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    Activation of proteolysis during oocyte In vitro maturation
    (John wiley and sons inc, 2025) Tepeköy, Filiz; Bulut, Berk
    In vitro maturation (IVM) is a form of assisted reproductive technology (ART) applied to obtain mature oocytes in culture. Decline in IVM success rates by age has led consideration of novel approaches based on cellular dynamics. Our aim was to achieve proteostasis in old bovine oocytes from 13 to 16-year-old bovine with a lower potential for fertilization. Lysosomal activation was achieved through increasing concentrations of proton pump activators PIP2 (0.1, 0.5, 1, and 5 mu M), PMA (0.1, 1, 10, and 50 mu M), and DOG (0.1, 1, 10, and 50 mu M) at 6, 12, 18, and 24 h of IVM in old bovine oocytes. Morphological analysis was performed and IVM rates were determined. DQ-Red BSA was applied to live oocytes to determine proteolytic activation while lysosome density was determined by Lysotracker probe. Protein carbonylation was detected through oxyblot analysis. Polar body extrusion (PBE), through which a haploid nonfunctional polar body is released in the perivitelline space after completion of the first meiotic division, was observed in PIP2-0.1 mu M, -0.5 mu M-6h; PIP2-5 mu M-12h; PMA-0.1 mu M-18h; PIP2-0.1 mu M, -0.5 mu M-24h groups. Oocyte diameter was the highest in DOG-1 mu M-6h, PMA-0.1 mu M-12h, PIP2-1 mu M-18h, and PIP2-0.5 mu M-24h groups. Morphological scores of oocytes were higher in young and old control groups. PIP2, PMA, and DOG affected oocyte quality positively after 6 h of IVM yielding in oocyte scores similar to the control group oocytes. However, they had a negative impact on the oocyte scores in longer periods of IVM, except for lower doses PMA (0.1 and 1 mu M) at 12 h and PIP2 (0.5 mu M) and PMA (0.1 mu M) at 18 h, which were able to maintain the scores relatively closer to the control oocytes. Proteolytic activation was achieved in all groups at 6 h of culture. At all other time points PIP2 and PMA groups showed a better response to proteolytic activation. Lysosome density was increased in PIP2-5 mu M-6h; PIP2-0.1 mu M, -1 mu M-12h; PIP2-1 mu M, -5 mu M-18h as well as PMA-0.1 mu M-6h; PMA-1 mu M, -10 mu M-12h; PMA-1 mu M-18h; DOG-50 mu M-6h and DOG-0.1 mu M-12h. Protein carbonylation was the lowest in PIP2-0.1 mu M groups at 12, 18, and 24 h. This study suggests that proton pump activators PIP2 and PMA was found to have a positive impact on IVM in terms of both morphological scores and proteolytic activation in a time and dose dependant manner.
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    Assessment of the effect of endocrine abnormalities on biomacromolecules and lipids by FT-IR and biochemical assays as biomarker of metabolites in early Polycystic ovary syndrome women
    (Elsevier, 2021) Güleken, Zozan; Bulut, Huri; Bulut, Berk; Depciuch, Joanna
    Polycystic ovary syndrome (PCOS) is a common endocrinopathy associated with metabolic disturbances.Both in the development and improvement of the disease, the structure of phospholipids and proteinsin the blood serum plays important role in the treatment of these disease. Herein, to investigate themetabolic process and the variations of biomacromolecules and lipids between young PCOS women andhealthy subjects, biochemistry and Fourier Transform InfraRed spectroscopy (FTIR) methods, were used.Moreover, partial least squares regression (PLS) and Principal component analysis (PCA) to researchdifferentiation of biomacromolecules, were performed. We obtained blood serum of of 100 individ-uals including 57 with PCOS and 43 healthy controls. The biochemical blood profile of PCOS womenwas presented by spectroscopic measurements, which is an analytical technique, as well as by labora-tory indexes and oxidative stress status measurements. There was a significant structural differentiationbetween studied groups in the number of functional groups and biomolecules differentiation dependingon the protein expression and oxidative stress status. Hence, FTIR spectroscopy and oxidative load can beeffectively utilized as tools for classifying quantitative and qualitative changes of biomolecules in PCOSsamples. PCOS samples did not correlate with luteinizing hormone (LH) level and proteins but had anegative correlation between carbohydrates and fatty acids, compared with control group.
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    Correlation between endometriomas volume and Raman spectra. Attempting to use Raman spectroscopy in the diagnosis of endometrioma
    (Elsevier B.V., 2022) Güleken, Z.; Bulut, Huri; Bulut, Berk; Paja, W.; Parlinska-Wojtan, M.; Depciuch, J.
