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Protein–protein interactions (PPIs) have great importance for intracellular signal transduction and sustaining the homeostasis of an organism. Thus, the identification of PPIs is necessary to better understand the downstream signaling functions of the proteins in healthy and pathological conditions. Förster resonance energy transfer (FRET) between fluorescent proteins (FPs) is a powerful tool for detecting PPIs in living cells. In literature, FRET analysis methods such as donor photobleaching (FLIM), acceptor photobleaching, spectral imaging, and the three-filter cube method (sensitized emission) are abundantly applied to investigate PPIs; however, they require various expensive instrumentations, and their calculation methods are very time consuming. Since confocal microscopy applications and live cell-based techniques of FRET are very costly, scientists sometimes prefer plate readers for FRET experiments. However, plate reader applications also have many disadvantages and considerations compared to confocal fluorescence microscopy, and complex calculation procedures should be performed. To overcome these problems, we propose a FRET-based high-throughput assay method with a standard monochromator-based microplate reader, which is generally available in most biochemistry laboratories, and an alternative calculation procedure. This rapid, low cost, and effective analysis method enables the scientists to prescreen PPIs in living cells as a preliminary study and quick glance at the experiment before preparing the whole experimental setup with the expensive instrumentations. Additionally, the alternative calculation procedure provides the FRET area comparison without complex bleed-through calculations in a non-conventional manner by shortening the analysis processes with this quick and uncomplicated spectral representation.

Type I Diabetes is a multisystem disease that causes alterations in carbohydrate, protein, and fat metabolisms due to hyperglycemia. It has an extensive pathology, especially the mechanism involving oxidative stress is still complex. Type I diabetes is correlated with increased formation of free radicals and decreased levels of antioxidant potential. Vitamin C (Vit C) is a powerful antioxidant that participates in antioxidant defense, protecting lipid membranes and proteins from oxidative damage by donating electrons to free radicals. The effect of type I diabetes and the recovery role of Vit C on the structure and composition of the biomolecular content of testicular tissue is still unknown. Therefore, the current study aimed to investigate the alterations in the biomolecules of rat testes due to Streptozotocin (STZ)-induced type I diabetes using Attenuated Total Reflectance (ATR)-Fourier Transform Infrared (FTIR) spectroscopy and histological staining. The results revealed that the biomolecular structure and composition of testicular tissue are highly affected due to the development of diabetes. We obtained decreased saturation levels and increased unsaturation index in the lipids indicating the presence of lipid peroxidation in the diabetic state. The elevated lipid peroxidation levels have been implicated in the pathogenesis of naturally occurring and chemically induced diabetes. On the other hand, the protein content of diabetic rat testicular tissue was shown to decrease considerably, indicating an increase in proteolysis processes. Supporting the ratio of protein structural and conformational change, protein secondary structural components were also found to alter substantially in the diabetic state. Diabetes was also shown to lead to a decrease in the content of nucleic acids compared to proteins. These diabetes-induced alterations were found to be substantially recovered with the administration of Vit C. Although different doses and administration types of Vit C have been reported in the literature, there is no consensus yet. Therefore, we used three different doses of Vit C in our study as high (100 mg/kg/day), medium (50 mg/kg/day) and low (15 mg/kg/day) doses intraperitoneally in the present study, and the medium dose was found to be the most effective in the recovery from the diabetes-induced structural damages on rat testicular tissue. Vit C may have a therapeutic effect to be used as a complementary therapy in the treatment of diabetes.

PubMed: 31983145 Obesity is associated with a higher risk of developing breast cancer and with worse disease outcomes for women of all ages. The composition, density, and organization of the breast tissue stroma are also known to play an important role in the development and progression of the disease. However, the connections between obesity and stromal remodeling are not well understood. We sought to characterize detailed organization features of the collagen matrix within healthy and cancerous breast tissues acquired from mice exposed to either a normal or high fat (obesity inducing) diet. We performed second-harmonic generation and spectral two-photon excited fluorescence imaging, and we extracted the level of collagen-associated fluorescence (CAF) along with metrics of collagen content, three-dimensional, and two-dimensional organization. There were significant differences in the CAF intensity and overall collagen organization between normal and tumor tissues; however, obesity-enhanced changes in these metrics, especially when three-dimensional organization metrics were considered. Thus, our studies indicate that obesity impacts significantly collagen organization and structure and the related pathways of communication may be important future therapeutic targets..

