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Introduction The O-[11C]methylated derivatives of the clinically used neprilysin inhibitor (NEPi) sacubitril ([11C]SacOMe, (2R,4S)-ethyl 5-([biphenyl]-4-yl)-4-(4-[11C]methoxy-4-oxobutanamido)-2-methylpentanoate) and LBQ657 ([11C]MeOLBQ, (2R,4S)-5-(biphenyl-4-yl)-4-[(3-carboxypropionyl)amino]-2-methylpentanoic acid [11C]methyl ester and [11C]LBQOMe, (2R,4S)-5-(biphenyl-4-yl)-4-[(4-[11C]methoxy-4-oxobutanamido)]-2-methylpentanoic acid) were evaluated to determine their potential as PET imaging tracers and investigate the effect of such labeling esterification on neprilysin (NEP) binding. Methods [11C]MeOLBQ, [11C]SacOMe and [11C]LBQOMe were synthesized by O-[11C]methylation using [11C]methyl triflate. Binding of these radiolabeled derivatives (5 nM) were assessed by autoradiography on rat neprilysin rich kidney slices with or without 10 μM NEPi (thiorphan or sacubitril) for 20 min at 37 °C. [11C]LBQOMe was further tested for binding selectivity in the presence of 10 μM of angiotensin-converting enzyme inhibitor (ACEi, captopril) or angiotensin II AT1 receptor blocker (AT1R, losartan). Radioligands were evaluated for their in vitro stability up to 20 min after incubation at 37 °C in rat and human plasma by reverse-phase column-switch HPLC. Non-radioactive SacOMe incubated in rat and human plasma was analyzed by HPLC-coupled with high resolution mass spectrometry (HRMS) to confirm the metabolites' identity. [11C]SacOMe main labeled metabolite was further analyzed by HPLC after incubation in rat kidney slices at 37 °C. Results The novel [11C]SacOMe and [11C]LBQOMe were produced in 32 ± 3% RCY and 15 ± 6% at EOS (decay-corrected from [11C]CO2, n = 3), high molar activity (407 ± 92 GBq/μmol and 260 ± 92 GBq/μmol), and high chemical (≥90%) and radiochemical (≥99%) purities in a total synthesis time of 31 and 34 min, respectively. High accumulation of [11C]SacOMe and [11C]LBQOMe in kidneys was completely blocked (>99.9%) by pre-incubation with NEPi, whereas [11C]MeOLBQ displayed negligible uptake in autoradiography studies. [11C]LBQOMe binding was not affected by saturating doses of losartan or captopril indicating binding selectivity for NEP. While [11C]SacOMe and [11C]LBQOMe were stable in human plasma (>92%) even after 20 min incubation at 37 °C, rat plasma analyses exhibited >95% biotransformation of [11C]SacOMe, 40% of [11C]LBQOMe and >80% loss of the 11C-methyl group of [11C]MeOLBQ after 5 min of incubation. Comparable results using the non-radioactive SacOMe were obtained by HPLC-HRMS. Radio-HPLC analysis of the extracted activity of rat kidney slices incubated with [11C]SacOMe demonstrated that >95% of the radioactive signal corresponded to [11C]LBQOMe as the main metabolite. Conclusion The desethyl active metabolite of [11C]SacOMe, [11C]LBQOMe, displayed stability in human plasma, binding selectivity for neprilysin over ACE or AT1R in rat kidney slices. Rapid plasmatic dealkylation at the 2-methylbutanoic acid position is in line with the necessity of incorporating the labeling group on oxobutanoic acid side in the strategy to develop a stable O-alkylated labeled derivative of sacubitril.

Supernova ejecta and stellar winds are believed to produce interstellar dust grains with relatively large sizes. Smaller grains can be produced via the shattering of large grains that have been stochastically accelerated. To understand this stochastic acceleration, we have implemented novel magnetohydrodynamic(MHD)-particle-in-cell(PIC) methods into the astrophysical fluid code RAMSES. We treat dust grains as a set of massive ``superparticles'' that experience aerodynamic drag and Lorentz force. We subject our code to a range of numerical tests designed to validate our method in different physical conditions, as well as to illustrate possible mechanisms by which grains can be accelerated. As a final test as well as a foundation for future work, we present the results of decaying dusty MHD turbulence simulations with grain parameters chosen to resemble 1-2 $\mu$m grains in typical cold neutral medium conditions. We find that in these conditions, these grains can be effectively accelerated to well beyond their shattering velocities. This is true for both electrically charged and neutral grains. While the peak of the gas-grain relative drift velocity distribution is higher for neutral grains, the drift velocity distribution of charged grains exhibits an extended exponential tail out to much greater velocities. Even so, the shapes of the distributions are such that the extra gas-grain coupling provided by the Lorentz force offers grains relative protection from shattering. We also discuss the connection between our simulations and the relatively pristine ~$\mu$m sized presolar grains that do not appear to have undergone significant wear in their lifetimes., Comment: 23 pages, 13 figures, Accepted for publication in MNRAS

