BACKGROUND: The aim of this study was to analyse the relationship between the first polar body (1st PB) morphology and the fertilization rate, cleavage rate, embryo quality, pregnancy and ...implantation rate. METHODS: This was a retrospective study on 167 consecutive cycles undergoing assisted reproduction with ICSI. The 1st PB morphology was evaluated at the moment of ICSI in the 596 injected oocytes and it was coded as intact or fragmented. The fertilization rate, cleavage rate, embryo quality (three grades), pregnancy rate, implantation rate and the time elapsed between oocyte retrieval and ICSI were evaluated. The 1st PB morphology was checked twice (denudation and ICSI) in a random sample of 180 oocytes in order to verify the effect of the in vitro culture. RESULTS: No significant relationship was found between the 1st PB morphology and the fertilization rate (P=0.703), cleavage rate (P=0.055), embryo quality (P=0.673), pregnancy rate (P=0.201) and implantation rate (P=0.511). A significant positive relationship (P=0.006) was found between the frequency of the 1st PB fragmentation and the time elapsed between denudation and ICSI. The pregnancy rate was significantly higher (P=0.008) when oocytes were injected between 5 and 7 h after retrieval rather than earlier or later. CONCLUSIONS: Our data suggest that the embryo quality, pregnancy rate and implantation rate are not related to the 1st PB fragmentation. The time which elapses between the oocyte retrieval and ICSI should be maintained at ∼6 h in order to obtain optimal results.
Background: In mild head injury, predictors to select patients for computed tomography (CT) and/or to plan proper management are needed. The strength of evidence of published recommendations is ...insufficient for current use. We assessed the diagnostic accuracy and the clinical validity of the proposal of the Neurotraumatology Committee of the World Federation of Neurosurgical Societies on mild head injury from an emergency department perspective. Methods: In a three year period, 5578 adolescent and adult subjects were prospectively recruited and managed according to the proposed protocol. Outcome measures were: (a) any post-traumatic lesion; (b) need for neurosurgical intervention; (c) unfavourable outcome (death, permanent vegetative state or severe disability) after six months. The predictive value of a model based on five variables (Glasgow coma score, clinical findings, risk factors, neurological deficits, and skull fracture) was tested by logistic regression analysis. Findings: At first CT evaluation 327 patients (5.9%) had intracranial post-traumatic lesions. In 16 cases (0.3%) previously undiagnosed lesions were detected after re-evaluation within seven days. Neurosurgical intervention was needed in 71 patients (1.3%) and an unfavourable outcome occurred in 39 cases (0.7%). The area under the ROC curve of the variables in predicting post-traumatic lesions was 0.906 (0.009) (sensitivity 70.0%, specificity 94.1% at best cut off), neurosurgical intervention was 0.926 (0.016) (sensitivity 81.7%, specificity 94.1%), and unfavourable outcome was 0.953 (0.014) (sensitivity 88.1%, specificity 95.1%). Interpretation: The variables prove highly accurate in the prediction of clinically meaningful outcomes, when applied to a consecutive set of patients with mild head injury in the clinical setting of a 1st level emergency department.
AGILE is a mission of the Italian Space Agency (ASI) Scientific Program dedicated to γ-ray astrophysics, and has operated in a low Earth orbit since 2007 April 23. It is designed to be a very light ...and compact instrument, capable of simultaneously detecting and imaging photons in the 18-60 keV X-ray energy band and in the 30 MeV-50 GeV γ-ray energy range with a good angular resolution ( 1◦ @ 1 GeV). The core of the instrument is the Silicon Tracker, supplemented by a CsI calorimeter and an AntiCoincidence system, which form the Gamma Ray Imaging Detector (GRID). Before launch, the GRID needed on-ground calibration with a tagged γ-ray beam to estimate its performance and validate the Monte Carlo simulation. The GRID was calibrated using a tagged γ-ray beam with energy up to 500 MeV at the Beam Test Facilities at the INFN Laboratori Nazionali di Frascati. These data are used to validate a GEANT 3-based simulation by comparing the data and the Monte Carlo simulation by measuring the angular and energy resolutions. The GRID angular and energy resolutions obtained using the beam agree well with the Monte Carlo simulation. Therefore the simulation, can be used to simulate the same performance on-flight with high reliability.
The mini-calorimeter of the AGILE satellite can observe the high-energy part of gamma-ray bursts with good timing-capability. We present the data of the 85 hard gamma-ray bursts observed by the ...mini-calorimeter since the launch (April 2007) until October 2009. We report the timing data for 84 and spectral data for 21 bursts.
