Dietary patterns and body adiposity in children in Brazil: a cross-sectional study.

OBJECTIVES: Many factors are associated with the increase in total and central body adiposity in children, especially eating habits. This has led to an increasing number of studies analysing food patterns, which consider the synergistic effect of food and nutrient intake on the nutritional status. The objective of this study was to identify the dietary patterns of children aged 4-7 years and associate these with different indicators of total and central body adiposity. STUDY DESIGN: Cross-sectional study METHODS: The study consisted of 403 children from a retrospective cohort in Minas Geraes, Brazil. Four indicators of body adiposity were evaluated: body mass index (BMI), waist-to-height ratio (WHtR) and percentages of total and central body fat (assessed by dual-energy X-ray absorptiometry). The dietary habits of the children were evaluated by identifying the dietary patterns using principal component analysis. The adjustment predictor variables were related to the socio-economic characteristics, lifestyle and duration of exclusive breastfeeding. Food patterns were identified by factor analysis. Linear regression was used to estimate the regression coefficient and the confidence interval, considering statistical significance of α = 5%. RESULTS: Five dietary patterns were identified, which explained 42.3% of the data variance: 'Traditional', 'Unhealthy', 'Milk and chocolate', 'Snack' and 'Healthy'. The multiple linear regression model showed that a greater adherence to the 'Traditional' and 'Unhealthy' patterns was related to higher BMI, WHtR, and total and central body adiposity. CONCLUSION: Children with a higher intake of food from the 'Traditional' and 'Unhealthy' patterns showed an increase in total and central body adiposity.

Breast-conservative surgery with and without radiotherapy in patients aged 55-75 years with early-stage breast cancer: a prospective, randomized, multicenter trial analysis after 108 months of median follow-up.

OBJECTIVES: Breast-conserving therapy (BCT), including postoperative whole breast irradiation (WBI), is generally accepted as the treatment of choice for most patients with early-stage breast cancer. The question whether WBI is mandatory in all patients remains one of the most controversial issues in BCT. To answer this question, a randomized, prospective, multicentre study was launched in January 2001. Primary endpoints of the study were to assess the cumulative incidence of in-breast-recurrences (IBR) and overall survival (OAS) after conservative surgery (BCS) with or without WBI. METHODS: From January 2001 until December 2005, 749 patients with unifocal infiltrating breast cancer up to 25 mm, 0-3 positive axillary lymph nodes, no extensive intraductal component or lymphvascular invasion from 11 centres in Italy, were randomly assigned to BCS+WBI (arm 1:373 patients) or BCS alone (arm 2:376 patients). Treatment arms were well balanced in terms of baseline characteristics. Systemic adjuvant therapy was administered according to the institutional policies. Kaplan-Meier method was used for survival analysis and log-rank test to evaluate the difference between the two arms. RESULTS (Last analysis 31.12.2012): After median follow-up of 108 months, 12 (3.4%) IBR were observed in arm 1 and 16 (4.4%) in arm 2. OAS was 81.4% in arm 1 and 83.7% in arm 2. There was no statistically significant difference regarding IBR and death in the two treatment groups. CONCLUSIONS: These data are promising and suggest that WBI after BCS can be omitted in selected patients with early stage breast cancer without exposing them to an increased risk of local recurrence and death. Longer follow-up is needed to further consolidate these results.

Arthroscopic treatment of glenohumeral instability in soccer goalkeepers.

The aim of this study was to report epidemiologic data and results of arthroscopic treatment of glenohumeral instability in soccer goalkeepers. We included 12 soccer goalkeepers with a mean age of 28.9 years (range 18-45 years) with acute or recurrent traumatic anterior instability who underwent an arthroscopic anatomic capsulolabral repair with bone anchors. Patients who underwent surgery within 4 weeks of the first episode of dislocation were classified as acute instability. The results were evaluated using the Rowe Scale and analyzed according to stability, range of motion and function. The mean follow-up was 3.8 years. The most common mechanism of injury (90% of the cases) was abduction, external rotation and extension. Associated injuries were present in 57.2% of recurrent cases and 20% of acute cases (p<0.293). Excellent or good results were observed in 80% of the cases of acute instability and in 57.2% of cases in the group with recurrent instability (p<0.586). From a total of 12 soccer goalkeepers who underwent the arthroscopic capsulolabral repair, good or excellent results were obtained in 66.6% of cases of glenohumeral instability. Surgical arthroscopic repair was possible in all cases of acute or recurrent instability based on well-established inclusion criteria, i. e., with well-defined exclusion criteria, such as HAGL lesion and significant glenohumeral bone loss, the arthroscopic capsulolabral repair can be carried out in soccer goalkeepers.