Neprilysin, also known as neutral endopeptidase, is a cell surface membrane metalo-endopeptidase that cleaves various peptides. Altered neprilysin expression has been correlated with various cancers and cardiovascular diseases. In this work, we present the radiosynthesis of the novel O-11 C-methylated derivative of LBQ657 (a potent neprilysin inhibitor). (2R,4S)-5-(Biphenyl-4-yl)-4-[(3-carboxypropionyl)amino]-2-methylpentanoic acid [11 C]methyl ester ([11 C]MeOLBQ) is an analog of sacubitril where the alkyl ester is a 11 C-methyl instead of an ethyl. [11 C]MeOLBQ was produced in a one-pot two-step synthesis. The O-11 C-methylation of the pentanoic acid part was done with [11 C]methyl triflate followed by the deprotection of the tert-butyl ester precursor in acidic conditions. [11 C]MeOLBQ ([11 C]7) was produced in 9.5 ± 2.5% RCY (25 ± 6% decay-corrected from [11 C]CO2 , n = 3) high molar activity 348 ± 100 GBq/μmol (9425 ± 2720 mCi/μmol) at EOS, in high chemical (>95%) and radiochemical (>99%) purities. The total synthesis time including HPLC purification and reformulation was 29 minutes. To our knowledge, this is the first PET-labeled analog of the clinically used NEP inhibitor sacubitril.

The O-[[The novel [The desethyl active metabolite of [

Neprilysin, also known as neutral endopeptidase, is a cell surface membrane metalo-endopeptidase that cleaves various peptides. Altered neprilysin expression has been correlated with various cancers and cardiovascular diseases. In this work, we present the radiosynthesis of the novel O

Resume Les telomeres sont des structures associant l’extremite de la molecule d’ADN de chaque bras chromosomique et plus d’une dizaine de proteines regulatrices. La sequence de l’ADN telomerique est constituee par la repetition de l’hexanucleotide TTAGGG. En raison des modalites de replication de l’ADN, les telomeres se raccourcissent progressivement a chaque cycle cellulaire, et ce phenomene est accelere par les stress oxydatifs. Une taille critique des telomeres active les programmes genetiques de senescence et/ou de mort cellulaire. La taille des telomeres leucocytaires est actuellement consideree comme un marqueur de l’âge biologique, du risque morbide (pathologies cardiovasculaires, metaboliques, demence) et de la mortalite. Un raccourcissement accelere a ete observe dans la schizophrenie, les troubles de l’humeur et l’exposition au stress psychique, suggerant un vieillissement premature. Aucune donnee n’est disponible sur le statut des telomeres dans le cortex de sujets deprimes. Nous avons etudie la taille des telomeres dans l’ADN du cortex occipital de 24 patients deprimes et 12 temoins (don du Stanley Research Institute). Aucune difference significative n’a ete mise en evidence. Ces resultats sont discutes a partir des donnees qui suggerent un role de l’inflammation et du stress oxydatif dans la depression, ainsi que l’existence d’un mecanisme de protection specifique des telomeres dans les neurones.

