Physiological aspect of apoptosis-regulating microRNAs expressions during fasting.

OBJECTIVE: Fasting is an activity that requires a certain calorie restriction without consuming food or drinks for a certain period of daytime. However, fasting triggers many complex events, including activating cellular stress response pathways, autophagy promotion, apoptosis pathways, and a change in hormonal balance. Among the many events affecting the regulation of apoptosis, the expression of microRNAs (miRNAs) plays an important role. Therefore, we aimed to investigate the levels and importance of miRNA expression in fasting. PATIENTS AND METHODS: The expressions of 19 miRNAs regulating different pathways from saliva samples, isolated by matching healthy university students (n = 34) as group 1 (fasting for 17 consecutive hours) and group 2 (testing 70 minutes after meal consumption), were examined using the real-time PCR method. RESULTS: In fasting, modulation of apoptotic pathways by miRNAs triggers anti-pathogenic effects, and the adaptation of abnormal cells in the body decreases. For this reason, vital diseases, such as cancer, can be treated by preventing the proliferation and growth of cancerous cells by increasing programmed cell death due to the downregulation expression mechanism of miRNAs. CONCLUSIONS: Our study aims to improve the knowledge about the mechanisms and functions of miRNAs in various apoptosis pathways during fasting and may be a model for further future physiological and pathological studies.

Does laparoscopic-guided transversus abdominis plane block have an effect on postoperative pain and recovery after sleeve gastrectomy?

OBJECTIVE: Postoperative pain management is thought to have an effect on patient comfort, morbidity, and mortality after bariatric surgery. Local anesthetic agents are frequently used for this purpose. Local anesthetics can be used in many different ways. In this study, we aimed to investigate the effect of transversus abdominis plane (TAP) block on postoperative pain by laparoscopic method. PATIENTS AND METHODS: A prospective randomized clinical trial was performed. While TAP block was applied to one group with bupivacaine, no action was taken for the other group. Postoperative analgesia was given to both patient groups with the "patient-controlled analgesia (PCA)" device. Demographic, operational, and postoperative clinical and pain data of the patients were recorded. RESULTS: TAP block and non-TAP block groups consisted of 30 patients each. Visual analog scale (VAS) scores of the patients at 6, 12, and 24 hours were lower in the TAP group compared to the non-TAP group (p=0.015, 0.018, 0.04, respectively). According to the PCA device data, the analgesic requirement was lower in the TAP group at 6, 12, and 24 hours (p <0.001). Rescue analgesia was required more in the non-TAP group (p=0.04). There was no statistically significant difference between the two groups in terms of gas discharge time (p=0.102), stool discharge occurred earlier in the TAP group (p=0.02). Oral intake times (p=0.554) and length of stay hospital (p=0.551) were similar. CONCLUSIONS: Laparoscopic TAP block using bupivacaine can be safely administered in morbidly obese patients and reduces postoperative analgesic requirements. Thus, side effects that may develop secondary to the use of analgesics are avoided.

Efficacy of Intravenous Paracetamol and Ibuprofen on Postoperative Pain and Morphine Consumption in Lumbar Disc Surgery: Prospective, Randomized, Double-Blind, Placebo-Controlled Clinical Trial.

OBJECTIVES: Effective postoperative pain management after lumbar disc surgery reduces complications and improves postoperative care. The purpose of this prospective, randomized, double-blind, placebo-controlled clinical study is to evaluate the effects of IV paracetamol and ibuprofen on postoperative pain, morphine consumption and side effects of morphine in patients who underwent lumbar disc surgery. MATERIALS AND METHODS: Seventy-five patients aged 18-85 years scheduled for lumbar disk surgery with a single level laminectomy included in this study. All patients received morphine with an IV patient-controlled analgesia device during the first postoperative 24hour. The patients were divided randomly and double-blinded into three groups (control, paracetamol and ibuprofen). The demographic characteristics and procedure data, VAS score, cumulative morphine consumption, opioid-related side effects were recorded. RESULTS: There was no significant difference regarding to demographic characteristics, comorbidities, and durations of anesthesia and surgery. There was a significant difference between all groups regarding to total morphine consumption (P<0.001). IV ibuprofen significantly reduced the total morphine consumption in comparison with control and paracetamol (P<0.001). Repeated measures ANOVA showed in all periods of the study that VAS score was significantly lower in ibuprofen (P<0.001), but not in paracetamol (P=0.394) in comparison with control. There was no difference between groups regarding postoperative heart rate, mean arterial pressure, nausea-vomiting, pruritus and urinary retention. CONCLUSIONS: This study showed that pain scores and morphine consumption, but not the side effects of patient-controlled analgesia during 24hours after the lumbar disk surgery, were significantly reduced by IV ibuprofen as a supplemental analgesic when compared with controls and paracetamols.

