The Importance of the FUT2 rs602662 Polymorphism in the Risk of Cardiovascular Complications in Patients after Kidney Transplantation.

The FUT2 gene encodes an enzyme called α-1,2-fucosyltransferase, which is involved in the formation of blood group antigens AB0(H) and is also involved in the processes of vitamin B12 absorption and its transport between cells. FUT2 gene polymorphisms are associated with vitamin B12 levels in the body. Vitamin B12 deficiency associated with hyperhomocysteinemia is a major risk factor for cardiovascular diseases (CVDs), which are one of the main causes of death in patients after kidney transplantation. The aim of our study was to determine the impact of the rs602662 (G>A) polymorphism of the FUT2 gene on the functionality of transplanted kidneys and the risk of CVD in patients after kidney transplantation. The study included 402 patients treated with immunosuppression (183 patients taking cyclosporine (CsA) and 219 patients taking tacrolimus (TAC)). The analysis of the FUT2 rs602662 (G>A) polymorphism was performed using real-time PCR. Patients with CsA were more likely to be underweight (1.64% vs. 0.91%) and obese (27.87% vs. 15.98%), while those taking TAC were more likely to be of normal weight (39.27%) or overweight (43.84%). No statistically significant differences were observed comparing the mean blood pressure, both systolic and diastolic. The renal profile showed a higher median urea nitrogen concentration in patients with CsA (26.45 mg/dL (20.60-35.40) vs. 22.95 mg/dL (17.60-33.30), p = 0.004). The observed frequency of rs602662 alleles of the FUT2 gene was similar in the analyzed groups. The A allele was present in 43.7% of patients with CsA and 41.1% of those taking TAC (OR = 0.898; 95% CI: 0.678-1.189; p = 0.453). In the group with CsA, the GG genotype was present in 32.2% of patients, the GA in 48.1% and the AA in 19.7%. A similar distribution was obtained in the TAC group: GG-33.8%, GA-50.2%, and AA-16.0%. An association of genotypes containing the G allele with a higher incidence of hypertension was observed. The G allele was present in 65% of people with hypertension and in 56% of patients with normal blood pressure (p = 0.036). Moreover, the evaluation of the renal parameters showed no effect of the FUT2 polymorphism on the risk of organ rejection because the levels of creatinine, eGFR, potassium, and urea nitrogen were prognostic of successful transplantation. Our results suggest that the rs6022662 FUT2 polymorphism may influence the risk of cardiovascular diseases.

Analysis of the Polymorphisms and Expression Levels of the BCL2, BAX and c-MYC Genes in Patients with Ovarian Cancer.

Ovarian cancer (OC) is one of the biggest problems in gynecological oncology and is one of the most lethal cancers in women worldwide. Most patients with OC are diagnosed at an advanced stage; therefore, there is an urgent need to find new biomarkers for this disease. Gene expression profiling is proving to be a very effective tool for exploring new molecular markers for OC patients, although the relationship between such markers and patient survival and clinical outcomes is still elusive. Moreover, polymorphisms in genes encoding both apoptosis-associated proteins and oncoproteins may serve as key markers of cancer susceptibility. The aim of our study was to analyze the polymorphisms and expressions of the BCL2, BAX and c-MYC genes in a group of 198 women, including 98 with OC. The polymorphisms and mRNA expressions of the BCL2, BAX and c-MYC genes were analyzed using real-time PCR. The analysis of the BAX (rs4645878; G>A) and c-MYC (rs4645943; C>T) polymorphisms showed no association with ovarian cancer risk. The BCL2 polymorphism (rs2279115; C>A) showed a significant difference in the frequency of genotypes between the studied groups (CC: 23.47% vs. 16.00%, AA: 25.51% vs. 37.00%; p = 0.046; OR = 1.61). Furthermore, the expression levels of the BCL2 and c-MYC genes showed a decrease at the transcript level for OC patients compared to the control group (BCL2: 17.46% ± 3.26 vs. 100% ± 8.32; p < 0.05; c-MYC: 37.56% ± 8.16 vs. 100% ± 9.12; p < 0.05). No significant changes in the mRNA level were observed for the BAX gene (104.36% ± 9.26 vs. 100% ± 9.44; p > 0.05). A similar relationship was demonstrated in the case of the protein expressions of the studied genes. These findings suggest that the CC genotype and C allele of the BCL2 polymorphism could be genetic risk factors for OC development. A gene expression analysis indicated that BCL2 and c-MYC are associated with OC risk.