The presence of tyrosinase mRNA in the peripheral blood cells of melanoma patients has been recently studied as a possible marker of haematogenous dissemination. However, considerable variations in the rates of detection have been noted. We determined the presence of tyrosinase mRNA-positive circulating cells using reverse transcriptase-polymerase chain reaction (RT-PCR) in 35 patients with stage I melanoma, two patients with stage II melanoma and two patients with stage III melanoma. Among the patients with stage 1, 13 were tested before and after surgery (< 1 h). Twenty healthy subjects served as negative controls. Out of the melanoma patients, the tyrosinase gene was expressed in three of the 52 samples tested. Tyrosinase mRNA was present in the circulating cells of only one patient with stage I melanoma after intra-congenital naevi resection. However, two other stage I patients developed rapidly lethal metastasis within the following 6 months, despite the lack of detectable tyrosinase mRNA. None of stage II patients were positive for the tyrosinase transcripts, while both patients with stage III melanoma showed enzyme expression. Our results confirm those of previous studies, showing that a small proportion of stage I melanoma patients have tyrosinase-positive circulating cells. Moreover, the lack of tyrosinase mRNA detection in the blood does not necessarily exclude metastatic progression. Therefore, this study indicates that the detection of tyrosinase mRNA-positive circulating cells by RT-PCR is not a predictive biomarker of a metastasis risk in patients with stage I melanoma.

IntroductionLe trouble dépressif majeur (TDM) est associé à des altérations de l’expression génique au niveau cérébral et en périphérie, dans les leucocytes sanguins. Chez les dépressifs, les études de neuroimagerie ont mis en évidence des anomalies structurales et fonctionnelles affectant deux régions clés du réseau frontocingulaire : le cortex préfrontal dorso-latéral (DLPFC) et le cortex cingulaire (CC). Actuellement, les mécanismes moléculaires permettant de faire le lien entre ces différents niveaux étiopathogéniques restent inconnus. Les mécanismes épigénétiques, situés à l’interface entre le génome et la réponse cellulaire, constituent des candidats potentiels.ObjectifExplorer le rôle de l’épigenèse dans le cadre du TDM en phase d’état.Matériel et méthodesNous avons mesuré l’expression de différents gènes impliqués dans les mécanismes épigénétiques dans les leucocytes du sang périphérique, le DLPFC et le CC, chez des patients dépressifs et des sujets contrôles appariés. Les niveaux des différents transcrits ont été mesurés par PCR quantitative en temps réel.RésultatsChez les patients dépressifs, nous avons mis en évidence une surexpression des gènes codant pour des enzymes intervenant dans la mise en place de marques chromatiniennes réprimant la transcription : HDACs 4-5-6-8 et DNMT3B dans le DLPFC, HDAC2 dans le CC et les leucocytes sanguins.ConclusionNos résultats retrouvent une activation des mécanismes épigénétiques réprimant l’expression génique dans le DLPFC et le CC chez les patients dépressifs. Il s’agit de la première fois qu’une telle dérégulation est décrite dans ces deux régions. Ces modifications pourraient participer aux dysfonctionnements affectant le DLPFC et le CC et participer à la physiopathologie du TDM. De plus, nous retrouvons une surexpression de HDAC2 dans les leucocytes sanguins des patients dépressifs, ce qui renforce son statut de potentiel biomarqueur périphérique.

Prospective studies of subjects at high genetic risk of psychosis (at least one first relative with schizophrenia) and retrospective studies of patients at the end of the first episode of delusion or hallucination have identified various subjective and objective symptoms which emerged months or years before the diagnostic of schizophrenia. The objective symptoms presently designed as prodromes are either transient or of infradiagnostic intensity. The objective signs have been used to define an ultrahigh risk (UHR) state, and have been operationalized by psychometric instruments, which also include criteria for genetic risk (schizotypal dimensions) and alteration of social functioning. The main instruments are the Comprehensive Assessment of At-risk Mental States (CAARMS) and the SIPS (Structured Interview for Prodromal Symptoms). The subjective symptoms, which consist exclusively in inner experiences, have been named basic symptoms, and are operationalized by the Bonn Scale for the Assessment of Basic Symptoms (BSABS) and the Schizophrenia Proneness Instrument (SPI-A, Adult version). Prospective studies of selected individuals with a psychiatric help demand have shown that signs of endogenicity (schizotypal traits), and severity (high symptomatic scores and poor functioning), are of major value to predict conversion. In contrast, the positive predictive value of isolated prodromal symptoms is rather poor (much less than the 80% required for clinical validation).This suggests that the population of subjects with prodromes is structured by two latent subgroups: those who express transient psychotic manifestations (which are quite frequent in the general population), and those with an active psychotic process, who are progressively evolving towards the categorical diagnostic of schizophrenia. However, results obtained by between groups comparison of mean psychometric variables are unable to distinguish between these two populations at baseline. Because of this, introduction of the risk of psychosis category in the DSM-V has been bitterly criticized. In accordance, therapeutic prevention assays using antipsychotics, antidepressants, or cognitive therapy have provided inconsistent results. Only the administration of ω-3 polyunsaturated fatty acids has produced a long term efficient effect.

