Two-step transplantation with adipose tissue-derived stem cells increases follicle survival by enhancing vascularization in xenografted frozen-thawed human ovarian tissue.

STUDY QUESTION: Do adipose tissue-derived stem cells (ASCs) enhance vascularization and follicle survival in xenografted ovarian tissue using a two-step transplantation approach? SUMMARY ANSWER: Higher rates of oxygenation and vascularization of ovarian tissue, as well as increased follicle survival rates, were detected in the early post-grafting period. WHAT IS KNOWN ALREADY: ASCs have multilineage differentiation potential, proangiogenic properties and enhance vascularization in a peritoneal grafting site. Some studies suggest that using ASCs may improve ovarian tissue quality by enhancing graft angiogenesis. STUDY DESIGN, SIZE, DURATION: A total of 15 severe combined immunodeficient (SCID) mice were intraperitoneally grafted with frozen-thawed human ovarian tissue (OT) from five different patients. A peritoneal transplantation site had been previously prepared in a first step using either empty fibrin (Fi+OT group [n = 5]) or ASC-loaded fibrin (Fi/ASCs+OT group [n = 5]) for 14 days prior to grafting. Five mice underwent the standard one-step transplantation procedure and served as controls (OT group). Lithium phthalocyanine (LiPc) crystals were inserted into all grafted human ovarian tissue before transplantation. Levels of partial pressure of oxygen (pO2) in grafts were monitored in vivo by electron paramagnetic resonance (EPR) oximetry on Days 3 and 7. Samples for histology and immunohistochemistry (IHC) were collected after euthanizing the mice on Day 7 following EPR. One piece of ovarian tissue per patient was fixed for analysis to serve as non-grafted controls. PARTICIPANTS/MATERIALS, SETTING, METHODS: Prospective experimental study conducted at the Gynecology Research Unit, Université Catholique de Louvain. All materials were used to perform pO2 measurements (EPR oximetry), histological (haematoxylin and eosin staining), immunohistochemistry (anti-mouse and human double CD34 and anti-human Ki-67) and TUNEL analyses. MAIN RESULTS AND THE ROLE OF CHANCE: A significant increase in pO2 was observed in all groups between Days 3 and 7 (P < 0.001). A significantly higher pO2 level was observed in the Fi/ASCs+OT group compared to the OT group on Day 7 (P = 0.028). Total CD34-positive vessel area on Day 7 was greater in the Fi/ASCs+OT group than in any other group (vs non-grafted group: P = 0.0014; vs OT group: P = 0.013; vs Fi+OT group: P = 0.018). Primordial follicle survival rates after grafting were higher in the Fi/ASCs+OT group than in the OT (P = 0.0059) or Fi+OT groups (P = 0.0307). TUNEL-positive follicle percentages after grafting were significantly lower in the Fi/ASCs+OT group than in any other grafted tissue (vs OT group: P = 0.045; vs Fi+OT group: P = 0.0268). Percentages of Ki-67-positive primordial follicles were significantly higher in all grafted groups compared to non-grafted tissue controls (P < 0.01). LIMITATIONS REASONS FOR CAUTION: As demonstrated by our results, the proposed two-step ovarian tissue transplantation procedure using ASCs enhances vascularization in the early post-grafting period, leading to increased follicle survival rates and decreased apoptosis. However, mechanisms involved in the proangiogenic behavior of ASCs remain to be elucidated. WIDER IMPLICATIONS OF THE FINDINGS: Our results suggest that the proposed transplantation procedure with ASCs is a promising step towards potentially solving the problem of massive follicle loss after ovarian tissue grafting. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS-PDR Convention T.0077.14, grant Télévie No. 7.6515.16 F to DDM and grant 5/4/150/5 awarded to MMD and CAA is research associate, FRS-FNRS), Fonds Spéciaux de Recherche, Fondation St Luc, and Foundation Against Cancer, and donations from the Ferrero family.

Evidence against solar influence on nuclear decay constants.

The hypothesis that proximity to the Sun causes variation of decay constants at permille level has been tested and disproved. Repeated activity measurements of mono-radionuclide sources were performed over periods from 200 days up to four decades at 14 laboratories across the globe. Residuals from the exponential nuclear decay curves were inspected for annual oscillations. Systematic deviations from a purely exponential decay curve differ from one data set to another and are attributable to instabilities in the instrumentation and measurement conditions. The most stable activity measurements of alpha, beta-minus, electron capture, and beta-plus decaying sources set an upper limit of 0.0006% to 0.008% to the amplitude of annual oscillations in the decay rate. Oscillations in phase with Earth's orbital distance to the Sun could not be observed within a 10-6 to 10-5 range of precision. There are also no apparent modulations over periods of weeks or months. Consequently, there is no indication of a natural impediment against sub-permille accuracy in half-life determinations, renormalisation of activity to a distant reference date, application of nuclear dating for archaeology, geo- and cosmochronology, nor in establishing the SI unit becquerel and seeking international equivalence of activity standards.

