Can inflammatory biomarkers based on first trimester complete blood count parameters predict placental abruption ?: A case-control study.

OBJECTIVES: Placental abruption (PA) is associated with adverse maternal and neonatal outcomes and has an etiological mechanism that is not yet fully understood. The prediction of PA, which has been the subject of numerous studies, remains a challenge. In particular, there is evidence that PA can be considered a chronic process. So, this study aimed to show inflammatory biomarkers based on complete blood count parameters may be used to predict PA. STUDY DESIGN: A sample of 110 cases (pregnant women with PA) and 110 controls (healthy pregnant women with spontaneous labor) were required the study. The present case-control study included a total of 220 pregnant women. Inflammatory makers were used to evaluate the PA prediction RESULTS: Increases in body mass index, mean corpuscular volume and paletelet lymphocyte ratio are considered protective factors, while increases in neutrophil, the systemic inflammatory response index, neutrophil lymphocyte ratio and the pan-immune inflammation score are considered risk factors. Each 1 unit increase in neutrophil count increases the risk of a PA diagnosis by 1.81 times. CONCLUSION: Recent studies indicate a strong heterogeneity of clinical courses leading to PA in premature and term births. In the present study, our results showed that, inflammation is associated with PA.

The effects of orlistat on oxidative stress, recognition memory, spatial memory and hippocampal tissue in experimentally induced obesity in rats.

This study investigates the impact of orlistat on oxidative stress, spatial memory, recognition memory, and hippocampal tissue in obese rats. The study groups were divided into control, high fat diet-induced obese (HFDIO), HFDIO+orlistat (HFDIO+ORL) groups, each consisting of 8 animals. While control fed with standart diet, HFDIO and HFDIO+ORL fed with high-fat diets for 8 weeks to induce obesity. Then, ORL treated 10 mg/kg for 7 weeks, while control and HFDIO get water. At 16th week, novel object recognition (NOR) and Morris water maze (MWM) tests were performed. TNF-alpha, IL-1beta levels in hippocampal tissue, and total/native thiol/disulphide levels in serum were measured. TNF-alpha level of HFDIO was higher than control, while lower in HFDIO+ORL compared to HFDIO as like IL-1beta level. On the contrary, serum total thiol level was lower in HFDIO than control and higher in HFDIO+ORL compared to the HFDIO, while disulphide level was opposite of the total thiol levels. While recognition index was higher in HFDIO+ORL, in MWM, latency of finding platform in HFDIO was higher than control and latency of HFDIO+ORL was very similar to control in 2-4 days. The HFDIO group demonstrated decrease in time spent in platform zone compared to control, whereas time spent of the HFDIO+ORL was higher than HFDIO. Our study demonstrates that orlistat administration exerts beneficial effects on oxidative stress, spatial memory, recognition memory, and hippocampal tissue in obese rats. It shows that orlistat may have potential therapeutic implications for obesity-related cognitive impairments and hippocampal dysfunction.

Investigation of cochlear microphonics recorded with different stimulus types.