Follicular-fluid anti-Müllerian hormone concentration is similar in patients with endometriosis compared with non-endometriotic patients.

This cross-sectional prospective study assessed follicular-fluid anti-Müllerian hormone (AMH) concentrations in infertile patients with mild/minimal endometriosis during natural IVF. Thirty-two women participated in the study. Patients were divided into two groups: tubal obstruction without endometriosis (control group) and minimal/mild endometriosis (study group). All patients underwent laparoscopy for assessment of infertility; at the same time, any foci of endometriosis found were cauterized. AMH concentration was measured in the follicular fluid of a single follicle when it achieved pre-ovulatory maturation by ultrasensitive enzyme-linked immunosorbent assay. Likewise, AMH, FSH and inhibin B content in serum was also measured. Age (30 ± 1.3 and 32 ± 0.8 years) and body mass index (22 ± 0.6 and 22 ± 0.5 kg/m(2)), day-3 antral follicle count (11.3 ± 1.7 and 10.7 ± 1.5), serum FSH concentrations (5.4 ± 0.6 and 5.0 ± 0.3 IU/ml) and follicular-fluid AMH concentrations (1.8 ± 0.3 and 1.5 ± 0.1 ng/ml, study and control group, respectively; mean difference 0.33, 95% CI -0.21 to 0.88) were similar in both groups. This study shows that infertile patients with minimal/mild endometriosis have a similar concentration of AMH in the follicular fluid after natural IVF as compared with control subjects.

Ezrin NH2-terminal domain inhibits the cell extension activity of the COOH-terminal domain.

Overexpression in insect cells of the full coding sequence of the human membrane cytoskeletal linker ezrin (1-586) was compared with that of a NH2-terminal domain (ezrin 1-233) and that of a COOH-terminal domain (ezrin 310-586). Ezrin (1-586), as well as ezrin (1-233) enhanced cell adhesion of infected Sf9 cells without inducing gross morphological changes in the cell structure. Ezrin (310-586) enhanced cell adhesion and elicited membrane spreading followed by microspike and lamellipodia extensions by mobilization of Sf9 cell actin. Moreover some microspikes elongated into thin processes, up to 200 microns in length, resembling neurite outgrowths by a mechanism requiring microtubule assembly. Kinetics of videomicroscopic and drug-interference studies demonstrated that mobilization of actin was required for tubulin assembly to proceed. A similar phenotype was observed in CHO cells when a comparable ezrin domain was transiently overexpressed. The shortest domain promoting cell extension was localized between residues 373-586. Removal of residues 566-586, involved in in vitro actin binding (Turunen, O., T. Wahlström, and A. Vaheri. 1994. J. Cell Biol. 126:1445-1453), suppressed the extension activity. Coexpression of ezrin (1-233) with ezrin (310-586) in the same insect cells blocked the constitutive activity of ezrin COOH-terminal domain. The inhibitory activity was mapped within ezrin 115 first NH2-terminal residues. We conclude that ezrin has properties to promote cell adhesion, and that ezrin NH2-terminal domain negatively regulates membrane spreading and elongation properties of ezrin COOH-terminal domain.

MitoP2: the mitochondrial proteome database--now including mouse data.

The MitoP2 database (http://www.mitop.de) integrates information on mitochondrial proteins, their molecular functions and associated diseases. The central database features are manually annotated reference proteins localized or functionally associated with mitochondria supplied for yeast, human and mouse. MitoP2 enables (i) the identification of putative orthologous proteins between these species to study evolutionarily conserved functions and pathways; (ii) the integration of data from systematic genome-wide studies such as proteomics and deletion phenotype screening; (iii) the prediction of novel mitochondrial proteins using data integration and the assignment of evidence scores; and (iv) systematic searches that aim to find the genes that underlie common and rare mitochondrial diseases. The data and analysis files are referenced to data sources in PubMed and other online databases and can be easily downloaded. MitoP2 users can explore the relationship between mitochondrial dysfunctions and disease and utilize this information to conduct systems biology approaches on mitochondria.

