Neslihan Toyran

Early alterations in myocardia and vessels of the diabetic rat heart: an FTIR microspectroscopic study

Diabetes mellitus is associated with a high incidence and poor prognosis of cardiovascular disease. The aim of the present study was to examine the effect of relatively short-term (5 weeks) Type I diabetes on the left ventricle, the right ventricle and the vessel (vein) on the left ventricle of the myocardium at molecular level by FTIR (Fourier-transform infrared) microspectroscopy. The rats were categorized into two groups: control group (for the left ventricle myocardium, n=8; for the right ventricle myocardium, n=9; for the vein, n=9) and streptozotocin-induced diabetic group (for the left ventricle myocardium, n=7; for the right ventricle myocardium, n=9; for the vein, n=8). Two adjacent cross-sections of 9 microm thickness were taken from the ventricles of the hearts in two groups of rats by using a cryotome. The first sections were used for FTIR microspectroscopy measurements. The second serial sections were stained by haematoxylin/eosin for comparative purposes. Diabetes caused an increase in the content of lipids, an alteration in protein profile with a decrease in alpha-helix and an increase in beta-sheet structure as well as an increase in glycogen and glycolipid contents in both ventricles and the vein. Additionally, the collagen content was found to be increased in the vein of the diabetic group. The present study demonstrated that diabetes-induced alterations in the rat heart can be detected by correlating the IR spectral changes with biochemical profiles in detail. The present study for the first time demonstrated the diabetes-induced alterations at molecular level in both ventricle myocardia and the veins in relatively short-term diabetes.

Chronic hypoperfusion alters the content and structure of proteins and lipids of rat brain homogenates: a Fourier transform infrared spectroscopy study

Arteriovenous malformations (AVMs), masses of abnormal blood vessels which grow in the brain, produce high flow shunts that steal blood from surrounding brain tissue, which is chronically hypoperfused. Hypoperfusion is a condition of inadequate tissue perfusion and oxygenation, resulting in abnormal tissue metabolism. Fourier transform infrared (FTIR) spectroscopy is used in this study to investigate the effect of hypoperfusion on homogenized rat brain samples at the molecular level. The results suggest that the lipid content increases, the protein content decreases, the lipid-to-protein ratio increases, and the state of order of the lipids increases in the hypoperfused brain samples. FTIR results also revealed that, owing to hypoperfusion, not only the protein synthesis but also the protein secondary structure profile is altered in favor of β-sheets and random coils. These findings clearly demonstrate that, FTIR spectroscopy can be used to extract valuable information at the molecular level so as to have a better understanding of the effect of hypoperfusion on rat brain.

Fourier transform infrared study of the effect of diabetes on rat liver and heart tissues in the CH region.

Diabetes mellitus is characterized by hyperglycemia, a relative lack of insulin. The metabolic disturbances in diabetic patients are often associated with cardiac and liver dysfunctions. Generally, experimental diabetic models in animals have been used to study diabetes-related changes in organ function, but the complexity of intact tissues can cause contradictory results. For this reason, different techniques have been used to understand the mechanisms of these dysfunctions in diabetic organs. The purpose of the present study is to investigate the effects of streptozotocin (STZ)-induced diabetes on rat liver and heart tissues at the molecular level by Fourier Transform Infrared (FTIR) spectroscopy. Wistar rats of both sexes, weighing 200-250 g, were made diabetic by a single injection of 50 mg kg(-1) intraperitoneal (i.p.) streptozotocin dissolved in 0.05 M citrate buffer (pH 4.5) and they were kept for 4-5 weeks. The diabetes status was checked by measuring the blood glucose level. In the complex FTIR spectra, the bands in the CH region for example the CH(2) antisymmetric and symmetric stretching, the CH(3) symmetric and asymmetric stretching vibrations due to lipids and proteins in the 3000-2800 cm(-1) region and CH(2) scissoring around 1464 cm(-1) and the CH(3) scissoring at 1454 cm(-1) were analyzed. Characteristic spectral bands of these diabetic samples were compared with those of control group to confirm the effect of diabetes on liver and heart tissues. The FTIR spectra revealed dramatic differences in the band positions and bandwidths, signal intensity values and signal intensity ratios between diabetic and control tissues. Similar differences were observed for diabetic liver and heart tissues. A significant increase in the bandwidth of the CH(2) symmetric and antisymmetric stretching and the CH(3) symmetric and asymmetric stretching bands has been observed for both tissue types. The wavenumber of the CH(3) asymmetric stretching band shifts to lower values, indicating an increase in the order in the deep interior part of the lipid chains. The ratio of the CH(2) symmetric to CH(3) symmetric stretching band (lipid/protein ratio) decreases by 13% for diabetic heart and liver tissues. A decrease is also detected in the ratio of the CH(2) scissoring to the CH(3) scissoring mode. The overall intensity of these bands is seen to increase for diabetic tissues.

Competitive effect of vitamin D2 and Ca2+ on phospholipid model membranes: an FTIR study.

The interaction of Ca(2+), with dipalmitoyl phosphatidylcholine (DPPC) model membranes was studied in the presence and absence of vitamin D(2) by using Fourier transform infrared spectroscopy. Addition of vitamin D(2) and/or Ca(2+) into pure DPPC liposomes shifts the phase transition to higher temperature, orders and decreases the dynamics of the acyl chains in both phases and does not induce hydrogen bond formation in the interfacial region. Moreover, the dynamics of the head group of the phospholipid decreases in both phases. The addition of vitamin D(2) into DPPC liposomes containing Ca(2+), decreases the effect of Ca(2+) at all the functional groups under investigation. Similarly, the effect of vitamin D(2) also decreases in the presence of Ca(2+). This behavior is dominant at high Ca(2+) concentrations. Our results show how simultaneous presence of vitamin D(2) and Ca(2+) alter the behavior of each other, which is reflected as a decrease in the interactions between the ions and vitamin D(2) within the membrane.