Same-admission laparoscopic cholecystectomy in acute cholecystitis: the importance of 72 hours and oxidative stress markers.

BACKGROUND: This prospective randomized study aims to compare outcomes between immediate laparoscopic cholecystectomy (LC) and same admission delayed LC in patients with acute cholecystitis and also to investigate the relation between oxidative stress markers and complication rates in the patients with AC. METHODS: This study included 64 patients with AC who were randomly divided into two groups. Patients in Group 1 (n=32) were immediately administered LC, while in Group 2 (n=32) patients underwent transient LC following medical treatment. All patients were operated on their first hospitalization. RESULTS: No statistically significant differences were observed between the groups for the comparison of complications, conversion rates, or operation durations (p>0.05). The length of postoperative hospital stay was found to be significantly shorter in group 1 compared to group 2 (1.75 vs 2.93 days; p=0.024). Only the total antioxidant status result was significantly higher in group 1 (p=0.017), but the finding was not correlated with complications. CONCLUSION: LC for AC was performed during the first admission was found to be safe, even beyond 72 hours following symptom onset. Pre-operative oxidative stress markers did not correlate with the complication rates.

Characterization of bevacizumab by dynamic light scattering while maintaining its native structure.

Bevacizumab, is a humanized monoclonal antibody and patents on Avastin® (Bevacizumab, Roche) will expire in the US in 2019 and in Europe in 2022. Therefore, bevacizumab is a popular target for biosimilar developers. One of the most common problems in the formulation of antibody drugs is protein aggregation. Dynamic light scattering (DLS) is a well-established method for the determination of hydrodynamic dimensions, aggregates, and aggregation points of proteins. In contradistinction to other techniques that require diluted samples or specific conditions, proteins and aggregates can maintain their native structure during DLS measurements. In recent studies, bevacizumab was characterized by DLS using diluted samples. In this study, we aimed at investigating the hydrodynamic dimensions, aggregates, and aggregation onset of bevacizumab (Altuzan®, Turkey, Roche) by DLS, while maintaining its native structure. The intensity, volume, and number-based particle size distribution profiles of the test samples were evaluated and the aggregation onset of the formulation was successfully determined against increasing temperature. It is shown that the preservation of the native structure of commercial formulations in DLS measurements provides an opportunity to the characterization of commercial products and development of biosimilars.

Nasotracheal Intubation in Children for Outpatient Dental Surgery: Is Fiberoptic Bronchoscopy Useful?

BACKGROUND: The aim of our study was to compare the hemodynamic responses and adverse events associated with nasotracheal intubation (NTI) using a fiberoptic bronchoscope (FOB) and a direct laryngoscope (DLS) in children undergoing general anesthesia for outpatient dental surgery. METHODS: Eighty children (aged 5-15 years) were scheduled to undergo outpatient dental surgery under general anesthesia and of these children those who required NTI were included. RESULTS: NTI was significantly longer in the FOB group (P = 0.03). In both groups, systolic blood pressure (SBP) and heart rate (HR) significantly decreased after the induction of anesthesia when compared with the baseline values. SBP was significantly higher in both groups at intubation and 1 and 3 min after intubation when compared with postinduction. SBP significantly increased in the DLS group compared with the FOB group at intubation and 1 min after intubation. HR was significantly increased at intubation and 1 min after intubation in the DLS group compared with the FOB group. Nose bleeding after intubation was significantly more frequent in the DLS group (30%) than in the FOB group (7.5%) (P = 0.034). The incidence of sore throat 24 h after surgery was 20% (8/40) in the DLS group and 2.5% (1/40) in the FOB group (P = 0.014). CONCLUSIONS: There are fewer hemodynamic responses and adverse events in the FOB group than in the DLS group; therefore, FOB can be safely used for NTI in children undergoing outpatient dental surgery, and FOB may be more successful than DLS for NTI.