BACKGROUND: Electrocochleography (ECochG), one of the first defined tests under auditory evoked potentials, is a total electrical response of inner and outer hair cells inside the cochlea and auditory nerve record technique to the presence of an acoustic stimulus. These records can be used in Meniere disease and auditory neuropathy spectrum disorder diagnosis, intraoperative monitoring. In addition, the presence of cochlear microphonics plays a crucial role in auditory neuropathy spectrum disorder diagnosis. In our study, healthy individuals were tested with extratimpanic electrocochleography record method via Click and LS CE-Chirp stimulus, and the results were compared to the age, sex, and noise sensitivity categories. METHODS: This study had executed at Baskent University, Faculty of Health Sciences Audiology laboratory. The study group consisted of 42 volunteers between 18 and 40 years old. To understand the suitability of volunteers, pure tone audiometry, tympanometry, and transient otoacoustic emission tests were performed. Individuals with no hearing loss were tested with 100 dBnHL intensity level via click and LS CE-Chirp stimulus. The obtained values were statistically evaluated in the SPSS 23.0 program in accordance with the data distribution. An independent sample t-test was used for data showing normal distribution, and Mann-Whitney U test was used for data not showing normal distribution. The level (p < 0.05) was considered statistically significant for all analyses performed. RESULTS: Cochlear microphonic amplitudes recorded with click and LS CE-Chirp stimuli were higher in males than in females (p = 0.051 and p = 0.001, respectively). When the age groups were evaluated, no difference was observed in the CM amplitudes obtained with both click and LS CE-Chirp stimuli. There was no correlation between age and CM amplitudes. Additionally, it was determined that the CM amplitudes recorded with the click stimulus in individuals with noise sensitivity were higher than those without noise sensitivity (p = 0.051). DISCUSSION: It is thought that the ECochG amplitudes of different gender, different age, and different noise sensitivity, which are the results of our study, can be used in the diagnosis of diseases such as auditory neuropathy spectrum disorder.

Fetuin-A expression in human umbilical vein endothelial cells and amnion cells of patients with gestational diabetes mellitus.

OBJECTIVES: To elucidate the link between fetuin-A expression in human umbilical vein endothelial cells (HUVECs) and amnion cells (ACs) and clinicopathological changes in patients with gestational diabetes mellitus (GDM) and newborns. METHODS: This retrospective cohort study included 82 pregnant patients (40 with GDM and 42 controls) between January 2019 and January 2022. The patients underwent a one-hour, 50 gram glucose challenge test (GCT) during the 24-28th weeks of pregnancy. Patients with positive GCTs immediately underwent a 3-hour, 100 gram oral glucose tolerance test. The expression level of fetuin-A in UVECs and ACs was evaluated by immunohistochemistry (IHC) and scored based on IHC staining in randomly selected slides. The IHC staining intensity was evaluated by the number of dots, which reflects the expression level of fetuin-A in both HUVECs and ACs. RESULTS: The GDM group displayed significantly higher fetuin-A expression in both HUVECs (p<0.0001) and ACs (p=0.0001) when compared with the control group. Fetuin-A expression in HUVECs was correlated with fetal macrosomia, neonatal hypoglycemia, and placental weight. However, there was no correlation with fetuin-A expression in ACs. CONCLUSION: There is a correlation between fetal macrosomia, neonatal hypoglycemia, placental weight, and fetuin-A expression of HUVECs in patients with GDM.

Effects of incubator oxygen and carbon dioxide concentrations on hatchability of fertile eggs, some blood parameters, and histopathological changes of broilers with different parental stock ages in high altitude.

The effects of incubator carbon dioxide (CO2) and oxygen (O2) concentrations with parental stock age (PSA) on embryonic deaths (ED), hatchability of fertile eggs (HFE), some blood parameters, and the tissue development of broilers were investigated. Four consecutive repetitions following the similar materials and methods were carried. From 3 different aged ROSS 308 broiler parental flocks 7,680 hatching eggs were obtained and classified as young (Y; 29 wk), middle (M; 37 wk) and old (O; 55 wk) as regards PSA, and randomly distributed. Four different incubator ventilation programs (IVP) as control (C; 0.67% CO2 and 20.33% O2), high CO2 (HC; 1.57% CO2 and 20.26% O2), high O2 (HO; 0.50% CO2 and 21.16% O2), and high CO2 + O2 (HCO; 1.17% CO2 21.03% O2) were applied with oxygen concentrator, and ED and HFE were investigated. Lung and heart tissues, hemoglobin value, packed cell volume, and red blood cell count, triiodothyronine, thyroxine, adrenocorticotropic hormone (ACTH) values of the chicks were analyzed. It was found that IVP affected ED and HFE. Higher rate of early ED (EED) was obtained from the HC than HCO, and higher middle+late stage+pipped but unhatched ED (MLPED) with a lower rate of HFE was observed in the C group than HO and HCO (P < 0.05). Association was found between PSA and IVP (P < 0.05), being more evident in EED for young PSA, in MLPED with HFE for Y and O PSA. From hematological values, no statistical difference in RBC, PCV, and Hb values were found among the treatment groups, ACTH concentration known as a response to stress was found to be higher than C in all groups, triiodothyronine concentration was higher in the HO group than C. In the histopathological examination, used IVPs were found to have negative effects on the lung and heart such as vacuolization, hemorrhage in all PSA groups except for C. Conclusively, PSA and IVP affected some hatching, blood and tissue development parameters of the broiler chicks.

