Heteromerous Bistricyclic Aromatic Enes: Dynamic Stereochemistry of Xanthylidene-Anthrones.

The heteromerous bistricyclic aromatic ene (BAE) 2,2'-dimethyl-10-(9H-xanthylidene)-9(10H)-anthrone (DMXA) was synthesized by a condensation of 10,10-dichloro-2-methylxanthene with 2-methylanthrone. X-ray crystallography of (E)-DMXA and xanthylidene-anthrone (XA) indicated that the molecules adopt anti-folded conformations with folding dihedral angles of 44°/44° and 39°/41°, respectively. The crystal structure of anti-folded (E)-DMXA does not indicate any xanthenylium-anthracenolate push-pull effect. E,Z-diastereomerization of DMXA was studied by (1) H-NMR coalescence-temperature measurements at different magnetic field strengths and by kinetic equilibration experiments. Free energy of activation for this process was 81.5 (±1.3) kJ/mol. B3LYP/6-311+G(d,p) calculations showed that anti-folded conformers of XA, (E)-DMXA, bianthrone (AA), and dixanthylene (XX) were global minima. The twisted conformers of XA, AA, and XX were local minima (ΔG298 = 16, 18, and 24 kJ/mol) with a substantial dipolar xanthenylium-anthracenolate dipolar contribution for XA. Chirality 27:919-928, 2015. © 2015 Wiley Periodicals, Inc.

Evolution of the global internal dynamics of a living cell nucleus during interphase.

Progress in cellular biology based on fluorescent microscopy techniques, shows that the spatial organization of the nucleus is dynamic. This dynamic is very complex and involves a multitude of phenomena that occur on very different time and size scales. Using an original light scattering experimental device, we investigated the global internal dynamics of the nucleus of a living cell according to the phases of the cell cycle. This dynamic presents two different and independent kinds of relaxation that are well separated in time and specific to the phase of the cell cycle.

Internal dynamics of a living cell nucleus investigated by dynamic light scattering.

Recent progresses in cellular biology have shown that the nucleus of a living cell is a structured integration of many functional domains with a complex spatial organization. This organization, as well as molecular and biochemical processes, is time regulated. In the past years many investigations have been performed using fluorescent microscopy techniques to study the internal dynamics of the nucleus of a living cell. These investigations, however, have never focussed on the global internal dynamics of the nucleus, which is still unknown. In this article we present an original light scattering experimental device that we built to investigate this dynamics during biological processes. By means of this experimental set-up, we investigated the global dynamics of the nucleus of a living cell treated with a DNA replication inhibitor. This dynamics presents different and independent kinds of relaxation well separated in time that vary as a function of the cell cycle phases.

BIRADS ultrasonography.

The fourth edition of the BIRADS mammography appeared in 2003 and is now associated with the first editions of the BIRADS ultrasound and MRI. BIRADS is a system of assistance to the drafting of the reports more and more used in the world and soon directly implemented on mammography and ultrasound units. The categories of evaluation of the BIRADS allow a clear synthesis of the descriptive data resulting from the use of the lexicon and invite the radiologist to a reasoned, objective and less intuitive step. They give an action to be taken and responsibility to the radiologist and the referring physicians in the assumption of the patients.

[Presentation of the French translation of the Breast Imaging Reporting System and Data System (BI-RADS)]. / Présentation de la traduction française du BI-RADS (Breast Imaging Reporting System and Data System).

BI-RADS is a system of assistance to the drafting of the reports more and more used in the world and soon directly implemented on mammography and ultrasound units. The categories of evaluation of the BI-RADS allow a clear synthesis of the descriptive data resulting from the use of the lexicon and invite the radiologist to a reasoned, objective and less intuitive step. They give an action to be taken and responsibility to the radiologist and the referring physicians in the assumption of the patients. The 4th edition of the BI-RADS mammography appeared in 2003, and is now associated with the first editions of the BI-RADS ultrasound and MRI.

[Dosimetry comparison of pelvimetry methods using conventional radiographs and CT]. / Comparaison dosimétrique des méthodes de pelvimétrie utilisant la radiologie conventionnelle et le scanner.

PURPOSE: To determine the fetal and maternal exposure to radiation by use of thermoluminescent dosimeters in order to compare conventional and CT-scan X-ray. Materials and methods. Dosimetry was performed with an anthropomorph phantom. Thermoluminescent dosimeters were positioned on the surface and in the depth of the phantom. Digital radiography of the pelvis was performed according to a standard technique. CT-scan of the pelvis was performed according to the Buthiau's technique. RESULTS: With CT, the dose reached 0.31 to 4.95 mGy, with a dose of 2.32 mGy for the fetal gonads. With standard technique, the doses reached 0.03 to 0.39 mGy, with a dose of 0.39 mGy for the fetal gonads. CONCLUSION: With CT the fetus and the mother were exposed to 1/10th of the total dose delivered using conventional X-rays and the dose distribution was more homogenous.

