GSK3ß polymorphisms, MAPT H1 haplotype and Parkinson's disease in a Greek cohort.

To determine whether polymorphisms in the microtubule-associated protein tau (MAPT) and/or glycogen synthase kinase-3ß (GSK3ß) genes underpin susceptibility to Parkinson's disease (PD), we conducted a case-control association study in a Greek cohort of 196 PD cases and 163 healthy controls. In our study, the MAPT H1 haplotype was found to be significantly associated with PD, no association was detected between the intronic rs6438552 (-157 T/C) GSK3ß polymorphism and PD, whereas the C/C genotype of the promoter rs334558 (-50 T/C) GSK3ß polymorphism was found to exert a protective role. The C/C genotype of the rs334558 GSK3ß polymorphism was also found to have an additional protective role in our MAPT H1/H1 PD subgroup. Haplotype analysis revealed that, the T-T haplotype of both GSK3ß polymorphisms was over-represented in PD patients compared to controls, and this association was independent of MAPT H1 haplotype.

Glutathione-S-transferase M1 and T1 and cytochrome P1A1 genetic polymorphisms and susceptibility to cervical intraepithelial neoplasia in Greek women.

The aim of the study was to determine the importance of genetic polymorphisms of glutathione-S-transferase T1 and M1 and cytochrome P1A1 genes in the development of cervical intraepithelial neoplasia in Greek women. This was a prospective, case-control study conducted by the Cervical Pathology and Colposcopy Unit of a University Ob/Gyn Department from 1999 to 2003. Cervical smears from 114 controls without any cytological and/or colposcopical evidence of cervical pathology and from 166 women with history of cervical intraepithelial neoplasia (56 CIN I, 54 CIN II and 56 CIN III) were examined with polymerase chain reaction for the above-mentioned genetic polymorphisms, taking also in mind their smoking attitudes. Statistical analysis was performed to detect any association between the null genotype of GSTM1 and GSTT1 genes and the CYP1A1 m1 polymorphism and the severity of cervical intraepithelial neoplasia. The distributions of the GSTT1 and GSTM1 wild-type genotypes were 57.48 and 39.75%, respectively. No woman with homozygous GSTT1 and GSTM1 null/null genotype was identified. CYP1A1 m1 polymorphism frequency was 24.49%. No woman with homozygous CYP1A1 m1/m1 genotype was detected as well. No significant difference in the frequencies of the GSTM1 and GSTT1 null alleles, and the CYP1A1 m1 polymorphism, was found between cases and controls. After application of Mantel-Haenszel chi procedure, there was no linear severity of the lesion and the frequency of these polymorphisms. According to our results, glutathione-S-transferase T1 and M1 and cytochrome P1A1 genetic polymporphisms do not appear to be a risk factor for cervical disease irrespective of smoking habits.

Expression of the HER family mRNA in breast cancer tissue and association with cell cycle inhibitors p21(waf1) and p27(kip1).

BACKGROUND: The HER family of the receptor tyrosine kinases epidermal growth factor receptor (EGFR), HER2, HER3 and HER4 are involved in the pathogenesis of many human malignancies. Although there is extensive literature on the expression of single HER-2 and EGFR receptors in breast cancer, little is known concerning the simultaneous expression at the mRNA level of these four receptors in human breast tissue and their influence in downstream signaling pathways that control cell cycle and proliferation. MATERIALS AND METHODS: The mRNA expression pattern of the four HER-receptors has been investigated and correlated with the expression of the cyclin-dependent kinase (CDK) inhibitors p21(Waf1) and p27(Kip1) in 67 breast cancer specimens. RESULTS: A positive correlation between HER-3 and HER-4 mRNA levels and a negative correlation between HER-2 and HER-3 was found. Compared to normal breast tissue, all four receptors were overexpressed in breast tumors and the strongest overexpression was found for HER-3 (p = 0.001). HER-4 expression was inversely related to histopathological grading (HPG), suggesting that elevated HER-4 mRNA expressions could be a biological marker of a more differentiated phenotype. The expression of p21(Waf1) protein was higher in HER-2-negative tumors, compared to HER-2-positive breast carcinomas. Compared to normal breast tissue, p21delta, the 19 kDa degraded form of p21 protein, was markedly expressed in breast cancer (p < 0.001). Conversely, p27(Kip1) was positively associated with HER-2 receptor and inversely associated with HER-3. CONCLUSION: The HER family members are overexpressed in breast cancer. Complex patterns of HER family expression were observed and the effect on cell cycle regulation was dependent on that pattern.