Combined Effects of Methyldopa and Flavonoids on the Expression of Selected Factors Related to Inflammatory Processes and Vascular Diseases in Human Placenta Cells-An In Vitro Study.

The aim of the study was to investigate combined effects of flavonoids (apigenin, baicalein, chrysin, quercetin, and scutellarin) and methyldopa on the expression of selected proinflammatory and vascular factors in vitro for prediction of their action in pregnancy-induced hypertension. The research was conducted on a trophoblast-derived human choriocarcinoma cell line and a primary human umbilical vein endothelial cell line. Cytotoxicity of compounds in selected concentrations (20, 40, and 100 µmol) was measured using the MTT test and the concentration of 40 µmol was selected for further analysis. Subsequently, their effects with methyldopa on the expression of selected markers responsible for inflammation (TNF-α; IL-1ß; IL-6) and vascular effects (hypoxia-inducible factor 1α-HIF-1α; placental growth factor-PIGF; transforming growth factor ß-TGF-ß; vascular endothelial growth factor-VEGF) at the mRNA and protein levels were assessed. It was found that every combined administration of a flavonoid and methyldopa in these cells induced a down-regulating effect on all tested factors, except PIGF, especially at the mRNA expression level. As hypertension generally raises TNF-α, IL-1ß, IL-6, HIF-1α, TGF-ß, and VEGF mRNA expression and/or protein levels, the results obtained in the studied model may provide a positive prognostic factor for such activity in vivo.

The rs1256044 polymorphism in the ESR2 gene and the risk for osteoporosis in Polish postmenopausal women.

Population association studies have demonstrated a strong association between ESR2 SNPs and BMD, indicating that ESR2 may influence attainment of bone mass. The aim of the study was to investigate the ESR2 gene, located on chromosome 14q linked with BMD, which demonstrates a correlation with changes in bone mass in healthy Caucasian women. The study included 675 unrelated Polish postmenopausal women, including 109 with osteopenia, 333 with osteoporosis and 233 healthy women. The women were classified into the following groups: osteopenia, osteoporosis and normal T-score. Analysis of genotype frequency for the ESR2 rs1256044 polymorphism revealed no statistically significant differences. No statistically significant differences were noted for the allele frequency. However, it is noticeable that the CT genotype occurred more often in women with osteopenia (50.4%, OR = 1.14) and osteoporosis (54.7%, OR = 1.33) than controls (47.7%). There were statistically significant differences between the clinical parameters and distribution of genotypes in patients with osteopenia but not osteoporosis. ESR2 polymorphisms demonstrate minimal influence on BMD changes in women. Identification of various genes with little impact on BMD, such as ESR2, might help design a screening panel for osteoporosis risk assessment in healthy subjects.

Expression profiling of genes modulated by rosmarinic acid (RA) in MCF-7 breast cancer cells.

OBJECTIVES: Cancer is the second most common cause of death, with breast cancer (BC) as the most frequently diagnosed neoplasm among females. The origin of BC is multifactorial and depends on environmental and genetic factors. The disease presents a significant challenge due to its drug resistance and frequent metastasis. Thus, new effective therapies and metastasis prevention are much needed. Rosmarinic acid (RA) is a natural polyphenol which possesses the ability to inhibit BC cell proliferation and demonstrates cytotoxic properties against those cells. In our study, we examined the effect of RA on the expression of ZEB1, MDM2, ABCB1, PTEN and TWIST1 genes in MCF-7 breast cancer cells. MATERIAL AND METHODS: MCF-7 cell cultures were treated with 0.2 µM doxorubicin (DOX) and 1.5, 15 or 50 µM of RA. Real-time PCR reaction was performed to analyze gene expression levels. RESULTS: PCR analysis showed a significant increase of the ZEB1 gene expression, which was about 3-fold for DOX 0.2 µM, 9-fold for 0.2 µM DOX + 1.5 µM RA and 0.2 µM DOX + 15 µM RA (p < 0.05), and about 6.5-fold for 0.2 µM DOX + 50 µM RA (p < 0.05). Furthermore, a decrease of the MDM2 gene expression was observed in all of the examined variants and was about 40-75% (p < 0.05). No influence of DOX and RA combined with DOX on the ABCB1, TWIST1 and PTEN genes was found. CONCLUSIONS: The results of our study suggest that RA might be used as an adjuvant therapeutic factor in BC treatment.