    The formation of the uterus lining, i.e. the endometrium, outside the uterus (ex. in the abdominal cavity, ovaries, or anywhere in the body) is called endometriosis. The presence of endometrial tissue present in the ovaries, thickens after menstruation, leading to menstrual-like bleeding and to the formation of chocolate cyst (Endometrioma) because of the accumulation of old, brown blood in the ovary. It is still unknown, what triggers the development of endometrioma. However, it leads to excessive bleeding during menstrual periods or abnormal bleeding between periods and infertility. Endometriosis is often first diagnosed in those who seek medical attention for infertility. Therefore, new markers of endometrioma as well as new methods of its diagnosis are sought. In this study we used Raman spectra of serum collected from 50 healthy women and 50 women suffering from endometriosis. The obtained Raman data were used in multivariate analysis to determine the Raman range, which can be used for endometrioma diagnostics. Partial Least Square (PLS), Principal Component Analysis (PCA) and Hierarchical Component Analysis (HCA) showed, that it is possible to distinguish between the serum collected from healthy and un-healthy women using the Raman range between 800 cm?1 and 1800 cm?1 and between 2956 cm?1 and 2840 cm?1, while the first range corresponds to the fingerprint region and the second one to lipids vibrations. Consequently, the Pearson correlation test showed a significant positive correlation between values of lipid intensity in Raman spectra and volume of endometriomas. Summarizing, Raman spectroscopy can be a helpful tool in endometrioma diagnosis and the lipid vibrations are candidates for being a spectroscopic marker of the disease being studied. © 2022 Elsevier B.V.
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    Effect of Taxifolin on Acrylamide-Related Oxidative Ovarian Damage, Infertility and Intrauterine Growth Retardation in Female Rats
    (Asian Network Scientific Information-Ansinet, 2023) Dinc, Kemal; Kiremitli, Tunay; Kiremitli, Sevil; Ozyurt, Ramazan; Bulut, Berk; Bulut, Seval; Yavuzer, Bulent
    [Abstract Not Available]
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    Identification of polycystic ovary syndrome from blood serum using hormone levels via Raman spectroscopy and multivariate analysis
    (Elsevier, 2022) Güleken, Z.; Bulut, Huri; Bulut, Berk; Paja, W.; Orzechowska, B.; Parlinska-Wojtan, M.; Depciuch, J.
    Polycystic ovarian syndrome (PCOS) is a disease, which causes infertility in women. The factors for the development of the disease are still not well understood and diagnostic methods need to be improved. Therefore, in this study, Raman spectroscopy as a potential diagnostic tool, was investigated and spectra of blood serum were collected from PCOS and healthy women. The obtained spectra showed distinct changes in intensities as well as shift of peaks for the blood serum collected from PCOS compared to healthy individuals. Partial Last Square (PLS) analysis and Principal Component Analysis (PCA) allowed to determine that Raman shifts of amides (1500 – 1700 cm?1) and CH2, CH3 lipid groups (2700 – 3000 cm?1), could be thus used as potential PCOS markers. Furthermore, the Pearson correlation test showed a strong correlation between hormones (lutropin (LH), prolactin (PRL), follicle-stimulating (FSH), dehydroepiandrosterone (DHEAS), thyroid-stimulating (TSH), Estradiol) and lipids, as well as between hormones and protein functional groups in PCOS women, compared to the control. These results show, that the lipid and protein balance could be potentially applied as a helpful PCOS marker in Raman spectra. © 2022 Elsevier B.V.
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    Preoperative factors associated with the need for the morcellation in total laparoscopic hysterectomy
    (Yuzuncu Yil Universitesi Tip Fakultesi, 2020) Kiyak H.; Karaaslan O.; Doga Seckin K.; Karacan T.; Ozyurek E.S.; Yilmaz G.; Bulut, Berk
    Removal of uterus during laparoscopic hysterectomy (LH) is occasionally challenging and therefore sometimes requires morcellation. Morcellation techniques for hysterectomy can spread the cancer cells which were presumed benign preoperatively in to the abdomen or pelvis. Probability of an undetected malignity should not be disregarded and accordingly must be shared with the patient prior to LH. The present study aimed to identify the demographic and the clinical factors associated with the need for morcellation in patients undergoing LH. A total of 153 patients who underwent LH for presumed benign causes were enrolled in this retrospective study. Subjects were divided into two groups according to the need for the morcellation during the LH: Morcellation group and intact vaginal removal (IVR) group (it was possible to deliver the uterine specimen vaginally). The two groups were compared with respect to demographic and clinical characteristics, indications for LH, preoperative ultrasonographic findings and postoperative complications. The frequency of the postmenopausal women was significantly higher in the morcellation group than the IVR group (p = 0.005). Preoperative uterus width (p < 0.001) and postoperative Uterus weight (p < 0.001) were significantly higher in the morcellation group compared to that of the IVR group. There were no significant differences between the groups regarding the complication rate. Uterine weight (OR: 1.020, 95% CI: 1.008-1.031, p = 0.001) and menopause (OR: 2.571, 95% CI: 1.328-4.980, p < 0.001) were predictive for the need of morcellation. A cut-off value of 287.5 gram was able to predict the need for morcellation with 87% sensitivity and 71% specificity. The present study demonstrates that uterine weight and presence of the menopause are predictive for the need of morcellation at the time of the total LH. These factors should be considered in preoperative planning and used to further guide surgeons in providing LH. © 2020, Yuzuncu Yil Universitesi Tip Fakultesi. All rights reserved.