Pre-implantation mouse embryos cultured In vitro under different oxygen concentrations show altered ultrastructures

Abstract Assisted Reproductive Technologies routinely utilize different culture media and oxygen (O2) concentrations to culture human embryos. Overall, embryos cultured under physiological O2 tension (5%) have improved development compared to embryos cultured under atmospheric O2 conditions (20%). The mechanisms responsible for this remain unclear. This study aimed to evaluate the effect of physiologic (5%) or atmospheric O2 (20%) tension on the microscopic ultrastructure of pre-implantation mouse embryos using Transmission Electron Microscopy (TEM). Embryos flushed out of the uterus after natural mating were used as the control. For use as the control, 2-cells, 4-cells, morulae, and blastocysts were flushed out of the uterus after natural fertilization. In vitro fertilization (IVF) was performed using potassium simplex optimized medium (KSOM) under different O2 tensions (5% and 20%) until the blastocyst stage. After collection, embryos were subjected to the standard preparative for light microscopy (LM) and TEM. We found that culture in vitro under 5% and 20% O2 results in an increase of vacuolated shaped mitochondria, cytoplasmic vacuolization and presence of multi-vesicular bodies at every embryonic stage. In addition, blastocysts generated by IVF under 5% and 20% O2 showed a lower content of heterochromatin, an interruption of the trophectodermal and inner cell mass cell membranes, an increased density of residual bodies, and high levels of glycogen granules in the cytoplasm. In conclusion, this study suggests that in vitro culture, particularly under atmospheric O2 tension, causes stage-specific changes in preimplantation embryo ultrastructure. In addition, atmospheric (20%) O2 is associated with increased alterations in embryonic ultrastructure; these changes may explain the reduced embryonic development of embryos cultured with 20% O2

Facility for validating technologies for the autonomous space rendezvous and docking to uncooperative targets

We present the latest advancements in the air-bearing facility installed at La Sapienza’s GN Lab in the School of Aerospace Engineering. This facility has been utilized in recent times to validate robust control laws for simultaneous attitude control and vibration active damping. The instrumentation and testbed have been restructured and enhanced to enable simulations of close proximity operations. Relative pose determination, accomplished through visual navigation as either an auxiliary or standalone system, is the first building block. Leveraging the acquired knowledge, optimal guidance and control algorithms can be tested for contactless operations (e.g. on-orbit inspection), as well as berthing and docking tasks

On the Origin of the Early Solar System Radioactivities. Problems with the AGB and Massive Star Scenarios

Recent improvements in stellar models for intermediate-mass and massive stars are recalled, together with their expectations for the synthesis of radioactive nuclei of lifetime $\tau \lesssim 25$ Myr, in order to re-examine the origins of now extinct radioactivities, which were alive in the solar nebula. The Galactic inheritance broadly explains most of them, especially if $r$-process nuclei are produced by neutron star merging according to recent models. Instead, $^{26}$Al, $^{41}$Ca, $^{135}$Cs and possibly $^{60}$Fe require nucleosynthesis events close to the solar formation. We outline the persisting difficulties to account for these nuclei by Intermediate Mass Stars (2 $\lesssim$ M/M$_\odot \lesssim 7 - 8$). Models of their final stages now predict the ubiquitous formation of a $^{13}$C reservoir as a neutron capture source; hence, even in presence of $^{26}$Al production from Deep Mixing or Hot Bottom Burning, the ratio $^{26}$Al/$^{107}$Pd remains incompatible with measured data, with a large excess in $^{107}$Pd. This is shown for two recent approaches to Deep Mixing. Even a late contamination by a Massive Star meets problems. In fact, inhomogeneous addition of Supernova debris predicts non-measured excesses on stable isotopes. Revisions invoking specific low-mass supernovae and/or the sequential contamination of the pre-solar molecular cloud might be affected by similar problems, although our conclusions here are weakened by our schematic approach to the addition of SN ejecta. The limited parameter space remaining to be explored for solving this puzzle is discussed.Comment: Accepted for publication on Ap

MEMS-based Inertial Navigation Systems onboard Balloons

Performances of low-cost inertial navigation sensors, usually poor, can often match different mission requirements by means of a careful signal and data processing and/or an augmentation by means of different observables. The paper presents the LOWCOINS navigation experiment, intended to fly onboard BEXUS balloon mission in late 2008. LOWCOINS has as the main component a low-cost three-axes inertial unit, integrating three accelerometers and three gyros. The slow dynamic typical of a balloon flight is deemed as an ideal test to verify the performances of the unit and to improve the knowledge on the data processing needed to obtain an accurate final navigation solution. In order to enlarge the set of available data, a cluster of magnetometers and a pressure sensor, always belonging to low-cost instrumentation range, are hosted on board. Measurements are both stored on board and downlinked to a ground station. Position and velocity components (both the onboard computed first guess and the post-flight calibrated solution) will be compared with the data gathered by a GPS receiver, which is a standard component of BEXUS balloon avionics. Substantial attention to thermal aspects has been requested in order to cope with environmental conditions prior of and all along the flight. The requested navigation unit case design is shortly reported

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

FRAT-up, a Web-based fall-risk assessment tool for elderly people living in the community.