Characterization of a de novo balanced 1;Y translocation in a phenotypically normal twin male infant.

OBJECTIVE: To report a translocation between an autosome and the Y chromosome. DESIGN: Amniocentesis of a fetus because of mother's advanced age followed by karyotype and PCR analysis. SETTING: Tertiary health center. PATIENT(S): A phenotypically normal twin male infant. INTERVENTION(S): Karyotype with G and Q banding and amplification of testis-specific protein 1-Y and of azoospermia factor (AZF) a, AZFb, AZFc, and distal AZFc regions of Y chromosome. MAIN OUTCOME MEASURE(S): Karyotype, PCR. RESULT(S): We report a phenotypically normal twin male infant with de novo 46,ChiY,t(1;Y)(p22;p11) that was found in amniocentesis. In genetic counseling, it was recommended that the fetus be monitored through a detailed prenatal ultrasonographic examination, which did not indicate any pathological findings. A phenotypically normal male baby was born who is now a 12-month-old healthy infant. The karyotype was confirmed in the peripheral blood with G and Q banding. Amplification of testis-specific protein 1-Y, AZFa, AZFb, AZFc, and distal AZFc regions of the Y chromosome did not reveal any deletions. CONCLUSION(S): We cannot predict whether this male infant will have oligospermia or azoospermia as an adult and, furthermore, whether in case of fertility there is a risk for unbalanced autosome;Y translocations in the offspring, with congenital malformations and dysmorphic features. This case illustrates the complexities in counseling for prenatally diagnosed de novo autosome;Y translocations and the need for additional cases to be reported.

Association of the Tau haplotype with Parkinson's disease in the Greek population.

We compared the distribution of the Tau H1 haplotype and related subhaplotypes in a group of clinically diagnosed Parkinson's disease patients (n = 133) and in control individuals (n = 113) from northern Greece. We were able to detect a statistically significant overrepresentation of the H1H1 genotype in our patient group (OR for H1H1 vs. H1H2 and H2H2: 1.73; 95% CI: 1.03-2.90; P = 0.037). The H1 subhaplotype significantly associated with the disease in our population was different from the one previously reported for a Norwegian population, suggesting that the nature of the association of Tau with Parkinson's disease is influenced by ethnic variation.

Synergistic effects of protein tyrosine kinase inhibitor genistein with camptothecins against three cell lines in vitro.

Genistein, a natural isoflavone product has been shown to induce cellular death and increase the apoptotic cell death induced by several DNA-damaging stimuli. We have explored the combined effect of genistein and camptothecins against three cell lines, HeLa (cervical cancer), OAW-42 (ovarian cancer) and L929 (normal fibroblasts). Combined effect was estimated in 96-well plates using the SRB method and median-effect analysis. Addition of genistein synergistically increased the antiproliferative affect of camptothecins, inhibiting the camptothecin-induced G2/M arrest and increasing the apoptotic cell population. In HeLa cells, genistein inhibited CDK1 phosphorylation after irinotecan treatment. Thus, abrogation of the G2/M checkpoint control by genistein may be a useful maneuver to increase cytotoxicity of agents that damage DNA and inhibit cell-cycle progression in the G2/M boundary.

Phospholamban gene mutations are not associated with hypertrophic cardiomyopathy in a Northern Greek population.

Hypertrophic cardiomyopathy (HCM) is a genetically transmitted cardiac disease characterized by unexplained myocardial hypertrophy and diverse clinical spectrum. Currently, more than 250 HCM-related mutations in 10 genes encoding contractile sarcomeric proteins have been identified. Phospholamban (PLN) is a modest modulator of intracellular Ca2+ homeostasis and may be a candidate gene responsible for cardiomyopathy. In this study 53 consecutive patients with HCM, coming from Northern Greece, were screened for mutations of PLN gene. The patients were evaluated by clinical history, physical examination, electrocardiogram and echocardiography. All PCR products were analyzed for mutation by both restriction analysis and sequencing. The systematic mutation screening did not reveal any mutation in exons 1 and 2 or in the promoter region of phospholamban gene. Additionally, no polymorphisms were detected in all patients. Therefore, PLN gene mutations were not found to be associated with HCM in a Northern Greece population.

