Effects of virtual tube current reduction and sparse sampling on MDCT-based femoral BMD measurements.

This study investigates the impact of tube current reduction and sparse sampling on femoral bone mineral density (BMD) measurements derived from multi-detector computed tomography (MDCT). The application of sparse sampling led to robust and clinically acceptable BMD measurements. In contrast, BMD measurements derived from MDCT with virtually reduced tube currents showed a considerable increase when compared to original data. INTRODUCTION: The study aims to evaluate the effects of radiation dose reduction by using virtual reduction of tube current or sparse sampling combined with standard filtered back projection (FBP) and statistical iterative reconstruction (SIR) on femoral bone mineral density (BMD) measurements derived from multi-detector computed tomography (MDCT). METHODS: In routine MDCT scans of 41 subjects (65.9% men; age 69.3 ± 10.1 years), reduced radiation doses were simulated by lowering tube currents and applying sparse sampling (50, 25, and 10% of the original tube current and projections, respectively). Images were reconstructed using FBP and SIR. BMD values were assessed in the femoral neck and compared between the different dose levels, numbers of projections, and image reconstruction approaches. RESULTS: Compared to full-dose MDCT, virtual lowering of the tube current by applying our simulation algorithm resulted in increases in BMD values for both FBP (up to a relative change of 32.5%) and SIR (up to a relative change of 32.3%). In contrast, the application of sparse sampling with a reduction down to 10% of projections showed robust BMD values, with clinically acceptable relative changes of up to 0.5% (FBP) and 0.7% (SIR). CONCLUSIONS: Our simulations, which still require clinical validation, indicate that reductions down to ultra-low tube currents have a significant impact on MDCT-based femoral BMD measurements. In contrast, the application of sparse-sampled MDCT seems a promising future clinical option that may enable a significant reduction of the radiation dose without considerable changes of BMD values.

Feasibility of opportunistic osteoporosis screening in routine contrast-enhanced multi detector computed tomography (MDCT) using texture analysis.

This study investigated the feasibility of opportunistic osteoporosis screening in routine contrast-enhanced MDCT exams using texture analysis. The results showed an acceptable reproducibility of texture features, and these features could discriminate healthy/osteoporotic fracture cohort with an accuracy of 83%. INTRODUCTION: This aim of this study is to investigate the feasibility of opportunistic osteoporosis screening in routine contrast-enhanced MDCT exams using texture analysis. METHODS: We performed texture analysis at the spine in routine MDCT exams and investigated the effect of intravenous contrast medium (IVCM) (n = 7), slice thickness (n = 7), the long-term reproducibility (n = 9), and the ability to differentiate healthy/osteoporotic fracture cohort (n = 9 age and gender matched pairs). Eight texture features were extracted using gray level co-occurrence matrix (GLCM). The independent sample t test was used to rank the features of healthy/fracture cohort and classification was performed using support vector machine (SVM). RESULTS: The results revealed significant correlations between texture parameters derived from MDCT scans with and without IVCM (r up to 0.91) slice thickness of 1 mm versus 2 and 3 mm (r up to 0.96) and scan-rescan (r up to 0.59). The performance of the SVM classifier was evaluated using 10-fold cross-validation and revealed an average classification accuracy of 83%. CONCLUSIONS: Opportunistic osteoporosis screening at the spine using specific texture parameters (energy, entropy, and homogeneity) and SVM can be performed in routine contrast-enhanced MDCT exams.

Both the frequency of HbA1c testing and the frequency of self-monitoring of blood glucose predict metabolic control: A multicentre analysis of 15 199 adult type 1 diabetes patients from Germany and Austria.

