Evaluation of maternal serum VEGF, TNF-alpha, IL-4, and IL-10 levels in differentiating placenta accreta spectrum from isolated placenta previa.

OBJECTIVE: Our study aimed to differentiate patients with placenta accreta spectrum (PAS) from those with placenta previa (PP) using maternal serum levels of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-4 (IL-4), and IL-10. METHODS: The case group consisted of 77 patients with placenta previa, and the control group consisted of 90 non-previa pregnant women. Of the pregnant women in the case group, 40 were diagnosed with PAS in addition to placenta previa and 37 had placenta previa with no invasion. The maternal serum VEGF, TNF-alpha, IL-4, and IL-10 levels were compared between the case and control groups. Then the success of these markers in differentiating between PP and PAS was evaluated. RESULTS: We found the VEGF, TNF-alpha, and IL-4 levels to be higher and the IL-10 level to be lower in the case group compared to the control group (p < 0.001). We observed a statistically significantly lower IL-10 level in the patients with PAS than those with PP (p = 0.029). In the receiver operating characteristic analysis, the optimal cut-off of IL-10 in the detection of PAS was 0.42 ng/mL). In multivariate analysis, the risk of PAS was significant for IL-10 (odds ratio (OR) 0.45, 95 % confidence interval (CI) 0.25-0.79, p = 0.006) and previous cesarean section (OR 2.50, 95 % Cl 1.34-4.66, p = 0.004). The model's diagnostic sensitivity and specificity, including previous cesarean section, preoperative hemoglobin (Hb), TNF-alpha, and IL-10 were 75 % and 72.9 %, respectively. CONCLUSION: The study showed that the IL-10 level was lower in patients with PAS than in those with PP. A statistical model combining risk factors including previous cesarean section, preoperative Hb, TNF-alpha, and IL-10 may improve clinical diagnosis of PAS in placenta previa cases. Cytokines may be used as additional biomarkers to the clinical risk factors in the diagnosis of PAS.

The influencer effect of Dexmedetomidine on radioiodine relevant to lacrimal gland impairment.

PURPOSE: To assess the potential influencing effects of Dexmedetomidine on impaired lacrimal glands after high-dose radioiodine treatment (RAI). METHODS: Thirty-six rats were arbitrarily separated into 3 groups: Sham, RAI, and Dexmedetomidine. Dexmedetomidine group was given Dexmedetomidine and RAI, the Sham group was given the same millimeters of saline, and the RAI group was given RAI only. All forms of lacrimal glands, including harderian glands (HG), extraorbital (EG), and intraorbital (IG) lacrimal glands, were evaluated for immunohistochemical, histopathologic assessments and also for tissue cytokines, oxidant and antioxidant levels. RESULTS: Dexmedetomidine significantly ameliorated histopathologic changes such as periacinar fibrosis, acinar atrophy, lymphocytic infiltration, ductal proliferation, lipofuscin-like accumulation, and nucleus changes caused by RAI in all lacrimal gland forms (p < 0.05 for all of the parameters). However, periductal fibrosis was improved significantly only in EG (p = 0.049), and mast cell infiltration was improved significantly only in IG (p = 0.038) in Dexmedetomidine groups. There was a significant decrease in the elevated caspase-3 and TUNEL levels after RAI administration in the Dexmedetomidine group in all lacrimal gland forms (p < 0.05 for all parameters). Dexmedetomidine attenuated NF-kb, TNF-α, and IL-6 levels significantly diminished total oxidant status and raised total antioxidant status levels (p < 0.05 for all parameters). CONCLUSIONS: The results of this study demonstrated that following RAI, Dexmedetomidine diminished inflammation, tissue cytokine levels, and apoptosis and ameliorated impaired histopathologic patterns of the lacrimal glands.

Evaluation of Midkine levels in pregnant women with preterm premature rupture of membranes: A cohort study from a tertiary hospital.