We report on a multiwavelength observation of the blazar 3C 454.3 (which we dubbed crazy diamond) carried out on November 2007 by means of the astrophysical satellites AGILE, International Gamma-Ray ...Astrophysics Laboratory (INTEGRAL), Swift, the Whole Earth Blazar Telescope (WEBT) Consortium, and the optical-NIR telescope Rapid Eye Mount (REM). Thanks to the wide field of view of the AGILE satellite and its prompt alert dissemination to other observatories, we obtained a long (three weeks), almost continuous gamma -ray coverage of the blazar 3C 454.3 across 14 decades of energy. This broadband monitoring allows us to study in great detail light curves, correlations, time lags, and spectral energy distributions (SEDs) during different physical states. Gamma-ray data were collected during an AGILE pointing toward the Cygnus Region. Target of Opportunity (ToO) observations were performed to follow up the gamma -ray observations in the soft and hard X-ray energy bands. Optical data were acquired continuously by means of a preplanned WEBT campaign and through an REM ToO repointing. 3C 454.3 is detected at a ~19 sigma level during the three-week observing period, with an average flux above 100 MeV of F E > 100 MeV = (170 ± 13) X 10-8 photons cm-2 s-1. The gamma -ray spectrum can be fitted with a single power law with photon index Gamma GRID = 1.73 ± 0.16 between 100 MeV and 1 GeV. We detect significant day-by-day variability of the gamma -ray emission during our observations, and we can exclude that the fluxes are constant at the 99.6% (~2.9 sigma ) level. The source was detected typically around 40 deg off-axis, and it was substantially off-axis in the field of view of the AGILE hard X-ray imager. However, a five-day long ToO observation by INTEGRAL detected 3C 454.3 at an average flux of about F 20-200 keV = 1.49 X 10-3 photons cm-2 s-1with an average photon index of Gamma IBIS = 1.75 ± 0.24 between 20-200 keV. Swift also detected 3C 454.3 with a flux in the 0.3-10 keV energy band in the range (1.23-1.40) X 10-2 photons cm-2 s-1 and a photon index in the range Gamma XRT = 1.56-1.73. In the optical band, both WEBT and REM show an extremely variable behavior in the R band. A correlation analysis based on the entire data set is consistent with no time lags between the gamma -ray and the optical flux variations. Our simultaneous multifrequency observations strongly indicate that the dominant emission mechanism between 30 MeV and 30 GeV is dominated by inverse Compton scattering of relativistic electrons in the jet on the external photons from the broad line region.
Abstract
STUDY QUESTION
Which is the best method for human ovarian tissue cryopreservation: slow freezing/rapid thawing (SF/RT) or vitrification/warming (V/W)?
SUMMARY ANSWER
The conventional SF/RT ...protocol used in this study seems to better preserve the morpho-functional status of human cryopreserved ovarian tissue than the used open carrier V/W protocol.
WHAT IS KNOWN ALREADY
Cryopreservation of human ovarian tissue is generally performed using the SF/RT method. However, reduction in the follicular pool and stroma damage are often observed. An emerging alternative procedure is represented by V/W which seems to allow the maintenance of the morphological integrity of the stroma.
STUDY DESIGN, SIZE, DURATION
This is a retrospective cohort study including six patients affected by oncological diseases and enrolled from January to December 2014.
PARTICIPANTS/MATERIALS, SETTING, METHODS
Ovarian tissue was laparoscopically harvested from the right and left ovaries and was cryopreserved using a routinary SF/RT protocol or a V/W method, involving tissue incubation in two solutions (containing propylene glycol, ethylene glycol and sucrose at different concentrations) and vitrification in an open system. For each patient, three pieces from each ovary were collected at the time of laparoscopy (fresh tissue) and after storage (SF/RT or V/W) and processed for light microscopy (LM) and transmission electron microscopy (TEM), to assess the morphological and ultrastructural features of follicles and stroma, and for laser scanning confocal microscopy (LSCM), to determine the functional energetic/redox stroma status. The preservation status of SF/RT and V/W ovarian tissues was compared with that of fresh ones, as well as between them.
MAIN RESULTS AND THE ROLE OF CHANCE
By LM and TEM, SF/RT and V/W samples showed cryodamage of small entity. Interstitial oedema and increased stromal cell vacuolization and chromatin clumping were observed in SF/RT samples; in contrast, V/W samples showed oocyte nuclei with slightly thickened chromatin and irregular shapes. The functional imaging analysis by LSCM revealed that the mitochondrial activity and intracellular reactive oxygen species levels were reduced both in SF/RT and in V/W samples compared with fresh samples. The study also showed progressive dysfunction of the mitochondrial activity going from the outer to the inner serial section of the ovarian cortex. The reduction of mitochondrial activity of V/W samples compared with fresh samples was significantly higher in the inner section than in the outer section.
LIMITATIONS, REASONS FOR CAUTION
The results report the bioenergetic and oxidative status assessment of fresh and cryopreserved human ovarian tissue by LSCM, a technique recently applied to tissue samples. The use of LSCM on human ovarian tissues after SF/RT or V/W is a new application that requires validation. The procedures for mitochondrial staining with functional probes and fixing are not yet standardized. Xenografting of the cryopreserved ovarian tissue in severe combined immunodeficient mice and in vitro culture have not yet been performed.
WIDER IMPLICATIONS OF THE FINDINGS
The identification of a cryopreservation method able to maintain the morpho-functional integrity of the ovarian tissue and a number of follicles comparable with those observed in fresh tissue might optimize results in clinical practice, in terms of recovery, duration of ovarian function and increased delivery outcomes after replanting. The SF/RT protocol allowed better morpho-functional tissue integrity than the V/W procedure.
STUDY FUNDING/COMPETING INTEREST(S)
Funding was provided by Fondazione del Monte di Bologna e Ravenna, Italy. Dr N.A.M. was granted by the project ONEV MIUR PONa3 00134-n.254/R&C 18 5 2011 and the project GR-2011-02351396 (Ministry of Health, Young Researchers Grant 2011/2012). There are no competing interests.
TRIAL REGISTRATION NUMBER
Clinical trial 74/2001/0 (approved:13 2 2002): ‘Pilot study on cryopreservation of human ovarian tissue: morphological and immunohistochemical analysis before and after cryopreservation’.
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