Analysis of a neutron-induced conversion electron spectrum of gadolinium.

A 100-nm-thick gadolinium layer deposited on a pixelated silicon sensor was activated in a neutron field to measure the internal conversion electron (ICE) spectrum generated by neutron capture products of 155Gd and 157Gd. The experiment was performed at the ISIS neutron and muon facility, using a bespoke version of the HEXITEC spectroscopic imaging camera. Signals originating from internal conversion electrons, Auger electrons, x rays and gamma rays up to 150 keV were identified. The ICE spectrum has an energy resolution of 1.8-1.9 keV at 72 keV and shows peaks from the K, L, M, N+ ICEs of the 79.51 keV and 88.967 keV 2+-0+ gamma transitions from the first excited states in 158Gd and 156Gd, respectively, as well as the K ICEs of the 4+-2+ transitions at 181.931 keV and 199.213 keV from the respective second excited states. Spectrum analysis was performed using a convolution of a Gaussian with exponential functions at the low and high energy side as the peak shaping function. Relative ICE intensities were derived from the fitted peak areas and compared with internal conversion coefficient (ICC) values calculated from the BrIcc database. Relative to the dominant L shell contribution, the K ICE intensity conforms to BrIcc and the M, N, O+ ICE intensities are somewhat higher than expected.

Air humidity and annual oscillations in 90Sr/90Y and 60Co decay rate measurements.

Parkhomov published decay rate measurements of 90Sr/90Y and 60Co beta decay sources with Geiger-Müller counters which showed annual cyclic deviations with less than 0.2% amplitude from a purely exponential slope. He investigated instrument instability induced by environmental parameters, yet did not find a clear coincidence with local temperature, atmospheric pressure, and relative humidity. Parkhomov hypothesised that gravitationally-focussed 'slow' cosmic neutrinos influenced beta decay. In the current work, environmental conditions in the Moscow area at the time of the experiment are presented. There appears to be a resemblance of the shape of the annual 90Sr/90Y decay rate anomalies with the inverse of the absolute air humidity, albeit with an apparent time shift of 0.05-0.15 year. Humidity may have influenced the range of beta particles in air, as well as geometric and electronic properties of the detection set-up, however causality could not be unambiguously demonstrated. The instabilities in the 60Co data were more difficult to correlate with environmental data, except for some similarities with temperature and external dew point.

Measurement of the 145Sm half-life.

The half-life of 145Sm has been measured by means of the reference source method with a HPGe detector. The long-lived radionuclide 44Ti was mixed into the source for reference. The time-dependency of the 145Sm/44Ti activity ratio was followed by assessing the count-rate ratio of their characteristic gamma-ray emissions at 61.2 keV (145Sm) and 67.9/78.3 keV (44Ti) in spectra recorded over periods of typically one day. In total, 220 measurements were performed over a period of 384 days or about one half-life period. The experiment and ensuing uncertainty budget are discussed in detail. Different error propagation is applied for random uncertainties, autocorrelated structures in the fit residuals, and potential systematic errors. The result for the 145Sm half-life, 345 (16) d, is compatible with the scarce literature values, however the experimental details of the old measurements were barely documented.

On the interpretation of annual oscillations in 32Si and 36Cl decay rate measurements.

The 32Si decay rate measurement data of Alburger et al. obtained in 1982-1986 at Brookhaven National Laboratory have been presented repeatedly as evidence for solar neutrino-induced beta decay. The count rates show an annual sinusoidal oscillation of about 0.1% amplitude and maximum at February-March. Several authors have claimed that the annual oscillations could not be explained by environmental influences on the set-up, and they questioned the invariability of the decay constant. They hypothesised a correlation with changes in the solar neutrino flux due to annual variations in the Earth-Sun distance, in spite of an obvious mismatch in amplitude and phase. In this work, environmental conditions at the time of the experiment are presented. The 32Si decay rate measurements appear to be inversely correlated with the dew point in a nearby weather station. Susceptibility of the detection set-up to local temperature and humidity conditions is a likely cause of the observed instabilities in the measured decay rates. Similar conclusions apply to 36Cl decay rates measured at Ohio State University in 2005-2012.

Derivation of an uncertainty propagation factor for half-life determinations.

An analytical equation is derived for the uncertainty propagation factor for a half-life determination from a least-squares fit to equidistant activity measurements performed with identical relative uncertainties. The obtained formula applies to a purely random statistical uncertainty component. It is equivalent to the solution published by Parker in Nucl. Instr. Meth. A 286, 502. A more general equation for weighted least-squares fitting is derived and presented in a compact manner. It is used as a benchmark to verify the applicability of Parker's solution to non-equidistant data with unequal uncertainties.