Coexistence of ACE (I/D) and PAI-1 (4G/5G) gene variants in recurrent miscarriage in Polish population.

OBJECTIVES: Recurrent miscarriage (RM) is one of the most common obstetric complications. Numerous studies have suggested that genetic variants leading to an impaired balance between coagulation and fibrinolysis may contribute to elevated risk of pregnancy loss. The aim of the study was to investigate a possible association between angiotensin-converting enzyme (ACE, rs1799752) I/D and plasminogen activator inhibitor type 1 (PAI-1, rs1799768) 4G/5G polymorphisms with RM among Polish women. MATERIAL AND METHODS: DNA was extracted from peripheral blood samples of 152 women with a history of ≥ 2 consecutive pregnancy losses before 22 weeks of gestation, and 180 healthy controls with at least 1 live birth at term and no history of pregnancy loss. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) were used to identify the polymorphisms. RESULTS: No statistically significant differences were found in genotype and allele frequencies of the studied polymorphisms. The most relevant difference between the study group and controls was found for the ID genotype distribution of the ACE gene (52.6 vs. 46.7%, OR = 1.27, p = 0.28). The analysis of genotype coexistence revealed a higher incidence of the combination of the ACE II and the PAI-1 4G/4G genotypes in the control group (10.0 vs.5.9% in control group; p = 0.17). CONCLUSIONS: The obtained results suggest no apparent association between the ACE I/D, PAI-1 4G/5G polymorphisms and increased RM susceptibility in the analyzed Polish population.

[The role of Wnt/ß-catenin pathway and LRP5 protein in metabolism of bone tissue and osteoporosis etiology]. / Rola szlaku sygnalowego Wnt/ß-katenina oraz bialka LRPS w metabolizmie tkanki kostnej oraz etiologii osteoporozy.

Osteoporosis is a metabolic bone disease, manifested by decreased bone mineral density microarchitectural disturbances of bone tissue, and increased risk of bone fractures. Owing to large-scale morbidity particularly among postmenopausal women, nowadays osteoporosis constitutes a significant global health problem. In recent years, much attention has been paid to the role of signaling Wnt/ß-catenin pathway and LRP protein in the pathomechanism of osteoporosis, indicating a possible contribution of polymorphic variants of the candidate LRP5 gene to disease development. The goal of our study is to present contemporary research on signaling Wnt/ß-catenin pathway and mechanism of LRP protein action in the process of bone tissue metabolism and etiology of osteoporosis.

DRD1 and DRD4 dopamine receptors in the etiology of preeclampsia.

INTRODUCTION: Recent reports have suggested an association between genetic polymorphisms of dopamine receptors and the development of an increased risk of chronic hypertension, as well as preeclampsia (PE). OBJECTIVES: The aim of the study was to evaluate the impact of the -48A>G DRD1 and -521C>T DRD4 polymorphisms in the etiology of PE among Polish women. MATERIAL AND METHODS: Ninety-eight preeclamptic women and 120 healthy pregnant controls were enrolled in the study The investigated polymorphisms of the DRD 1 and DRD4 genes were identified using PCR/RFLP methods. RESULTS: As far as the -48A>G DRD 1 polymorphism is concerned, the mutated -48GG genotype was more often found in controls (14.2%) than in the PE group (10.2%, ns), and the subgroup with severe PE (8.2%). Also, the frequency of the mutated -48G allele was higher in controls (39.6%) than in the PE group (33.2%, ns), and in the subgroup with severe PE (31.6%, ns). As for the -521C>TDRD4 polymorphism, a similar occurrence of the mutated -521 TTgenotype and the -521T allele in all of the investigate groups was observed. Lower serum concentrations of total protein (5.59 g/L and 5.57 g/L vs. 6.17 g/L in carriers of the -52100 genotype, p=0.02) were noted in patients with the mutated homozygous -521 TT genotype and heterozygous -521CT genotype of DRD4. CONCLUSIONS: The obtained results suggest a possible protective role of the mutated -48G DRD1 allele in the etiology of preeclampsia, especially its severe form. The presence of the mutated -521 T DRD4 allele could influence the decrease of total blood protein in preeclamptic patients. The observed frequency of dopamine DRD1 and DRD4 polymorphisms is similar to the distribution of these variants in other Caucasian populations.