Effect of stereotactic radiosurgery on lipids and proteins of normal and hypoperfused rat brain homogenates: A Fourier transform infrared spectroscopy study

Purpose: The effect of stereotactic radiosurgery on lipids and proteins of normal and hypoperfused rat brain was investigated to see if hypoxic areas are really more resistant to radiation effects or not. Materials and methods: Rat brain samples from contol, stereotactically irradiated and chronically hypoperfused plus stereotactically irradiated groups were homogenized separately with saline phosphate buffer, and centrifuged at 125,000 g for 15 min. Membrane rich parts (pellet) of these homogenates were used for Fourier Transform Infrared (FTIR) spectroscopy studies. Mann-Whitney U tests were performed on the groups, two by two, to test the significance of the differences between the control group and stereotactically irradiated group as well as the control group and chronically hypoperfused plus stereotactically irradiated group. Results: After a single high dose of X-rays to healthy rat brain, the lipid concentration increased slightly, protein content decreased significantly (p < 0.05) and protein-to-lipid ratio decreased slightly. The secondary structure of the proteins was altered in the irradiated brain samples such that the content of α-helical structure decreased significantly (p < 0.01) and random coil increased dramatically (p < 0.05). The effect of radiation on the content of α-helical structure was not found to be significant in the hypoperfused group, but the decrease in the content of random coil was significant (p < 0.01). Conclusion: Stereotactic radiosurgery of the brain increased the lipid concentration, decreased the protein concentration and consequently resulted in a decrease in the protein to lipid ratio compared to unirratiated brain. Radiation also altered the secondary structure of protein. The variations in lipid and protein content and the resulting lipid to protein ratio imply that chronically hypoperfused brain is more vulnerable to radiation than non-hypoperfused brain and suggests chronic hypoperfusion does not prevent cerebral damage caused by irradiation. However, irradiation of hypoperfused brain resulted in less alteration in protein structure than in non-hypperfused brain, suggesting higher resistance to irradiation using this endpoint.

Vitamin D2 at high and low concentrations exert opposing effects on molecular order and dynamics of dipalmitoyl phosphatidylcholine membranes

Fourier Transform Infrared spectroscopic studies show that low concentrations of vitamin D2 (1 and 3m ol %) does not induce significant change in the overall shape of the thermotropic profile of dipalmitoyl phosphatidylcholine (DPPC) membrane. In contrast, at higher concentrations of vitamin D2 (9 and 12 mol %), the phase transition shifts to lower temperatures and a significant broadening in the phase transition curve is also observed. Low concentration of vitamin D 2 decreases the frequency of the CH2 stretching mode, implying an ordering effect, whilst high concentration of vitamin D 2 disorders the system. Furthermore, at low and high concentrations, vitamin D2 causes opposing effect on membrane dynamics. It decreases the bandwidth of the CH2 stretching modes at low concentrations while increasing it at high concentrations. We have also observed different actions of vitamin D2 at low and high concentrations in the deep interior and interfacial region of the membrane, by monitoring the frequency of the CH3 stretching band and C=O stretching bands, respectively.

Infrared Spectroscopic Studies on the Dipalmitoyl Phosphatidylcholine Bilayer Interactions with Calcium Phosphate: Effect of Vitamin D2

In the present work, the interaction of calcium-phosphate with DPPC (dipalmitoyl phosphatidylcholine) model mem- branes has been studied in the presence and absence of vitamin D2 by using Fourier Transform Infrared (FTIR) spectroscopy. Calcium and phosphorus are the most abundant elements in the body. They combine in the form of calcium phosphate salt, called hydroxyapatite. Hydroxyapatite is the major structural component of the bone. Calcium phosphate assists with the diges- tion and absorption of food and is vitally important for the building of sturdy bone and body structures and a robust constitution. Phosphorus is extracted from foods and its use is controlled by vitamin D and calcium. FTIR spectral analysis results suggested that, calcium-phosphate complex, which is the major component of the bones, decreases the phase transition temperature to lower values, causes a loss in cooperativity of the acyl chains, decreases the order of the membrane in both phases and decreases the dynamics of the membrane in the liquid crystalline phase, increases the flexibility of the chains in the center of the bilayer in both phases, and increases the mobility of the head group of DPPC in the gel phase. The effect of calcium-phosphate on DPPC liposomes diminishes with the addition of vitamin D2 into the liposomes. Our results suggest how calcium-phosphate and/or vitamin D2, which have indispensable role for the functioning of the bone tissue, affect the thermal behaviour of DPPC liposomes at molecular level.

Investigation of diabetes-induced effect on apex of rat heart myocardium by using cluster analysis and neural network approach: An FTIR study

Diabetes mellitus (DM) is a progressive chronic disorder, which affects people belonging to all age groups of the population. This disease is accompanied by a greatly increased risk of cardiovascular death. In the present study, the effects of streptozotocin (STZ)-induced type 1 diabetes on apex myocardium of the rat heart have been investigated using Fourier Transform Infrared (FTIR) spectra. The cluster analysis has been applied to FTIR spectra to differentiate the diabetic samples from the normal controls. In addition, the protein secondary structures of diabetic and normal tissues were predicted by neural networks based on the amide I band of the FTIR spectra. The findings mainly suggest that 5 weeks of diabetes alters the lipid and protein profile of normal rat heart apex myocardium, which might have an important role in understanding the molecular mechanism of diabetes-related heart diseases.