Bartosz Adam Frycz

Decreased expression of ten-eleven translocation 1 protein is associated with some clinicopathological features in gastric cancer.

A decrease in ten-eleven translocation 1 (TET1) transcript and 5-Hydroxymethylcytosine (5hmC) levels has recently been demonstrated in primary gastric cancer (GC). However, little is known about TET1 protein levels in gastric tumoral and nontumoral tissue. Therefore, using reverse transcription, real-time quantitative polymerase chain reaction and western blotting analysis, we determined the TET1 transcript and protein levels in tumoral and nontumoral tissue from 38 patients with GC. We also assessed the association between the decrease in TET1 transcript and protein levels and some clinicopathological features in primary GC. We found significantly decreased levels of TET1 transcript (P=0.0023) and protein (P=0.00024) in primary tumoral tissues as compared to nontumoral tissues in patients with GC. Moreover, we also observed significantly lower amounts of TET1 transcript (P=0.03) and protein (P=0.00018) in tumoral tissues in patients aged>60. We also found significant lowered TET1 protein levels in male patients (P=0.0014), stomach (P=0.044) and cardia (P=0.013) tumor localization, T3 depth of invasion (P=0.019), N1 (P=0.012) and N3 lymph node metastasis (P=0.013) and G3 histological grade (P=0.0012). There were also significant decreases in TET1 transcript levels in female patients (P=0.042), intestinal histological types (P=0.0079) and T4 depth of invasion (P=0.037). Our results demonstrated that a decrease in TET1 transcript and protein levels is associated with some clinicopathological features in GC.

Thymic emigration patterns in patients with type 2 diabetes treated with metformin

Recent data suggest that thymic output, which provides the naive T cells necessary for the normal functioning of T‐cell‐dependent immunosurveillance cellular immunity including anti‐cancer protection, can be disturbed in the course of type 2 diabetes. Metformin, an anti‐diabetic drug commonly confirmed as an agent with many potential anti‐cancer activities, might be helpful in this immune correction. The profile of thymic output was evaluated in the current study on the basis of the signal‐joint T‐cell receptor excision circle (sjTREC) concentration in peripheral blood polymorphonuclear cells and thymic emigrant content in peripheral blood evaluated from CD127 and/or CD132 antigen expression. It was revealed that recent thymic emigrants and more differentiated CD127+ CD132+ cell populations were decreased among naive T cells and CD8+ T cells, whereas RTE count was increased in CD4+ T cells, and the CD127+ CD132+ cell population was less numerous than in non‐diabetic participants. Terminally differentiated thymic emigrants, i.e. CD127− CD132+ cells, were increased in naive T cells and in CD8+ T cells. Metformin affects mainly the early phases of thymic export, increasing CD127+ CD132− and CD127+ CD132+ cell populations in naive T cells and the CD127+ CD132− population in CD4+ T lymphocytes. It could be concluded that type 2 diabetes deteriorates thymic immunostasis. The decreased thymic output could be compensated by metformin, especially with regard to CD4+ naive T cells. It is the first time that therapy with metformin has been documented by us as particularly useful in the control and normalization of thymus function, regarding correction of early populations of thymic emigrants.

Expression of 17β-hydroxysteroid dehydrogenase type 2 is associated with some clinicopathological features in gastric cancer

In most populations, gastric cancer (GC) incidence is higher in men than in women, which may suggest the role of sex steroid hormones in gastric cancerogenesis. Both, androgens and estrogens can be synthetised in peripherial tissues. This process is controlled by expression of steroidogenic enzymes. Therefore, we evaluate the 17β-hydroxysteroid dehydrogenase type 2 (HSD17B2) transcript and protein levels in gastric tumoral and nontumoral tissue. We also determined the association between HSD17B2 transcript and protein levels and some clinicopathological features in GC. We found significantly decreased levels of HSD17B2 transcript (P=0.00072) and protein (P=0.00017) in primary tumoral tissues of GC patients, as compared to nontumoral tissues. In patients above 60 years of age the amounts of HSD17B2 transcript (P=0.00044) and protein (P=0.00027) were significantly lower in tumoral than nontumoral tissues. Similarly, lower HSD17B2 levels, both in terms of the transcript and protein, were observed in tumoral tissues of male (P=0.013, P=0.0014), patients stomach (P=0.0062, P=0.045) and cardia (P=0.02, P=0.02) site of tumor, T3 (P=0.018, P=0.014) depth of invasion, N0 (P=0.017, P=0.045) lymph node metastasis, G3 (P=0.0027, P=0.014) malignancy grade. We also observed significantly reduced level of HSD17B2 transcript in tumoral tissue specimens of females (P=0.014), T4 depth of invasion (P=0.02), N3 lymph node metastasis (P=0.037) and G2 malignancy grade (P=0.045). Furthermore, diffuse GC histological types were associated with lower HSD17B2 protein level (P=0.024) than nontumoral tissues. We demonstrated that HSD17B2 transcript and protein levels are linked to some clinicopathological features in GC.

