Vitamin D-neutralizing CYP24A1 expression, oncogenic mutation states and histological findings of human papillary thyroid cancer.

OBJECTIVE: The aims of the present study were to examine gene and protein expression of the vitamin D-inactivating 24-hyroxylase (CYP24A1) and the activating 1-alpha-hydroxylase (CYP27B1) enzyme in human papillary thyroid cancer (PTC), furthermore, to investigate the association between CYP24A1 expression and numerous clinical, histological parameters and somatic oncogene mutation status of thyroid tumor tissues. MATERIALS AND METHODS: Gene expression analysis was carried out in 100 Hungarian thyroid samples, both normal and papillary tumor tissue sections of the same patient. The specific mRNA to the selected genes was analyzed by TaqMan probe-based quantitative real-time RT-PCR. The somatic oncogene mutation states of BRAF, NRAS, HRAS and KRAS were also tested. RESULTS: CYP24A1 mRNA expression was markedly increased in 52 cases (52%) of the examined papillary cancers compared with that of normal thyroid tissue. There was a tendency toward difference in the distribution of high-level CYP24A1 in the PTC accompanied with somatic oncogene mutation. Positive correlation was seen between increased CYP24A1 expression rate and a group of variables reflecting tumor malignity (mainly vascular invasion, lymph node metastasis, tumor size, hypothyreosis) by principal components analysis. No significant alteration was seen in CYP27B1 gene expression between neoplastic and normal tissues. CONCLUSIONS: A definite alteration was seen in vitamin D3-inactivating CYP24A1 gene activity in PTC compared to their normal tissues on a relatively large patient population. Our findings raise the possibility that CYP24A1 may also directly be involved in thyroid carcinogenesis.

Long term efficacy of radioiodine treatment in hyperthyroidism.

OBJECTIVES: Radioiodine is the mainstay of the treatment of thyroid hyperfunction. However, it is difficult to apply the appropriate amount of radioidone to achieve optimal efficacy with the least possible adverse effects. Results of the investigation on the efficacy of a relatively new protocol for radioiodine treatment of hyperthyroidism are reported. DESIGN: A retrospective evaluation of data from 326 patients with a mean average follow-up of 5.7 (1.0-11.7) years was performed. 64% of these patients suffered from Graves' disease and 36% had uni- or multinodular toxic goitre. RESULTS: In Graves' disease, the recurrence rate was 5% 1 year after the treatment, and that remained the same after 5 years. In toxic goitre, these rates were 6 and 7%, respectively. After 5 years 70% of the patients with autonomous adenomas were euthyroid, while 78% of the Graves patients developed hypothyroidism and 17% showed euthyroid state. A relationship between the lack of normalisation of thyroid-stimulating hormone levels after radioiodine treatment and the increased recurrence of late hyperthyroidism has also been established in patients with Graves's disease. CONCLUSION: Compared to the available data published in the literature, the success rate of the treatment is fairly high confirming the effectiveness of our protocol.

New approach to analyze genetic and clinical data in bisphosphonate-induced osteonecrosis of the jaw.

OBJECTIVES: Osteonecrosis of the jaw (ONJ) is a major complication associated with long-term use of bisphosphonates (BP). We aimed to investigate the effect of CYP2C8 rs1934951 SNP and its relationship to a number of clinical and biochemical factors in 46 Hungarian subjects with bisphosphonate-induced ONJ. METHODS: Blood samples were collected from each subject and genomic DNA was extracted. SNP analysis of CYP2C8 gene was carried out by predesigned TaqMan primer/probe sets. The genetic data together with clinical and biochemical variables were evaluated by chi-square test, logistic regression, and principal component analysis (PCA). RESULTS: The risk of mandibular localization of ONJ was 19.2-fold higher in subjects with AG genotype than in normal GG genotype. PCA revealed strong positive correlations between maxillar localization of ONJ and a group of variables including intravenous BP application and serum lipid markers. Mandibular localization of ONJ was correlated positively with serum calcium, 25-hydroxy-vitamin D and PTH levels, oral BP application, and the length of BP therapy. The degree of the disease and the number of recurrences were correlated with the application of hormone-deprivation therapy for breast cancer patients. CONCLUSION: The statistical approach applying PCA to our data may contribute to the better understanding of factors playing role in the development of bisphosphonate-induced ONJ.

LCT 13910 C/T polymorphism, serum calcium, and bone mineral density in postmenopausal women.