Early detection of cancer: ideas for a debate.

Even if the overall number of cancer is increasing, the mortality has started to decrease in the Western World. The role of early detection in this decrease is a matter of debate. To assess its impact on mortality it is important to distinguish between diagnosis of cancer in symptomatic patients, and early detection in asymptomatic individuals who may self-refer or who may be offered ad hoc or systematic screening. The policies for early detection and screening vary greatly between European countries, despite many similarities in their cancer burden, and this partly reflects the uncertainties surrounding asymptomatic testing for cancer. A Task Force of European expert, held in Azzate (VA), Italy, established to address these issues, acknowledged the need for more research in the field of individual risk assessment since general statistics are more and more perceived as inadequate to design personal early detection plans. The group also recognised that combinations of early detection and screening will enforce the effectiveness of new treatments in curbing mortality curves, although policies will vary with different cancers.

Influence of DNA repair polymorphisms on biomarkers of genotoxic damage in peripheral lymphocytes of healthy subjects.

DNA repair polymorphisms may represent susceptibility factors affecting DNA integrity, and possibly cancer risk, in human population. In order to elucidate the influence of a few widely studied DNA repair polymorphisms on individual levels of DNA damage and their possible interaction with lifestyle and environmental exposures, 171 subjects from a well-characterized human population enrolled in a previous study on genetic effects of air pollution were genotyped for the XRCC1 Arg280His and Arg399Glu, XRCC3 Thr241Met and ERCC2 Lys751Gln polymorphisms. The association between DNA repair genotype, alone or in combination with metabolic genotype, on the levels of SCE, micronuclei and tail moment values in peripheral lymphocytes was evaluated. A significant influence of the ERCC2 genotype on SCE frequency was observed. Subjects with ERCC2 751 Gln/Gln genotype had significantly higher risk of high (above the median) SCE/cell with respect to Lys/Lys referents (OR 4.55, 95% CI 1.48-13.99). A non-significantly elevated OR was also observed in Gln/Lys heterozygotes, suggesting a gene dosage effect. When subjects were categorized by smoking habits and professional exposure, the variant ERCC2 751 Gln/Gln genotype was associated with elevated SCE rates in non-smokers and in exposed subjects, but not in smokers. The results of this study support the hypothesis that some DNA repair polymorphisms exert a modifying effect on individual levels of DNA damage in healthy subjects, possibly also modulating cancer risk.

MitoP2, an integrated database on mitochondrial proteins in yeast and man.

The aim of the MitoP2 database (http://ihg.gsf.de/mitop2) is to provide a comprehensive list of mitochondrial proteins of yeast and man. Based on the current literature we created an annotated reference set of yeast and human proteins. In addition, data sets relevant to the study of the mitochondrial proteome are integrated and accessible via search tools and links. They include computational predictions of signalling sequences, and summarize results from proteome mapping, mutant screening, expression profiling, protein-protein interaction and cellular sublocalization studies. For each individual approach, specificity and sensitivity for allocating mitochondrial proteins was calculated. By providing the evidence for mitochondrial candidate proteins the MitoP2 database lends itself to the genetic characterization of human mitochondriopathies.

Intravitreal aflibercept for macular oedema secondary to central retinal vein occlusion in patients with prior treatment with bevacizumab or ranibizumab.