Using PVA and TPGS as combined emulsifier in nanoprecipitation method improves characteristics and anticancer activity of ibuprofen loaded PLGA nanoparticles.

In the preparation of nanoparticles (NPs) by the nanoprecipitation method, emulsifiers play a key role for NPs' characteristics. The present study aimed to investigate the combined emulsifier effect on ibuprofen loaded poly(lactic-co-glycolic acid) (PLGA) NPs' characteristics and anticancer activity. Ibuprofen loaded PLGA NPs were prepared by nanoprecipitation using different concentrations of PVA (poly(vinyl alcohol)) or PVA-TPGS (d-α-tocopherol polyethylene glycol 1000 succinate) combination as emulsifier. It was found that encapsulation efficiencies of NPs varied between 17.9 and 41.9 % and the highest encapsulation efficiency was obtained with 0.5% PVA + 0.1% TPGS (coded as PLGA PVA/TPGS NPs). PLGA PVA/TPGS NPs were characterized and compared with PLGA PVA NPs, which was obtained by 0.5% PVA alone. Polydispersity index of PLGA PVA/TPGS and PLGA PVA NPs were found to be 0.08 and 0.15, respectively. Incorporation of TPGS with PVA slightly decreased the initial ibuprofen release. Transmission electron microscopy analyses demonstrated a nearly uniform particle size distribution and spherical particle shape of the PLGA PVA/TPGS NPs. Additionally, PLGA PVA/TPGS NPs were significantly more cytotoxic than PLGA PVA NPs on the MCF-7 (human breast adenocarcinoma cells) and Caco-2 (human epithelial colorectal adenocarcinoma) cells (p<0.05). Also PLGA PVA/TPGS NPs were not cytotoxic on normal cells (L929, mouse healthy fibroblast cells) (p>0.05). In conclusion, these results indicated that using a combination of TPGS and PVA as an emulsifier in nanoprecipitation could be a promising approach for preparing ibuprofen loaded PLGA NPs because of their improved characteristics and anticancer activity.

The effects of luteolin on the intestinal ischemia/reperfusion injury in mice.

The purpose of this study is to investigate the potential protective effect of the flavonoid Luteolin on ischemia-reperfusion (IR) injury in mouse intestine, which has not previously been studied. Twenty-four female C57BL/6 mice were randomly assigned to four groups, each consisting of 6 mice: a sham group (laparotomy, but no IR injury), a sham + Luteolin group (no IR, and Luteolin was administered intraperitoneally 30 min after laparotomy), IR group (30 min occlusion of the superior mesenteric artery (SMA) then 2 hr' reperfusion), IR + Luteolin (30 min occlusion of the SMA then 2 hr' reperfusion; Luteolin was administered intraperitoneally before reperfusion). Intestine tissues were harvested from the mice for histopathological and biochemical analysis. Total oxidant status (TOS) and total antioxidant capacity (TAC) of the intestinal tissues were measured using Erel's method. Oxidative stress index (OSI) was calculated using the TOS/TAC ratio. Intestinal histological changes were significantly decreased in the IR + Luteolin group compared with the IR group (p = .037). TOS tissue levels were also significantly decreased in the IR + Luteolin group compared with the IR group (p = .005). TAC levels did not increase significantly in the IR treatment group and were not affected by Luteolin treatment (p > .05). The results of this study show that Luteolin administration provides considerable protection against IR injury in the mouse intestine.

Association between emotional and behavioral problems and metabolic control in children and adolescents with Type 1 diabetes.

OBJECTIVE: We aimed to investigate the differences in emotional and behavioral problems and to explore the association between the level of psychiatric problems and the metabolic control in Type 1 diabetes. METHODS: The children with Type 1 diabetes (no.=42) and the "healthy" control group (no.=42), their parents and endocrinology specialist completed the forms prepared for the study. The parents completed the Child Behavioral Checklist (CBCL/4- 18). RESULTS: The groups had significant differences in CBCL activities (p<0.001), social competence (p<0.001), total competences (p<0.001), withdrawal (p=0.036), anxiety/depression (p=0.033), social problems (p=0.009), and aggressive behavior (p=0.04) scores. We did not find significant differences in CBCL scores between the groups with good, moderate and bad metabolic control (p>0.05). DISCUSSION: The parents of children with Type 1 diabetes reported emotional and behavioral problems significantly more. We did not find any significant association between the level of metabolic control and the emotional and behavioral problems.