SUMMARY: LCT 13910 CC genotype is associated with lactose intolerance, a condition often resulting in reduced milk intake. Women with the CC genotype were found to have decreased serum calcium and reduced bone mineral density. INTRODUCTION: The CC genotype of the 13910 C/T polymorphism of the LCT gene is linked to lactose intolerance and low calcium intake. METHODS: We studied 595 postmenopausal women, including 267 osteoporotic, 200 osteopenic, and 128 healthy subjects. Genotyping, osteodensitometry, and laboratory measurements were carried out. RESULTS: Frequency of aversion to milk consumption was 20% for CC genotype and 10% for TT + TC genotypes (p = 0.03). The albumin-adjusted serum calcium was 2.325 +/- 0.09 mmol/L for CC genotype and 2.360 +/- 0.16 mmol/L for TT + TC genotypes (p = 0.031). Bone mineral density (BMD; Z score) was lower in the CC than TT + TC genotypes, respectively, at the radius (0.105 +/- 1.42 vs 0.406 +/- 1.32; p = 0.038), at the total hip (-0.471 +/- 1.08 vs -0.170 +/- 1.09; p = 0.041), and at the Ward's triangle (-0.334 +/- 0.87 vs -0.123 +/- 0.82; p = 0.044). CONCLUSION: LCT 13910 C/T polymorphism is associated with decreased serum calcium level and lower BMD in postmenopausal women.

CYP3A7*1C polymorphism, serum dehydroepiandrosterone sulfate level, and bone mineral density in postmenopausal women.

The CYP3A7 enzyme metabolizes some steroid hormones, including dehydroepiandrosterone sulfate (DHEAS). The age-related decline of serum DHEAS levels is believed to contribute to osteoporosis. Previously, the CYP3A7*1C polymorphism has been shown to cause a persistent high CYP3A7 enzyme activity, resulting in lower levels of DHEAS in men. We hypothesized that the CYP3A7*1C polymorphism might contribute to bone loss through decreased levels of serum DHEAS in postmenopausal women. Postmenopausal women (n = 319) were divided into two subgroups: 217 with osteoporosis and 102 healthy controls. Genotyping, serum DHEAS measurement, and osteodensitometry of the lumbar spine and femoral neck were carried out in all subjects. Homozygous CYP3A7*1C carriers had significantly lower BMD at the lumbar spine compared to wild types (T score -3.27 +/- 1.02 in CYP3A7*1C homozygous mutants vs. -1.35 +/- 1.53 in wild types, P = 0.041). This association remained significant after adjustment for menopausal age, serum DHEAS level, alcohol consumption, steroid intake, smoking habits, and previous fractures. No association was found between genotypes and serum DHEAS levels in the total study population or in the subgroups. Serum DHEAS levels correlated positively with bone mineral density at the lumbar spine (r = 0.59, P = 0.042) after correction for age. Our data suggest that the CYP3A7 polymorphism might have an influence on bone mass at the lumbar spine independently of serum DHEAS concentrations.

Inhibition of C1q-beta-amyloid binding protects hippocampal cells against complement mediated toxicity.

Activation of complement by beta-amyloid (A beta) contributes to the pathology of Alzheimer's disease (AD). Here, we show that C1-Inhibitor (C1-Inh) protects cultured rat hippocampal cells against beta-amyloid induced complement lysis indicating a classical pathway-mediated activation mechanism. We report on screening of compound libraries to identify compounds that inhibit C1q binding to beta-amyloid. Characterization of these compounds revealed that C1q possessed distinct binding sites for beta-amyloid and antibodies. One selected compound protected cultured hippocampal cells against complement-dependent beta-amyloid toxicity. These results provide evidence that complement has the potential to damage hippocampal cells, and C1q is a relevant target to suspend this deleterious mechanism in Alzheimer's disease.

Cost-effectiveness of enoxaparin as thromboprophylaxis in acutely ill medical patients in Spain.

OBJECTIVE: The objective of this study was to determine the cost-effectiveness of thromboprophylaxis with enoxaparin versus no thromboprophylaxis in patients with acute medical illness in Spain from the society perspective. METHODS: Markov process analysis techniques were used to model the health economic outcomes. Clinical data were derived mainly from the MEDENOX trial, while health-care utilization was derived from Delphi panels. RESULTS: An analysis over the MEDENOX trial period shows that the cost per event avoided is currency 432, while the cost per life saved is currency 1527. The cost per event includes all medical resource utilization costs associated with the event. The lifetime model, which assumes no higher risk for recurrence of venous thromboembolism (VTE) and mortality in asymptomatic patients, shows that the use of enoxaparin leads a cost per event avoided of currency 270 and cost per life-year gained of currency 71. If the lifetime model assumes a higher risk for recurrence of VTE in asymptomatic patients, enoxaparin is dominant over no thromboprophylaxis. CONCLUSION: The results showed that the favorable clinical benefit of enoxaparin as thromboprophylaxis in patients with acute medical illness, which was observed in the MEDENOX trial, results in a positive health economic benefit in both the short term and the long term in the health-care setting of Spain.