PurposeTo report the visual and anatomic outcomes in eyes with macular oedema (MO) secondary to central retinal vein occlusion (CRVO) that were switched from either intravitreal bevacizumab or ranibizumab to intravitreal aflibercept.MethodsTwo-center retrospective chart review. Eyes with MO secondary to CRVO that received a minimum of three intravitreal injections of bevacizumab or ranibizumab and were switched to intravitreal aflibercept for persistent or recurrent MO not responding to either bevacizumab and/or ranibizumab.ResultsIn all 42 eyes of 42 patients were included in the study. The median visual acuity before the switch was 20/126, 1 month after the first injection of aflibercept 20/89 (P=0.0191), and at the end of the follow-up 20/100 (P=0.2724). The median CRT before the switch was 536 µm, 1 month after the first injection of aflibercept 293.5 µm (P=0.0038), and at the end of the follow-up 279 µm (P=0.0013 compared to before the switch). The median number of weeks between injections before the switch was 5.6 and after the switch was 7.6 (P<0.0001).ConclusionConverting eyes with refractory MO due to CRVO to aflibercept can result in stabilization of the vision, improved macular anatomy, and extension of the injection interval.

Identification of a phosphatidylinositol-4,5-bisphosphate-binding domain in the N-terminal region of ezrin.

Purified human recombinant ezrin cosediments with large liposomes containing phosphatidylserine (PS). This interaction is optimal at low ionic strength. At physiological ionic strength (130 mM KCl) ezrin interacts strongly with liposomes containing > or = 5% phosphatidylinositol-4,5-bisphosphate (PIP2), the residual being phosphatidylcholine (PC). When PIP2 is replaced by phosphatidylinositol-4-monophosphate (PIP), phosphatidylinositol (PI) or PS, the interaction is markedly reduced. Furthermore we show, that a purified N-terminal glutathione S-transferase (GST) fusion protein of ezrin (1-309) still has retained the capacity to interact with PIP2-containing liposomes, whereas a C-terminal fusion protein (310-586) has lost this ability.

Structure/function studies on cytoskeletal proteins in Dictyostelium amoebae as a paradigm.

The actin cytoskeleton in motile non-muscle cells is being regulated by a large number of actin-binding proteins. A deeper insight into the complex nature of the dynamic rearrangements of the microfilament system during cell movement requires an experimental system that allows the combined application of biochemical, biophysical, cell biological and molecular methods. Dictyostelium amoebae are well suited especially for a genetic approach because they are amenable to gene disruption, antisense and gene tagging techniques. The actin-binding proteins profilin, hisactophilin and protovillin are being described in this context as typical examples that either bind to G-actin, or anchor the actin cytoskeleton to the plasma membrane, or are structurally similar to vertebrate proteins but distinct in their functions.

Prediction of rodent carcinogenicity of further 30 chemicals bioassayed by the U.S. National Toxicology Program.

Recently the U.S. National Toxicology Program (NTP) sponsored a comparative exercise in which different prediction approaches (both biologically and chemically based) were challenged for their predictive abilities of rodent carcinogenicity of a common set of chemicals. The exercise enjoyed remarkable scientific success and stimulated NTP to sponsor a second challenging round of tests, inviting participants to present predictions relative to the rodent carcinogenicity of a further 30 chemicals; these are currently being tested. In this article, we present our predictions based on structure-activity relationship considerations. In our procedure, first each chemical was assigned to an activity mechanism class and then, with semiquantitative considerations, was assigned a probability carcinogenicity score, taking into account simultaneously the hypothesized action mechanism and physical chemical parameters.

Relationships among in vitro mutagenicity assays: quantitative vs. qualitative test results.

In previous investigations, we studied the relationships between the profiles of the qualitative responses of in vitro short-term tests (mutation in Salmonella typhimurium, chromosomal aberrations in CHO cells, sister chromatid exchanges in CHO cells, and mutation in mouse lymphoma cells) and common sets of chemicals. In this paper, we address the study of the quantitative responses (potency). We show that two analyses point to similar patterns of relationships: the mutation in mouse lymphoma cells assay is most similar to the CHO sister chromatid exchange assay, and the Salmonella assay is most similar to the CHO chromosomal aberrations assay.

QSAR models for both mutagenic potency and activity: application to nitroarenes and aromatic amines.