BACKGROUND: The objective of this study was to examine the association between metabolic control and frequency of haemoglobin A1c (HbA1c ) measurements and of self-monitoring of blood glucose, as well as the interaction of both. METHODS: Data of 15 199 adult type 1 diabetes patients registered in a standardized electronic health record (DPV) were included. To model the association between metabolic control and frequency of HbA1c testing or of self-monitoring of blood glucose, multiple hierarchic regression models with adjustment for confounders were fitted. Tukey-Kramer test was used to adjust P values for multiple comparisons. Vuong test was used to compare non-nested models. RESULTS: The baseline variables of the study population were median age 19.9 [Q1; Q3: 18.4; 32.2] years and diabetes duration 10.4 [6.8; 15.7] years. Haemoglobin A1c was 60.4 [51.5; 72.5] mmol/mol. Frequency of HbA1c testing was 8.0 [5.0; 9.0] within 2 years, and daily self-monitoring of blood glucose frequency was 5.0 [4.0; 6.0]. After adjustment, a U-shaped association between metabolic control and frequency of HbA1c testing was observed with lowest HbA1c levels in the 3-monthly HbA1c testing group. There was an inverse relationship between self-monitoring of blood glucose and HbA1c with lower HbA1c associated with highest frequency of testing (>6 daily measurements). Quarterly HbA1c testing and frequent self-monitoring of blood glucose were associated with best metabolic control. The adjusted Vuong Z statistic suggests that metabolic control might be better explained by HbA1c testing compared to self-monitoring of blood glucose (P < .0001). CONCLUSION: This research reveals the importance of quarterly clinical HbA1c monitoring together with frequent self-monitoring of blood glucose in diabetes management to reach and maintain target HbA1c .

Implementation of a spark plasma sintering facility in a hermetic glovebox for compaction of toxic, radiotoxic, and air sensitive materials.

Spark plasma sintering (SPS) is a rapidly developing method for densification of powders into compacts. It belongs to the so-called "field assisted sintering techniques" that enable rapid sintering at much lower temperatures than the classical approaches of pressureless sintering of green pellets or hot isostatic pressing. In this paper, we report the successful integration of a SPS device into a hermetic glovebox for the handling of highly radioactive material containing radioisotopes of U, Th, Pu, Np, and Am. The glovebox implantation has been facilitated by the replacement of the hydraulic system to apply pressure with a compact electromechanical unit. The facility has been successfully tested using UO2 powder. Pellets with 97% of the theoretical density were obtained at 1000 °C for 5 min, significantly lower than the ∼1600 °C for 5-10 h used in conventional pellet sintering.

De-targeting by miR-143 decreases unwanted transgene expression in non-tumorigenic cells.

MicroRNA dysregulation often results in the development and progression of cancer. miR-143 is ubiquitously expressed in most human and murine tissues but downregulated in many cancer types. This differential miRNA expression can be utilized for targeted cancer gene therapies. Multiple copies of the miR-143 complementary target sequence were inserted into the 3'UTR of plasmid vectors encoding either for different reporter genes or for the therapeutic gene TNFα. With these transgenes, we analyzed the miR-143-dependent gene expression in cancer cells and normal cells. Moreover, we investigated miR-143-regulated luciferase expression in an NMRI nude/HUH7 xenograft mouse model using a nonviral carrier system for in vivo transfections. We showed low and high levels of miR-143 in cancer cells and normal cells, respectively, leading to a differential gene expression of the reporters and the therapeutic TNFα. According to the miR-143 levels, the luciferase reporter gene expression was silenced in the mouse lungs but not in HUH7 tumors. Thus, we utilized the differential miR-143 expression in healthy and cancerous tissues to de-target the lung by specifically targeting the tumor in an in vivo HUH7 xenograft mouse model. The use of an miR-143-regulated therapeutic transgene may present a promising approach for cancer gene therapy.

[Intervention of psychological and ethical professionals of human science in obstetrical morbidity and mortality conferences]. / L'intervention des sciences humaines en psychologie et en éthique dans les revues de morbi-mortalité en obstétrique.