Modeling the cost-effectiveness and budgetary impact for subpopulations.

There is currently a trend to increasing demand for health-economic and budgetary-impact data in the decision-making process in Europe. A parallel development is the tendency to restrict the prescription of new drugs to subpopulations that may depend on the results of the above health-economic analysis and financial analysis. We present modeling techniques for determining the optimal subpopulation considering the cost-effectiveness and budgetary impact of a new drug. The methodology consists of incorporating confounding variables into the Markov health states by means of health state specific regression equations for costs and utilities. The strategy is applied to a hypothetical Markov model for new product in Parkinson's disease. The results of the presented analyses suggest that within the registered range of indications a further restriction in the application for a new drug can be made from the point of view of cost-effectiveness and budgetary impact. These results can also be considered in the decision-making process.

Effect of transforming growth factor-beta1 on microglial MHC-class II expression.

In the present report, the effects of IFN-gamma and transforming growth factor beta1 (TGF-beta1) on major histocompatibility complex class II (MHC-II) gene expression in isolated mouse brain microglial cells, in the MH-S macrophage cell line and in the primary mouse macrophage cultures were examined. IFN-gamma is a potent inducer of MHC-II gene and this induction was further elevated in microglia by TGF-beta1, while TGF-beta1 inhibited IFN-gamma, induction in macrophages. The enhancing effect of TGF-beta1 was also detected in microglia at the protein level. Transient transfection of microglia with 5' deletional mutants of the MHC-II IAalpha promoter linked to the chloramphenicol acetyltransferase reporter gene demonstrated that TGF-beta1 acts at the transcriptional level to enhance the MHC-II expression induced by IFN-gamma.

The E249K mutator mutant of DNA polymerase beta extends mispaired termini.

The DNA polymerase beta mutant enzyme, which is altered from glutamic acid to lysine at position 249, exhibits a mutator phenotype in primer extension assays and in the herpes simplex virus-thymidine kinase (HSV-tk) forward mutation assay. The basis for this loss of accuracy was investigated by measurement of misincorporation fidelity in single turnover conditions. For the four misincorporation reactions investigated, the fidelity of the E249K mutant was not significantly different from wild type, implying that the mutator phenotype was not caused by a general inability to distinguish between correct and incorrect bases during the incorporation reaction. However, the discrimination between correct and incorrect substrates by the E249K enzyme occurred less during the conformational change and chemical steps and more during the initial binding step, compared with pol beta wild type. This implies that the E249K mutation alters the kinetic mechanism of nucleotide discrimination without reducing misincorporation fidelity. In a missing base primer extension assay, we observed that the mutant enzyme produced mispairs and extended them. This indicates that the altered fidelity of E249K could be due to loss of discrimination against mispaired primer termini. This was supported by the finding that the E249K enzyme extended a G:A mispair 8-fold more efficiently than wild type and a C:T mispair 4-fold more efficiently. These results demonstrate that an enhanced ability to extend mispairs can produce a mutator phenotype and that the Glu-249 side chain of DNA polymerase beta is critical for mispair extension fidelity.

Differential regulation of major histocompatibility complex class II expression and nitric oxide release by beta-amyloid in rat astrocyte and microglia.

Astrocytes and microglial cells were examined for expression of two immunologically important molecules, major histocompatibility complex class II (MHC-II) and nitric oxide (NO) following treatment with IFN-gamma and beta-amyloid (betaA) peptides, betaA(1-42) and betaA(25-35). IFN-gamma is a potent inducer of both MHC-II gene expression and NO production. The induction of MHC-II was inhibited by both betaA peptides in astrocytes but they had little or no effect in microglia. betaA peptides had no effect on NO release in astrocytes but on microglia betaA(1-42) synergistically induced NO release with IFN-gamma. Transient transfection of astrocytes with 5' deletional mutants of MHC-II IAalpha promoter linked to the chloramphenicol acetyl transferase reporter gene (IAalpha-CAT), demonstrated that betaA acts at the transcriptional level to downregulate IFN-gamma induced MHC-II gene expression in astrocytes. In previous studies, the induction of MHC-II on glial cells were suggested to be involved in the pathogenesis of neurodegenerative diseases and MHC-II(+) microglial cells were observed at much higher frequency than astrocytes. This study provides information on the regulation of the MHC-II gene expression in astrocytes and in microglial cells by betaA and this pathway may be critically involved in the immune/inflammatory regulation within the central nervous system.