OBJECTIVE: To assess midkine (MK) levels in pregnant women with preterm premature rupture of membranes (PPROM) and compare them to healthy pregnant women. We also assessed the performance of the maternal serum MK level in predicting neonatal intensive care unit (NICU) requirement in the PPROM group. METHODS: Forty pregnant women who presented to our clinic at 24-37 gestational weeks and were diagnosed with PPROM were included in the study group. During the same period, 40 healthy pregnant women at similar gestational weeks were randomly selected as the control group. Clinical characteristics, inflammatory markers, and serum MK levels were compared between the groups. The same parameters were then compared between the PPROM cases with and without NICU requirement. Finally, the receiver operating characteristic (ROC) analysis was performed to assess the predictive value of MK for NICU requirement. RESULTS: The PPROM and control groups were similar in terms of demographics. The MK level of the pregnant woman with PPROM was significantly higher than that of the controls. No statistically significant difference was found between the MK levels of the cases with and without NICU requirement in the PPROM group. In the ROC analysis, the optimal cut-off value of was found to be 0.287, at which it had 63 % sensitivity and 65 % specificity (area under the curve(AUC): 0.78, 95 % confidence interval(CI): 0.683-0.881, p < 0.001) for the prediction of NICU requirement in cases with PPROM. In the same analysis performed for the prediction of PPROM, when the optimal cut-off value was taken as 0.298, MK had 56 % sensitivity and 60 % specificity (AUC: 0.65, 95 % CI: 0.522-0.770, p = 0.037). CONCLUSION: Serum MK seems to be associated with complicated inflammatory processes leading to PPROM, and this novel marker has the potential to predict NICU requirement in PPROM cases.

Comparison of VEGF-A levels in women with threatened abortion, early pregnancy loss and uncomplicated healthy pregnancies.

INTRODUCTION: To estimate the possible role of VEGF-A in predicting poor early pregnancy outcomes including threatened abortion and early pregnancy loss. METHODS: We conducted a prospective case-control study with three groups of pregnant women diagnosed with threatened abortion, early pregnancy loss, and uncomplicated healthy pregnancies between 01 March 2023 and 15 March 2023. Maternal serum VEGF-A concentration was measured using the Sandwich-ELISA method in accordance to the commercial kit's instructions. There were 30 patients in each 3 group and the gestational age of the patients was between 6 and 14 weeks. The Kruskal-Wallis test was performed for comparing the median values between the groups. Mann-Whitney U test was conducted for pairwise comparisons. RESULTS: VEGF-A levels were compared between 3 groups and a statistically significant difference was found (p = 0.007). There was a moderately significant correlation between VEGF-A levels and poor early pregnancy outcomes. For poor early pregnancy outcomes, the area under the curve (AUC) was 0.75 (95% CI: 0.64-0.85). The best balance of sensitivity/specificity in ROC curves was 0.60 (63.3% sensitivity, 74.3% specificity). DISCUSSION: In conclusion, this study pointed out the increased VEGF concentrations in pregnant women with threatened miscarriage and early pregnancy loss. VEGF-A may be a potential biomarker for the indication of poor early pregnancy outcomes.

Maternal serum midkine level in fetal growth restriction: a case-control study.

OBJECTIVES: To compare maternal serum midkine (MK) level in pregnant women with idiopathic fetal growth restriction (FGR) and healthy. In addition, we assessed the value of maternal serum MK level in predicting neonatal intensive care unit (NICU) admission. METHODS: A total of 144 pregnant women were included, 72 with idiopathic FGR and 72 healthy in this study. The control group was matched for the mother's age, parity, gestational age, and pre-pregnancy body mass index (BMI) with the idiopathic FGR group at the time of recruitment into the study and sample collection. RESULTS: Serum MK level is higher in the idiopathic FGR than the control group (0.24 ng/mL (0.19-0.32) vs. 0.18 ng/mL (0.14-0.23), p<0.001). In addition, we compared the maternal serum MK level of those with and without NICU admission in the FGR group (0.25 ng/mL (0.19-0.37) vs. 0.21 ng/mL (0.18-0.28), p=0.014). We performed ROC curve analysis to serum MK level predicting NICU admission in the FGR group (AUC: 0.668, %95 CI [0.550, 0.785], p=0.014). A sensitivity of 63% and a specificity of 62% for the serum MK level were achieved with a cut-off value of 0.22 for NICU admission. CONCLUSIONS: To the best of our knowledge, this study is the first to compare maternal serum MK level in pregnant women with idiopathic FGR and healthy. We showed that maternal serum MK level was significantly elevated in pregnant women with FGR than healthy.