Polymorphisms of collagen 1A1 (COL1A1) gene and their relation to bone mineral density in postmenopausal women.

OBJECTIVES: The goal of this study was to evaluate the frequency of Sp1 +1245G>T (rs 1800012) and -199 7G>T (rs 1107946) COL1A1 gene polymorphisms in postmenopausal women with osteoporosis and osteopenia as well as assessing their relations with the clinical parameters and parameters of bone turnover. STUDY DESIGN: The study included 538 (236 postmenopausal and 302 healthy reproductive) Polish women. The postmenopausal group included women with osteoporosis (n = 90), osteopenia (n = 90), as well as healthy individuals (n = 56). All women of reproductive age were healthy BMD was marked in the L2-L4 lumbar region of the spine using dual energy X-ray absorptiometry (DXA). Genomic DNA was isolated from peripheral blood, the genotype frequency of investigated polymorphisms was determined by PCR-RFLP technique. RESULTS: The frequency of Sp1 +1245G>T and -1997G>T polymorphisms of COL1A1 gene showed no statistically significant differences between group with osteoporosis, osteopenia and correct T-score and women of reproductive age. In postmenopausal women it was found that osteopenia and osteoporosis were correlated with age, birth weight, age of last menses occurrence, height, body weight and BMI value. Clinical parameters in all groups of women did not show any statistically significant correlation with frequency of Sp1 +1245G>T and -1997G>T COL1A1 polymorphisms. CONCLUSIONS: An evaluation of Sp1 +1245G>T (rs1800012) and -1997G>T(rs 1107946) COL1A1 polymorphisms showed any influence of these genetic variants on osteoporosis development in Polish postmenopausal women. The presented correlation between osteoporosis and age, birth weight, age of last menses occurrence, height, body weight and BMI value confirms the important role of environmental factors in disease etiology.

Analysis of -11391G>A and +45>G polymorphisms of ADIPOQ gene in women with excessive weight gain during pregnancy.

OBJECTIVES: The aim of our study was to evaluate the frequency of genotypes and alleles of the -11391G>A and +45T>G polymorphisms of the ADIPOQ gene in Polish women with excessive weight gain during pregnancy. A possible correlation between these polymorphisms and selected clinical and anthropometric parameters has been analyzed. MATERIAL AND METHODS: A total of 153 pregnant Caucasian women of Polish origin with normal pre-pregnancy body mass were analyzed: 78 women with excessive weight gain (study group) and 75 women with normal weight gain during pregnancy (control group). The analysis of the polymorphisms was performed by PCR/RFLP. RESULTS: The influence of the -11391G>A polymorphism on body mass and BMI values at the end of pregnancy (p < 0.05) was observed. We also detected a correlation of the +45T>G polymorphism with body mass at the end of pregnancy and pre-pregnancy WHR values (p < 0.05). CONCLUSIONS: The observed effect of the -11391G>A polymorphism on the parameters assessed at the end of pregnancy (BMI and body mass), suggests a protective role of the -11391A genetic variant in excessive weight gain. It is claimed that the mutated +45G allele of the +45T>G ADIPOQ polymorphism shows a possible connection with higher pre-pregnancy WHR values and body mass at the end of pregnancy Our findings suggest a possible contribution of the -11391G>A and +45T>G polymorphisms of the ADIPOQ gene to the pathomechanism of excessive weight gain in pregnant women from the Polish population. This observation should be confirmed in a larger sample size study

The importance of rs1021737 and rs482843 polymorphisms of cystathionine gamma-lyase in the etiology of preeclampsia in the Caucasian population.