We studied the molecular determinants that discriminate between mutagenic and inactive compounds for: a) aromatic and heteroaromatic amines; b) nitroarenes. Mutagenic activity (data from literature) had been previously assessed in Salmonella typhimurium and Escherichia coli (SOS repair). The Quantitative Structure-Activity Relationships (QSAR) found were compared with those obtained in the laboratory of Professor C. Hansch for the mutagenic potency of the same compounds. It appears that there is a dramatic difference between the QSARs for potency, and those for yes/no activity: hydrophobicity played a major role in determining the potency of the active compounds, whereas mainly electronic factors differentiated the actives from the inactives. The electronic factors were those expected on the basis of the hypothesized metabolic pathways of the chemicals. Our interpretation is that the electronic factors (together with size/shape, possibly) determine the minimum requirement for the chemicals to be metabolized, whereas the hydrophobicity determines the extent of activity of chemicals that can be metabolized (actives). Moreover, the different QSARs found for the Salmonella strains TA98 and TA100 were discussed in the light of recent progress in the understanding of the molecular mechanisms of mutagenicity in these organisms. It is concluded that the nonlinear relationship observed for these chemicals between the two types of QSAR should be taken into account both when planning QSAR studies, and when using mutagenicity data for risk assessment.

Toxicology of halogenated aliphatic hydrocarbons: structural and molecular determinants for the disturbance of chromosome segregation and the induction of lipid peroxidation.

The induction of mitotic chromosome malsegregation, mitotic arrest and lethality by a set of 55 halogenated hydrocarbons was investigated. To this aim, genetic assays in the mould Aspergillus nidulans, able to provide precise quantitative information on the end-points studied, were used throughout the work. The experimental data obtained were used to develop QSAR models for the induction of aneuploidy, which pointed to a major role of electrophilicity as molecular determinant for the aneugenic potential of the halogenated hydrocarbons investigated. Within the hypothesis of a link between the electrophilicity of haloalkanes and their propensity to undergo a reductive biotransformation, with production of free radical species, a subset of 27 compounds was also tested for the ability to induce lipid peroxidation in rat liver microsomes in vitro. The results obtained indicate a partial coincidence between the abilities to initiate lipid peroxidation and to disturb chromosome segregation at mitosis. The data base obtained was also used to investigate the relationship between chemical structure and peroxidative potential. The analysis indicated that electronic and structural parameters related to the ease of homolitic cleavage of the carbon-halogen bond play a pivotal role as determinants for the peroxidative character of haloalkanes.

Rodent carcinogenicity and toxicity, in vitro mutagenicity, and their physical chemical determinants.

In this paper, we considered rodent carcinogenicity and toxicity, and four in vitro mutagenicity systems, and we made a global comparison between their different response profiles to a common set of 297 chemicals. This analysis is complemented with a study of the physical chemical properties of active and inactive compounds in the different systems. A clearcut separation between the different classes of toxicological end-points (carcinogenicity, in vivo toxicity, in vitro carcinogenicity) was evident. The observed lack of association between carcinogenicity and toxicity supports the validity of the rodent bioassays; this is contrary to the position that the positive results obtained are due mainly to the use of excessive doses that exert cytotoxic effects. We found substantial consistency in the responses of the in vivo toxicity systems (maximum tolerated dose and LD50), but we also found that remarkable differences exist between the in vitro mutagenicity assay systems. The study of the structure-activity relationships showed that: (a) the hydrophobic-electronic properties of the chemicals influence rodent carcinogenicity, with the tendency of carcinogens to be more electrophilic and more hydrophobic than non-carcinogens; (b) steric effects are implied in in vitro mutagenicity, bulkier molecules being less mutagenic than smaller molecules; (c) no clear association between in vivo toxicity and physical chemical properties was apparent. The differences between carcinogenicity and in vitro mutagenicity may hypothetically be related to their different experimental procedures. The relatively short treatment of in vitro mutagenicity requires that chemicals penetrate easily into the cells, and are well dissolved into the aqueous medium, size and hydrophilicity thus being critical for the action of the chemicals. The size of the molecules is not critical in the long-term rodent carcinogenicity experiments, where other factors, like bioaccumulation (hydrophobicity) and electronic reactivity, become essential.

Detection of DNA damage in human lymphocytes by alkaline single cell gel electrophoresis after exposure to benzene or benzene metabolites.