OBJECTIVE: To identify the defence mechanisms manifested by medical staff which could disturb the decision making, revealed by professionals of human science (PHS) in morbidity and mortality conferences (MMC). MATERIALS AND METHODS: Application of two methods of psychological intervention in MMC, conducted between March 1st, 2009 and November 30, 2010, in 20 randomized maternity among five perinatal networks: the method of inter-active problem solving targeted at the functioning of the teams and the method for developing professional practice centred on individual. The data collection was realized during analyse of case in MMC, with note-taking by two pair PHS. The oral expressions of RMM' participant were secondarily re-written, analyzed and classed by theme. RESULTS: Fifty-four MMC were performed. The mechanisms of defence have been identified by PHS intervention in MMC: denial of situation, pact of denegation, rift and overprotection. They were be identified by two PHS intervention methods, this consolidates these results. This intervention began staff medical to transformation at different level, in particular to improve the capacity of cooperation. CONCLUSION: The identification of the mechanisms of defence in MMC enables staff medical to improve communication and quality relationship between healthcare professionals. This could constitute an actual factor of practices improvement. However, complementary studies must be performed to confirm this hypothesis.

Progression of atherosclerosis in ApoE-deficient mice that express distinct molecular forms of TNF-alpha.

TNFalpha (TNF) critically regulates inflammation-driven atherosclerosis. Because the transmembrane (tmTNF) and soluble (sTNF) forms of TNF possess distinct immuno-modulatory properties, we hypothesized that they might differentially regulate atherosclerosis progression. Three groups of male ApoE(-/-) mice were studied: one expressing wild-type TNF (WT-TNF); one expressing exclusively a mutated non-cleavable form of TNF (KI-TNF); and one deficient in TNF (KO-TNF). Mice aged 5 weeks were fed the high-fat diet for 5 (T5) and 15 weeks (T15) or a standard chow diet for 15 weeks. At T5, in mice fed the high-fat diet, no significant differences in lesion area were observed among the three groups, either in valves or in aortas. At T15, lesion areas in valves were significantly lower in KO-TNF mice compared with those in WT-TNF mice, whereas in KI-TNF mice, they were intermediate between KO- and WT-TNF mice but not significantly different from these two groups. In aortas, lesions in KI-TNF were comparable to those of KO-TNF, both being significantly lower than those in WT-TNF. Theses differences were not linked to circulating lipids, or to macrophage, actin, and collagen contents of lesions. At T15, in mice fed the chow diet, lesion areas in valves and the aortic arch were not significantly different between the three groups. Levels of IL-6, IFNgamma, IL-10, and Foxp3 mRNAs in spleens and production of IL-6, IL-10, MCP-1, RANTES, and TNFR-2 by peritoneal macrophages at T15 of the high-fat diet showed a decrease in pro-inflammatory status, more marked in KO-TNF than in KI-TNF mice. Apoptosis was reduced only in KO-TNF mice. In conclusion, these data show that TNF effects on atherosclerosis development are detectable at stages succeeding fatty streaks and that wild-type TNF is superior to tmTNF alone in promoting atherosclerosis. TNF-dependent progression of atherosclerosis is probably linked to the differential production of pro-inflammatory mediators whether tmTNF is preponderant or essentially cleaved.

Predicting within-family variability in juvenile height growth of Salix based upon similarity among parental AFLP fingerprints.

Willow is being developed as a crop for biomass plantations in the Northeast and North-central United States, but has only recently been the subject of controlled breeding to generate improved genotypes. Maximizing variability among progeny within full-sib families produced by controlled pollination may increase the probability of producing willow clones exhibiting desirable extreme phenotypes. Yet, predicting combinations of parents yielding highly variable progeny is not currently possible. Controlled pollinations were completed among 15 Salix eriocephala clones and the resulting progeny were vegetatively propagated and planted in a greenhouse progeny test. Heights of rooted cuttings were measured after 4 months of growth. Genetic similarity among parents was estimated based on 77 polymorphic AFLP bands. Strong negative correlation ( r = -0.88) was detected between mean female-parent similarity indices and the standard deviation of height among half-sib progeny from those females. Parent combinations that had relatively low similarity indices tended to produce progeny that had greater variability in height. This negative relationship suggests that AFLP fingerprints of S. eriocephala parents may be useful for predicting parent combinations that will yield families with large variability.

