[sFRP1 Expression Induces miRNAs That Modulate Wnt Signaling in Chronic Myeloid Leukemia Cells].

Chronic myeloid leukemia is a clonal hematopoietic stem cell disease characterized by myeloid expansion. The hallmark of the disease is the Philadelphia chromosome, which results in the formation of the BCR-ABL oncogene, a tyrosine kinase that is involved in many signaling pathways including Wnt signaling. The latter has a unique role in chronic myeloid leukemia progression; activated signaling leads to the establishment of an additional leukemic stem cell population derived from granulocyte-macrophage progenitors. sFRP1 is an inhibitor of Wnt signaling and its expression is important for differentiation and maintenance of hematopoietic stem cells. In this study, we aimed to investigate miRNAs that target Wnt signaling by being co-induced along with the expression of sFRP1 in K562 cells. We present a detailed analysis of hsa-mir-221 -3p, hsa-mir-222-3p, hsa-mir-106b-5p, hsa-let-7f-5p, hsa-mir-182-5p, hsa-mir-191-5p, and hsa-mir-183-5p and their target genes and their involvement in Wnt signaling.

Comparison of Single Antigen Bead-Based, Cell-Based, and Solid Phase-Based Crossmatch Methods.

AIM: Antibody assessment during pretransplantation term is important to detect donor specific antibodies. These donor-specific antibodies are determined by various crossmatch methods (flow cytometric [FCXM], complement-dependent cytotoxic [CDCXM], and Luminex [LMXM]). Recently, single antigen bead (SAB) assays have been used for the assessment of hypersensitized patients. The aim of this study was to compare sensitivity and specificity of the 3 crossmatch methods in reference to the SAB method. METHOD: In this study, 69 hypersensitized patients with high class I and/or class II panel reactive antibodies were tested using the flow cytometric SAB method. Serum samples were cross-matched by 3 crossmatch methods with the cells of a volunteer healthy individual, and the results were evaluated according to HLA and cross-reactive epitope groups (CREGs). RESULTS: Sensitivity was found to be better with T FCXM (0.91) and class I LMXM (0.87). Specificity of peripheral blood lymphocyte CDCXM method (1.0) was found to be better than the other 2 methods (0.33 and 0.57, respectively). Sensitivity of class II LMXM (0.88) was found to be better than the others (0.42 for B CDCXM and 0.82 for B FCXM, respectively). The specificity of the B CDCXM, B FCXM, and class II LMXM was similar (0.44, 0.44, and 0.33, respectively). CREGs results were similar to HLA results. CONCLUSION: Although CDCXM has high specificity for the detection of anti-HLA antibodies, it has low sensitivity. To increase sensitivity, FCXM or LMXM methods may be used with the CDCXM test. These results will be beneficial for laboratories and clinicians during graft survival and patient health assessment.

A Comparative Study: Flow Cytometry, Complement-dependent Cytotoxicity, and Luminex Methods.

AIM: The cell-based flow cytometric and bead-based Luminex crossmatch methods have been used alongside the standard complement-dependent cytotoxic crossmatch (CDCXM) test to detect donor specific anti-HLA antibodies. In this study, it was aimed to compare flow cytometric crossmatch (FCXM), CDCXM, and Luminex donor-specific crossmatch (LM-XM) tests for pre-transplant assessment of patients who applied to Tepecik Education and Research Hospital for kidney transplantation from related or deceased donors. METHOD: HLA tissue typing of 1120 patients were tested using the sequence specific oligonucleotide probe method with low resolution. FCXM and LM-XM were performed according to the manufacturer's instructions. The CDCXM test was performed according to the standard procedure. The results were analyzed using SPSS version 21.0 software (IBM, Armonk, NY, United States). P < .05 was accepted as statistically significant. RESULTS: FCXM, CDCXM, and LM-XM tests were performed on 58.2% (n = 652), 91% (n = 1019), and 55.4% (n = 620) of 1120 patients. There were statistically significant differences between FCXM/CDCXM, LM-XM/CDCXM, and FCXM/LM-XM (P < .0001), although there was also a moderate correlation between them (for class I, r = .599, r = .693, and r = .507; for class II, r = .546, r = .471, and r = .495, respectively). The results obtained according to donor type were compatible with the total study group. CONCLUSION: The utilization of FCXM and/or LM-XM tests together with the CDCXM method before kidney transplantations from related and/or deceased donor may facilitate the determination of target cells of donor-specific antibodies or their antibody class, which may increase the success of transplantation.

Assessment of Interleukin 2 Cytokine Expression Levels After Renal Transplantation.