3'-Azido-3'-deoxythymidine-resistant mutants of DNA polymerase beta identified by in vivo selection.

We developed an in vivo selection to identify 3'-azido-3'-deoxythymidine (AZT)-resistant mutants of rat DNA polymerase beta (pol beta). The selection utilizes pol beta's ability to substitute for Escherichia coli DNA polymerase I (pol I) in the SC18-12 strain, which lacks active pol I. pol beta allows SC18-12 cells to grow, but they depend on pol beta activity, so inhibition of pol beta by AZT kills them. We screened a library of randomly mutated pol beta cDNA for complementation of the pol I defect in the presence of AZT, and identified AZT-resistant mutants. We purified two enzymes with nonconservative mutations in the palm domain of the polymerase. The substitutions D246V and R253M result in reductions in the steady-state catalytic efficiency (Kcat/Km) of AZT-TP incorporation. The efficiency of dTTP incorporation was unchanged for the D246V enzyme, indicating that the substantial decrease in AZT-TP incorporation is responsible for its drug resistance. The R253M enzyme exhibits significantly higher Km(dTTP) and Kcat(dTTP) values, implying that the incorporation reaction is altered. These are the first pol beta mutants demonstrated to exhibit AZT resistance in vitro. The locations of the Asp-246 and Arg-253 side chains indicate that substrate specificity is influenced by residues distant from the nucleotide-binding pocket.

[Accidents of the elderly]. / Idóskori balesetek.

The topic of accidents in elderly population has been studied on 2055 injured older than 65 years. The severity of injuries indicates the importance of accidents in this age-group. The proportion of fractures in the elderly population is twice of the average-in females over 75 years of age they take the half of the total number of injuries. While the incidence rate of males in the whole material with all of the age groups is twice of that of females, in elderly population it turns into equal. The rate of home-accidents grows with ageing. The elderly people specially the males suffer a high number of road traffic accidents as pedestrians or riding their bicycles. The severity of injuries and the broken health status in elderly people is a major problem both for the health care and the whole society.

Structure of the cysteine-rich intestinal protein, CRIP.

LIM domains are Zn-binding arrays found in a number of proteins involved in the control of cell differentiation, including several developmentally regulated transcription factors and a human proto-oncogene product. The rat cysteine-rich intestinal protein, CRIP, is a 76-residue polypeptide which contains a LIM motif. The solution structure of CRIP has been determined by homonuclear and 1H-15N heteronuclear correlated nuclear magnetic resonance spectroscopy. Structures with individual distance violations of < or = 0.03 angstrom and penalties (squared sum of distance violations) of < or = 0.06 angstrom2 were generated with a total of 500 nuclear Overhauser effect (NOE)-derived distance restraints (averaging 15.6 restraints per refined residue). Superposition of backbone heavy atoms of ordered residues relative to mean atom positions is achieved with pairwise rms deviations of 0.54(+/-0.14) angstrom. As observed previously for a peptide with the sequence of the C-terminal LIM domain from the avian cysteine-rich protein, CRP (cCRP-LIM2), CRIP binds two equivalents of zinc, forming N-terminal CCHC (Cys3, Cys6, His24, Cys27) and C-terminal CCCC (Cys30, Cys33, Cys51, Cys55) modules. The CCHC and CCCC modules in CRIP contain two orthogonally-arrayed antiparallel beta-sheets. The C-terminal end of the CCHC module contains a tight turn and the C terminus of the CCCC module forms an alpha-helix. The modules pack via hydrophobic interactions, forming a compact structure that is similar to that observed for cCRP-LIM2. The most significant differences between the structures occur at the CCHC module-CCCC module interface, which results in a difference in the relative orientations of the modules, and at the C terminus where the alpha-helix appears to be packed more tightly against the preceding antiparallel beta-sheet. The greater abundance of NOE information obtained for CRIP relative to cCRP-LIM2, combined with the analysis of J-coupling and proton chemical shift data, have allowed a more detailed evaluation of the molecular level interactions that stabilize the fold of the LIM motif.