INTRODUCTION: Recently an increasing number of reports indicate the participation of genetic factors in the pathogenesis of preeclampsia (PE). The genes involved in the synthesis of nitric oxide that participates in the vasolidation, may play an important role in the development of this disorder. Hydrogen sulfide (H2S) which is produced by cystathionine gamma-lyase exhibits a similar effect to nitric oxide. It is suggested that certain polymorphisms of the CTH gene may participate in the development of chronic hypertension and preeclampsia. AIM OF THE STUDY: To evaluate the frequency of genotypes and alleles of rs1021737 and rs482843 polymorphisms of CTH gene in women with preeclampsia from Wielkopolska region. MATERIAL AND METHODS: The study group consisted of 60 patients with diagnosed preeclampsia, into the control group 120 healthy pregnant women were enrolled. The examined rs1021737 and rs482843 polymorphisms of CTH gene were determined using PCR-RFLP method. RESULTS: Analysis of rs482843 polymorphism in the CTH gene showed a statistically significant difference in the prevalence of mutated GG genotype (p<0.000001) and mutated G allele (p<0.000001) in the group of pregnant women with PE compared to the control group. There was no such correlation for the rs1021737 polymorphism. Furthermore, there are also no relationship between studied polymorphisms and selected clinical and biochemical parameters. CONCLUSIONS: The results of rs482843 polymorphism analysis suggest that mutated GG genotype predisposes to preeclampsia occurrence. There was no such relationship for the rs1021737 polymorphism of CTH gene. Hence, further studies based on the determination of CSE expression level in women with PE may confirm the observed relationship between the rs482843 polymorphism and the risk of preeclampsia.

Evaluation of anti-inflammatory and analgesic activities of extracts from herb of Chelidonium majus L.

The aim of the study was to evaluate analgesic activity ("hot plate" test), anti-inflammatory activity (carrageenan-induced paw edema) and locomotor activity in rats under the influence of three fractions of Chelidonium majus herb extract: full water extract (FWE), protein enriched fraction (PEF), and non-protein fraction (NPF). Effects of the fractions on the level of chosen cytokines and their mRNA levels were also assessed using lipopolysaccharide (LPS) administration as a proinflammatory cue. All fractions and diclofenac did not affect the locomotor activity of rats in comparison with the control group. FWE and PEF three hours after administration showed statistically significant analgesic activities comparable to morphine (p < 0.05). A slight reduction in rat paw edema was observed after three (comparable with diclofenac) and six hours in the NPF group. FWE revealed a statistically significant pro-inflammatory effect after three hours in comparison with the control group. Peripheral IL-1 and IL-4 cytokine concentrations were reduced under FWE and NPF, PEF fractions. The combination of FWE, PEF and NPF together with LPS showed only the effects of LPS. We suggest that protein enriched fraction (PEF) produced centrally mediated (morphine-like) analgesic action, whereas the anti-inflammatory potential was shown only after LPS-induced inflammation. The precise mechanisms involved in the production of anti-nociceptive and anti-inflammatory responses of studied fractions are not completely understood, but they may be caused rather by the presence of protein more than alkaloids-enriched fraction. This fraction of the extract could be used as an alternative therapy for the prevention of inflammatory-related diseases in the future, but further studies are needed.

The APOB gene polymorphism in the pathogenesis of gallstone disease in pre- and postmenopausal women.

AIM OF THE STUDY: The decrease in estrogen levels in the postmenopausal period changes the lipid profile by the expression of hepatic genes related to metabolism of cholesterol and bile acid synthesis that could be important in the pathogenesis of cholelithiasis. The aim of the study was to determine the APOB gene 7673C>T and 12669G>A polymorphisms in the pathogenesis of gallstones and analysis of the composition of gallstones in pre- and postmenopausal women. MATERIAL AND METHODS: The study group consisted of 94 women qualified to the laparoscopic cholecystectomy while the control group consisted of 81 women in whom gallstones and other changes in the bile ducts were excluded. Gallstones composition analysis was performed using commercially available assays. The prevalence of the APOB gene polymorphisms was determined using the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). RESULTS: When assessing the composition of gallstones in pre- and postmenopausal women, we observed differences in the studied parameters. Analysis of genetic variants of APOB gene 7673C>T and 12669G>A polymorphisms showed no significant statistical differences between studied groups and controls. CONCLUSIONS: Analysis of 7673C>T and 12669G>A polymorphisms showed no relationship between specific genetic variants and the risk of gallstones in pre- and postmenopausal women, pointing to the fact that the investigated polymorphisms are not relevant as prognostic factors in gallstone disease in the Caucasian population. Because of the possible contribution of a variety of factors in gallstones pathogenesis the studies are required to take account of additional environmental factors, what may indicate different occurrence between investigated polymorphisms, gallstone disease development and gallstones composition in Caucasians.