Weak gravitational lensing provides a unique way of mapping directly the dark matter in the Universe. The majority of lensing analyses use the two‐point statistics of the cosmic shear field to constrain the cosmological model, a method that is affected by degeneracies, such as that between σ8 and Ωm which are respectively the rms of the mass fluctuations on a scale of 8 Mpc/h and the matter density parameter, both at z = 0. However, the two‐point statistics only measure the Gaussian properties of the field, and the weak lensing field is non‐Gaussian. It has been shown that the estimation of non‐Gaussian statistics for weak lensing data can improve the constraints on cosmological parameters. In this paper, we systematically compare a wide range of non‐Gaussian estimators to determine which one provides tighter constraints on the cosmological parameters. These statistical methods include skewness, kurtosis, and the higher criticism test, in several sparse representations such as wavelet and curvelet; as wel...

Telomeres are complex structures formed by the end of the DNA molecule at the tip of chromosomal arms. The telomeric sequence, which results from the repetition of the hexanucleotide TTAGGG, is partly single strand and is associated with more than ten proteins, including the enzyme telomerase. Because of the characteristics of the DNA replication process, only telomerase is able to elongate the telomeric sequence. Since the telomerase gene is repressed in virtually all the somatic cells, telomeres progressively shorten at each S phase of the cell cycle, and this shortening is accelerated by oxidative stress. A critically shortened telomere activates the genetic program of cell senescence and/or apoptosis. The telomere length measured in peripheral blood leucocytes is considered a reliable marker of biological age, mortality risk and exposure to various pathological conditions, including cardiovascular disease, dementia and metabolic syndrome. Telomere erosion has been observed in psychiatric disorders including schizophrenia and mood disorders, suggesting an accelerated aging of 10 to 20 years. Whether this peripheral dynamic is reflected by a similar pattern in the brain remains unknown. To address this issue, we have measured the telomere length in the occipital DNA cortex of 24 patients with major depressive disorder and 12 controls (donated by the Stanley Research Institute).The mean telomere length has been evaluated by a real time quantitative PCR technique, which amplified the telomere sequence and a reference single copy sequence. Results have been expressed by the ratios of Ct obtained for the two amplification curves.The mean Ct values were strictly identical (0.79 ± 0.001) and the 36 PCR curves were coincident.This study demonstrates for the first time that there is no shortening of telomeres in the cortex of patients with depressive disorder. Previous results have shown that in normal tissues telomeres length is inversely correlated to age, even in non proliferating tissues, but that the change is minimal in the brain. Thus, although consistent evidence for the role of a systemic and brain inflammation associated oxidative stress in depression has been provided, it must be concluded that the cerebral state of telomeres is not affected by the mechanism operating in the leucocytes. This observation raises the issue of the relation between the psychiatric pathological process and the peripheral telomere marker. It suggests the existence of specific telomere stabilizing factors in the cortex cells.

This chapter discusses the structure formation with numerical simulations. The impact of computational cosmology in understanding of the galaxy formation is discussed. The chapter stresses the importance of having both high-resolution simulations with as realistic physical ingredient as possible. Numerical simulations have been used to determine a fundamental statistical tool in cosmology, the mass function (MF) of dark matter haloes. The chapter presents a powerful analytical approach, that is, the halo model. The halo model is the most successful approach to describe the gravitational clustering in the non-linear regime. It relies on numerical simulations, as most of the model parameters are fitted to them, but gives a good understanding of the main physical processes at the origin of the hierarchical model. Both numerical and analytical approaches shed light on the poorly understood processes leading to the formation of galaxies in the universe, and ultimately to our own galaxy, the Milky Way.