[Injuries during adolescence]. / Sérülések a serdülökorban.

The incidence of injuries reaches the highest level in the adolescent age-group, as proved also in our survey of accidents in a period of one year in County Vas, Hungary. The incidence is extremely high in boys: 241 per 1000 heads of this age-group while no increase can be observed in girls due to an earlier puberty. The risk-rate of boys appears high at every type of injuries even at those that were caused intended or in an intoxicated state. As to the rest of indices the age-group of adolescents shows a transition between the childhood and the adult age, meanwhile the injuries of the adolescents stay nearer to those of the young adults.

Common metal ion coordination in LIM domain proteins.

The LIM motif is a cysteine- and histidine-rich sequence that was first identified in proteins involved in control of gene expression and cell differentiation. In order to characterize structural features of the LIM domain, we have carried out biophysical studies on two polypeptides that display LIM domains: the cysteine-rich intestinal protein (CRIP) and a fragment of the cysteine-rich protein (CRP). Bacterial expression vectors were constructed for the intact CRIP molecule and the C-terminal half of CRP, designated LIM2, such that each expressed protein contained a single LIM domain. Both proteins were recovered as soluble, Zn(II)-containing proteins. The metal coordination properties of these two distinct LIM domain proteins were highly similar, suggesting that a common structural architecture may exist in LIM domain proteins. Both proteins exhibit a maximum of two tetrahedrally bound Zn(II) ions per molecule. Electronic spectroscopy of Co(II) complexes and 113Cd NMR of Cd(II) complexes of CRIP and LIM2 revealed a similar ligand field pattern with one tetrathiolate (S4) site and one S3N1 site for divalent metal ions. The nitrogen ligand was shown to arise from a histidyl imidazole by heteronuclear multiple quantum coherence NMR. The eight conserved residues within the LIM domains of CRIP and LIM2 include seven cysteines and one histidine. It is likely that these conserved residues generate the S4 and S3N1 Zn(II)-binding sites. Metal binding to the two sites within a single LIM domain is sequential, with preferential occupancy of the S4 site. Slow metal ion exchange occurs between sites within an LIM domain, and metal exchange with exogenous metal ions is observed, with exchange at the S3N1 site being kinetically more facile. In the absence of metal binding both proteins appear to be substantially unfolded. Metal binding stabilizes a tertiary fold containing appreciable secondary structural elements. The common metal ion coordination in CRIP and LIM2 suggests that the LIM motif may constitute a structural module with conserved features.

[Significance of home accidents]. / Az otthoni balesetek jelentösége.

Authors analyse the incidence, demographic relations and data of hospitalization of the home accidents. The significance of the home accidents is emphasized. It is stated that the home accident is the most frequent type of accidents and its ratio is increasing. The incidence was found 45 on 1000 inhabitants, the home accidents give nearly the half of all accidents and more than 1/3 of the hospitalized injured. Attention is called to the possibilities of unutilized possibilities of the prevention of home accidents.

[Use of preserved homograft for covering burn injuries]. / Konzervbör-homograft alkalmazása égési sérülteken.

Authors describe the considerations supporting the use of cadaveric skin homografts. The surgical relations of the burns, the site and advantages of the application of the homografts in the treatment of the burned patients are summarized.

[Childhood accidents]. / Gyermekbalesetek.

A survey of accidents in a period of one year in County Vas, Hungary was performed. The injury of children in the age-group of 0-14 was found in case 5841 i.e. in 22.5% of the total number of the injured. The incidence was 101/1000 head of puerile population. The rate of accidents appeared to be low in early childhood while it showed a significant increase in the school age. The home accidents accounted for the most of the accidents especially in the early childhood. Over the age of 7 the school and sport/playground accidents exceeded. The roads were involved as a high risk factor for each of the age groups. The high (21.5%) rate of fractures indicates severity of the injuries although the number of the most severe injuries remained low. The accident conditions of County Vas are at an advantage compared with international data meanwhile the high rate of accidents in the school age group calls for prevention and educational work.

[Emergency treatment of severe burns in daily practice]. / Súlyos égési sérültek elsö ellátása a napi gyakorlatban.

The paper examines the initial treatment of the severely burned patient to be transferred. It is stated that the aspirations for a uniform expert treatment have not yet brought a complete success. The suggestions concerning this problem, described in the Methodological Letter "Treatment of Thermic Injuries" issued by the Nat. Institute of Traumatology are discussed and repeated.