Serum midkine level might be a diagnostic tool for COVID19 disease in pregnancy: From the disease severity, hospitalization and disease progression respects.

BACKGROUND: New biomarkers for diagnosis and monitoring the COVID-19 disease are the most important topics to be studied recently. We aimed to investigate the association between midkine levels and disease severity in pregnant women with COVID-19. METHODS: Totally 186 pregnant women were participated in this study. 96 of them were healthy pregnant women, 90 of them were pregnant women with COVID19. Pregnant women were evaluated according to their trimesters. Serum midkine level, biochemical profile clinical and disease severity outcomes of pregnant women were obtained. RESULTS: Our results showed that pregnant women with COVID19 have significantly increased serum midkine level compared to healthy pregnant women (1.801 ± 0.977 vs 0.815 ± 0.294 ng/dL). According to the data among each trimester, it was shown that there were significant increase in serum midkine level during all pregnancy trimesters (1st trimester Control Group: 0.714 ± 0.148, COVID-19 group 1.623 ± 0.824, p < 0.0001; 2nd trimester Control Group: 0.731 ± 0.261, COVID-19 group 2.059 ± 1.146, p < 0.0001; 3rd trimester Control Group: 1.0 ± 0.35, COVID-19 group 1.723 ± 0.907, p = 0.001). Serum midkine levels were significantly different between disease severity subgroups of pregnant women with COVID19; moderate and severe/critic groups had significantly higher serum midkine level than mild group. There was also significant correlation between serum midkine level and severity status (p:0.0001, r: 0.468). The most striking results of serum midkine levels were corelation between length of hospitalization (p: 0.01, r: 0.430) and O2 saturation (p < 0.0001, r: -0.521). ROC curve analysis showed that serum midkine level might be a tool for predicting COVID-19 in pregnant women with COVID-19 (AUC: 0.912, 95% CI: [0.871, 0.952], p < 0.0001) CONCLUSION: Our data showed that there is an obvious relation between COVID19 progression and serum midkine level for the first time which might be used for monitoring the disease process.

Maternal serum midkine level increases in pregnant women with diabetes mellitus: a case-control study.

OBJECTIVE: We aimed to compare maternal serum midkine level in pregnant women with different types of diabetes mellitus (DM) and healthy pregnant women. We also assessed maternal serum midkine level performance to predict adverse neonatal outcomes in the DM group. METHODS: The study included 57 pregnant women diagnosed with gestational diabetes mellitus (GDM) and 41 pregnant women with preexisting DMThe control group consisted of 98 healthy pregnant women. RESULTS: Serum midkine level is higher in the DM group than healthy ones (0.93 ± 0.8 vs. 0.23 ± 0.2, p<.001). When the diabetic groups were compared, the highest serum midkine level was found in GDM, followed by Type 1 DM and Type 2 DM (1.33 ± 0.9 ng/ml, 0.58 ± 0.5 ng/ml vs. 0.30 ± 0.2, respectively). Maternal serum midkine level was higher in the DM group with adverse perinatal outcomes than those without adverse outcomes, but there was no statistical difference (0.97 ± 0.91vs. 0.87 ± 0.73, p=.571). CONCLUSIONS: Serum midkine level was significantly higher in pregnant women with GDM, Type 1, and 2 DM than healthy ones. Serum midkine level did not predict adverse neonatal outcomes in the DM group.

Maternal and Placental Zinc and Copper Status in Intra-Uterine Growth Restriction.