Reconstitution of hybrid proteasomes from purified PA700-20 S complexes and PA28alphabeta activator: ultrastructure and peptidase activities.

The activity of the proteasome, the major non-lysosomal proteinase in eukaryotes, is stimulated by two activator complexes, PA700 and PA28. PA700-20 S-PA700 proteasome complexes, generally designated as 26 S proteasomes, degrade proteins, whereas complexes of the type PA28-20 S-PA28 degrade only peptides. We report, for the first time, the in vitro reconstitution of previously identified hybrid proteasomes (PA700-20 S-PA28) from purified PA700-20 S proteasome complexes and PA28 activator. In electron micrographs, the hybrid appears as a corkscrew-shaped particle with a PA700 and a PA28 activator each bound to a terminal alpha-disk of the 20 S core proteasome. The multiple peptidase activities of hybrid proteasomes are not different from those of PA28-20 S-PA28 or PA700-20 S-PA700 complexes.

Uptake of the ATP-binding cassette (ABC) transporter Ste6 into the yeast vacuole is blocked in the doa4 Mutant.

Previous experiments suggested that trafficking of the a-factor transporter Ste6 of Saccharomyces cerevisiae to the yeast vacuole is regulated by ubiquitination. To define the ubiquitination-dependent step in the trafficking pathway, we examined the intracellular localization of Ste6 in the ubiquitination-deficient doa4 mutant by immunofluorescence experiments, with a Ste6-green fluorescent protein fusion protein and by sucrose density gradient fractionation. We found that Ste6 accumulated at the vacuolar membrane in the doa4 mutant and not at the cell surface. Experiments with a doa4 pep4 double mutant showed that Ste6 uptake into the lumen of the vacuole is inhibited in the doa4 mutant. The uptake defect could be suppressed by expression of additional ubiquitin, indicating that it is primarily the result of a lowered ubiquitin level (and thus of reduced ubiquitination) and not the result of a deubiquitination defect. Based on our findings, we propose that ubiquitination of Ste6 or of a trafficking factor is required for Ste6 sorting into the multivesicular bodies pathway. In addition, we obtained evidence suggesting that Ste6 recycles between an internal compartment and the plasma membrane.

Influence of PAI-1 on adipose tissue growth and metabolic parameters in a murine model of diet-induced obesity.