The effect of MDR1 (ABCB1) polymorphism on the pharmacokinetic of tacrolimus in Turkish renal transplant recipients.

There is marked interindividual variability in trough blood levels of tacrolimus (TRL) following standard dosing. TRL is a substrate for P-glycoprotein (P-gp), the product of the multidrug resistance-1 (MDR1)(ABCB1) gene. P-gp acts as a membrane efflux pump, which affects TRL absorption from the gut. Some of the single nucleotide polymorphisms (SNP) of ABCB1 gene are associated with pharmacokinetic characteristics of TRL. The objective of this study was to determine the role of ABCB1 C3435T polymorphism on TRL dose requirements, trough values and dose-adjusted trough TRL concentrations among Turkish renal transplant recipients. Renal transplant recipients receiving TRL (n=92) were genotyped for ABCB1. TRL daily doses, trough concentrations, dose-adjusted trough concentrations, demographic features, and clinical data were obtained at 1, 6, and 12 months after renal transplantation. The frequency of the ABCB1 3435 CC genotype was 30.4%, whereas 47.8% of patients were 3435 CT and 21.7% of patients were 3435 TT. TRL daily doses were significantly lower among patients with the 3435 TT genotype at months 1 and 6. At 6 and 12 months after transplantation patients who were homozygous for the ABCB1 3435 CC showed significantly lower dose-adjusted trough TRL concentrations compared with subjects of 3435 TT and CT genotypes. Knowledge of ABCB1 genotype may be useful to adjust the optimal dose of TRL in transplant patients, thereby rapidly achieving target concentrations.

Assessment and comparison of endothelial function between dialysis and kidney transplant patients.

Dialysis and kidney transplant patients display endothelial dysfunction. Previous studies concerning comparisons of endothelial function in dialysis and kidney transplant patients included subjects with cardiovascular risk factor(s) that alone may lead to endothelial dysfunction. In this study, we compared endothelial function between dialysis and transplant patients who did not show known cardiovascular risk factors that lead to endothelial dysfunction. We studied age- and gender-matched cohorts: 30 hemodialysis (HD), 30 peritoneal dialysis (PD), and 30 kidney transplant patients. We also included 20 age- and gender-matched healthy controls. We assessed the endothelial function of patients and controls by a noninvasive technique. Serum biochemistry profiles of patients were also similar to controls in terms of lipid profile and fasting blood glucose level. Although mean FMD% levels of HD and PD patients were similar (6.6% +/- 3.1% vs 6.8% +/- 3.0%, P > .05), the mean percent of flow-mediated endothelium-dependent dilatation (FMD%) level in transplant patients was higher than those in HD or PD patients (10.50% +/- 3.0% vs 6.6% +/- 3.1% and 6.8% +/- 3.0%, respectively; P < .01). In addition, the mean FMD% level in healthy controls was higher than those in HD, PD, and transplant patients (14.0% +/- 2.3% vs 6.6% +/- 3.1%, 6.8% +/- 3.0% and 10.50% +/- 3.0%; P < .01, respectively). In conclusion, endothelial functions in transplant patients were better than those in dialysis patients.

Relation of C-2 monitoring with serum lipid profile in stable renal transplant patients.

C-2 monitoring has been proposed as a more effective strategy than C-0 to predict the risk of acute rejection in the early stages posttransplantation. However, cyclosporine (CsA) is associated with posttransplant dyslipidemia. The aim of this retrospective study was to evaluate the correlations of C-0 and C-2 levels with atherogenic risk factors in the first 6 months versus after 6 months posttransplantation. We evaluated the data from 127 stable renal transplant recipients (89 males, 38 females) of mean age 38.10 +/- 12.79 years who received Neoral-based immunosuppression to investigate the relation of C-2 levels to serum lipid profile compared with C-0 values in the early and late posttransplantation periods. Receiver operating characteristic (ROC) analyses were performed to define a C-2 cutoff level that identified subjects with hypercholesterolemia, defined as a total cholesterol (TC) >200 mg/dL. There were significant positive correlations between both C-0 and C-2 levels and TC as well as the ratio of total cholesterol/HDL cholesterol (TC/HDL) in the late period. When the C-2 levels in the late posttransplantation period were stratified, serum TC concentrations showed statistically significant differences between the groups. Whole blood C-2 levels above 850 ng/mL were associated with increased serum TC concentrations; the C-2 cutoff level leading to hypercholesterolemia was 888 ng/mL. Maintenance immunosuppressive therapy under the proposed whole blood C-2 level of 888 ng/mL seemed to preserve graft function while preventing atherogenic risks for cardiovascular diseases in the late posttransplantation period.