BACKGROUND AND AIM: Zinc and copper are essential trace elements for cell growth and proliferation. Their deficiency may contribute to intrauterine growth restriction (IUGR). We aimed to determine the zinc and copper status of maternal serum and placenta samples of pregnant women with fetal IUGR and age-matched pregnant women without IUGR. METHOD: Serum and placenta samples obtained from 37 IUGR and 21 healthy pregnant women were analyzed at delivery. RESULTS: Placenta zinc concentrations and placenta zinc/copper ratio were significantly lower in the IUGR group compared to controls (p < 0.05). Placenta zinc concentrations correlated with birth weight (p: 0.01, r: 0.31). Maternal levels of zinc and copper were similar between pregnant women with IUGR and controls. CONCLUSIONS: Lower placental zinc and zinc/copper ratio levels in pregnancies with IUGR may indicate that placenta zinc and placental zinc/copper status might be involved in IUGR.

Maternal Serum and Umbilical Cord Brain Natriuretic Peptide Levels in Fetuses with Intrauterine Growth Restriction.

Aim: We compared maternal serum and fetal cord blood proBNP levels in healthy pregnancies to those with intrauterine growth restriction (IUGR). Methods: Prospectively, maternal and cord blood samples at childbirth of 40 pregnant women with isolated IUGR and 40 healthy pregnant women were evaluated for ProBNP levels. Results: The mean serum ProBNP level was significantly higher in newborn cord blood with IUGR than in the control group (181.28 ± 145.37 vs. 91.41 ± 49.77 pg/mL, p = <0.01). Mean serum ProBNP level trended higher in women with IUGR compared to the controls, but was not statistically significant (124.21 ± 113.32 vs. 88.73 ± 85.18 pg/mL, p= >0.05). Conclusion: Third trimester mean proBNP in fetal cord blood are increased in IUGR fetuses at term birth compared to pregnancies with normal fetal growth.

Comparison of VEGF-A values between pregnant women with COVID-19 and healthy pregnancies and its association with composite adverse outcomes.

The aim is to compare VEGF-A values between pregnant women with coronavirus disease 2019 (COVID-19) and healthy controls. Furthermore, the association of inflammation parameters, disease severity, and obstetric complications with VEGF-A was investigated. This prospective case-control study was conducted on pregnant women who were admitted to Ankara City Hospital between June 14, 2020 and August 28, 2020. Pregnant women with COVID-19 (n = 95) were compared with a control group of healthy pregnant women (n = 92) with similar clinical and demographic characteristics. Demographic features, clinical characteristics, laboratory test results, VEGF-A values were compared between the groups. A correlation analysis was performed between VEGF-A levels, inflammation parameters, and clinical characteristics of the cases for pregnant women with COVID-19. VEGF-A levels were also compared between patients with composite adverse outcome and patients without any complication in the COVID-19 group. The two groups were similar except for obstetric complications (p > .05). The obstetric complication rate was higher in the COVID-19 group (p =.02). The two groups were comparable in terms of neutrophil to lymphocyte ratio and VEGF-A values. VEGF-A values were slightly different between the trimesters. A negative moderate statistically significant correlation was found between the neutrophil and VEGF-A values (r = -0.231, p =.02). VEGF-A values were similar between patients with and without composite adverse outcomes (p > .05). VEGF-A values were similar between pregnant women with COVID-19 and healthy controls.

Evaluation of maternal serum afamin and vitamin E levels in pregnant women with COVID-19 and its association with composite adverse perinatal outcomes.

To evaluate the maternal serum afamin and vitamin E levels in pregnant women with coronavirus disease 2019 (COVID-19) and to investigate their association with composite adverse perinatal outcomes. This prospective, case-control study consisted of 60 pregnant women with COVID-19 infection and 36 age-matched pregnant women without any defined risk factors. Demographic features, laboratory test results, afamin and vitamin E levels were compared between the groups. A receiver operating characteristic (ROC) curve was used to assess the relationship of afamin and vitamin E levels in predicting composite adverse perinatal outcomes. A correlation analysis was performed between afamin and C-reactive protein (CRP) levels in pregnant women with COVID-19. The obstetric complication rate was higher in the COVID-19 group (13.3% vs. 2.8%) (p = .01). Afamin levels were higher and vitamin E levels were lower in the COVID-19 group (p = .02 and p < .001, respectively). Vitamin E levels were lower in the COVID-19 group for the all trimesters (p < .001, p < .001, and p = .004, respectively). Afamin levels were higher in the COVID-19 group for the all trimesters without reaching statistical significance (p > .05). The values in the ROC curves with the best balance of sensitivity/specificity for afamin and vitamin E were 0.424 mg/l (70.6% sensitivity, 44.3% specificity) and 3.150 µg/ml (76.5% sensitivity, 58.2% specificity), respectively. A positive moderate statistically significant correlation was found between afamin and CRP levels (r = .264, p = .009). Higher afamin and lower vitamin E levels may support the elevated oxidative stress in the etiopathogenesis of COVID-19 and the relationship with composite adverse perinatal outcomes.