The alkaline single cell gel electrophoresis (Comet) assay was applied to study the occurrence of DNA damage in peripheral lymphocytes of human subjects with occupational exposure to low levels of benzene (twelve gasoline station attendants, with average benzene exposure of 0.3 mg/m3, 8 h TWA). The results obtained show a significant excess of DNA damage in lymphocytes of exposed workers, compared to matched unexposed controls (p = 0.028, Mann-Whitney U-test). Averaged tail moment values, based on 100 cells/individual, were 1.900 microns in the exposed and 0.936 micron in the unexposed group. In addition, exposed subjects showed a clearcut excess of heavily damaged cells, with tail moments > 90th percentile of the overall distribution (13.5 vs. 6.5%, p = 0.013, Mann-Whitney U-test). No correlation was found between the extent of DNA damage and the ages or smoking habits of the subjects. In order to assess the plausibility of the involvement of benzene in the results of the ex vivo study, further experiments were performed treating in vitro peripheral lymphocytes from unexposed donors with benzene metabolites hydroquinone, benzoquinone and benzenetriol. In these experiments, all benzene metabolites exerted a marked effect on resting lymphocytes, the lowest effective concentrations being below 1 microgram/ml. Conversely, far greater concentrations were required for the induction of significant DNA damage in parallel experiments with hydroquinone on mitogen stimulated lymphocytes. Addition of the DNA repair inhibitor cytosine arabinoside (Ara-C, 1-10 micrograms/ml) partially restored the sensitivity of stimulated cells to hydroquinone, an indication of the active processing of induced DNA lesions in growing cells. These results are discussed also in relation to the role of peripheral lymphocytes as target tissue in the biomonitoring of human exposure to genotoxic agents.

DNA damage by hydroquinone in human white blood cells: analysis by alkaline single-cell gel electrophoresis.

The genotoxicity of hydroquinone (HQ) in human white blood cells was investigated by means of alkaline single-cell gel electrophoresis (SCGE). The exposure of purified lymphocytes to HQ (0.5-50 microg/ml) produced significant and dose-related increases in DNA migration; conversely, no induction of DNA damage was observed in leukocytes after in vitro treatment of whole blood samples (100-500 microg/ml). Similar differences in DNA damage between whole blood samples and purified lymphocytes were observed after treatments with hydrogen peroxide (H2O2, 50 microM). The DNA damaging activity of HQ was significantly (p<0.001, U-test) inhibited by exogenous catalase (250 U/ml), indicating the generation of peroxides in the mechanism of genotoxicity of HQ. Parallel experiments using the standard SCGE protocol, and an acellular method entailing the lysis of cells before HQ treatment, provided fairly similar results, indicating that HQ oxidation does not require endogenous metabolism. Experiments to compare the effectiveness of HQ in the induction of single-strand breaks and alkali-labile sites in resting cells and micronuclei in cytokinesis-blocked cells indicate that despite the extensive DNA damage detected by SCGE immediately after treatment, a significant excess of micronuclei is not observed after stimulation and in vitro cultivation. These data explain the apparent discrepancy between the high DNA damaging potential of HQ in human lymphocytes, as revealed by SCGE, and the relatively low activity reported in most cytogenetic assays with HQ on the same cell type.

The induction of mitotic chromosome malsegregation in Aspergillus nidulans. Quantitative structure activity relationship (OSAR) analysis with chlorinated aliphatic hydrocarbons.

The biological activity of 24 chlorinated aliphatic hydrocarbons has been studied in the mold Aspergillus nidulans. The ability to induce chromosome malsegregation, lethality and mitotic growth arrest has been experimentally determined for each chemical. These data, together with those of 11 related compounds previously investigated, generated a data base which was used for quantitative structure-activity relationship (QSAR) analysis. To this aim, both physico-chemical descriptors and electronic parameters of each compound have been calculated and included in the analysis. The QSAR analysis indicated that toxic effects induced by chlorinated aliphatics in A. nidulans are mainly dependent on steric factors, as indicated by the correlation with molar refractivity (MR). Conversely, the ease with which they accept electrons, parametrized by LUMO (energy of the lowest unoccupied molecular orbital), plays a prevailing role in determining the aneuploidizing properties. An involvement of free radicals, generated by the reductive metabolism of haloalkanes, is hypothesized as an explanation of the data.