Detection and typing of human papillomavirus DNA in uterine cervices with coexistent grade I and grade III intraepithelial neoplasia: biologic progression or independent lesions?

OBJECTIVE: To examine the HPV type infection of cervical cone specimens with coexistent CIN1 and CIN3 lesions, in order to define if coexistence of low- and high-grade lesions in the same cervix represent different stages of evolution in a continuing process that is caused by a single viral type or independent lesions induced by different HPV types. STUDY DESIGN: The examined material included 43 cases with coexistent CIN1 and CIN3 in the cone biopsy specimen. Detection and typing of HPV was made by RFLP-PCR. RESULTS: All CIN1 lesions were HPV positive, while three CIN3 lesions were HPV-negative. The proportion of agreement of the HPV type in the two lesions, excluding negative cases (n = 40), was 60% (95% confidence interval: 43.3-75.1). HPV 16 was the most common type in both CIN3 (56.8%) and CIN1 (46.5%). CONCLUSIONS: The so-called morphologic progression of CIN is not always synonymous with biologic progression, since many coexistent CIN lesions are caused by different HPV types, and so represent different cell clones. Clonality of coexistent CIN lesions may be implicated in the evolution of CIN as other recent studies have shown.

Cholesteryl ester transfer protein TaqI B and lipoprotein lipase Ser447Ter gene polymorphisms are not associated with ischaemic stroke in Greek patients.

Cholesteryl ester transfer protein (CETP) and lipoprotein lipase (LPL) are both key players in plasma lipoprotein homeostasis and, as such, genetically induced alterations in their respective activities may affect susceptibility to cerebrovascular diseases. In this study, we examined the distribution of two common polymorphisms, namely CETP TaqI B and LPL Ser447Ter in a cohort of Greek clinically diagnosed late-onset ischaemic stroke patients (n = 98) and an ethnicity-, age- and sex-matched control group with no manifestations of vascular disease (n = 100). Our study revealed no statistically significant differences with respect to the distribution of either polymorphism, examined separately or in combination, between the two groups.

The association of p53 mutations and p53 codon 72, Her 2 codon 655 and MTHFR C677T polymorphisms with breast cancer in Northern Greece.

The aim of this study was to explore a possible association between p53 codon 72, Her 2 codon 655 and MTHFR C677T polymorphisms and breast cancer in Northern Greece. We examined 42 women with breast cancer and 51 controls. A total of 42 women with breast cancer as well as healthy controls were investigated and results showed that p53 codon 72 polymorphism is statistically significantly associated with breast cancer (OR for Arg/Arg to non-Arg/Arg was 6.66, P = 0.0001 at 95% CI 2.63-16.9), but not Her 2 and MTHFR polymorphisms are associated with breast cancer (OR for Ile/Ile to non-Ile/Ile was 1.33, P = 0.54 at 95% CI 0.52-3.38 and OR for T/T versus non-T/T was 1.07, P = 0.89 at 95% CI 0.35-3.25). All subjects were examined for p53 exons 5-8 mutations. Three novel sequence variations in exons 7 and 8 of TP53 gene were found in three patients. One of them induces an amino acid change at Ser 241Gly, the second is a silent mutation Gly244Gly, and the third one results in a premature stop codon 294 (Glu294stop) and a truncated p53 protein.

An association study of the tumor necrosis factor alpha C-850T polymorphism and childhood acute lymphoblastic leukemia in a population from northern Greece.

Excessive production of tumor necrosis factor alpha (TNFalpha) may influence the risk and/or progression of hematologic malignancies and has been associated with febrile episodes at diagnosis. We have examined the putative association of the C-850T polymorphism of the human TNFalpha gene with childhood acute lymphoblastic leukemia (ALL) and with clinical and laboratory findings at diagnosis, in 58 childhood ALL patients from northern Greece. No statistically significant associations have emerged between this polymorphism and either the risk for childhood ALL or presence of fever, anemia, leukocytosis and leukopenia at diagnosis, in this study.