63rd Annual Meeting of the Biophysical-Society -- MAR 02-06, 2019 -- Baltimore, MD WOS:000460779802840 Radiotherapy is widely used to cure cancer. During treatment, ionizing radiation is applied by a linear accelerator to the area that is being treated. During this process normal cells can also be damaged by radiotherapy. Biophys Soc

WOS:000351405900019 PubMed: 25692183 Obesity is a heterogeneous disorder which increases risks for multiple metabolic diseases, such as type 2 diabetes. The current study aims to characterize and compare visceral and subcutaneous adipose tissues in terms of macromolecular content and investigate transdifferentiation between white and brown adipocytes. Regarding this aim, Fourier transform infrared (FTIR) microspectroscopy and uncoupling protein 1 (UCP1) immunohistological staining were used to investigate gonadal (visceral) and inguinal (subcutaneous) adipose tissues of male Berlin fat mice inbred (BFMI) lines, which are spontaneously obese. The results indicated a remarkable increase in the lipid/protein ratio, accompanied with a decrease of UCP1 protein content which might be due to the transdifferentiation of brown adipocytes to white adipocytes in obese groups. It has been widely reported that brown adipose tissue has a strong effect on fatty acid and glucose homeostasis and it could provide an opportunity for the therapy of obesity. When the amount of brown adipose tissue was decreased, lower unsaturation/saturation ratio, qualitatively longer hydrocarbon acyl chain length of lipids and higher amount of triglycerides were obtained in both adipose tissues of mice lines. The results also revealed that subcutaneous adipose tissue was more prone to obesity-induced structural changes than visceral adipose tissue, which could originate from it possessing a lower amount of brown adipose tissue. The current study clearly revealed the power of FTIR microspectroscopy in the precise determination of obesity-induced structural and functional changes in inguinal and gonadal adipose tissue of mice lines. Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [SBAG-110S235] This work was supported by The Scientific and Technological Research Council of Turkey (TUBITAK) with a grant number SBAG-110S235. This work was carried out as a part of the COST Action (BM0901)-SYSGENET.

The excess deposition of triglycerides in adipose tissue is the main reason of obesity and causes excess release of fatty acids to the circulatory system resulting in obesity and insulin resistance. Body mass index and waist circumference are not precise measure of obesity and obesity related metabolic diseases. Therefore, in the current study, it was aimed to propose triglyceride bands located at 1770-1720 cm-1 spectral region as a more sensitive obesity related biomarker using the diagnostic potential of Fourier Transform Infrared (FTIR) spectroscopy in subcutaneous (SCAT) and visceral (VAT) adipose tissues. The adipose tissue samples were obtained from 10 weeks old male control (DBA/2J) (n = 6) and four different obese BFMI mice lines (n = 6 per group). FTIR spectroscopy coupled with hierarchical cluster analysis (HCA) and principal component analysis (PCA) was applied to the spectra of triglyceride bands as a diagnostic tool in the discrimination of the samples. Successful discrimination of the obese, obesity related insulin resistant and control groups were achieved with high sensitivity and specificity. The results revealed the power of FTIR spectroscopy coupled with chemometric approaches in internal diagnosis of abdominal obesity based on the spectral differences in the triglyceride region that can be used as a spectral marker.

Lipid accumulation and storage of lipids in adipocytes during obesity cause structural and functional changes in adipose tissue conformation. The expansion of visceral (VAT) and subcutaneous (SCAT) adipose tissue mass in the body is the main reason of obesity and many times it results in disturbed lipid and glucose metabolism. Gender is an important factor for the research in obesity and other metabolic diseases because it leads to different fat distribution and the pathophysiology. This study aims to determine gender effect on the structural and functional parameters on VAT and SCAT. To achieve this, FTIR microspectroscopic imaging technique and UCP1 immunohistological staining have been used. FTIR microspectroscopy is a rapid and effective technique to monitor molecular alterations in biological tissues induced by different conditions such as disease, chemical treatment and variations in the environmental factors. UCP1 protein content gives information about the amount of brown adipose tissue (BAT), and therefore about the transdifferentiation of BAT to the white adipose tissue (WAT). The results of FTIR imaging study revealed a decrease in unsaturation level of lipids and an increase in the amount of triglycerides in adipose tissue samples. Furhermore, the longer hydrocarbon acyl chain length was obtained in the lipids of obese samples. All of these spectral paramaters could be used as biomarkers in obesity. The results of the present study showed that, these obesity indicators are more significant in SAT of female mice, whilst, they are more significant inVAT of male mice. The amount of UCP1 protein which is a marker of the transdifferentiation, showed a decrease in male samples rather than females. Consequently, obesity has adverse effects on health of both genders but especially on men.