Archeops is a balloon--borne experiment widely inspired by the Planck satellite and by its High Frequency Instrument (HFI). It is mainly dedicated to measure the Cosmic Microwave Background (CMB) temperature anisotropies at high angular resolution (about 12 arcminutes) over a large fraction of the sky (around 30 %) in the millimetre and submillimetre range at 143, 217, 353 and 545 GHz. Further, the Archeops 353 GHz channel consists of three pairs of polarized sensitive bolometers designed to detect the polarized diffuse emission of Galactic dust. We present in this paper the update of the instrumental setup as well as the inflight performance for the last Archeops flight campaign in February 2002 from Kiruna (Sweden). We also describe the processing and analysis of the Archeops time ordered data for that campaign which lead to the measurement of the CMB anisotropies power spectrum in the multipole range l=10-700 (Benoit et al. 2003a, Tristram et al. 2005) and to the first measurement of the dust polarized emission at large angular scales and its polarized. We present maps of 30 % of the sky of the Galactic emission, including the Galactic plane, in the four Archeops channels at 143, 217, 353 and 545 GHz and maps of the CMB anisotropies at 143 and 217 GHz. These are the firstever available sub--degree resolution maps in the millimetre and submillimetre range of the large angular-scales Galactic dust diffuse emission and CMB temperature anisotropies respectively., A full resolution version of the paper including figures is available at http://www.archeops.org/Archeops_Publication/Pub/Processing.pdf 39 pages, 40 figures

The objective was to relate the response of the HSP70 and P53 genes to the cessation and the recovery of cardiac muscle cell functions when submitted to ischemia-reperfusion. We have measured the electromechanical activity, the released enzymes and HSP70 RNA and protein levels in cultured neonatal rat cardiomyocytes (CM) in a substrate-free, hypoxia-reoxygenation model of ischemia-reperfusion. In parallel the expression of the two genes P53 (the key apoptosis regulator gene) and P21/Waf1 (the P53 target gene) has been evaluated. The functional recovery during post-'ischemic' reoxygenation was associated with an overexpression of HSP70 and P53 lasting until the functional parameters reverted back to the normal, prehypoxic values. In contrast, extending the substrate-free hypoxic treatment worsens the dysfunction of the cardiac muscle cell and, in these conditions, reoxygenation failed to restore cell functions and to activate HSP70. Finally, in the conditions of reversible 'ischemic' cell injury, an early and transitory activation of P53 was associated with the functional recovering process of the CM submitted to simulated ischemia. These observations are suggestive of a contributive role of both HSP70 and P53 to a cytoprotective program activated by reoxygenation in post-'ischemic' CM.

Mutations of the p53 gene are the most common genetic alteration in malignant human tumors. A cyclin-dependent kinase inhibitor, p21WAF1/CIP1, is thought to be an important mediator of p53-induced cell cycle arrest. Although numerous studies have reported p53 expression and mutation in colorectal cancer few of them have correlated p53 expression with that of its downstream effector p21 and with the proliferation index as measured by expression of the Ki67 nuclear antigen. We studied p53, p21 and Ki67 expression by immunohistochemistry and molecular biology in 35 colorectal carcinomas. We compared these findings with each other and with clinical factors. Sixty three percent of tumors expressed p53 whereas seventy one percent expressed p21WAF1/CIP1. In adenocarcinomas, p21 staining was heterogeneous: p21-reactive cells were seen in the most differentiated areas. There was no correlation between p21WAF1/CIP1 and p53 expression, p53 mutation, Ki67 expression or clinical factors such as sex or location of the tumor. On the other hand, there was a statistical relationship between p21 expression and survival: our results indicated an association between high p21 expression and lower stages p21WAF1/CIP1 appears to be induced independently of p53 in these tumors and may be associated with differentiation rather than proliferation.

The cytogenetic analysis of 67 meningiomas (58 intracranial and 9 spinal tumors) identified chromosomal abnormalities in 63% of cases. When chromosomes involved in numerical and structural changes with a frequency of more than one standard deviation above the mean were considered, distinct cytogenetic patterns could be identified according to sex, anatomical location and histology. The chromosomes more frequently affected were 1, 2, 3, 4, 8, 14, 15, 19, 22, Y. No conclusion could be drawn regarding the prognostic significance of these karyotypic alterations.

To assess the effect of a single chromosomal translocation on the nuclear phenotype of human cells, seven diploid adenomas and five diploid carcinomas of the thyroid gland were studied using quantitative nuclear morphometry. Four adenomas and three carcinomas were cytogenetically normal, whereas three adenomas and two carcinomas had a unique chromosomal translocation. A densitometric parameter discriminated adenomas from carcinomas (skewness of the optical density histogram, SODH), and tumours with and without chromosomal translocation (standard deviation of the optical density, SDODH). These results demonstrate that single chromosomal structural rearrangements produce quantifiable alterations of nuclear organisation, but that other nuclear features which do not express an aneuploid DNA content or an abnormal karyotype differentially characterise benign and malignant conditions.