Screening of trace elements in hair of the female population with different types of cancers in Wielkopolska region of Poland.

BACKGROUND: Cancer constitutes a major health problem worldwide. Thus, search for reliable and practical markers of the disease process remains the key issue of the diagnostic process. OBJECTIVES: The study aims at linking the trace element status of an organism, assessed by hair analysis, with the occurrence of cancer diseases. MATERIAL AND METHODS: Hair samples were collected from 299 patients with cancer diseases confirmed by a histopathological test and from 100 controls. Cancer patients were divided into three groups, depending on cancer type: hormone-dependent cancer, cancer of the alimentary tract, and cancer with high glycolytic activity. Mineral element analysis of hair was performed using an atomic emission spectrophotometer with inductively coupled plasma (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: Statistically significantly lower concentrations of selenium, zinc, copper, germanium and boron, iron, and magnesium were observed in the three groups of cancer patients. Disturbance in the axis glucose-insulin and changes in concentrations of heavy metals and toxic elements were also noted. CONCLUSIONS: It seems safe to conclude that our results confirmed usefulness of hair element analysis in screening tests for the assessment of the biomarker of various cancer diseases in a female population.

The -323P0/P10 factor VII gene polymorphism and the risk of recurrent miscarriage.

OBJECTIVES: Genetically determined disturbances in the activity of coagulation factor VII may lead to obstetric complications. The aim of the study was to evaluate the correlation between -323P0/P10 factor VII gene polymorphism and the risk of recurrent miscarriage. MATERIAL AND METHODS: The study group consisted of 152 women with a history of > or = 2 miscarriages. The control group comprised 180 women with no history of miscarriage and > or = 1 pregnancy who gave birth to a healthy newborn at term. The study group was further subdivided twice into two subgroups: 1174 patients with a history of 2 miscarriages and 38 subjects with a history of > or = 3 miscarriages, and 123 patients with miscarriages < 13 gw. and 29 with miscarriages < 21 gw. Genetic analysis was performed with the use of PCR/RFLP. RESULTS: Overrepresentation of P0/P0 genotype and lower frequency of P0/P10 genotype was noted in the study group as compared to controls (P0/P0: 80.26 vs. 76.67%, p = 0.25; P0/P10: 18.42 vs. 22.78%, p = 0.20). A higher presentation of P0/P0 genotype and P0 allele, lower frequency of P0/P10 genotype and P10 allele was observed in the subgroup of women with > or = 3 miscarriages as compared to controls (P0/P0: 8.84 vs. 76.67%, p = 0.12; P0: 93.42 vs. 88.06%, p = 0.12; P0/P10:13.16 vs. 22.78%, p = 0.13; P10: 11.94 vs. 6.58%, p = 0.12). CONCLUSIONS: The obtained results suggest a probable protective role of -323P10 allele against the risk of miscarriage in women with > or = 3 recurrent pregnancy losses.

[Modern diagnostics of osteoporosis based on the use of biochemical markers of bone turnover]. / Nowoczesna diagnostyka osteoporozy w oparciu o wykorzystanie biochemicznych markerów obrotu kostnego.

Osteoporosis is a disease with low bone mass and disorganization of the internal microarchitecture of bone tissue. Determination of biochemical markers allows for early diagnosis of changes in bone tissue metabolism. The search for a marker whose biological function could be directly connected with bone metabolism, clearly indicating a connection between its concentration and risk fracture as well as response to treatment, continues. Currently measurement of collagen-derived markers of bone resorption is used in the majority of cases. They are, first of all, telopeptides of collagen type 1 localized on the amino end-N-terminal telopeptide of type 1 collagen (NTX), as well as on the carboxy end-C-telopeptide of type 1 collagen (CTX) of collagen molecule. Among markers of bone synthesis, special attention is paid to the procollagen type 1 carboxy-terminal propeptide (POCP) and procollagen type 1 amino-terminal propeptide (P1NP). Simultaneous application of bone synthesis and resorption markers allows for a full imaging of the bone remodeling process and application of biochemical markers in the diagnosis and therapy of osteoporosis.

The significance of TNF-alpha gene polymorphisms in preterm delivery.