Gender-related effects of vitamin D metabolites on cartilage and bone.

Sex steroid hormones are known to have gender-dependent effects on bone and cartilage in vivo and in vitro. To investigate whether this is a general property of steroids, or is specific to the sex steroid hormones, we examined whether the effects on bone of 1,25-(OH)2D3 and 24,25(OH)2D3, the two active metabolites of vitamin D, are also gender-dependent. One-month-old male and female rats were treated for 1 month with various doses of 1,25-(OH)2D3, 24,25-(OH)2D3, or a combination of both metabolites. The direct effects of both metabolites on the skeleton of the treated animals were similar in male and female rats. 24,25-(OH)2D3 alone or in combination with 1,25-(OH)2D3 increased bone calcium and phosphorus, while 1,25-(OH)2D3 slightly decreased bone mineral content. 24,25-(OH)2D3 also enhanced the differentiation of cartilage in the growth plate, increasing the size of the hypertrophic zone. In addition, an increased metaphyseal bone volume was observed following 24,25-(OH)2D3 treatment in rats of both sexes, but not with 1,25-(OH)2D3. Vitamin D metabolites affected the weight gain of the experimental animals in a gender-dependent manner; 1,25-(OH)2D3 increased weight gain of male rats and 24,25-(OH)2D3 decreased weight gain of female rats. In addition, 1,25-(OH)2D3 increased bone weight and ash weight in male animals. These gender-dependent effects of vitamin D metabolites may occur indirectly via effects of sex steroid hormones, the latter being a sex-related effect.

Changes in the nature of inpatient medical services--impact on medical education and patient care.

One hundred sequential admissions to an internal medicine department in a 765-bed teaching hospital in 1973 were compared to 100 admitted in 1987. Mean age in 1973 was 50.3 years as compared to 57.4 in 1987. Length of stay shortened from 8.8 days in 1973 to 4.7 days in 1987. The overwhelming majority of the admissions during both periods had circulatory and/or respiratory disease. The number of diagnoses on admission increased from 2.3 in 1973 to 3.8 in 1987. In 1973, 22% of the patients received no drugs as compared to 6.3% in 1987. X-ray studies per hospital day doubled and invasive procedures more than quintupled. Intravenous fluids were given on 2.5% of days in 1973 and on 22.3% in 1987. Thus medical patients are now older and sicker, yet stay for much shorter periods of time. This radical change in the intensivity and tempo of work raises serious questions about the appropriateness of these sites as major loci for undergraduate teaching.

A stable Escherichia coli-Mycobacterium smegmatis plasmid shuttle vector containing the mycobacteriophage D29 origin.

A plasmid shuttle vector for Escherichia coli and mycobacteria was constructed from an E. coli plasmid containing the ColE1 origin, a 2.6-kb PstI fragment from bacteriophage D29 that grows in numerous mycobacterial species, and the kanamycin resistance gene either of Tn903 or of Tn5. The resultant plasmid is 7.63 kb and can be introduced via transformation into Mycobacterium smegmatis with high efficiency. In M. smegmatis the plasmid is stable and apparently present in multiple copies. Bioluminescence (luxA and luxB of Vibrio harveyi and fischeri) has been expressed in M. smegmatis from the aminoglycoside transferase promoter of Tn5. The D29 fragment should carry an origin of replication and some associated genes that act on it since various mutations destroy the ability of this fragment to replicate in M. smegmatis. The fragment was localized on the D29 genome map.

Citrate synthase from Mycobacterium smegmatis. Cloning, sequence determination and expression in Escherichia coli.

A Mycobacterium smegmatis PstI library was constructed by cloning these fragments downstream from the lac promoter of the expression vector pHG171. Three identically sized clones were isolated by complementation of an Escherichia coli strain (chi 2338) deficient in citrate synthase. One insert (pBL265) was used in hybridization experiments with DNA from E. coli and M. smegmatis and it was demonstrated that the clones were indeed from M. smegmatis. The transcription of the M. smegmatis citrate synthase gene in E. coli relied upon the lac promoter. In translation experiments performed in vitro pBL265 gave rise to a novel protein of about 42 kDa. This band was not seen in 'opposite-orientation' subclones. Various subclones in which the 5'-end was shortened nevertheless complement E. coli chi 2338 and produce the 42 kDa protein. This demonstrates that the M. smegmatis citrate synthase gene uses its own ribosome-binding site in E. coli. The relevant 1.8 kb of the 2.8 kb insert was sequenced. A consensus E. coli ribosome-binding site was found centred precisely 10 bp upstream of the methionine codon. Other interesting features revealed by the sequence are discussed. Citrate synthase activity was assayed in vitro and the mycobacterial enzyme was found to be similar to those of the Gram-positive bacteria.