Maternal selenium status plays a crucial role on clinical outcomes of pregnant women with COVID-19 infection.

Adequate maternal selenium level is essential for immune response and healthy pregnancy. This study aimed to shed light on the selenium status of pregnant women with COVID-19 and the effects of potential deficiency in serum selenium levels. Totally 141 pregnant women, 71 of them were COVID-19 patients, in different trimesters were included in the study. Maternal serum selenium levels, demographic and clinical parameters were determined. Serum selenium levels of pregnant women in the second (p: .0003) and third (p: .001) trimesters with COVID-19 were significantly lower than in the healthy group. Maternal selenium level was found to be negatively correlated with gestational week (p < .0001, r: -.541), D-dimer (p: .0002, r: -.363) and interleukin-6 (IL-6) level (p: .02, r: -.243). In the second trimester, serum selenium level positively correlated with white blood cell (p: .002, r: .424), neutrophil (p: .006, r: .39), lymphocyte (p: .004, r: .410) count and hemoglobin (p: .02, r: .323), hematocrit (p: .008, r: .38) status. In the third trimester, it was found that maternal selenium level positively correlated with monocyte (p: .04, r: .353) and negatively correlated with C-reactive protein level (p: .03, r: -.384). Serum selenium level was gradually decreased during the pregnancy period, however, this natural decrease was enhanced together with COVID-19 infection. The reason might be increased selenium needs depended on the immune response against infection. The decrease in maternal selenium level was found to be related to IL-6 and D-dimer levels, which indicate selenium's role in disease progression.

The impact of COVID-19 infection on the cytokine profile of pregnant women: A prospective case-control study.

OBJECTIVE: To compare the levels of various cytokines between pregnant women with confirmed coronavirus disease (COVID-19) infection and pregnant women without any defined risk factor. MATERIALS AND METHODS: Pregnant women with confirmed COVID-19 infection (study group)(n = 90) were prospectively compared to a gestational age-matched control group of pregnant women without any defined risk factors (n = 90). Demographic features, clinical characteristics, laboratory parameters, interferon-gamma (IFN γ), interleukin (IL-2), IL-6, IL-10, and IL-17 levels were compared between the groups. Additionally, a correlation analysis was performed in the study group for the assessment of IFN γ, IL-2, IL-6, IL-10, and IL-17 levels with disease severity and CRP levels. RESULTS: Study group had significantly higher pregnancy complication rate, erythrocyte sedimentation rate, C-reactive protein, procalcitonin, ferritin, D-dimer, lactate dehydrogenase, IFN γ, and IL-6 values (p < 0.05). On the other hand, the control group had significantly higher hemoglobin, leukocyte, platelet, lymphocyte, IL-2, IL-10, and IL-17 values (p < 0.05). Statistically significant differences were found between the groups for IFN γ, IL-2, IL-10, and IL-17 values between the trimesters (p < 0.05). Statistically significant positive correlations were found for IFN γ and IL-6 with disease severity (r = 0.41 and p < 0.001 for IFN γ and r = 0.58 and p < 0.001 for IL-6). On the other hand, a moderate negative correlation for IL-2 and a weak negative correlation for IL-10 were present (r = -0.62 and p < 0.001 for IL-2 and r = -0.19 and p = 0.01 for IL-10). A statistically significant positive moderate correlation was found between IL-6 and CRP (r = 0.40 and p < 0.001) CONCLUSION: COVID-19 infection seems to have an impact on the cytokine profile of pregnant women varying according to pregnancy trimesters and cytokine levels seem to be correlated with disease severity.