An increased plasma plasminogen activator inhibitor-1 (PAI-1) level is a risk factor for myocardial infarction, particularly when associated with visceral obesity. Although the link between PAI-1 and obesity is well documented, little is known about the physiological relevance of PAI-1 production by adipose tissue. Therefore, we have compared adipose tissue development and insulin resistance plasma parameters in PAI-1-deficient mice (PAI-1(-/-)) and wild-type littermates (PAI-1(+/+)) in a model of nutritionally induced obesity. After 17 weeks of consuming a high-fat diet (HFD), PAI-1(+/+) mice showed marked obesity, with a 52% increase in body weight compared with mice that were kept on a standard fat diet (P<0.0001). This weight gain was accompanied by adipocyte hypertrophy and an increase in the number of stroma cells in the gonadal fat pad, expressed as stroma cells/adipocytes (0.67+/-0.05 versus 0.43+/-0. 02; P<0.001). In plasma, the HFD induced a marked increase in PAI-1 antigen (5.1+/-0.56 versus 2+/-0.22 ng/mL; P<0.001), fasting insulinemia (1.1+/-0.21 versus 0.21+/-0.04 ng/mL; P<0.001), and glycemia (7.4+/-0.5 versus 5+/-0.3 mmol/L; P<0.001), whereas plasma triglyceride levels were not affected. When we compared PAI-1(-/-) and PAI-1(+/+) mice on the HFD, PAI-1(-/-) mice gained weight faster than did PAI-1(+/+) mice, with a significant difference in body weight between 3 and 8 weeks of the diet (32+/-1.7 versus 26+/-1.6 g at 6 weeks; P<0.05). After 17 weeks of the HFD, its effect on weight gain and the number and size of adipocytes was similar in PAI-1(+/+) and PAI-1(-/-) mice. By contrast, the increase in the number of stroma cells presented by PAI-1(+/+) mice was not observed in PAI-1(-/-) mice. In obese PAI-1(-/-) mice, tissue-type PA activity and antigen levels in the gonadal fat pad were significantly higher than in obese PAI-1(+/+) mice (230+/-50 versus 47+/-20 arbitrary units/g, P<0.01; 40+/-13 versus 17+/-13 ng/g, P<0.05, respectively), whereas urokinase-type PA activity and antigen levels were similar in both groups. In plasma, nonobese PAI-1(-/-) mice displayed 62% higher insulin levels (P<0.05) than did PAI-1(+/+) mice. Obese PAI-1(-/-) mice displayed 68% higher triglyceride levels (P<0.01) and 21% lower glucose levels (P<0.05) than did PAI-1(+/+) mice. These data support an effect of PAI-1 on weight gain and adipose tissue cellularity in the induction of obesity in mice. Moreover, PAI-1 influences glucidolipidic metabolism. The elevated expression of PAI-1 observed in human obesity could be involved in mechanisms that control adipose tissue development.

In situ hybridization and immunohistochemistry study of thyroid peroxidase expression in thyroid tumors.

Malignant thyroid tumors reportedly exhibit an anomaly in thyroid peroxidase (TPO) resulting in a lower affinity for monoclonal antibody 47 (mAb 47) in immunohistochemistry studies. The purpose of the present study was to compare TPO immunostaining in normal, benign, and malignant thyroid tissue with expression of mRNA sequences in four exons of the molecule including the epitope of mAb 47. TPO immunostaining was performed using mAb 47 and a polyclonal antibody (pAb). Messenger RNA expression was investigated by in situ hybridization using probes specific for mRNA sequences in exons 2, 12 (epitope of mAb), 15, and 17. As expected, pAb immunostaining was significantly positive on all benign tumors and 50% of carcinomas. With mAb 47, little or no immunostaining was observed in 16 of 17 carcinomas while significantly positive immunostaining was found in normal tissue and benign tumors. In situ hybridization showed a decrease and heterogeneity in the expression of all mRNA sequences in carcinomas as compared to normal tissue and benign tumors. Unlike the other three probes, the probe specific for exon 12 hybridized strongly with benign tumors but poorly with most carcinomas. Poor hybridization was usually correlated with defective mAb 47 immunostaining. These results confirm that TPO is expressed in thyroid carcinomas but in smaller amounts than in normal tissue and benign tumors. In malignant tumors, qualitative changes in TPO may also impede mAb 47 immunostaining. In situ hybridization showed a concomitant decrease in the corresponding TPO mRNA sequence. These changes could be due to abnormalities in the maturation of TPO mRNA leading to a different splicing variant.

Identical subunit topographies of human and yeast 20S proteasomes.

The arrangement of subunits in human 20S proteasomes was recently determined by us by immunoelectron microscopy and chemical cross-linking. The positions of 4 of the 14 subunits differed from those found in the yeast proteasome by X-ray crystallography. Double labeling of human 20S proteasomes with antibodies to subunits C2 and C5 has now shown that these subunits are nearest neighbors. The result contradicts our published model for the human proteasome but is in accordance with the subunit arrangement in yeast proteasomes, suggesting that yeast and human proteasomes most probably have identical subunit arrangements. Immunoelectron microscopy also showed that the C-terminal extension at the human C2 subunit is flexible but takes up a well-defined position in the proteasome.