Fusion of LuxA and LuxB and its expression in E. coli, S. cerevisiae and D. melanogaster.

Luciferase from Vibrio harveyi is encoded by two adjacent genes, luxA and luxB. The two genes were fused by replacing a segment extending from near the end of luxA into the N-terminal end of luxB by a synthetic oligonucleotide. The construction removed the TAA stop codon at the end of luxA, the intervening region of 26 base pairs, and the initial methionine of luxB. A Smal site was included at the junction between the two genes and an AatII site was created near the end of luxA without altering its amino acid sequence. In Escherichia coli the fused luxAB gene could be expressed to produce functional luciferase that gave about 20% of the activity in cells without the fusion. An out-of-frame ATG exists close to and preceding the ATG of the luxA gene. This was removed and the entire fused gene bracketed by several restriction enzyme sites. The fused luxAB gene was successfully expressed in Saccharomyces cerevisiae and Drosophila melanogaster by transferring it to appropriate plasmid vectors.

Translational signals of a major head protein gene of bacteriophage lambda.

The D gene of bacteriophage lambda which codes for a major head protein is expressed at a high level during lytic growth. We have constructed a set of D-lacZ gene fusions in order to examine the factors determining the high efficiency of the D translational initiation signals. It was found that an integral sequence, 300 bp long and upstream of the ATG initiation codon, is required for maximal protein synthesis.

Isolation of the nuclear yeast genes for citrate synthase and fifteen other mitochondrial proteins by a new screening method.

To isolate nuclear genes specifying imported mitochondrial proteins, a yeast genomic clone bank was screened by an RNA hybridization-competition assay. This assay exploited the fact that mRNAs for imported mitochondrial proteins are enriched in polysomes which are bound to the mitochondrial surface in cycloheximide-inhibited yeast cells. Clones selectively hybridizing to these enriched mRNAs were further screened by hybrid-selected translation and immunoprecipitation with monospecific antisera against individual mitochondrial proteins. Thirty-six clones were isolated which contained complete or partial copies of 16 different genes for imported mitochondrial proteins. Several of these clones caused expression of the corresponding precursor polypeptide in Escherichia coli or over-expression of the corresponding mature protein in yeast. The gene for the matrix enzyme citrate synthase was sequenced; the derived amino acid sequence of the precursor polypeptide revealed an amino-terminal extension containing basic but no acidic residues.

Spectrophotometric quantitation of silver grains eluted from autoradiograms.

Autoradiograms or fluorograms can be quantified by eluting the silver grains from the developed film with 1 M NaOH and measuring the absorbance of the eluate in a conventional spectrometer. In contrast to direct scanning of the film, the present method is rapid, requires only simple equipment, and permits accurate quantitation of even heavily exposed bands.

Import of proteins into mitochondria. Translatable mRNAs for imported mitochondrial proteins are present in free as well as mitochondria-bound cytoplasmic polysomes.

In order to assess the role of different polysomal populations in mitochondrial protein import, yeast spheroplasts were treated with cycloheximide to prevent polysome "run-off" and fractionated into free polysomes, polysomes bound to the mitochondrial outer surface, and polysomes bound to the endoplasmic reticulum. These polysomes were analyzed for translatable mRNAs coding for 8 cytosolic and 12 imported mitochondrial proteins. The mitochondrial proteins included 7 proteins of the inner membrane, 2 proteins of the matrix, 2 proteins of the intermembrane space, and 1 protein of the outer membrane. Of the mRNAs for imported mitochondrial proteins, 8 were enriched in mitochondria-bound polysomes, 3 were enriched in free polysomes, and 1 was enriched in neither. All mRNAs for cytosolic proteins were enriched in free polysomes. Polysomes bound to the endoplasmic reticulum lacked significant levels of translatable mRNAs for either cytosolic or mitochondrial proteins. Even though mRNAs for imported mitochondrial proteins were enriched in mitochondria-bound polysomes, these polysomes represented only 12-18% of the total cytoplasmic polysomes. As a consequence, none of the translatable mRNAs for imported mitochondrial proteins tested was predominantly associated with mitochondria-bound polysomes. While mitochondria-bound polysomes may contribute to mitochondrial protein import, they do not appear to be obligatory for this process.