Number of cells in parathyroid tissue in primary hyperparathyroidism cases and its relationship with serum calcium value.

BACKGROUND: The relationship between serum calcium (Ca) level to serum parathyroid hormone (PTH), phosphorus (P) levels and tissue properties of the parathyroid gland is unknown in primary hyperparathyroidism cases. Revealing this relationship may be useful for understanding the etiopathogenesis of primary hyperparathyroidism and determining the time of treatment. METHODS: Ninety patients (71 females, 19 males, age range; 27-73 years, average age; 46) who underwent single gland excision with the diagnosis of primary hyperparathyroidism were studied. The patients were divided into 2 groups as serum Ca level <12 and serum Ca level ≥12. Age and sex of the patients, mean cell number of the gland, mean volume of the gland, serum levels of PTH, P, and histopathologic type of hyperplasia were evaluated. RESULTS: The mean cell number per cubic centimeter is 22.9 (10-220 range) million in all glands. Serum Ca level was <12 in 82 (91.1%) of the patients, and ≥12 in 8 (8.9%) cases. Mean cell number of the gland, mean volume of the gland, existence of cystic hyperplasia of the gland, serum levels of PTH and P were statistically significant between the 2 groups (P < .001, P < .001, P < .05, P < .001, P < .05 respectively). CONCLUSION: In primary hyperparathyroidism cases serum Ca level is not related to age and sex but directly related to proportionals to the cell number and volume of the gland and serum levels of PTH, inversely related to cystic hyperplasia and serum levels of P. Early surgical intervention should be planned since the serum Ca level will be high in large adenomas with a noncystic radiological appearance.

Assessment of tebuconazole exposure on bovine testicular cells and epididymal spermatozoa.

This study is the first to investigate the effects of tebuconazole (TEB) on the physiological functions of bovine testicular cells and epididymal spermatozoa. Motility and plasma membrane integrity of spermatozoa exposed to TEB (0.001-100 µM) were evaluated at different incubation times (0-6 h), while TEB-induced spermiotoxicity was assessed after 24 h in cell cultures. Testicular cells, obtained from the parenchyma of bovine testes, were seeded at 1.0 × 104 and 1.5 × 106 cells/well in 96- and 12-well culture plates and incubated for 48 h in culture media containing TEB (0.001-100 µM) to evaluate cytotoxicity and hormone release, respectively. TEB did not affect the motility and plasma membrane integrity. However, significant spermiotoxicity occurred at higher TEB (1-100 µM) concentrations (P < 0.05) compared to control and lower doses. Although no dose caused cytotoxicity in testicular cells (P > 0.05), 1 and 100 µM TEB caused a significant increase in testosterone secretion (P < 0.05). As a result, high doses of TEB (1-100 µM) had slightly suppressive effects on spermatozoa; however, these doses had stimulatory effects on testosterone secretion by testicular cells. It appears that the disruption of hormonal homeostasis of testicular cells after TEB exposure may result in metabolic and especially reproductive adverse effects in bulls.