The distribution of 271 chromosomal breakpoints involved in 203 clonal structural non "specific" rearrangements identified in 82 human primary solid tumors of various histologies has been analyzed. According to the "mean + 1 SD" criterion the normalized breakpoint frequency was significantly higher for chromosomes 1p, 3p, 3q, 4q, 6q, 7q, and 11p. Clusters of 3 or more breakpoints were assigned to only 24 of the 329 bands (7%) of the standard karyotype, and 9 high density breakpoint segments have been identified. These results indicate that a nonrandom pattern of chromosomal rearrangements can be extracted from the complex karyotypic changes observed in solid tumors, which suggest that the involved regions play a general role in tumorigenesis.

The systematic study of the karyotype in Adult Psychiatry reveals among the group of periodical psychoses the existence of the 3-D Syndrome, marked by long clinical cycles, a course towards a dementia, the presence of a X-dysgonosomy with mosaicism, a relative and better clinical response with the use of lithium than with the use of tricyclic antidepressants. These characteristics discriminate the 3-D syndrome from the periodical manic depressive psychosis, which neither includes the course towards a dementia nor does it include until now identified cellular abnormalities. From the anatomical point of view the brain is quasi-normal, when the ultimate phase is a dementia. The 3-D Syndrome seems to be linked with a phenomenon of chromosomal instability. The caryotype takes a part among the usual biological tests in psychiatry.

A cytogenetic study was performed in 63 thyroid tumors after a monolayer short-term culture. Clonal chromosomal changes were found in 47% of carcinomas and 31% of adenomas. Chromosome 7 was altered in 40% of cytogenetically abnormal tumors. The modal DNA index measured in 26 tumors was consistent with the chromosomal mode in 88% of cases. A quantitative morphometric analysis of nuclear features differentiated between diploid thyroid adenomas with or without a single translocation, which suggests that they have different biological properties. Clonal chromosomal changes were observed in 78% of carcinomas with an aggressive behavior, but only 28% of those had no risk factors. The two patients who died early had abnormalities of chromosome 7.

A cytogenetic study was performed in 18 renal cell carcinomas using a culture method previously described. The significance of chromosome involvement in clonal aberrations was evaluated according to the mean + 1 SD objective criterion. Chromosomes 3, 7, 9, and 17 were preferentially involved in both numerical and structural changes. The cytogenetic data have been correlated with the clinical staging, but in contrast to a previous study, abnormalities of chromosome 3 were not associated with a higher clinical stage.

The frozen tissue was sliced and then homogenized at 20 °C in LiCl, 2 m; lauryltrimethyl ammonium chloride (K & K No. 4484), 5 %; pronase, B grade (Calbiochem), 1 mg/ml and Tris-HCl 10 mM pH 7.5. The homogenate was left to stand 15 min at 20 °C with occasional shaking. After centrifugation at 35 000 × g for 30 min at 0 °C, the supernatant containing the crude DN A was purified by filtration on Ultrogel A 2 (LKB, Sweden). The Ultrogel A 2 column (2.5 × 45 cm) was equilibrated with a solution containing NaCl 2 m, EDTA 2.5 mM, Tris-HCl 10 mM pH 7.5. The flow rate was 3 ml cm−2 h−1. Five ml of the supernatant were placed on the column. The first peak contained highly polymerized (as demonstrated by ultracentrifugation) pure DNA (A260/A230 = 3.19; A260/A280 = 1.82). The yield was 2.26 mg of DNA/g of fresh liver.

Total HMW-RNAs were prepared by three different methods (method with phenol, method with NaCIO4, method without phenol using Ultrogel AcA 22 filtration). Giant RNAs were obtained in the void volume by filtration on Sepharose 2B.The giant RNAs/total HMW-RNA ratio is higher (6.77%) with the gel filtration method than with phenol or NaCIO4 methods (1.41% and 1.00% respectively).The nucleotide composition of these RNAs is DNA-like and the sedimentation constants are approximately 70-100 S.