INTRODUCTION: Nowadays the strong genetic background of preterm delivery (PTD) in connection with immune answer has been indicated. The purpose of the study was the assessment of frequency of TNF-alpha -238G>A, -308G>A, -376G>A gene polymorphisms in the etiology of preterm delivery MATERIAL AND METHODS: The study group consisted of 150 women with PTD (22+0 - 36+6 gw.), the controls of 150 women who delivered at term (> 37 gw). PTD group was divided into subgroups: a/delivery between 22-28 gw, b/28-32 gw., and c/32-36+6 gw. Genetic analysis was performed by PCR/RLFP method. RESULTS: Overrepresentation of -238GA genotype (12.7 vs. 4.7%, p = 0.011) and -238A allele (7.7 vs. 2.3%, p = 0.002) in PTD group has been observed. In PTD 28-32 gw. subgroup, higher frequency of -238GA genotype (31.6 vs. 4.7%, p = 0.00095), and mutated -238A allele (21.1 vs. 2.3%, p = 0.00004) was noted. Moreover in PTD 28-32 gw. subgroup we have noted higher presence of heterozygous -376GA genotype (10.5 vs. 1.3%, p = 0.063) and mutated -376A allele (5.3 vs. 0.7%, p = 0.064). Analysis of TNF-alpha polymorphisms co-occurrence showed statistically significant overrepresentation of genotypes containing mutated -238A allele in PTD group (-238GA/-308GG/-376GG: 8.0 vs. 2.7%, p = 0.035). Haplotype analysis revealed statistically significant difference between PTD and controls in the incidence of -376G/-308G/-238A haplotype containing mutated -238A allele (0.063067 vs. 0.016634, p = 0.030). CONCLUSION: The study indicated the strong association of mutated -238A allele of TNF-alpha gene with increased risk of PTD. Analysis of genotypes and alleles prevalence in PTD women divided according to gestational age suggests the possible role of mutated variants of -238G>A and -376G>A TNF-alpha polymorphisms in Polish women delivering between 28 and 32 gw.

The influence of soybean extract on the expression level of selected drug transporters, transcription factors and cytochrome P450 genes encoding phase I drug-metabolizing enzymes.

OBJECTIVE: Soybean phytoestrogens, such as genistein and daidzein, reduce climacteric symptoms and the risk of certain chronic diseases such as cancer and cardiovascular diseases. Despite their widespread use in functional foods and dietary supplements, there is very little data available on their safety and herb-drug interactions, especially with antineoplastic agents. Hence, the aim of our study was to assess the effects of soybean extracts on the expression level of CYP genes and their transcriptional factors. We also investigated the effect of soybean on the mRNA level of transporters, such as P-glycoprotein (MDRI) and multidrug resistance-associated proteins (MRP1, MRP2). MATERIALS AND METHODS: Wistar rats were fed a standardized soybean extract (100 mg/kg, p.o.). cDNA was synthesized from total RNA isolated from different tissues (liver and intestinal epithelium) using reverse transcription. Gene expression level was analyzed by RT-PCR method. RESULTS: We demonstrated a significant increase of CYP1A1 mRNA level (by 89%, p = 0.002 and 125%, p = 0.004) as compared with the control group. An increase of AHR and CAR expression after 10 days was also observed (by 60%, p < 0.001 and 52%, p > 0.05, respectively). Additionally an inductive effect for CYP2D1 by 22% (p = 0.008), Mdr1a by 267% (p < 0.0001), Mdr2b by 86% (p < 0.00001), Mrp1 by 9-fold (p < 0.0001), Mrp2 by 83% (p < 0.0001) in the liver and for Mrp2 by 35% (p < 0.001) in the intestinal epithelium, was evaluated. A significant decrease of mRNA level was observed for CYP3A1 (human CYP3A4) in the liver and Mdr1b in the intestinal epithelium. Moreove, we also showed a slight decrease in the amount of mRNA for CAR, PXR and ARNT after 3 days. CONCLUSIONS: Our results suggest that Glycine max may change the expression level of CYPs, especially CYP3A4 and CYP1A 7, involved in biotransformation of xenobiotics (drugs, procarcinogens) and may participate in clinically significant interactions with drugs metabolized by these enzymes. Moreover an increase of CYP1A1 (homologue to human CYPIA 1) mRNA level may not only reduce the carcinogenicity of foreign compounds, but may also activate some compounds to their carcinogenicity In case of transporters, it is considered that an increase of their expression in the body may lead to increased fetoprotection. Also, it may reduce both, the exposure of sensitive tissues (e.g. brain, placenta) to xenobiotics and treatment effectiveness of certain diseases. Hence, the search for a safe substance that could effectively modulate transporter activity especially in the treatment of certain hormone -dependent disorders, e.g. osteoporosis and breast cancer occurring mainly in postmenopausal period, continues.

