Freeze/thaw stress induces organelle remodeling and membrane recycling in cryopreserved human mature oocytes

Purpose: Our aim was to evaluate the ultrastructure of human metaphase II oocytes subjected to slow freezing and fixed after thawing at different intervals during post-thaw rehydration. Methods: Samples were studied by light and transmission electron microscopy. Results: We found that vacuolization was present in all cryopreserved oocytes, reaching a maximum in the intermediate stage of rehydration. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates decreased following thawing, particularly in the first and intermediate stages of rehydration, whereas mitochondria-vesicle (MV) complexes augmented in the same stages. At the end of rehydration, vacuoles and MV complexes both diminished and M-SER aggregates increased again. Cortical granules (CGs) were scarce in all cryopreserved oocytes, gradually diminishing as rehydration progressed. Conclusions: This study also shows that such a membrane remodeling is mainly represented by a dynamic process of transition between M-SER aggregates and MV complexes, both able of transforming into each other. Vacuoles and CG membranes may take part in the membrane recycling mechanism

Prostasome-like particles in stallion semen.

Human semen contains membranous vesicles called prosta- somes. They are secreted by the prostate gland and contain large amounts of cholesterol, sphingomyelin, and Ca2. Prostasomes enhance the motility of ejaculated spermatozoa and are in- volved in a number of additional biological functions. No prostasome-like vesicles have been described in horse se- men up to now. We have demonstrated the presence of pros- tasome-like vesicles in the equine semen and characterized them as to size, morphology, and lipid composition; we have found that they are similar to human prostasomes in many re- spects. We propose that these vesicles might be important for the fecundity of horse semen. This is of interest since the success of artificial insemination is limited by the fact that stallion sperm barely survive cryopreservation

Revision of the 15N(p,{\gamma})16O reaction rate and oxygen abundance in H-burning zones

The NO cycle takes place in the deepest layer of a H-burning core or shell, when the temperature exceeds T {\simeq} 30 {\cdot} 106 K. The O depletion observed in some globular cluster giant stars, always associated with a Na enhancement, may be due to either a deep mixing during the RGB (red giant branch) phase of the star or to the pollution of the primordial gas by an early population of massive AGB (asymptotic giant branch) stars, whose chemical composition was modified by the hot bottom burning. In both cases, the NO cycle is responsible for the O depletion. The activation of this cycle depends on the rate of the 15N(p,{\gamma})16O reaction. A precise evaluation of this reaction rate at temperatures as low as experienced in H-burning zones in stellar interiors is mandatory to understand the observed O abundances. We present a new measurement of the 15N(p,{\gamma})16O reaction performed at LUNA covering for the first time the center of mass energy range 70-370 keV, which corresponds to stellar temperatures between 65 {\cdot} 106 K and 780 {\cdot}106 K. This range includes the 15N(p,{\gamma})16O Gamow-peak energy of explosive H-burning taking place in the external layer of a nova and the one of the hot bottom burning (HBB) nucleosynthesis occurring in massive AGB stars. With the present data, we are also able to confirm the result of the previous R-matrix extrapolation. In particular, in the temperature range of astrophysical interest, the new rate is about a factor of 2 smaller than reported in the widely adopted compilation of reaction rates (NACRE or CF88) and the uncertainty is now reduced down to the 10% level.Comment: 6 pages, 5 figure

Protocol for the DeFOG trial: A randomized controlled trial on the effects of smartphone-based, on-demand cueing for freezing of gait in Parkinson's disease

Background: Freezing of gait (FOG) is a highly incapacitating symptom that affects many people with Parkinson's disease (PD). Cueing triggered upon real-time FOG detection (on-demand cueing) shows promise for FOG treatment. Yet, the feasibility of implementation and efficacy in daily life is still unknown. Therefore, this study aims to investigate the effectiveness of DeFOG: a smartphone and sensor-based on-demand cueing solution for FOG. Methods: Sixty-two PD patients with FOG will be recruited for this single-blind, multi-center, randomized controlled phase II trial. Patients will be randomized into either the intervention group or the active control group. For four weeks, both groups will receive feedback about their physical activity using the wearable DeFOG system in daily life. In addition, the intervention group will also receive on-demand auditory cueing and instructions. Before and after the intervention, home-based assessments will be performed to evaluate the primary outcome, i.e., “percentage time frozen” during a FOG-provoking protocol. Secondary outcomes include the training effects on physical activity monitored over 7 days and the user-friendliness of the technology. Discussion: The DeFOG trial will investigate the effectiveness of personalized on-demand cueing in a controlled design, delivered for 4 weeks in the patient's home environment. We anticipate that DeFOG will reduce FOG to a greater degree than in the control group and we will explore the impact of the intervention on physical activity levels. We expect to gain in-depth insight into whether and how patients control FOG using cueing methods in their daily lives. Trial registration: Clinicaltrials.gov NCT03978507

Effects of the pesticide lindane on granulosa cell ultrastructure

The excessive exposure to pesticides in the Aral Sea area was correlated to the increased reproductive pathologies in those regions. One of the principal chemical employed was the gamma-hexachlorocycloexane herbicide Lindane (L), a persistent organochlorine that may induces alterations in granulosa cell (GCs) survival. However, a comprehensive experimental study on the L-induced dose-effect morphological alterations, has not yet addressed. Therefore, we studied by means of transmission and scanning electron microscopy, the morphological changes of mouse GCs, matured in vitro with increasing concentrations of L. GCs showed several dose-dependent changes, in respect to controls. In particular, we observed significant reduction of GC microvilli and decrease of cytoplasmic processes between adjacent GCs. In addition, peripheral aggregation of chromatin under the nuclear membrane, extensive plasma membrane blebbing, abundant GC remnants and cellular debris were also present. Mitochondria, endoplasmic reticula and Golgi apparatuses did not show significant changes. In conclusion, our results showed a dose-dependent toxicity of L on GCs, associated to morphological signs of apoptosis. Alterations of GCs may be associated to impaired oocyte competence and sterility

