The effects of CLP-induced sepsis on proliferation and apoptosis of granulosa and theca cells in rat ovary: A histochemical and ultrastructural study.

Sepsis is defined as a systemic inflammatory response to infection. This study is aimed to evaluate the effects of experimental sepsis on the proliferation and apoptosis of granulosa and theca cells in the rat ovary. 28-day-old immature Wistar-Albino female rats were treated with pregnant mare serum gonadotrophin to develop the first generation of preovulatory follicles. Sepsis was induced by cecal ligation and puncture (CLP). Following in vivo 5-Bromo-2-deoxyuridine (BrdU) labeling, animals were sacrificed and ovaries were embedded in paraffin and Epon. Besides electron microscopic evaluation, BrdU, cleaved caspase-3, p27 immunostaining, and TUNEL labeling were performed. In CLP-operated animals, cleaved caspase-3 immunoreactivity was significantly increased in Graafian follicles. TUNEL and BrdU labeling in the ovarian follicles were not statistically different between CLP and sham-operated rats. In septic animals, p27 immunoreactivity was increased significantly in the nuclei of oocytes and decreased in the cytoplasm of granulosa and theca cells in multilaminar primary follicles compared to the sham group. In ultrastructural evaluation, increased apoptosis was observed in theca interna and granulosa cells in both the early and late stages of follicles in the CLP group. In conclusion, experimentally-induced sepsis leads to apoptosis in ovarian follicles at advanced stages of development. Our data suggest that although sepsis may not cause a potential threat to developing follicles at least in the short term, more severe damage may occur during advanced stages of follicle development.

The effects of magnesium sulfate on cyclophosphamide-induced ovarian damage: Folliculogenesis.

Cyclophosphamide (CYP) is one of the alkylating chemotherapeutic agents and its adverse effects on folliculogenesis in the ovary are well-known due to the previous scientific research on this topic. Magnesium has various effects in organisms, including catalytic functions on the activation and inhibition of many enzymes, and regulatory functions on cell proliferation, cell cycle, and differentiation. In this study, the effects of magnesium sulfate (MgSO4) on CYP induced ovarian damage were investigated. Immature Wistar-Albino female rats of 28-days were treated with pregnant mare serum gonadotrophin (PMSG) to develop the first generation of preovulatory follicles. Rats of the experimental groups were then treated with either CYP (100 mg/kg, i.p) and MgSO4 (270 mg/kg loading dose; 27 mg/kg maintenance doseX12, i.p) solely or in combination. Following in-vivo 5-bromo-2-deoxyuridine (BrdU) labeling, animals were sacrificed and ovaries were embedded in paraffin and Epon. In the ovaries, added to the evaluation of general morphology and follicle count; BrdU and TUNEL-labeling, cleaved caspase-3 and p27 (cyclin-dependent kinase inhibitor) staining was also performed immunohistochemically and an ultrastructural evaluation was performed by transmission electron microscopy (TEM). The number of primordial follicles were decreased and multilaminar primary and atretic follicles were increased in CYP group. After MgSO4 treatment, while primordial follicle pool were elevated, the number of atretic follicles were decreased. Additionally, decreased BrdU-labeling, increased cleaved caspase 3 immunoreactivity and increased TUNEL labeling were observed in CYP group. In CYP treated animals, observations showed that while MgSO4 administration caused no alterations in BrdU proliferation index and caspase-3 immunoreactivity, it significantly reduced the TUNEL labeling. It was also observed that, while p27 immunoreactivity significantly increased in the nuclei of granulosa and theca cells in the CYP group; MgSO4 treatment significantly reduced these immunoreactivities. The ultrastructural observations showed frequent apoptotic profiles in granulosa and theca cells in both early and advanced stages of follicles in the CYP group and the MgSO4 treatment before the CYP application led to ultrastructural alleviation of the apoptotic process. In conclusion, our data suggest that MgSO4 may provide an option of pharmacologic treatment for fertility preservation owing to the beneficial effects of on chemotherapy-induced accelerated follicular apoptotic process, and the protection of the primordial follicle pool.

Cost of Healthcare Associated With Deep Vein Thrombosis in Patients Treated With Warfarin in Turkey: 2010-2013 Database Analysis of a Tertiary Care Center.

OBJECTIVES: To evaluate the cost of healthcare with respect to the quality of anticoagulation in patients with deep vein thrombosis (DVT) treated with warfarin in daily practice via the database analysis of a tertiary care center in the period 2010 to 2013. METHODS: Of 258 307 records in total, 42 582 unique patients with DVT and 32 012 patients with international normalized ratio (INR) measurements were included. Overall, 6720 unique patients with DVT diagnosis and one or more INR measurements were identified, and the records of 4377 out of 6720 unique patients were validated and included in the analysis data set. The cost analysis was based on direct medical costs from the payer's perspective. Cost items were related to healthcare resource utilization (inpatient and outpatient services) during the study period, which provided a basis for calculation of per-patient, outpatient, inpatient, and total direct medical costs. RESULTS: Mean outpatient, inpatient, and total hospital admission costs were $578, $2195, and $2785, respectively, for patients with time in the therapeutic range of 70% or more, whereas the same costs were $571, $2163, and $3192, respectively, for patients with time in the therapeutic range of less than 70%. CONCLUSIONS: Our findings for a retrospective cohort of patients with DVT undergoing warfarin therapy reveal that patients spent 70% or more, as opposed to less than 70%, of follow-up time within the therapeutic INR range and that outpatient care, as opposed to inpatient care, was associated with lower healthcare costs. Given the significant contribution that hospital stay makes to the cost burden of DVT, our findings also highlight the association between poor warfarin anticoagulant control and increased hospitalization costs.