Effects of mesh surgery on sexual function in pelvic prolapse and urinary incontinence

ABSTRACT Purpose: We aimed to determine pre-operative and post-operative sexual function scores of patients who underwent four-arm polypropylene mesh implantation surgery to treat urinary incontinence and pelvic organ prolapse. Materials and Methods: A prospective study from January 2011 to November 2015 including patients (n: 72) submitted to surgical mesh implantation (four-arm anterior mesh implant (Betamix POP4®, Betatech Medical, Turkey) questioned the patients with Female Sexual Function Index evaluation form. The questionnaire was applied to all patients at pre-operative, post-operative 3rd month and post-operative 1st year periods. Results: The mean age of the patients was 47.2±7.1 years. The mean Body Mass Index (kg/m2) was 28.7±3.7. The average of incontinence duration (year) was 4.6±2.6 and the average for operation time (min) was 35.7±2.1. After the urinary incontinence and pelvic organ prolapse surgery, it was observed that incontinence complaints of patients reduced. Furthermore, there was a positive change in quality of life and sexual function of patients at the post-operative period. There was a statistically significant increase according to Female Sexual Function Index score among all three periods (16%, 86% and 100% respectively, p=0.001) and improvement of sexual functions was observed. Conclusions: Transvaginal mesh use in the surgical treatment of pelvic organ prolapse improves quality of life. However, risk factors such as transvaginal mesh usage indication, surgical technique and experience of the surgeon, suitability of the material, the current health status of the patient and postoperative personal care of the patient may affect the success of operations.

Effects of mesh surgery on sexual function in pelvic prolapse and urinary incontinence.

PURPOSE: We aimed to determine pre-operative and post-operative sexual function scores of patients who underwent four-arm polypropylene mesh implantation surgery to treat urinary incontinence and pelvic organ prolapse. MATERIALS AND METHODS: A prospective study from January 2011 to November 2015 including patients (n: 72) submitted to surgical mesh implantation (four-arm anterior mesh implant (Betamix POP4R, Betatech Medical, Turkey) questioned the patients with Female Sexual Function Index evaluation form. The questionnaire was applied to all patients at pre-operative, post-operative 3rd month and post-operative 1st year periods. RESULTS: The mean age of the patients was 47.2±7.1 years. The mean Body Mass Index (kg/m2) was 28.7±3.7. The average of incontinence duration (year) was 4.6±2.6 and the average for operation time (min) was 35.7±2.1. After the urinary incontinence and pelvic organ prolapse surgery, it was observed that incontinence complaints of patients reduced. Furthermore, there was a positive change in quality of life and sexual function of patients at the post-operative period. There was a statistically significant increase according to Female Sexual Function Index score among all three periods (16%, 86% and 100% respectively, p=0.001) and improvement of sexual functions was observed. CONCLUSIONS: Transvaginal mesh use in the surgical treatment of pelvic organ prolapse improves quality of life. However, risk factors such as transvaginal mesh usage indication, surgical technique and experience of the surgeon, suitability of the material, the current health status of the patient and postoperative personal care of the patient may affect the success of operations.

Antioxidant effects of melatonin in heart tissue after induction of experimental periodontitis in rats.

The aim of this study was to evaluate the effects of melatonin on the oxidative stress in heart tissues after induction of experimental periodontitis in rats. Thirty Wistar Albino male rats were divided into four groups as follows: healthy + saline solution (Hs, n = 7), healthy + melatonin (Hm, n = 7), periodontitis + saline solution (Ps, n = 8), and periodontitis + melatonin (Pm, n = 8). Experimental periodontitis was induced using a ligature placed at the gingival margin of the maxillary second molars. Melatonin was applied intraperitoneally (10 mg/kg) every day for 2 weeks. After sacrificing the rats, serum levels of malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) levels, and melatonin levels were evaluated. The Pm group exhibited lower alveolar bone loss than the Ps group. Melatonin levels increased in the periodontitis groups, and the Pm group had lower MDA levels and higher GSH-Px levels than the Ps group. These findings suggest that melatonin administration reduces MDA and increases GSH-Px levels in heart tissue, and these effects may be due to its antioxidant properties. Further studies are needed to understand the effects of melatonin on the association between periodontitis and cardiovascular disease.