Effect of Epilobium angustifolium and Serenoa repens extracts on regulation of non-genomic signaling pathway of kinases.

OBJECTIVES: Changes of kinase activity of non-genomic cellular signaling pathway may influence the effectiveness of pharmacotherapy in case of hormone-dependent tumors. Our study investigated a possible interaction at the molecular level between an aqueous herbal extract of Epilobium angustifolium as well as a lipid-sterolic fruit extract of Serenoa repens and synthetic drugs used in the treatment of hormone-dependent cancers. MATERIAL AND METHODS: E. angustifolium and Serenoa repens extracts were orally administered to testosterone-induced rats for 21 days. Changes of RafA/Mapk3/Mapk1 mRNA levels were analyzed by real-time quantitative PCR using target specific primers. RESULTS: The level of RafA mRNA slightly increased in rats receiving Epilobium angustifolium (p = 0.076) and Serenoa repens (p = 0.016) extracts. Administration of these extracts resulted in significantly elevated Mapk1 and Mapk3 transcripts in the investigated animals (p < 0.05 for each extract). The levels of Mapk1 and Mapk3 mRNA strongly increased (p < 0.05 for each extract) in animals receiving concomitantly testosterone and the extracts, while RafA transcription slightly decreased (p < 0.05), as compared to controls. CONCLUSIONS: The results of our study may indicate a potential effect of S. repens and E. angustifolium extracts on the functioning of non-genomic cellular signaling kinases pathway. We investigated safety of these extracts to detect possible drug interactions between synthetic drugs used in the treatment of proliferative changes in hormone-dependent reproductive organs and herbal preparations.

The importance of 8993C>T (Thr399Ile) TLR4 polymorphism in etiology of osteoporosis in postmenopausal women.

INTRODUCTION: Toll-like receptors (TLR) may play a key role in initiating cellular signaling pathways by increasing the levels of inflammatory cytokines which, cooperating with osteoclasts, influence bone turnover Numerous research articles focused on the genetic background of this condition, among others on polymorphic variants in TLR genes. The aim of the study was to examine the role of 20877G>A (Arg753GIn) in TLR2 gene and 8993C>T (Thr399lle) in TLR4 gene in the etiopathogenesis of postmenopausal osteoporosis in Polish women. MATERIAL AND METHODS: This study included 180 postmenopausal women (t-score < or = -2.5), 153 postmenopausal women with osteopenia (t-score between -2.5 and -1), and 91 postmenopausal healthy women with correct t-score (t-score >-1). The 20877G>A TLR2 and 8993C>T TLR4 polymorphisms were determined by PCR/RFLP analysis. RESULTS: The analysis did not reveal statistically significant differences in the distribution of genetic variants of 20877G>A TLR2 polymorphism between the investigated groups of women. The most interesting results were connected with 8993C>T TLR4 polymorphism. Comparison of the group with osteoporosis and controls revealed overrepresentation of heterozygous 8993CT genotype (13.3 vs. 5.5%, OR=2.65, p=0.03). Also, mutated 8993T allele was overrepresented in the group with osteoporosis (6.7 vs. 2.7%, OR=2.52, p=0.04). Higher frequency of heterozygous 8993CT genotype (13.3 vs. 4.6%, OR=3.21, p=0.004) and mutated 8993T allele (6.7 vs. 2.3%, OR=3.05, p=0. 005) was noted in osteoporotic women as compared to the group with osteopenia. Higher frequency of heterozygous 8993CT genotype (13.3% vs. 5.3%, OR=2.73, p=0.003) and mutated 8993T allele (6.7 vs. 2.7%, OR=2.67, p=0.004) was observed in the group with osteoporosis as compared to women with osteopenia and with correct t-score. CONCLUSIONS: Results of our study suggest an important role of mutated 8993T allele of 8993C>T TLR4 polymorphisms in the etiology of postmenopausal osteoporosis. Nevertheless, this observation requires further investigation with larger sample size comprised of Polish women.