Retrospective cohort study for evaluating the INR monitoring patterns in patients with deep vein thrombosis in daily practice: Analysis of 2010-2013 database of a tertiary care center.

OBJECTIVE: To evaluate the international normalized ratio (INR) monitoring patterns in patients with deep vein thrombosis. METHODS: Of 32,012 patients with ≥1 outpatient INR measurement and 42,582 patients with confirmed deep vein thrombosis diagnosis registered to our hospital between 1 January 2010 and 31 December 2013, 6720 records were identified to have both deep vein thrombosis and international normalized ratio measurement, and 4.377 out of 6.720 single patient records were determined to be statistically analyzable. RESULTS: Median INR measurement frequency was 6.47 times/year and patients had INR levels of 2-3 in 34.3% of follow-up time. Having ≥70% vs. <70% of follow-up time within therapeutic range was associated with lower hospital admission frequency (9.7 vs. 10.3 times/year). CONCLUSION: Our study revealed only one-third of the follow-up time to be spent within therapeutic INR, association of INR therapeutic range with lesser number of hospital admissions and INR monitoring frequency of 6.47 times/year despite lack of stable INR control in most of the deep vein thrombosis patients.

The Effect of Left Ventricular Assist Device Implantation on Serum Albumin, Total Protein and Body Mass: A Short-Term, Longitudinal Follow-Up Study.

BACKGROUND: Cardiac cachexia and low serum albumin levels are poor prognostic signs in advanced heart failure, while overweight patients or patients who gain weight after treatment have more favourable outcomes. Weight gain following LVAD implantation is common, while the dynamic changes in body mass or serum proteins have not been studied adequately. Our aim was to study short-term changes in serum albumin, total protein and body weight following LVAD implantation and to compare these changes with heart failure patients treated medically. MATERIALS AND METHODS: A total of 15 patients scheduled for LVAD implantation and 15 patients receiving medical treatment were prospectively enrolled. Anthropometric and laboratory data for the patients were obtained at baseline and at first and sixth months after LVAD implantation. RESULTS: Anthropometric, demographic and clinical characteristics between two groups were similar at baseline. Both serum albumin (3.59±0.71 vs. 4.17±0.46g/dl, p=0.01) and total protein (6.45±0.80 vs. 7.12±0.35g/dl, p<0.01) levels were significantly lower in LVAD group at baseline. Both total protein and serum albumin levels increased significantly in LVAD group (final total protein 7.60±0.62g/dl and serum albumin 4.20±0.46g/dl; p<0.01 for both), while there was a nonsignificant small decrease in serum albumin in medical group. The change in serum albumin, but not total protein was significantly different between LVAD and medical groups at the sixth month. Body weight initially decreased in LVAD group at first month but was nonsignificantly higher compared to baseline and medical group at the sixth month. There was a moderate correlation between the percentage weight gain and percentage increase in serum albumin in LVAD group at six months (r=0.44). CONCLUSIONS: In suitable patients with advanced heart failure, LVAD treatment can correct hypoalbuminaemia associated with heart failure within six months after implantation.

Systemic and myocardial inflammation in traditional and off-pump cardiac surgery.

In this study, we attempted to determine the role of off-pump coronary artery bypass grafting (CABG) in the myocardial and systemic inflammatory responses. Twenty patients who underwent elective CABG were enrolled in this study. Ten patients underwent on-pump CABG, and 10 patients underwent off-pump CABG. There were no differences between patients in preoperative clinical variables. We took systemic venous blood samples for the measurement of tumor necrosis factor-alpha, the MB isoenzyme of creatine kinase (CK-MB), and cardiac troponin I, and we took myocardial biopsies from the interventricular septum for chemiluminescence assay of reactive oxygen species (hydroxyl, hydrogen peroxide, hypochlorite, and superoxide). There was no significant difference in the myocardial tissue release of hydrogen peroxide, hydroxyl, hypochlorite, and superoxide between the 2 groups (P > 0.05). The systemic tumor necrosis factor-alpha levels in the off-pump group were significantly lower than in the on-pump group (P <0.01). The cardiac troponin I and creatine kinase-MB levels at 6, 12, and 24 postoperative hours were not statistically different between the 2 groups (P >0.05). We conclude that off-pump CABG appears to reduce systemic inflammation, without reducing myocardial oxidative stress and inflammation.