Manganese superoxide dismutase, glutathione peroxidase and catalase gene polymorphisms and clinical outcomes in acute kidney injury.

INTRODUCTION: The aim of this study was to evaluate the potential association of single gene polymorphisms of manganese superoxide dismutase (MnSOD), glutathione peroxidase 1 (GPX1) and catalase (CAT) with clinical outcomes of acute kidney injury (AKI). MATERIALS AND METHODS: Ninety AKI patients and 101 healthy volunteers were included in the study. Determination of MnSOD rs4880, GPX1 rs1050450 and CAT rs769217 polymorphisms was performed using real-time polymerase chain reaction amplification. The duration of hospitalization of AKI patients, dialysis and intensive care requirements, sepsis, oliguria and in-hospital mortality rates were assessed. RESULTS: The MnSOD, GPX1 and CAT genotypes and allele frequencies of AKI patients did not differ significantly from those of healthy controls. In patients with a T allele in the ninth exon of the CAT gene, intensive care requirements were greater than those of patients with the CC genotype (p = 0.04). In addition, sepsis and in-hospital mortality were observed significantly more frequently in patients with a T allele in the ninth exon of the CAT gene (p = 0.03). Logistic regression analysis determined that bearing a T allele was the primary determinant of intensive care requirements and in-hospital mortality, independent of patient age, gender, presence of diabetes and dialysis requirements (OR 6.10, 95% CI 1.34-27.81, p = 0.02 and OR 10.25, 95% CI 1.13-92.80, p = 0.04, respectively). CONCLUSION: Among AKI patients in the Turkish population, hospital morbidity and mortality were found to be more frequent in patients bearing a T allele of the rs769217 polymorphism of the CAT gene.

The prophylactic effect of Viscum album in streptozotocin-induced diabetic rats.

OBJECTIVE: Viscum album (VA) is a species of mistletoe in the family Santalaceae that is thought to have therapeutic properties for several diseases, including diabetes. In the present study, conventional experimental rat model was used with diabetes induced with streptozotocin (STZ) to evaluate effect of VA on lipid peroxidation and antioxidant system. METHODS: Total of 32 adult, male Sprague-Dawley rats were divided into 4 groups of 8 rats: Control group, STZ group, VA group, and group administered VA+STZ. VA extract was 100 mg/kg preparation delivered once a day by oral gavage for 10 days. Single dose of 55 mg/kg STZ citrate buffer (0.1 M, pH 4.5) was administered intraperitoneally to induce diabetes. Fasting blood glucose level was measured and recorded. Animals were sacrificed, and catalase (CAT), malondialdehyde (MDA), and protein present in liver and kidney tissue samples were measured. Activity of CAT, an antioxidant enzyme, was studied according to the Aebi method. MDA, a product of lipid peroxidation, was analyzed using Draper and Hadley spectrophotometric procedure. Protein level was determined using supernatant and extract of tissue homogenates according to Lowry method. Data were assessed using one-way analysis of variance and pairwise comparisons between groups. Post-hoc analysis included Dunnet test, Duncan test, and least significant difference test. P<0.05 was considered significant probability value. RESULTS: Oxidative stress is associated with diabetic complications. VA administered to diabetic rats reduced oxidative stress and improved their general condition. CONCLUSION: Further studies are needed to enhance understanding of potential antidiabetic and antioxidant effects of VA.

Genetic Polymorphism of Manganese Superoxide Dismutase in Behçet's Disease.