Subunit arrangement in the human 20S proteasome.

In human 20S proteasomes two copies of each of seven different alpha-type and seven different beta-type subunits are assembled to form a stack of four seven-membered rings, giving the general structure alpha(1-7), beta(1-7), beta(1-7), alpha(1-7). By means of immunoelectron microscopy and chemical crosslinking of neighboring subunits, we have determined the positions of the individual subunits in the proteasome. The topography shows that for the trypsin-like, the chymotrypsin-like, and the postglutamyl cleaving activities, the pairs of beta type subunits, which are thought to form active sites, are nearest neighbors.

Screening for molecules interacting with proteasomes in Thermoplasma acidophilum.

Thermoplasma acidophilum cell extracts were fractionated by gel filtration. Proteasomes were eluted as two major peaks. The first one (molecular mass(r) about 2 MDa) contained proteasomes associated with DNA/protein complexes. Proteasomes eluted in the other peak were partially resolved into three subpeaks and based on their preferential hydrolysis of casein, Z-GGL-MCA, and suc-LLVY-MCA, were designated C, L and Y, respectively. Further purification of proteasomes from peak Y resulted in a homogenous enzyme preparation, whereas proteasomes purified from peak C contained a homomultimeric protein composed of 20 kDa subunits. Thus, association of proteasomes with this protein seems to be responsible for the observed increase in molecular mass and for inhibition of caseinolytic activity by Ca2+-ions.

The human proteasome subunit HsN3 is located in the inner rings of the complex dimer.

Subunit HsN3 of the human proteasome is a beta-type subunit homologous to PRE4 from yeast, X1 beta from Xenopus and RN3 from the rat. Using electron microscopy, the binding sites of a monoclonal antibody with specificity for subunit HsN3 have been located in the two juxtaposed inner rings of the human proteasome. Subunit HsN3 was present in two copies, one in each ring, in accordance with our concept of two identical halves making up the complete human proteasome. The subunit is involved in the trypsin-like as well as the peptidylglutamyl-peptide cleavage activities.

Structural properties of native and sonicated cinerean, a beta- (1-->3) (1-->6)-D-glucan produced by Botrytis cinerea.

Cinerean, the extracellular beta-(1-->3) (1-->6)-D-glucan of the fungus Botrytis cinerea was studied. Electron micrographs of the native polysaccharide revealed quasi-endless fibrils with an estimated diameter of ca. 1.5 nm. A particle mass of 10(9)-10(10) daltons was determined from dilute solutions by low-angle laser light scattering. Sonication of increasing duration led to fragmentation of the native polymer with an approximately exponential decrease of mass in the range of average molecular masses between 250,000 and 50,000 daltons. Shadowed by platinum, cinerean fibril fragments with a weight-average molecular mass of 172,000 +/- 3000 daltons could be characterized from electron micrographs as a distribution of rods of most probable length of 45 nm and an average length of 72 nm. Small-angle X-ray scattering confirmed the fibrillar structure of the native cinerean and the rodlike structure of sonicated cinerean. A rod diameter of 1.9 +/- 0.2 nm and a mass per unit length of 2250 +/- 490 daltons/nm were found. The latter is in agreement with the value of 1830 daltons/nm calculated from the length distribution determined from the electron micrographs. These data-especially the mass per unit length-suggest a quaternary structure for the polysaccharide. Such a structure would explain the rigidity of the rods which, in turn, is responsible for the characteristic phase separation behaviour in aqueous solutions observed by nephelometry and viscometry.

Representativeness of wind measurements with a cw Doppler lidar in the atmospheric boundary layer.

The representativity problem of laser Doppler anemometer wind measurements in the boundary layer under different atmospheric conditions has been investigated theoretically and experimentally. The calculations of the mean wind-velocity measurement errors for the surface layer under different types of thermal stratification and for the boundary layer under neutral conditions have been carried out. The theoretical conclusions are confirmed by the experimental results.