Associations of the calcium-sensing receptor gene CASR rs7652589 SNP with nephrolithiasis and secondary hyperparathyroidism in haemodialysis patients.

Nephrolithiasis, secondary hyperparathyroidism (sHPT), and cardiovascular complications are associated with disturbances in Ca handling and contribute to morbidity/mortality during haemodialysis (HD). Calcimimetics, activators of the calcium-sensing receptor (CaSR), provide an effective means of reducing parathyroid hormone (PTH) secretion in sHPT. Polymorphism in CaSR gene (CASR) influences Ca-related parameters, however it was not shown in HD patients for CASR rs7652589. The minor allele at this polymorphism modifies the binding sites of transcription factors and CaSR expression. We hypothesized that CASR rs7652589 variants may also influence CaSR in end stage renal disease (ESRD). We aimed to determine the associations of rs7652589 with nephrolithiasis-related ESRD, Ca, P, ALP, PTH, response to treatment with cinacalcet, prevalence of coronary artery disease, and all-cause/cardiovascular mortality in HD patients (n = 1162). Healthy individuals (n = 918) were controls. This study shows that the A allele of rs7652589 is a risk allele for nephrolithiasis-related ESRD. The AA genotype is associated with more severe sHPT (higher Ca and PTH concentrations). The A allele is associated with reduced CaSR transcript level in peripheral blood mononuclear cells. According to computational analysis, potential binding sites for GLI3, AHR and TP53 are removed by the A allele, whereas binding sites for SOX18 and TP63 are created.

Transcript level of AKR1C3 is down-regulated in gastric cancer

Steroid hormones have been shown to play a role in gastric carcinogenesis. Large amounts of steroid hormones are locally produced in the peripheral tissues of both genders. Type 5 of 17β-hydroxysteroid dehydrogenase, encoded by the AKR1C3 gene, plays a pivotal role in both androgen and estrogen metabolism, and its expression was found to be deregulated in different cancers. In this study we measured AKR1C3 transcript and protein levels in nontumoral and primary tumoral gastric tissues, and evaluated their association with some clinicopathological features of gastric cancer (GC). We found decreased levels of AKR1C3 transcript (p < 0.0001) and protein (p = 0.0021) in GC tissues compared with the adjacent, apparently histopathologically normal, mucosa. Lower levels of AKR1C3 transcript were observed in diffuse and intestinal types of GC, whereas AKR1C3 protein levels were decreased in tumors with multisite localization, in diffuse histological type, T3, T4, and G3 grades. We also determined the effect of the histone deacetylase inhibitor sodium butyrate (NaBu) on AKR1C3 expression in EPG 85-257 and HGC-27 GC cell lines. We found that NaBu elevates the levels of both AKR1C3 transcript and protein in the cell lines we investigated. Together, our results suggest that decreased expression of AKR1C3 may be involved in development of GC and can be restored by NaBu.

New arguments for beneficial effects of alpha-lipoic acid on the cardiovascular system in the course of type 2 diabetes

Purpose Alpha‐lipoic acid (ALA), widely known as an antioxidant, modifies also serum levels of angiogenic factors in type 2 diabetic patients. These pharmacological activities may influence the status of the cardiovascular system. Taking into consideration that diabetes is related to the increased cardiovascular risk we investigated several effects of ALA on angiogenic factors in the myocardium and in the aortal wall using a rat model of type 2 diabetes. Methods Diabetes was induced in Wistar rats by a fat‐rich diet and by intraperitoneal injection of a small dose of streptozotocin (30 mg/kg). Animals were divided into 3 groups: ALA‐treated type 2 diabetes rat model, placebo‐treated type 2 diabetes rat model and placebo‐treated non‐diabetic rats. ALA was administered orally once a day, 20 mg/kg, for 8 consecutive weeks. mRNA VEGF, VEGF‐R1 and VEGF‐R2 expression was measured in the myocardium and the aortal wall, simultaneously with circulating VEGF and circulating endothelial cells (cEC) and endothelial progenitor cells (cEPC). Results ALA induced pro‐angiogenic effect in the myocardium of rats with diabetes increasing mRNA VEGF expression and decreasing mRNA VEGFR‐1 expression, while in the aortal wall ALA increased mRNA VEGFR‐2 and VEGFR‐1 expression. cVEGF in the ALA‐treated group was higher comparing to both control groups. It was revealed that cEC percentage in the ALA‐treated group was decreased with no effect on the percentage of cEPC. Conclusions In summary, the current data provide novel findings about potential beneficial effects of ALA on angiogenic factors in the cardiovascular system, especially on myocardium, in the course of type 2 diabetes. Graphical abstract Figure. No caption available.