Relaxation times T1 and T2 of water protons for stimulated and unstimulated parotid and submandibular saliva from healthy patients were measured in the conventional way using the Bruker PC 20 Minispec. Some measurements of dispersion were also carried out. An interpretation of the average values for T1 and T2, observed in the 20 MHz range is proposed for these types of saliva in relation to biochemical composition (total proteins, amylase activity, sialic acids and hexosamins) and viscosity. For parotid saliva T−12−T−11 correlates significantly with amylase activity. In the submandibular saliva T−12−T−11 correlates significantly with both mucins and α-amylase activity. © 1989 Academic Press, Inc.

An investigation of the liquefaction of human semen was performed using a technique of nuclear magnetic resonance, by measuring the time of spin-spin proton relaxation. Measurements made at 24 degrees C over a 15-day period showed that significant degradation processes and protein transformations occurred during the first 5 days. Different phases of these processes were identified.

Total HMW-RNAs were prepared by three different methods (method with phenol, method with NaClO4, method without phenol using Ultrogel AcA 22 filtration). Giant RNAs were obtained in the void volume by filtration on Sepharose 2B. The giant RNAs/total HMW-RNA ratio is higher (6.77%) with the gel filtration method than with phenol or NaClO4 methods (1.41% and 1.00% respectively). The nucleotide composition of these RNAs is DNA-like and the sedimentation constants are approximately 70-100 S.

An investigation of the liquefaction of human semen was performed using the technique of Nuclear Magnetic Resonance, by measuring the time of spin-spin proton relaxation. Measurements which were made at 24 degrees C and 37 degrees C over a 15-day period showed that significant degradation processes and protein transformations occurred during the first 3-5 days. Different phases of these processes were identified.

A simple, precise and rapid pulse nuclear magnetic resonance technique for measuring the rate of water exchange across the erythrocyte membrane is presented. The effects of cell volume on cell water lifetime may be eliminated by the introduction of a cell water diffusion coefficient. This parameter is applied to the study of eleven blood samples.

The present knowledge of the physiology of H-Y antigen has been reviewed. It would appear that the results that have been obtained contradict one another and this brings into doubt the role that this antigen plays in testicular differentiation and the link it has with the Y chromosome.

136 patients admitted into a psychiatric unit underwent a routine caryotype examination, without any previous selection, 9 anomalies were noticed, including one case of X-tetrasomia, reported elsewhere, and eight mosaics. The patients concerned do not present any morphological anomaly or intellectual deficiency. The authors consider those--mostly gonosomial--mosaics as a minimal brain damage, favourising the appearance of the particular psychiatric syndrome.

The frozen tissue was sliced and then homogenized at 20 degree C in LiCl, 2 M; lauryl-trimethyl ammonium chloride (KK No. 4484), 5%; pronase, B grade (calbiochem), 1 mg/ml and Tris-HCl 10 mM pH 7.5. The homogenate was left to stand 15 min at 20 degree C with occasional shaking. After centrifugation at 35 000 X g for 30 min at 0 degree C, the supernatant containing the crude DNA was purified by filtration on Ultrogel A 2 (LKB, Sweden). The Ultrogel A 2 column (2.5 X 45 cm) was equilibrated with a solution containing NaCl 2 M, EDTA 2.5 mM, Tris-HCl 10 mM pH 7.5. The flow rate was 3 ml cm-2 h-1. Five ml of the supernatant were placed on the column. The first peak contained highly polymerized (as demonstrated by ultracentrifugation) pure DNA (A260/A230 = 3.19; A260/A280 = 1.82). The yield was 2.26 mg of DNA/g of fresh liver.

During February 1979 to December 1982 we studied the karyotype of 706 psychiatric adult patients without making a clinical selection. We found a chromosomial mosaïc aberration in 37 cases, concerning one leukocyte mitosis for ten studied cells : so a proportion of 5.2% cases. There do not exist statistics concerning the general population's mosaïc aberrations but in the same population the proportion of the homogeneous aberrations is near 0.5%, so 10 times lower than the proportion that we observed. The largest number of aberrations found concern the gonosomes, essentially some monosomics 45,XO or some trisomics 47,3X, among women who present a maniac depressive psychosis or another type of depression.

The differentiation between inflammatory and non inflammatory states has been performed using Nuclear Magnetic Resonance (NMR) in vitro by measuring relaxation times T1 and T2 in 84 synovials fluids obtained from various rheumatologic diseases. The results show that the T1/T2 ratio is more sensitive to distinguish these two situations rather than the isolated T1 or T2 values. In particular, high values of T1/T2 ratio are found in septic arthritis.