An association study of the cholesteryl ester transfer protein TaqI B polymorphism with late onset Alzheimer's disease.

Cholesteryl ester transfer protein (CETP) is reportedly able to affect the amount of cholesterol available for deposition and/or removal from peripheral tissues, in its capacity to mediate the transfer of cholesterol from high density lipoprotein (HDL) to very low density lipoprotein, in exchange for triacylglycerols from the latter. The TaqI B polymorphism of the human CETP gene has been associated with decreased CETP mass and an increase in HDL-cholesterol. While many studies have addressed the atherogenic or anti-atherogenic potential of this polymorphism, little is known about its effect on neurodegeneration, despite the fact that CETP is expressed in the brain and the disturbance of cholesterol homeostasis appears to be an important factor in the pathogenesis of Alzheimer's disease (AD). In this report, we have compared the distribution of the TaqI B polymorphism in an independent population of 102 clinically diagnosed late onset AD patients and a spousal control group of 97 individuals. We have also examined the possible interaction between this polymorphism and two other polymorphisms suspected of affecting cholesterol flux, namely apolipoprotein E APOE epsilon4, and lipoprotein lipase LPLS447X. No statistically significant differences have emerged with respect to either genotype or allele frequencies between the AD and control populations. CETP TaqI B did not interact significantly with either APOE epsilon4 or LPLS447X, in this study.

An association study of a functional catalase gene polymorphism, -262C-->T, and patients with Alzheimer's disease.

According to the oxidative stress hypothesis which has been proposed as one of a number of possible mechanisms underlying pathogenesis of Alzheimer's disease (AD), accumulation of hydrogen peroxide in the brain of affected individuals, due to overproduction and/or insufficient detoxification, can trigger a cascade of neurotoxic events, thus contributing to the neuronal damage characteristic of the disease. The upregulation of enzymes that are able to neutralize hydrogen peroxide (catalase, peroxidases) would then be conceivably able to offer at least some protection from the damaging effects of this agent. In this study we examined the distribution of a functional polymorphism in the gene for catalase, -262C-->T, in an independent population of 137 AD patients and 130 control individuals. The presence of the polymorphism, which results in the elimination of a SmaI restriction site, was tested with a PCR amplification/SmaI digestion-based assay. No significant difference has emerged from the comparison of either genotype or allele frequencies (P>0.5). We conclude that the catalase gene -262C-->T polymorphism does not confer a protective effect with respect to AD.

Subspecies variation in Greek strains of Chlamydophila abortus.

The Greek chlamydial strains FAS, FAG, VPG and LLG, isolated from aborted sheep or goat foetuses, had been previously characterized as divergent on the basis of mouse cross-protection experiments, with LLG and its homologous POS significantly different from the rest in inclusion morphology, polypeptide profiles and reactivity with monoclonal antibodies. To determine the genetic basis of their divergence the 16S-23S ribosomal intergenic spacer was analysed by RFLP analysis of PCR 16SF2/23R amplicons. Using the restriction enzymes BfaI, SfcI, HpaI, BclI, DdeI and AclI, the strains were classified as Chlamydophila abortus. However, digestion with RsaI made it possible to differentiate strains FAS, FAG and VPG from strains LLG and POS, generating DNA fragments of 530/55 and 585bp, respectively. By subsequent sequence analysis of the 23S domain I rRNA gene only strain FAS was identical to reference strain A22 of C. abortus. Strains FAG and VPG presented an identical nucleotide deviation at position 593 of signature sequences. Strains LLG and POS presented three identical nucleotide deviations at positions 156, 186 and 307. Variation within the domain I signature sequences for the examined abortion strains was < or =0.69%. In conclusion, substantial genetic and biological diversity among strains of C. abortus was demonstrated, suggesting that subspecies variation status for certain strains may be applicable. Our findings suggest that differentiation may be possible at a subspecies level by RFLP analysis.