The effect of Topotecan on oxidative stress in MCF-7 human breast cancer cell line.

PURPOSE: Topotecan, a semisynthetic water-soluble derivative of camptothecin exerts its cytotoxic effect by inhibiting topoisomerase I and causes double-strand DNA breaks which inhibit DNA function and ultimately lead to cell death. In previous studies it was shown that camptothecin causes ROS formation. The aim of this study was to investigate if Topotecan like camptotecin causes oxidative stress in MCF-7 human breast cancer cell line. Determining the oxidant effect of Topotecan may elucidate a possible alternative mechanism for its cytotoxicity. EXPERIMENTAL DESIGN: MCF-7 cells were cultured and exposed to Topotecan for 24 h at 37 degrees C. The viability of the cells (% of control) was measured using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Lipid peroxidation (TBARS), protein oxidation (carbonyl content), sulfhydryl, glutathione (GSH) levels, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities were determined in MCF-7 cells with and without Topotecan incubation. RESULTS: We found the IC(50) concentration of Topotecan as 0.218 microM in MCF-7 cells. This concentration of Topotecan was used in the incubations of the cells. Our data indicated increased oxidative status, as revealed by increased lipid peroxidation and protein oxidation, and decreased GSH and sulfhydryl levels in MCF-7 cells exposed to Topotecan compared to control cells. In contrast, there was a slight increase in SOD and a significant increase in GPx and catalase activity in MCF-7 cells incubated with Topotecan compared to the control. CONCLUSIONS: These results support our hypothesis that Topotecan increases oxidative stress in MCF-7 cells.

Effect of pentylenetetrazol-induced epileptic seizure on the antioxidant enzyme activities, glutathione and lipid peroxidation levels in rat erythrocytes and liver tissues.

OBJECTIVES: In order to clarify whether oxidative stress accompanies epilepsy, we examined the effects of pentylenetetrazol (PTZ)-induced epilepsy on the lipid peroxidation and antioxidant enzyme activities in erythrocytes and liver tissues of adult Wistar rats. MATERIALS AND METHODS: The activities of antioxidative enzymes (glucose-6-phosphate dehydrogenase (G-6-PD)), copper, zinc-superoxide dismutase (Cu,Zn-SOD), catalase (CAT), selenium-dependent glutathione peroxidase (Se-GSH-Px) and the levels of reduced glutathione (GSH) and thiobarbituric acid-reactive substances (TBARS) were measured in erythrocytes and liver tissues of pentylenetetrazol (PTZ)-induced epileptic adult Wistar rats. RESULTS: Single PTZ treatment in a convulsive dose of 50 mg/kg significantly reduced the erythrocyte Cu,Zn-SOD, CAT enzyme activities and GSH levels compared to controls (P < 0.001, P < 0.001, P < 0.05, respectively). Erythrocyte and liver tissue TBARS levels in the epileptic group were significantly higher than controls (P < 0.0001). There was a significant decrease in liver tissue Cu,Zn-SOD activity and GSH levels in the epileptic group (P < 0.0001), whereas significantly higher activities of G-6-PD and Se-GSH-Px were found in the epileptic group. CONCLUSIONS: Our results demonstrate a generalized diminished antioxidant activity and increased TBARS level indicating enhanced oxidative stress in the liver and erythrocytes of epileptic rats. Increased oxidative stress in the liver of epileptic rats might be due to the activation of the recently found glutamate receptors in the liver. These findings suggest that the use of antioxidants with antiepileptic drugs and new drugs such as type-5 metabotropic glutamate receptor (mGlu5) antagonist (MPEP) might protect erythrocytes and liver tissue against anoxic damage and oxidative stress.

Effects of some hematological parameters on whole blood tacrolimus concentration measured by two immunoassay-based analytical methods.