Relative indoleamine 2,3-dioxygenase transcript level concerning anti-HBs titers in response to HBV vaccination in hemodialysis patients

ABSTRACT Background: Indoleamine 2,3-dioxygenase (IDO) is a negative regulator of the immune system. To approach reasons of variability in the generation of anti-HBs antibodies in response to HBV vaccination among hemodialysis (HD) subjects, we aimed to investigate whether the IDO gene (IDO1) transcript levels are associated with post-vaccination anti-HBs production and IDO1 polymorphic variants. Methods: The IDO1 transcript was determined by qRT-PCR analysis in 110 HD patients. IDO1 (rs3739319, rs9657182) genotyping was carried out by HRM analysis. Results: The relative IDO1 transcript levels were not associated with IDO1 polymorphic variants. There were 16 non-responders (not able to produce anti-HBs >10 IU/L), 74 patients with anti-HBs 10–999 IU/L, and 20 hyperactive responders (anti-HBs ≥1000 IU/L). IDO1 transcript levels were different among these groups (0.832, 0.423–4.373; 1.114, 0.317–6.582; 0.680, 0.164–3.014; respectively, Kruskal-Wallis P = 0.024). Significance in IDO1 transcript was shown between anti-HBs titers 10–999 IU/L and ≥1000 IU/L (P = 0.020). IDO1 transcript level <0.743 indicated 3.38 (1.17–9.72) higher probability of hyperactive immunization (adjusted P = 0.005). Conclusion: In HD patients, ability to generate anti-HBs is not associated with IDO1 transcript levels. Hyperactive anti-HBs responses occur in patients showing lower IDO1 transcript. The latter cannot be predictable by genotyping IDO1 rs3739319 or rs9657182.

Simvastatin attenuates the aberrant expression of angiogenic factors induced by glucose variability

AIM Over the last few years, studies have indicated that fluctuant hyperglycemia is very likely to increase the risk of cardiovascular complications of diabetes. Statins are widely used in diabetes for the prevention of cardiovascular complications, but it is still not clear whether simvastatin could also prevent glycaemic variability - induced aberrant angiogenesis which plays a significant role in the development of atherosclerosis. METHODS Wistar rats were divided into four groups: (1) simvastatin-treated (20 mg/kg for 8 consecutive weeks) type 2 diabetes rat model with daily glucose excursions, (2) placebo-treated type 2 diabetes rat model with daily glucose excursions, (3) placebo-treated stable well-controlled type 2 diabetes rat model and (4) placebo-treated non-diabetic rats. Daily glucose fluctuations and several angiogenic factors: cVEGF, mRNA VEGF, VEGF-R1, VEGF-R2, TGF-beta expression, circulating endothelial and progenitor endothelial cells were measured in all groups. RESULTS Simvastatin decreased several factors enhanced by glucose excursions: circulating VEGF, mRNA TGF-beta expression in the myocardium and mRNA VEGFR-2 expression in the aorta. Simvastatin increased some factors attenuated by glucose fluctuations: mRNA VEGF expression and mRNA VEGFR-1 expression in the myocardium and in the aorta. In the simvastatin-treated group with glycaemic variability, the percentage of circulating endothelial cells was lower and the percentage of progenitor endothelial cells in peripheral blood was higher than in the placebo-treated rats with glucose-fluctuations. CONCLUSIONS Simvastatin used in the rat model of type 2 diabetes with glucose variability reduces glucose variability and limits glucose fluctuations-induced changes in the expression of angiogenic factors in the cardiovascular system.

mRNA expression of steroidogenic enzymes, steroid hormone receptors and their coregulators in gastric cancer

Epidemiological and experimental findings suggest that the development of gastric cancer (GC) is regulated by steroid hormones. In postmenopausal women and older men, the majority of steroid hormones are produced locally in peripheral tissue through the enzymatic conversion of steroid precursors. Therefore, using reverse transcription-quantitative polymerase chain reaction analysis, the mRNA expression of genes encoding steroidogenic enzymes, including steroid sulfatase (STS), hydroxy-delta-5-steroid dehydrogenase 3 beta- and steroid delta-isomerase 1 (HSD3B1), 17β-hydroxysteroid dehydrogenase type 7 and aromatase (CYP19A1), was investigated in primary tumoral and adjacent healthy gastric mucosa from 60 patients with GC. Furthermore, the mRNA levels for estrogen receptor α, estrogen receptor β (ESR2) and androgen receptor (AR), along with their coregulators, including proline, glutamate and leucine rich protein 1, CREB binding protein, nuclear receptor coactivator 1 (NCOA1), nuclear receptor corepressor 1 (NCOR1) and nuclear receptor subfamily 2 group F member 1 (NR2F1), were investigated. Additionally, the association between the mRNA expression of these genes and the clinicopathological features of patients with GC was examined. Significantly decreased levels of STS, HSD3B1, ESR2, AR, NCOA1 and NCOR1 mRNA, in addition to significantly increased levels of CYP19A1 mRNA were demonstrated in tumoral tissue samples compared with adjacent healthy gastric tissue samples. Deregulated expression of these genes in the analyzed tissue samples was associated with certain clinicopathological features of GC, such as age and localization of the tumor. The results of the current study suggest that all of the genes analyzed are expressed in tumoral and adjacent healthy gastric mucosa. In addition, the results indicate that abnormal expression of STS, ESR2, AR, NCOA1 and NCOR1 may serve a role in the development and progression of GC, and may be associated with specific clinicopathological features in patients with GC.