Is ProBNP a New Marker for Predicting Intrauterine Growth Restriction?

PURPOSE: To evaluate the usability of first-trimester maternal serum ProBNP levels in the prediction of intrauterine growth restriction (IUGR). Methods In this prospective study, blood samples taken from 500 women who applied to our polyclinic for routine serum aneuploidy screening between the 11-14th gestational weeks were centrifuged. The obtained plasma samples were placed in Eppendorf tubes and stored at -80+°C. For the final analysis, first-trimester maternal serum ProBNP levels of 32 women diagnosed with postpartum IUGR and 32 healthy women randomly selected as the control group were compared. FGR was defined as estimated fetal weight below the 10th percentile for the gestational age. RESULTS: The mean ProBNP levels were statistically and significantly higher in the women with intrauterine growth restriction (113.73±94.69 vs. 58.33±47.70 pg/mL, p<0.01). At a cut-off level of 50.93, ProBNP accurately predicted occurrence of IUGR (AUC+= 0.794 (95% confidence interval 0.679-0.910), p+= 0.001) with sensitivity and specificity rates of 78.1 and 69.0%, respectively. Conclusion First-trimester serum ProBNP level was significantly higher in women who developed IUGR compared to healthy controls. First-trimester ProBNP level can be used as a potential marker to predict the development of IUGR in pregnant women.

[The Role of Vitamin D3 on the Immune Responses of Monocytes]. / Vitamin D3'ün Monositlerin Immün Yaniti Üzerindeki Rolü.

The active form of vitamin D (Vit D), 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3), is important for cell functions and immunity, as well as its role in bone metabolism. Monocytes/macrophages initiate innate immune response, and is considered to be the cell that first comes into contact with the pathogen. They play effective roles in innate immune and inflammatory responses by intercellular relations and inflammatory mediator secretion. Human THP-1 leukemia cells are frequently used for the in vitro determination of the signal pathways, and the functions of monocyte/macrophages. Nuclear factor-kappa B (NF-κB) are complex networks of signaling pathways that regulate many important cellular behaviors, especially in inflammation, cell death, cell differentiation or proliferation. Midkine (MK) is a cytokine and growth factor that is one of the regulators of inflammatory processes, immune cell functions, proliferation and autoimmunity. The effects of Vit D3 on inflammation and MK secretion in hyperglycemia is still unknown. In this study, it was aimed to determine the dose-dependent effects of Vit D3 on the lipopolysaccharide (LPS) stimulated pro/ anti-inflammatory cytokine, NF-κB and MK responses of THP-1 monocyte cells under normo and hyperglycemic conditions. For this purpose, THP-1 monocyte cells stimulated with LPS (Escherichia coli, 0111, 1 µg/ml) under normoglycemic (glucose 100 mg/dl)/hyperglycemic (glucose 500 mg/dl) conditions, were incubated for 24 hours in the presence and absence of 10-50-100 IU/ml Vit D3. MK, TNF-α, IL-8, IL-10 cytokine levels in the supernatants collected from the wells at the end of the incubation periods, and NF-κB levels in the obtained cell lysates were detected by ELISA method. LPS stimulation induced higher levels of TNF-α, IL-8 and MK responses in hyperglycemic conditions. IL-10 secretions were found to be decreased under hyperglycemia. Vit D3 modulates TNF-α, IL-10 and MK secretions in hyperglycemic conditions. The MK and TNF-α levels were determined to be correlated with NF-κB and IL-10. The results obtained in the study showed that Vit D3 can play a role in immune modulation by regulating NF-κB and cytokine/ chemokine-like molecule MK suppression and proinflammatory/anti-inflammatory cytokine balance. The mechanism of the action of Vit D3 under different conditions should be examined in detail.

Relationship between aldose reductase enzyme and the signaling pathway of protein kinase C in an in vitro diabetic retinopathy model.

