimulation or by PEMF treatment. The results of this study showed that PEMF treatment suppressed IL-1β-mediated upregulation of MMP-9 protein levels in a dual effect manner. This finding may offer new perspectives in the therapy of bone cancer.

Nonlinear analysis of heart rate variability of healthy subjects and patients with chronic obstructive pulmonary disease

Abstract Chronic obstructive pulmonary disease (COPD) is one of the causes of mortality worldwide with an increasing prevalence. Heart rate variability (HRV) reflects the regulation mechanism of the cardiac activity by the autonomic nervous system. The assessment of HRV by using nonlinear methods is more sensitive for the detection of complexity when compared to linear methods. This study aims to get information about the autonomic dysfunction occurred in patients with COPD by analysing the complexity of HRV. Electrocardiogram signals recorded from healthy subjects, patients with moderate COPD and severe COPD (eight subjects per group) were analysed. The HRV signals were acquired from ECG signals. Signals were reconstructed in the phase space and largest Lyapunov exponent (LLE), correlation dimension, Hurst exponent and approximate entropy (ApEn) values were calculated. It has seen that for the patients with COPD LLE, correlation dimension, Hurst exponent and ApEn values were less than control group. According to this, HRV complexity decreases in the presence of COPD. However, there is no significant difference between COPD groups and the severity of COPD has no effect on the chaoticity of the system. The results revealed that autonomic dysfunction occurred in patients with COPD is associated with reduced HRV complexity.