AIM: End-stage renal disease is a disease in which the kidney is not able to perform its functions. Kidney transplantation is the most effective treatment and cost-effective modality of renal replacement therapy for patients with end-stage renal disease. However, the most important problem in end-stage renal disease patients is the unpredictability of immunologic response after transplants. In this study, it was aimed to investigate the possible association between the interleukin 2 (IL-2) expression level and an organ rejection or rejection episode. MATERIALS AND METHODS: Lymphocytes were isolated from peripheral blood obtained from 21 end-stage renal disease-diagnosed patients prior to transplant and at the sixth month after transplant. CD4+ T cells were separated from lymphocytes by the magnetic cell-sorting method. The purity of these cells were controlled by a flow cytometer. After total RNA isolation from CD4+ T cells, IL-2 was examined by the real-time polymerase chain reaction (RT-PCR) method. RESULTS: Among nonrejection patients (n = 18), the IL-2 expression level decreased in 12 patients in post-transplant time, and 3 of these were statistically significant (P < .05). The level was the same in 1 of 18 patients; it increased in 5 patients, and 1 of them was significant (P < .05). The IL-2 expression level also increased in 3 patients who had a rejection episode, and the increase was statistically significant in 2 samples (P < .05). CONCLUSION: When the patients were evaluated individually, it was observed that there might be a relationship between IL-2 expression levels in CD4+ T cells and rejection episodes. The clinical data of the patients, the immunosuppressive therapies, and post-transplant evaluation of cytokines should be considered together.

Comparison of Anti-HLA Antibody Production According to Gestational Periods in Pregnant Women.

BACKGROUND: To evaluate paternal anti-HLA antibody profiles, sera samples were collected from pregnant women in different trimesters and the panel-reactive antibody (PRA) specificities were identified. METHODS: From 2013 to 2015, serum samples were obtained from 41 pregnant women who had registered at the Izmir Tepecik Education and Research Hospital Gynecology Clinic. Anti-HLA antibodies were screened by using the panel reactive antibody screening and identification tests. Sera samples were obtained at the first, second, and third trimesters. The primary outcome was to determine the anti-HLA antibody production term during pregnancy; the secondary outcome was identification of anti-HLA antibodies. RESULTS: None of the women had a sensitization history except during pregnancy. We observed that 54% of the women produced paternal antibodies, either class I or II. Class I PRA positivity of the women who had a first or second pregnancy was the same in all 3 trimesters, whereas class II positivity was increased in the third trimester. Class II and both class I and II positivity increased in the third trimester; class I positivity was decreased in the third trimester. PRA positivity could be affected by the history of pregnancy and could be raised, but no impact was observed from the history of abortion and miscarriage (odds ratios, 1.9, 0.4, and 0.5 [95% confidence intervals, 0.5-7.8, 0.1-2.0, and 0.3-0.7], respectively; P > .05). The most frequently detected antibodies were A2, B7, DR7, DR4, DR11, DR13, DQ2, and DQ8. CONCLUSIONS: Anti-HLA antibodies against paternal HLA antigens were detected more in multiparous women than in primiparous women. Anti-HLA antibody detection ratios did not change until the third trimester and were followed by a specific increase in class II anti-HLA antibody production.

Comparision of anti-HLA antibodies of kidney transplant candidates with chronic renal failure by two different methods: Flow-PRA and Luminex PRA.

OBJECTIVE: The aim of the study was to compare anti-HLA antibodies examined as panel-reactive antibody (PRA) in kidney transplant candidates with chronic renal failure (CRF) with the use of 2 methods: Flow-PRA and Luminex-PRA. METHODS: CRF patients displaying class I PRA (n = 34) and/or class II PRA (n = 41) were tested by the 2 different methods from April 2012 to September 2012, using antigen-coated beads. RESULTS: Eleven (32.3%) 34 patients tested for class I PRA were female and 23 (67.7%) male; 17 (41.5%) 41 patients tested for class II PRA were female and 24 (58.5%) male. Only 2 patients were preemptive, the others had been subjected to dialysis. The concordance ratio of class I PRA test results between Flow-PRA and Luminex PRA was 67.6%. Whereas 13 samples (38.2%) were positive by Flow-PRA, 22 (64.7%) were positive by Luminex-PRA. Two of the 3 patients not previously immunized were found to be positive only by Luminex PRA; 1 was noted to be positive only by Flow-PRA. Regarding class II PRA screening, the concordance between Flow-PRA and Luminex PRA was 70.7%. Whereas 14 (34.1%) samples were positive for Flow-PRA; 24 (58.5%) were positive for Luminex-PRA. The 2 patients not previously immunized were positive only in Luminex PRA. CONCLUSIONS: We speculated that the reason for the low concordance ratios was due to the use of sera that had been previously found to be indeterminate in PRA tests. We also speculated that the low concordance ratios were due to the coating procedure for the beads, which may cause changes in antigenic epitopes and decrease concordance between Flow-PRA and Luminex-PRA.