Protein kinase C (PKC) and aldose reductase (AR) enzyme activities are increased in diabetes and complications are include retinopathy, nephropathy, and neuropathy. However, the relationship between PKC and AR and the underlying molecular mechanisms is still unclear. We aimed to evaluate the relationship between these two enzymes and clarify the underlying molecular mechanisms by the related signaling molecules. The effects of hyperglycemia and oxidative stress on AR and PKC enzymes and the signaling molecules such as nuclear factor-kappa B (NF-κB), inhibitor kappa B-alpha (IkB-α), total c-Jun, phospho c-Jun, and stress-activated protein kinases (SAPK)/Jun amino-terminal kinases (JNK) were evaluated in human retinal pigment epithelial cells (ARPE-19). AR, PKC protein levels, and related signaling molecules increased with hyperglycemia and oxidative stress. The AR inhibitor sorbinil decreased PKC expression and activity and all signaling molecule protein levels. Increased AR expression during hyperglycemia and oxidative stress was found to be correlated with the increase in PKC expression and activity in both conditions. Decreased expression and activity of PKC and the protein levels of related signaling molecules with the AR inhibitor sorbinil showed that AR enzyme may play a key role in the expression of PKC enzyme and oxidative stress during diabetes.

Antioxidant effects of vitamin D on lacrimal glands against high dose radioiodine-associated damage in an animal model.

PURPOSE: To evaluate antioxidant effects of active vitamin D (calcitriol) against high-dose radioiodine (RAI) therapy-associated damage of lacrimal gland. MATERIALS AND METHODS: Wistar albino rats were used and divided into three groups randomly (n = 12/group). The first group was appointed as the negative control group and received no RAI or medication. The second group was appointed as the positive control group that only received 3 mCi/kg (111 MBq/kg) RAI via gastric gavage and the last group was the treatment group that received 3 mCi/kg RAI via same method and calcitriol (200 ng/kg/day) via intraperitoneal administration. Seven days after RAI administration, bilateral intraorbital (IG), extraorbital (EG) and Harderian (HG) glands were removed for the evaluations of histopathologic, tissue cytokine, total oxidant status (TOS) and total antioxidant status (TAS). RESULTS: RAI led to significant increase in tissue TOS, TNF-α, IL-6 levels and significant decrease in IL-10 and TAS levels (p < 0.05 for each). Addition of adjunctive calcitriol reversed all these parameters significantly (p < 0.05 for each).The following histopathologic parameters were seen more frequently in positive control group than the other groups: Abnormal lobular pattern, perivascular infiltration, periductal infiltration, lipofuscin-like accumulation, acinar atrophy, periductal and periacinar fibrosis in all lacrimal gland types (p < 0.05), acinar fibrosis in EG (p = 0.049), periductal fibrosis in EG and HG (p = 0.049 and 0.038, respectively), abnormal cell outlines in EG and HG (p = 0.020 and 0.011, respectively) and variation in cell size in the IG and the HG (p = 0.003 and 0.049 respectively). CONCLUSIONS: RAI caused significant oxidative stress and inflammation in lacrimal glands. Vitamin D demonstrated potent anti-inflammatory, antioxidant and radio-protective effects on lacrimal glands in histopathologic, tissue cytokine and oxidant/antioxidant level evaluations.

Optimization of hyaluronic acid production and its cytotoxicity and degradability characteristics.

In the present study, culture conditions of Streptococcus equi was optimized through Box-Behnken experimental design for hyaluronic acid production. About 0.87 gL-1 of hyaluronic acid was produced under the determined conditions and optimal conditions were found as 38.42 °C, 24 hr and 250 rpm. The validity and practicability of this statistical optimization strategy were confirmed relation between predicted and experimental values. The hyaluronic acid obtained under optimal conditions was characterized. The effects of different conditions such as ultraviolet light, temperature and enzymatic degradation on hyaluronic acid produced under optimal conditions were determined. 118 °C for 32 min of autoclaved HA sample included 63.09 µg mL-1 of d-glucuronic acid, which is about two-fold of enzymatic effect. Cytotoxicity of hyaluronic acid on human dermal cells (HUVEC, HaCaT), L929 and THP-1 cells was studied. In vitro effect on pro or anti-inflammatory cytokine release of THP-1 cells was determined. Although it varies depending on the concentration, cytotoxicity of hyaluronic acid is between 5 and 30%. However, it varies depending on the concentration of hyaluronic acid, TNF-α release was not much increased compared to control study. Consequently, purification procedure is necessary to develop and it is worth developing the bacterial hyaluronic acid.