Algorithms for walking speed estimation using a lower-back-worn inertial sensor: A cross-validation on speed ranges

Walking/gait speed is a key measure for daily mobility characterization. To date, various studies have attempted to design algorithms to estimate walking speed using an inertial sensor worn on the lower back, which is considered as a proper location for activity monitoring in daily life. However, these algorithms were rarely compared and validated on the same datasets, including people with different preferred walking speed. This study implemented several original, improved, and new algorithms for estimating cadence, step length and eventually speed. We designed comprehensive cross-validation to compare the algorithms for walking slow, normal, fast, and using walking aids. We used two datasets, including reference data for algorithm validation from an instrumented mat (40 subjects) and shanks-worn inertial sensors (88 subjects), with normal and impaired walking patterns. The results showed up to 50% performance improvements. Training of algorithms on data from people with different preferred speeds led to better performance. For the slow walkers, an average RMSE of 2.5 steps/min, 0.04 m, and 0.10 m/s were respectively achieved for cadence, step length, and speed estimation. For normal walkers, the errors were 3.5 steps/min, 0.08 m, and 0.12 m/s. An average RMSE of 1.3 steps/min, 0.05 m, and 0.10 m/s were also observed on fast walkers. For people using walking aids, the error significantly increased up to an RMSE of 14 steps/min, 0.18 m, and 0.27 m/s. The results demonstrated the robustness of the proposed combined speed estimation approach for different speed ranges. It achieved an RMSE of 0.10, 0.18, 0.15, and 0.32 m/s for slow, normal, fast, and using walking aids, respectively

Noncanonical Fungal Autophagy Inhibits Inflammation in Response to IFN-γ via DAPK1

Defects in a form of noncanonical autophagy, known as LC3-associated phagocytosis (LAP), lead to increased inflammatory pathology during fungal infection. Although LAP contributes to fungal degradation, the molecular mechanisms underlying LAP-mediated modulation of inflammation are unknown. We describe a mechanism by which inflammation is regulated during LAP through the death-associated protein kinase 1 (DAPK1). The ATF6/C/EBP-β/DAPK1 axis activated by IFN-γ not only mediates LAP to Aspergillus fumigatus but also concomitantly inhibits Nod-like receptor protein 3 (NLRP3) activation and restrains pathogenic inflammation. In mouse models and patient samples of chronic granulomatous disease, which exhibit defective autophagy and increased inflammasome activity, IFN-γ restores reduced DAPK1 activity and dampens fungal growth. Additionally, in a cohort of hematopoietic stem cell-transplanted patients, a genetic DAPK1 deficiency is associated with increased inflammation and heightened aspergillosis susceptibility. Thus, DAPK1 is a potential drugable player in regulating the inflammatory response during fungal clearance initiated by IFN-γ

Synchronization and directed percolation in coupled map lattices

We study a synchronization mechanism, based on one-way coupling of all-or-nothing type, applied to coupled map lattices with several different local rules. By analyzing the metric and the topological distance between the two systems, we found two different regimes: a strong chaos phase in which the transition has a directed percolation character and a weak chaos phase in which the synchronization transition occurs abruptly. We are able to derive some analytical approximations for the location of the transition point and the critical properties of the system. We propose to use the characteristics of this transition as indicators of the spatial propagation of chaoticity.Comment: 12 pages + 12 figure

Gemcitabine and docetaxel in relapsed and unresectable high-grade osteosarcoma and spindle cell sarcoma of bone

BACKGROUND: Few new compounds are available for relapsed osteosarcoma. We retrospectively evaluated the activity of gemcitabine (G) plus docetaxel (D) in patients with relapsed high-grade osteosarcoma and high-grade spindle cell sarcoma of bone (HGS). METHODS: Patients receiving G 900 mg/m(2) d 1, 8; D 75 mg/m(2) d 8, every 21 days were eligible. Primary end-point: progression-free survival (PFS) at 4 months; secondary end-point: overall survival (OS) and response rate. RESULTS: Fifty-one patients were included, with a median age of 17 years (8–71), 26 (51 %) were pediatric patients. GD line of treatment: 2nd in 14 patients, ≥3rd in 37. 25 (49 %) patients had metastases limited to lungs, 26 (51 %) multiple sites. Histology: 40 (78 %) osteosarcoma, 11 (22 %) HGS. Eight (16 %) patients achieved surgical complete response (sCR2) after GD. Four-month PFS rate was 46 %, and significantly better for patients with ECOG 0 (ECOG 0: 54 % vs ECOG 1: 43 % vs ECOG 2: 0 %; p = 0.003), for patients undergoing metastasectomy after GD (sCR2 75 % vs no-sCR2 40 %, p = 0.02) and for osteosarcoma (osteosarcoma 56 % vs HGS 18 %; p = 0.05), with no differences according to age, line of treatment, and pattern of metastases. Forty-six cases had RECIST measurable disease: 6 (13 %) patients had a partial response (PR), 20 (43 %) had stable disease (SD) and 20 (43 %) had progressive disease (PD). The 1-year OS was 30 %: 67 % for PR, 54 % for SD and 20 % for PD (p = 0.005). CONCLUSIONS: GD is an active treatment for relapsed high-grade osteosarcoma, especially for ECOG 0 patients, and should be included in the therapeutic armamentarium of metastatic osteosarcoma