OBJECTIVES: Tacrolimus (FK506) is a potent immunosuppressive drug used for prevention of rejection following transplantation. Several methods including immunoassays have been used for monitoring tacrolimus levels. The purpose of the present study was to compare the effects of various hematological parameters on whole blood tacrolimus concentrations which were measured with two different analytical methods, namely the microparticle enzyme immunoassay (MEIA II) and enzyme multiplied immunoassay technique (EMIT). DESIGN AND METHODS: The effects of hematological variables, namely hematocrit (Htc), hemoglobin (Hb), red blood cell (RBC), mean cell volume (MCV), mean cell hemoglobin (MCH), mean cell hemoglobin concentration (MCHC), red cell distribution width (RDW) and platelet (PLT) counts on tacrolimus concentrations (n = 2430 measurements) measured with EMIT (n = 1171) and MEIA II (n = 1259) methods in whole blood samples from kidney or liver or combined kidney-pancreas transplant patients (n = 162) during a 2-year post-transplantation period were compared. RESULTS: The whole blood tacrolimus concentrations measured with MEIA II method were affected much more significantly by hematological parameters than those measured with EMIT method. In MEIA II method, RDW (r = 0.479, P < 0.01) showed a stronger correlation with tacrolimus concentration than Htc (r = -0.239, P < 0.01) in all patients. A negative significant correlation (r = -0.468, P < 0.01) was also observed between the Htc and tacrolimus concentration in patients with Htc values < or =25% in MEIA II method. CONCLUSIONS: The results of the present study suggest that EMIT method might be preferred to MEIA II in determination of whole blood tacrolimus concentrations in anemic transplant patients. For better therapeutic drug monitoring, physicians should be aware of these assay differences. Evaluation of hematologic factors that affect the whole blood concentrations of tacrolimus may be helpful in deciding the dosage of this drug.

Mannose-binding lectin levels in children with asthma.

Mannose-binding lectin (mbl), one of the important components of innate immunity, can activate the lectin pathway of the complement system. After binding mannose containing carbohydrate structures of foreign antigen, mbl initiates and regulates the inflammatory responses. Asthma is a complex inflammatory disease of the lung involving many components of the immune system. Our objective was to investigate the serum mbl levels of asthmatic children in comparison with healthy controls. Serum mbl levels were determined by nephelometric assay in 72 asthmatic children (5-15 yr old) and 30 healthy age-matched controls. Mbl levels of asthmatic children were measured both during acute attack and after complete remission. There was no significant difference between the mbl levels during acute attack (median 4.1 mg/l) or quiescence of symptoms (median 3.6 mg/l). Serum mbl levels both during acute attack or quiescence of symptoms was significantly higher in asthmatic children than in the healthy controls (median 2.8 mg/l, p < 0.0001 for each). Furthermore, mbl levels of asthmatic children positively correlated with peripheral blood eosinophils (r = 0.377, p < 0.001), which is a systemic component of airway inflammation in asthma. Our findings indicate that mbl may be implicated in the pathogenesis of asthma by contributing to airway inflammation or by increasing the risk of developing asthma.

The effect of quercetin on topotecan cytotoxicity in MCF-7 and MDA-MB 231 human breast cancer cells.

BACKGROUND: Topotecan, which is a Camptothecin derivative, shows a large spectrum in anti-tumor activity. Topotecan exerts its cytotoxic effect on tumor cells mainly by inhibition of topoisomerase I activity resulting in double-strand DNA breaks. In our study, we investigated the combined cytotoxic action of Topotecan and Quercetin in MCF-7 and MDA-MB 231 human breast cancer cells. To examine the possible relation between the cytotoxic activity of Topotecan and oxidative stress, we measured ROS and nitrite levels in both human breast cell lines. MATERIALS AND METHODS: MCF-7 and MDA-MB 231 cells were exposed to Topotecan, Quercetin, or a combination of both agents for 24 h at 37 degrees C. The viability of the cells was measured using the colorimetric MTT (3-(4,5)-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. We determined reactive oxygen species and nitrite levels as indicators of oxidative stress in both cell lines with and without Topotecan and/or Quercetin incubations using fluorometric dichlorofluorescin diacetate (DCFH-DA) and diaminonaphtalene (DAN) assay. RESULTS: The IC(50) concentration of Topotecan was 100 ng/ml in MCF-7 cell line and 160 ng/ml in MDA-MB231 cell line. Treatment with Quercetin enhanced cytotoxicity of Topotecan as 1.4-fold in MCF-7 and 1.3-fold in MDA-MB-231 cell line. A significant increment on ROS and nitrite levels was found in MCF-7 and MDA-MB-231 cells following Topotecan incubation. CONCLUSIONS: Our results suggest that Topotecan has cytotoxic activity against both of the breast cancer cell lines in vitro. A combination with Quercetin increases efficacy of Topotecan in the treatment of breast cancers. Our results indicate that increased oxidative stress plays a role in the cytotoxic action of Topotecan.