Comparison of complement-dependent cytotoxic and flow-cytometry crossmatch results before cadaveric kidney transplantation.

AIM: The presence of HLA donor-specific antibodies (DSA) before kidney transplantation decreases graft survival. In this study, we compared crossmatch results of kidney transplantation candidates, for cadaveric renal donation between March 10, 2012, and September 7, 2012. MATERIAL AND METHOD: The 47 kidney transplantation candidates tested for crossmatch included 10 for cadaveric donor organs. Two crossmatch methods were performed: complement-dependent cytotoxic crossmatch (CDCXM) and flow cytometry crossmatch (FCXM). Spleen cells were used as the source of lymphocytes for all crossmatch tests. RESULTS: The T and B cell ratios isolated from spleen were 38.8% and 34.8%, respectively. The concordance ratio of the two methods was 76.6% with 23.4% discordant results. Regarding the discordant results, 4.2% were positive CDCXM but negative FCXM; 191%, negative CDCXM but positive FCXM. All patients displaying positive crossmatches had a previous immunization history. As a result, we speculated that the positive CDCXM but negative FCXM results were due to the washing procedures in the FCXM disturbing antigen-antibody complexes. We suggest at least two different methods to be performed for crossmatch tests before kidney transplantation. CDCXM detects immunoglobulin G1 (IgG1) and IgG3, which are critical for rejection. FCXM is able to detect all IgG subgroups because of its high sensivity. As a result we suggest that both CDCXM and FCXM are preferrable strategies to detect DSAs.

The effects of ochratoxin A on lipid peroxidation and antioxidant enzymes: a protective role of melatonin.

Various studies indicate that the mycotoxin ochratoxin A (OTA) is a carcinogenic, genotoxic, teratogenic, immunotoxic, and nephrotoxic agent. In the present study, the activities of some enzymes in the serum and liver of rats with ochratoxicosis and the effects of melatonin on these enzymes were investigated. Rats were divided into three equal groups, each consisting of eight rats; control, OTA (289 microg/kg per day) and OTA + melatonin groups for this study. In the OTA treated group, the level of lipid peroxidation (LPO) and the activities of glutathione peroxidase (GSH-Px) were increased in the liver and serum in comparison with the control group. The activities of catalase (CAT) and superoxide dismutase (SOD) were significantly changed in the serum when compared with controls. Our results showed structural tissue damage in the liver in OTA-treated rats. Melatonin decreased the OTA-induced damage to support the antioxidant defense system and/or with free radical scavenger action.

Histopathologic changes in liver and renal tissues induced by Ochratoxin A and melatonin in rats.

Nephrotoxicity and hepatotoxicity induced by Ochratoxin A (OTA) and ameliorating effects of melatonin were investigated in rats exposed to OTA. Experimental groups were as follows: control; OTA-treated; and OTA plus melatonin (MEL)-treated (OTA+MEL). The rats in the control group were administered with only a daily oral administration of 0.5 M NaHCO3. OTA was administered with a dose of 289 microg/kg in the same way. OTA and MEL were administered orally with OTA (289 microg/kg) and melatonin (10 mg/kg) in two different periods of time during the same day. The histopathologic changes in the liver and kidney tissues of control, OTA and OTA+MEL-treated rats were examined. There were no significant changes in the kidney and liver tissues of the control rats. Significant histopathologic changes were found in the kidney and liver tissue of rats treated with OTA. These were granular or vacuolated degeneration and necrosis of the liver cells, sinusoidal and central vein dilatation, bile duct proliferation, enlargement of periportal areas with mononuclear cell inflammatory infiltration and mild degree fibrous tissue proliferation, tubular epithelial cells degeneration, necrosis, proliferation and karyomegaly in the epithelial cells nuclei and peritubular and periglomerular lymphocyte infiltration, stromal fibrous tissue proliferation, hyperemic vessels. The severity of the lesions was significantly reduced by administration of melatonin. These results revealed that OTA induced significant histopathologic changes in liver and kidney tissue advocating OTA toxicity (P < 0.001), and administration of MEL+OTA significantly reduced the toxic effect of OTA on kidney and liver tissue of rats (P > 0.05).