OBJECTIVES: This study aims to investigate the genetic association between single nucleotide mutation in mitochondrial manganese superoxide dismutase and a Behçet's disease (BD) population by using molecular techniques. PATIENTS AND METHODS: Ninety-three BD patients (45 males, 48 females; mean age 33.15±8.99 years; range 17 to 65 years) and 125 controls (58 males, 67 females; mean age 28.33±7.31 years; range 18 to 62 years) were genotyped by polymerase chain reaction-restriction fragment length polymorphism method. The genotypic distributions in BD patients and controls were consistent with the Hardy-Weinberg equilibrium. RESULTS: Significant differences were observed between BD patients and controls in terms of genotypic distribution. Frequencies of alanine (Ala)/Ala, Ala/valine (Val), and Val/Val were 14.0% (n=13), 45.2% (n=42), and 40.9% (n=38) in BD patients and 21.6% (n=27), 53.6% (n=67), and 24.8% (n=31) in controls, respectively (p=0.033). CONCLUSION: The Val/Val genotype of the manganese superoxide dismutase gene is associated with the physiopathology of BD in a group of Turkish patients.

Malondialdehyde and nitric oxide levels and catalase, superoxide dismutase, and glutathione peroxidase levels in maternal blood during different trimesters of pregnancy and in the cord blood of newborns.

BACKGROUND/AIM: To determine whether levels of malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) change during the 3 trimesters of pregnancy, and to compare the third trimester of pregnancy with newborn cord blood in respect to the levels of MDA, NO, and antioxidant enzymes. MATERIALS AND METHODS: Maternal blood samples were collected at 9-13, 22-26, and 36-40 weeks of gestation. Cord blood was collected at the time of delivery. RESULTS: NO levels and GPx activity were higher in the second and third trimesters than in the first trimester. MDA level was lower in the third trimester and SOD and CAT activities were lower in the second and third trimesters than the first trimester. In cord blood, MDA and NO levels were lower while CAT and GPx activities were higher than in the third trimester of pregnancy. CONCLUSION: These results suggest that the balance of free radical and antioxidant production during pregnancy ensures healthy fetus growth and development.

Effects of pentoxifylline and alpha lipoic acid on methotrexate-induced damage in liver and kidney of rats.

The aim of the current study was to investigate the probable protective effects of Pentoxifylline (PTX) and Alpha Lipoic Acid (ALA), which display anti-oxidative efficacy against hepatotoxicity and nephrotoxicity, those being the major side effects of Methotrexate (MTX). Rats were divided into four groups: a control group; MTX (20mg/kg/day) group; MTX+PTX (20mg/kg/day+50mg/kg/day) group; and an MTX+ALA (20mg/kg/day+100mg/kg/day) group. At the end of the experiment, biochemical, histochemical and immunohistochemical analyses were performed on liver and kidney tissues of rats. We determined Glutathione Peroxidase (GSH-Px), Superoxide Dismutase (SOD), Catalase (CAT), Malondialdehyde (MDA), Nitric Oxide (NO) and Xanthine Oxidase (XO) levels in the liver and kidney. Moreover, Gamma Glutamyl Transferase (GGT), Direct Bilirubin (DBil), Blood Urea Nitrogen (BUN), and urea levels were measured in the serum. The histochemical evaluation revealed a significant decrease in MTX caused damage in the PTX- and ALA-treated groups (especially in ALA group). On the other hand, the immune staining of iNOS and TNF-α were observed most densely in the MTX group, while the density decreased in the PTX- and ALA-administered groups. We determined increased GGT, BUN, urea and levels of CAT, MDA, NO, and XO values in both groups, while GSH-Px (an increase in liver tissue) and DBil levels were decreased in the group that received MTX. However, we determined decreased SOD levels in liver tissue. In the PTX and ALA groups, the levels of GGT, BUN and urea as well as the levels of CAT, MDA, NO and XO decreased (SOD increased in the liver tissue), and the levels of GSH-Px and DBil increased. In conclusion, it can be stated that, although ALA is more effective in preventing the toxic effects of MTX on the liver and kidney, PTX also has a preventive effect. As a result, we can readily suggest that ALA and PTX can have protective effects by decreasing MDA, NO, BUN and urea values as antioxidants against MTX-induced damage in liver and kidney of rats.

Paliperidone regulates intracellular redox system in rat brain: Role of purine mechanism.