Influence of inflammation on the relation between markers of iron deficiency in renal replacement therapy.

Iron deficiency is an important factor in the management of anemia in both dialysis and transplant patients. Serum ferritin and transferrin saturation (TS) may be influenced by the presence of inflammation. Recently, the soluble transferrin receptor (s-TfR) has been considered to be a marker of functional iron stores. In this study, parameters of the iron state were investigated in terms of agreement (assessed by kappa) with the diagnosis of iron deficiency and with inflammation. The study was performed in 38 hemodialysis, 31 continuous ambulatory peritoneal dialysis, and 21 anemic renal transplant patients. CRP and amyloid A protein (AAP) were studied as markers of inflammation. Iron deficiency was defined as ferritin <100 mg/L, TS <20%, or s-TfR >1.76 mg/mL. We observed that s-TfR levels were significantly related to both dialysis duration (r = 0.28 in dialysis and r = 0.60 in transplant patients, both P <.05) and PTH levels (r = 0.23 in dialysis and r = 0.55 in transplant patients, both P <.05). Among the transplant group, ferritin and TS, as well as TS and s-TfR were significantly related (r = 0.84 and r = -0.64, respectively), but not s-TfR and ferritin. Among the dialysis group, ferritin and TS, and also TS and s-TfR, were significantly related (r = 0.35 and r = -0.30, respectively), whereas s-TfR and ferritin were not. In the transplant group, the kappa value for agreement between ferritin and TS in the diagnosis of iron deficiency was 0.76 (P =.006), and 0.33 (P =.04), respectively. Among patients with CRP levels <0.3 mg/L or AAP levels <6.4 mg/L, the relation between parameters of iron state was more robust. The kappa value for agreement between ferritin and s-TfR was 0.49 (P =.006) in the dialysis group and 1 (P =.002) for that between ferritin and TS in the transplant group. Our results suggest that PTH levels may influence s-TfR levels. Discordance between ferritin, TS, and s-TfR as markers of iron deficiency might be explained by the effects of inflammation.

Evaluation of the new EMIT tacrolimus assay in kidney and liver transplant recipients.

Tacrolimus (FK506) is a potent macrolide immunosuppressant used for prevention of organ transplant rejection following transplantation. Monitoring of blood tacrolimus concentrations is essential to assess organ rejection and toxicity, because of the agent's narrow therapeutic range, wide inter- and intraindividual pharmacokinetic variability as well as drug interactions mediated by alteration in cytochrome P450. Several methods have been developed to monitor tacrolimus; immunoassays, bioassays, and HPLC/MS. The purpose of this study was to compare two analytical methods: the well-established MEIA II tacrolimus immunoassay using the IMx analyzer and the new EMIT 2000 tacrolimus immunoassay on the Cobas Integra 400 system. Tacrolimus results obtained using the two methods have been compared on 180 whole blood samples from kidney and liver transplant patients. The analytical sensitivities of both methods were defined as 1.2 ng/mL for EMIT and 1.5 ng/mL for MEIA II. The within-run CVs (n = 15) obtained with four-level controls were 9.08%, 9.41%, 5.23% and 4.4% for EMIT 2000. The comparison showed the following relationship between two methods: MEIA = 1.08.EMIT + 0.20 (r =.893). In conclusion, the EMIT 2000 tacrolimus immunoassay is a reliable alternative for the MEIA II method to monitor tacrolimus in organ transplant recipients. It provides a valid quantitative measurement of tacrolimus with comparable % CVs in quality-control as well as patient blood samples. Additionally, the EMIT 2000 method provides a rapid analysis of a large number of samples in one run with a low turnaround time and possibilities to reanalyze critical samples.