Peut-on optimiser la congélation des spermatozoïdes testiculaires? L’expérience du Centre Hospitalier de Poissy Saint-Germain

50% or more of non-obstructive azoospermic men have no spermatozoa in their testicular tissue, and no non-invasive predictor of spermatogenesis is yet available. For this reason, we therefore performed all TESE (74 TESE for non-obstructive azoospermia and 37 TESE for obstructive azoospermia) prior to initiating ovarian stimulation. 34% (25/74) of TESE performed for non-obstructive azoospermia were successful. Spermatozoa were retrieved in 100% of cases of obstructive azoospermia. When TESE were positive, spermatozoa were frozen in 25–50 μl micro-droplets (several straws). 60 ICSI cycles (25 couples) were treated for non obstructive azoospermia. The clinical pregnancy rate per ICSI cycle was 18%, and the implantation rate per embryo transferred was 9.2%. 81 ICSI cycles (37 couples) were treated for obstructive azoospermia. The fertilization rate was 54%, and embryo transfer was performed in 89% (72/81) of cycles. The clinical pregnancy rate per embryo ICSI cycle was 26%, and the implantation rate per embryo transferred was 16%. This management of azoospermic patients, including TESE and multiple testicular tissue freezing in micro-droplets prior to ovarian stimulation, avoids ova pick-up cancellation and multiple TESE, as several ICSI can be performed after a single TESE. Our results show that this micro-technique for freezing testicular tissue is effective not only for obstructive azoospermia, but also for non-obstructive azoospermia when only very few spermatozoa can be extracted from the testis.     

Polycyclic nitrogen heterocycles as potential thymidine phosphorylase inhibitors: synthesis,biological evaluation,and molecular docking study

New polycyclic heterocycles were synthesised and evaluated as potential inhibitors of thymidine phosphorylase (TP). Inspired by the pharmacophoric pyrimidinedione core of the natural substrate, four series have been designed in order to interact with large empty pockets of the active site: pyrimidoquinoline-2,4-diones (series A), pyrimidinedione linked to a pyrroloquinoline-1,3-diones (series B and C), the polycyclic heterocycle has been replaced by a pyrimidopyridopyrrolidinetetraone (series D). In each series, the tricyclic nitrogen heterocyclic moiety has been synthesised by a one-pot multicomponent reaction. Compared to 7-DX used as control, 2d, 2l, 2p (series A), 28a (series D), and the open intermediate 30 showed modest to good activities. A kinetic study confirmed that the most active compounds 2d, 2p are competitive inhibitors. Molecular docking analysis confirmed the interaction of these new compounds at the active binding site of TP and highlighted a plausible specific interaction in a pocket that had not yet been explored.     

The Notch pathway attenuates interleukin 1β (IL1β)-mediated induction of adenylyl cyclase 8 (AC8) expression during vascular smooth muscle cell (VSMC) trans-differentiation

Vascular smooth muscle cell (VSMC) trans-differentiation, or their switch from a contractile/quiescent to a secretory/inflammatory/migratory state, is known to play an important role in pathological vascular remodeling including atherosclerosis and postangioplasty restenosis. Several reports have established the Notch pathway as tightly regulating VSMC response to various stress factors through growth, migration, apoptosis, and de-differentiation. More recently, we showed that alterations of the Notch pathway also govern VSMC acquisition of the inflammatory state, one of the major events accelerating atherosclerosis. We also evidenced that the inflammatory context of atherosclerosis triggers a de novo expression of adenylyl cyclase isoform 8 (AC8), associated with the properties developed by trans-differentiated VSMCs. As an initial approach to understanding the regulation of AC8 expression, we examined the role of the Notch pathway. Here we show that inhibiting the Notch pathway enhances the effect of IL1β on AC8 expression, amplifies its deleterious effects on the VSMC trans-differentiated phenotype, and decreases Notch target genes Hrt1 and Hrt3. Conversely, Notch activation resulted in blocking AC8 expression and up-regulated Hrt1 and Hrt3 expression. Furthermore, overexpressing Hrt1 and Hrt3 significantly decreased IL1β-induced AC8 expression. In agreement with these in vitro findings, the in vivo rat carotid balloon-injury model of restenosis evidenced that AC8 de novo expression coincided with down-regulation of the Notch3 pathway. These results, demonstrating that the Notch pathway attenuates IL1β-mediated AC8 up-regulation in trans-differentiated VSMCs, suggest that AC8 expression, besides being induced by the proinflammatory cytokine IL1β, is also dependent on down-regulation of the Notch pathway occurring in an inflammatory context.     