The Philadelphia chromosome (Ph) is an abbreviated chromosome number 22 resulting, in the majority of cases, from a balanced translocation between the 9 and 22 long arms. It is considered a marker of chronic myeloid leukemia and its diagnostic and prognostic value in this disease has been demonstrated. It is also present in a number of patients with acute leukemias of poor prognosis. The Ph arises in a bone marrow progenitor cell and seems to allow the clonal expansion of the malignant cells. The typical 9;22 translocation results in the transposition of the cellular oncogene Abelson in close proximity of chromosome 22 linked critical sequences. This structural change leads to the transcription of an hybrid mRNA coding for an abnormal protein with a tyrosine kinase activity which could play a major role in the leukemogenesis process.

The sex-differentiation H-Y antigen was studied in 5 males subjects with Y-chromosome polysomy. All had gonadal deficiency and significant decrease of H-Y antigen as compared with 77 normal 46, XY male controls. Men with additional Y-gonosomes may be hypogonadic or, more often, fertile, as was probably the case with the 3 patients with very high H-Y antigen titers previously described by other research workers. The phenotypic polymorphism of these subjects therefore seems to be parallel with their H-Y antigen polymorphism and there may be, in our 5 subjects, a relationship between abnormal spermatogenesis and H-Y antigen deficiency.

A cytogenetic study performed on 6 human renal cell carcinomas after short-term culture on extracellular matrix with serum-free medium showed aneuploidy without structural changes in 2 tumors and a rearrangement of the short arm of chromosome 3 in 4 tumors, including deletions and a translocation involving the 3p14 and 3p21 bands. Chromosomal in situ hybridization with a c-raf 1 probe demonstrated that in 2 renal cancers with del3(p14 or 21) the cellular oncogene had shifted from 3p25 to 3p14 as a result of an interstitial deletion.

Human ovarian adenocarcinoma cells IGROV1 were induced to become resistant in vitro to Vincristine (VCR) by continuous and discontinuous stepwise exposure to the drug. Both sublines exhibited similar indexes of resistance (approx. 800). The acquisition of VCR resistance was associated with changes of morphology and with cross-resistance to Adriamycin, VP16 and actinomycin D. Addition of verapamil enhanced sensitivity of resistant cells to VCR. This multi-drug-resistant (MDR) phenotype was associated with high levels of human MDR1 gene transcripts (25- to 100-fold). In contrast to other reports, no significant amplification of the locus could be observed. Reactivity with MRK16, a monoclonal antibody (MAb) directed against the MDR 170 kDa protein, was found to be related to the level of MDR1 transcription. In addition, both OV1/VCR sublines exhibited a marked loss of tumorigenicity. Besides the chromosomal markers of OV1/p, the two resistant sublines revealed a common cytogenetic rearrangement: a deletion of the short arm of one chromosome 11: del(11)(p12). We suggest that the acquisition of this marker might be associated with the non-tumorigenic phenotype of OV1/VCR-resistant sublines.

A fifth case of rec(18) resulting from recombination of a paternal pericentric inversion is described. The propositus' complement includes a chromosome 18 with partial deletion of the long arm, and partial duplication of the short. The recombination risk is evaluated at 5%. The eventuality of deleterious effects of pericentric inversions is discussed.

A cytogenetic study performed on 6 human renal cell carcinomas after short-term culture on extracellular matrix with serum-free medium showed aneuploidy without structural changes in 2 tumors and a rearrangement of the short arm of chromosome 3 in 4 tumors, including deletions and a translocation involving the 3p14 and 3p21 bands. Chromosomal in situ hybridization with a c-raf 1 probe demonstrated that in 2 renal cancers with del3(p14 or 21) the cellular oncogene had shifted from 3p25 to 3p14 as a result of an interstitial deletion.

A cytogenetic study was performed on 48 primary human benign, borderline malignant, and malignant solid tumors from more than 10 different tissues. An improved cell culture method using an extracellular matrix substrate and a polyvalent serum-free medium was applied, which ensured a success rate of about 68%. The results disclosed a nonrandom involvement of chromosomes 1, 3, and 7 (especially duplication of chromosome 7) in numerical changes and a clustering of breakpoints on chromosomes 1, 3, 7, and 11. These data indicate that some chromosomal changes are shared by various types of tumors, suggesting some common genetic clonal evolution pathways.