Background: About 30% of people over 65 are subject to at least one unintentional fall a year. Fall prevention protocols and interventions can decrease the number of falls. To be effective, a prevention strategy requires a prior step to evaluate the fall risk of the subjects. Despite extensive research, existing assessment tools for fall risk have been insufficient for predicting falls. Objective: The goal of this study is to present a novel web-based fall-risk assessment tool (FRAT-up) and to evaluate its accuracy in predicting falls, within a context of community-dwelling persons aged 65 and up. Methods: FRAT-up is based on the assumption that a subject\u2019s fall risk is given by the contribution of their exposure to each of the known fall-risk factors. Many scientific studies have investigated the relationship between falls and risk factors. The majority of these studies adopted statistical approaches, usually providing quantitative information such as odds ratios. FRAT-up exploits these numerical results to compute how each single factor contributes to the overall fall risk. FRAT-up is based on a formal ontology that enlists a number of known risk factors, together with quantitative findings in terms of odds ratios. From such information, an automatic algorithm generates a rule-based probabilistic logic program, that is, a set of rules for each risk factor. The rule-based program takes the health profile of the subject (in terms of exposure to the risk factors) and computes the fall risk. A Web-based interface allows users to input health profiles and to visualize the risk assessment for the given subject. FRAT-up has been evaluated on the InCHIANTI Study dataset, a representative population-based study of older persons living in the Chianti area (Tuscany, Italy). We compared reported falls with predicted ones and computed performance indicators. Results: The obtained area under curve of the receiver operating characteristic was 0.642 (95% CI 0.614-0.669), while the Brier score was 0.174. The Hosmer-Lemeshow test indicated statistical significance of miscalibration. Conclusions: FRAT-up is a web-based tool for evaluating the fall risk of people aged 65 or up living in the community. Validation results of fall risks computed by FRAT-up show that its performance is comparable to externally validated state-of-the-art tools. A prototype is freely available through a web-based interface. Trial Registration: ClinicalTrials.gov NCT01331512 (The InChianti Follow-Up Study); http://clinicaltrials.gov/show/NCT01331512 (Archived by WebCite at http://www.webcitation.org/6UDrrRuaR)

Adoptive cell therapy of triple negative breast cancer with redirected cytokine-induced killer cells

Cytokine-Induced Killer (CIK) cells share several functional and phenotypical properties of both T and natural killer (NK) cells. They represent an attractive approach for cell-based immunotherapy, as they do not require antigen-specific priming for tumor cell recognition, and can be rapidly expanded in vitro. Their relevant expression of Fc\u3b3RIIIa (CD16a) can be exploited in combination with clinical-grade monoclonal antibodies (mAbs) to redirect their lytic activity in an antigen-specific manner. Here, we report the efficacy of this combined approach against triple negative breast cancer (TNBC), an aggressive tumor that still requires therapeutic options. Different primitive and metastatic TNBC cancer mouse models were established in NSG mice, either by implanting patient-derived TNBC samples or injecting MDA-MB-231 cells orthotopically or intravenously. The combined treatment consisted in the repeated intratumoral or intravenous injection of CIK cells and cetuximab. Tumor growth and metastasis were monitored by bioluminescence or immunohistochemistry, and survival was recorded. CIK cells plus cetuximab significantly restrained primitive tumor growth in mice, either in patient-derived tumor xenografts or MDA-MB-231 cell line models. Moreover, this approach almost completely abolished metastasis spreading and dramatically improved survival. The antigen-specific mAb favored tumor and metastasis tissue infiltration by CIK cells, and led to an enrichment of the CD16a+ subset. Data highlight the potentiality of this novel immunotherapy strategy where a nonspecific cytotoxic cell population can be converted into tumor-specific effectors with clinical-grade antibodies, thus providing not only a therapeutic option for TNBC but also a valid alternative to more complex approaches based on chimeric antigen receptor-engineered cells. List of abbreviations: ACT, Adoptive Cell Transfer; ADCC, Antibody-Dependent Cell-mediated Cytotoxicity; ADP, Adenosine diphosphate; BLI, Bioluminescence Imaging; CAR, Chimeric Antigen Receptor; CIK, Cytokine Induced Killer cells; CTX, Cetuximab; DMEM, Dulbecco\u2019s Modified Eagle Medium; EGFR, Human Epidermal Growth Factor 1; ER, Estrogen; FBS, Fetal Bovine Serum; FFPE, Formalin-Fixed Paraffin-Embedded; GMP, Good Manufacturing Practices; GVHD, Graft Versus Host Disease; HER2, Human Epidermal Growth Factor 2; HRP, Horseradish Peroxidase; IFN-\u3b3, Interferon-\u3b3; IHC, Immunohistochemistry; IL-2, Interleukin-2; ISO, Irrelevant antibody; i.t., intratumoral; i.v., intravenous, mAbs, Monoclonal Antibodies; mIHC, Multiplex Fluorescence Immunohistochemistry; MHC, Major Histocompatibility Complex; NK, Natural Killer; NKG2D, Natural-Killer group 2 member D; NSG, NOD/SCID common \u3b3 chain knockout; PARP, Poly ADP-ribose polymerase; PBMCs, Peripheral Blood Mononuclear Cells; PBS, Phosphate-buffered saline; PDX, Patient-derived xenograft; PR, Progesterone; rhIFN-\u3b3, Recombinant Human Interferon-\u3b3; RPMI, Roswell Park Memorial Institute; STR, Short tandem Repeat; TCR, T Cell Receptor; TNBC, Triple Negative Breast Cancer; TSA, Tyramide Signal Amplification