[Investigation of cytokine and midkine responses of human THP-1 leukemia cells induced by phorbol-12-Myristate-13-Acetate (PMA) at different concentrations and times]. / Degisik konsantrasyonlarda ve sürelerde Phorbol-12-Myristate-13-Acetate ile uyarilan insan THP-1 lösemi hücrelerinin sitokin ve midkin yanitlarinin arastirilmasi.

Macrophages are accepted as cells that initially contact with the pathogens and initiate the innate immune response. They play effective roles in innate immune and inflammatory responses by intercellular relations and inflammatory mediator secretion. Human THP-1 leukemia cells are frequently used for the in vitro determination of the signal pathways, and the functions of macrophages. Phorbol-12-Myristate-13-Acetate (PMA) is commonly used to induce macrophage differentiation of monocytic cell lines but the extent of differentiation in comparison to primary tissue macrophages is unclear. Midkine acts as a cytokine and growth factor which organizes proliferation, differentiation, survival, adhesion and migration of immune cells. The aim of this study was to observe the differences in the secretion of midkine, TNF-α, IL-10 and IFN-γ of macrophages differentiated from monocytes when stimulated with different doses of PMA for different durations. For this purpose, THP-1 monocytic cells were proliferated by PMA at 24, 48 and 72 hours by using the concentrations of 10 ng/ml, 20 ng/ml, 40 ng/ml and 60 ng/ ml. Midkine, TNF-α, IL-10 and IFN-γ cytokine levels were determined by ELISA in the supernatants of the cells collected at the end of incubation times. PMA stimuli initiated changes that were indicative of differentiation in the cell morphology. Differentiation of cells by PMA induced midkine, TNF-α, IL-10 and IFN-γ secretions in monocytic cells even at the lowest dosage (10 ng/ml). PMA caused cytotoxicity in the cells when the dosages were increased (> 20 ng/ml). THP-1 cells have a basal secretion of midkine and are increased by dosage dependent with PMA stimulation. Midkine secretion has shown changes dependent with dosage and time. A difference was also observed in the cytokine profile of PMA stimulated cells at different doses. The results indicated that the differentiation of THP-1 monocytes to macrophages requires optimization to ensure that this in vitro macrophage model more precisely reflects real in vivo physiologic conditions. As a conclusion the results have shown that a modified PMA differentiation protocol (20 ng/ml and 48 hours incubation) might enhance macrophage differentiation of THP-1 cells without induced cell death (viability 92.2%) and cytokine secretion and midkine responses were the important discriminators of the level of macrophage differentiation.

The effect of midkine on growth factors and oxidative status in an experimental wound model in diabetic and healthy rats.

Wound healing is important for longevity. Midkine is a cytokine involved in controlling tissue repair and new tissue development, and in regulating inflammation. We investigated the effect of midkine on wound healing in rats. In total, 108 Wistar albino rats were used: 12 as healthy and diabetic controls; 96 were split into 4 groups: healthy, saline treated; healthy, midkine (10 ng/kg, 48 h intervals) treated; diabetic, saline treated; and diabetic, midkine treated. Following wound creation, 6 rats per group were euthanized on days 3, 7, 14, and 28; the wounded skin was removed. Levels of epidermal growth factor (EGF), matrix metalloproteinase-8 (MMP-8), transforming growth factor beta (TGF-ß), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), and thiobarbituric acid reactive substances (TBARS) were measured. MMP-8 and PDGF levels fluctuated in all groups; TGF-ß fluctuated in the diabetic groups and was significantly higher in the HM group than other groups after 14 days. EGF and VEGF levels were increased in the HM group after 3 days. TBARS levels were highest in the diabetic groups. Macroscopically, the midkine-treated groups healed better. Midkine can accelerate wound healing by influencing growth factors and oxidative status in wound tissues.