Long-Term Survival of Legionella pneumophila in the Viable But Nonculturable State After Monochloramine Treatment

Legionella pneumophila, a facultative intracellular human pathogen, can persist for long periods in natural and artificial aquatic environments. Eradication of this bacterium from plumbing systems is often difficult. We tested L. pneumophila survival after monochloramine treatment. Survival was monitored using the BacLight Bacterial Viability Kit (Molecular Probes), ChemChrome V6 Kit (Chemunex), quantitative polymerase chain reaction and culturability on buffered charcoal-yeast extract agar. In nonculturable samples, regain of culturability was obtained after addition of the amoeba Acanthamoeba castellanii, and esterase activity and membrane integrity were observed after >4 months after treatment. These results demonstrate for the first time that L. pneumophila could persist for long periods in biofilms into the viable but nonculturable (VBNC) state. Monitoring L. pneumophila in water networks is generally done by enumeration on standard solid medium. This method does not take into account VBNC bacteria. VBNC L. pneumophila could persist for long periods and should be resuscitated by amoeba. These cells constitute potential sources of contamination and should be taken into account in monitoring water networks.     

CRL2LRR-1 E3-Ligase Regulates Proliferation and Progression through Meiosis in the Caenorhabditis elegans Germline

The ubiquitin-proteolytic system controls the stability of proteins in space and time. In this study, using a temperature-sensitive mutant allele of the cul-2 gene, we show that CRL2^LRR-1 (CUL-2 RING E3 ubiquitin-ligase and the Leucine Rich Repeat 1 substrate recognition subunit) acts at multiple levels to control germline development. CRL2^LRR-1 promotes germ cell proliferation by counteracting the DNA replication ATL-1 checkpoint pathway. CRL2^LRR-1 also participates in the mitotic proliferation/meiotic entry decision, presumably controlling the stability of meiotic promoting factors in the mitotic zone of the germline. Finally, CRL2^LRR-1 inhibits the first steps of meiotic prophase by targeting in mitotic germ cells degradation of the HORMA domain-containing protein HTP-3, required for loading synaptonemal complex components onto meiotic chromosomes. Given its widespread evolutionary conservation, CUL-2 may similarly regulate germline development in other organisms as well.     

From Oscillatory Transcranial Current Stimulation to Scalp EEG Changes: A Biophysical and Physiological Modeling Study

Both biophysical and neurophysiological aspects need to be considered to assess the impact of electric fields induced by transcranial current stimulation (tCS) on the cerebral cortex and the subsequent effects occurring on scalp EEG. The objective of this work was to elaborate a global model allowing for the simulation of scalp EEG signals under tCS. In our integrated modeling approach, realistic meshes of the head tissues and of the stimulation electrodes were first built to map the generated electric field distribution on the cortical surface. Secondly, source activities at various cortical macro-regions were generated by means of a computational model of neuronal populations. The model parameters were adjusted so that populations generated an oscillating activity around 10 Hz resembling typical EEG alpha activity. In order to account for tCS effects and following current biophysical models, the calculated component of the electric field normal to the cortex was used to locally influence the activity of neuronal populations. Lastly, EEG under both spontaneous and tACS-stimulated (transcranial sinunoidal tCS from 4 to 16 Hz) brain activity was simulated at the level of scalp electrodes by solving the forward problem in the aforementioned realistic head model. Under the 10 Hz-tACS condition, a significant increase in alpha power occurred in simulated scalp EEG signals as compared to the no-stimulation condition. This increase involved most channels bilaterally, was more pronounced on posterior electrodes and was only significant for tACS frequencies from 8 to 12 Hz. The immediate effects of tACS in the model agreed with the post-tACS results previously reported in real subjects. Moreover, additional information was also brought by the model at other electrode positions or stimulation frequency. This suggests that our modeling approach can be used to compare, interpret and predict changes occurring on EEG with respect to parameters used in specific stimulation configurations.     

Does land use within the home range drive the exposure of roe deer (Capreolus capreolus) to two abortive pathogens in a rural agro-ecosystem?

The recent increase in interactions between wildlife and domestic animals has heightened the risks of transmission of pathogens between wildlife, livestock, and humans. Our objective was to better understand how the spatial behavior of wildlife impacts their exposure to pathogens. We carried out a 5-year serological survey of Toxoplasma gondii and Chlamydia abortus in a roe deer population (Capreolus capreolus) living in a rural landscape and monitored using GPS technology. We tested whether the apparent seroprevalence of these pathogens was related to the characteristics of roe deer’s home range, indirectly reflecting the probability of contact between deer and domestic hosts (cats for T. gondii and ruminants for C. abortus). We controlled for the effects of temperature and rainfall, as local weather can affect pathogen survival in the environment. Overall, apparent seroprevalences were 43.7 and 15.8 % for T. gondii (N?=?135) and C. abortus (N?=?133), respectively. The apparent seroprevalence to T. gondii increased with the proportion of human dwellings (a proxy of cat presence) within the roe deer’s core home range and was higher during mild and wet years. This result illustrates how the spatial behavior of wild animals can modulate their exposure to pathogens through the degree of spatial overlap with a domestic host. By contrast, apparent seroprevalence to C. abortus increased with the annual number of frosty days but not with the proportion of meadows occupied by domestic ruminants within the core home range. This suggests that the cycle of C. abortus in wild animals could be somewhat independent from that in livestock.