OBJECTIVE: The treatment of schizophrenia is multifactorial, with antipsychotic medications comprising a major part of treatment. Paliperidone is a newly commercialized antipsychotic whose formulation includes the principal active metabolite risperidone, 9-hydroxyrisperidone. Ever since the relationship between schizophrenia and oxidative stress was first demonstrated, many studies have been conducted in order to probe the potential protective effects of antipsychotic drugs on the oxidant-antioxidant system and lipid peroxidation. The basic aim of this study is to determine the effects of the newly marketed drug paliperidone on the activities of the enzymes adenosine deaminase (ADA), xanthine oxidase (XO), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) as well as on malondialdehyde (MDA) and nitric oxide (NO) levels in rat brain tissues. METHODS: Twenty male Sprague-Dawley rats were used for the study, which were divided into two equal groups. The first was the control group (n = 10) and the second was the paliperidone group (n = 10). Saline was administered once daily for 14 days in the control group. In the paliperidone group, paliperidone was administered once daily with a dose of 1 mg/kg for 14 days. All rats were sacrificed at the end of the fourteenth day. Brain samples were collected and then analyzed. RESULTS: Our results demonstrated that paliperidone significantly decreased the activities of ADA (P = 0.015), XO (P = 0.0001), and CAT (P = 0.004) while insignificantly increasing the activity of SOD (P = 0.49), MDA (P = 0.71), and NO (P = 0.26) levels in rat brain tissues. In addition, paliperidone insignificantly decreased the activity of GSH-Px (P = 0.30) compared to the control group in rat brain tissues. DISCUSSION: In conclusion, the data obtained in this study suggest that paliperidone can positively alter antioxidant status and, accordingly, can offer positive outcomes in the treatment of schizophrenia by reducing activity in the enzymes ADA and XO, which are associated with purine metabolism. We believe that such a comprehensive approach used with other antipsychotic drugs warrants further study.

Effects of cholesterol and cAMP on progesterone production in cultured luteal cells isolated from pseudopregnant cat ovaries.

The present study was designed to incubate luteal cells isolated from pseudopregnant cats and to investigate the effects of cholesterol and cAMP on luteal progesterone production. Corpora lutea were collected from the cats on days 10 and 15 of pseudopregnancy. Luteal cells were isolated from the ovaries by collagenase digestion. Steroidogenic luteal cells were stained for 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activity. Cells (2 x 10(4)) staining positive for 3beta-HSD were cultured for up to 7 days. The cells were treated with 22(R)-hydroxycholesterol (22R-HC) and dibutyryl cyclic AMP (dbcAMP) on days 1, 3 and 7. Treatment of cells with 22R-HC resulted in a dose-dependent increase (p<0.001) in progesterone production. When 22R-HC was used at a concentration of 10 microg/ml, it resulted in 2.7- and 5.1-fold increases in progesterone production on days 3 and 5, respectively. When the dose was doubled (20 microg/ml), treated cells produced four times more progesterone on days 3 and 7, and three times more on day 5. By day 7, progesterone production increased up to 9.1 times more than the control. Incubation of cells with both concentrations of dbcAMP (0.1 mM and 1 mM) resulted in significant stimulations of progesterone on days 5 and 7 (p<0.001). However, on day 3, only higher doses of dbcAMP (1 mM) resulted in significant stimulation (p<0.05). Progesterone production was increased up to 2- and 2.9-fold of the control when cells were treated with lower concentration of dbcAMP (0.1 mM) on days 5 and 7, respectively. Incubation of cells with 1 mM concentrations of dbcAMP induced a 3.2-fold increase on day 5 and a 5-fold increase on day 7. In conclusion, a successful incubation was performed for long-life culturing of luteal cells collected from pseudopregnant cats. The method works well and allows for optimal growth and development of cells in the culture. The present study also demonstrated that incubating cat luteal cells with 22R-HC and dbcAMP induces a significant increase in luteal progesterone synthesis.