Automated optical distance measurements for counting at a defined solid angle.

Two optical distance measurement devices have been compared for accurately defining the source-detector geometry of alpha-particle counters. The first consists of a travelling microscope with unifocal lens, to which a linear distance gauge is attached. The focusing is done by human eye. The second is a modern 3D coordinate measurement machine, equipped with a CMOS colour camera as the sensor. Focusing is performed automatically by software. With both devices, distance measurements of various drop-deposited sources on glass and stainless steel disks have been performed. There is a good mutual agreement of results. Even though the new device can sample more reference points, the accuracy of the measurements has not improved drastically due to the imperfect flatness of the sources. The main advantages of the new system are its ease of use and the lower health risk involved due to the larger distance between the performer and the source.

Conversion electron spectroscopy of the 59.54 keV transition in 241Am alpha decay.

A windowless Peltier-cooled silicon drift detector (SDD) was used to measure internal conversion electron (ICE) spectra of thin 241Am sources. The ICE peaks associated with the 59.54 keV gamma transition in 237Np were deconvoluted and relative ICE intensities were derived from the fitted peak areas. Corrections were made for energy dependence of the full-energy-peak counting efficiency, based on Monte Carlo simulations. As expected for this anomalous E1 transition, a significant discrepancy was found with the theoretical internal conversion coefficient (ICC) values calculated from the BrIcc database. Penetration effects are known to cause such anomalies in highly retarded transitions. The measured ICE intensities are in good agreement with a specific combination of literature data obtained with magnetic spectrometers.

The 55Fe half-life measured with a pressurised proportional counter.

The half-life of 55Fe has been measured accurately by following the decay curve of three sources with a large pressurised proportional counter. An argon(90%)-methane(10%) mixture was used as counter gas, at atmospheric pressure (∼1 × 105 Pa) and at enhanced pressures of 5 × 105 Pa and 8 × 105 Pa (for 1 source), respectively. The first measurements were performed in 2001, but the experiment was executed more systematically between 2005 and 2018, covering a period of about 5 half-lives. The residuals from an exponential decay curve were of the order of 0.1% to 0.2% at 1 × 105 Pa, and 0.03% at 5 × 105 Pa and 8 × 105 Pa. The gain of stability with increased gas pressure was due to asymptotically reaching the maximum counting efficiency, resulting in lower sensitivity to pressure variations. The deduced half-life value of T1/2(55Fe) = 1006.70 (15) d or 2.7563 (4) a is more accurate than other data in literature, which are mutually discrepant. It is consistent with previous measurements at JRC with an X-ray defined solid angle counter.

Alpha-particle emission probabilities of 231Pa derived from first semiconductor spectrometric measurements.

The nuclide 231Pa is a member of the 235U decay chain. It is a complex alpha emitter with 25 identified alpha emissions. Formerly published alpha-particle emission probabilities were derived from measurements taken with magnetic spectrometers. This work presents the first measurements made with semiconductor detectors. High-resolution alpha-particle spectrometry was carried out at CIEMAT and JRC using ion-implanted planar silicon detectors. Alpha-particle emission probabilities of 23 transitions were derived from deconvolutions of the spectra. For the major lines, uncertainties are lower than 1%, a significant improvement to existing data. The new data set will allow a more accurate evaluation of the decay scheme of 231Pa.

Measurement of absolute γ-ray emission probabilities in the decay of 227Ac in equilibrium with its progeny.

The emission probabilities of γ rays produced in the 227Ac decay series were determined by high-resolution γ-ray spectrometry of sources with standardised activity. The sources were prepared quantitatively on glass discs by drop deposition of a solution with 227Ac in radioactive equilibrium with its daughter nuclides. Their activity was measured by a primary standardisation technique based on alpha-particle counting at a defined low solid angle. Four laboratories performed γ-ray spectrometry and derived absolute γ-ray intensities. Mean values were calculated and compared with literature data and the currently recommended evaluated data. New values on certain γ-ray emission probabilities are proposed.

Cascades of pile-up and dead time.

Count loss through a cascade of pile-up and dead time is studied. Time interval density-distribution functions and throughput factors are presented for counters with a series arrangement of pile-up and extending or non-extending dead time. A counter is considered, where an artificial dead time is imposed on every counted event, in order to control the length and type of dead time. For such a system, it is relatively easy to determine an average count-loss correction factor via a live-time clock gated by the imposed dead-time signal ('live-time mode'), or otherwise to apply a correction factor based on the inversion of the throughput function ('real-time mode'). However, these techniques do not account for additional loss through pulse pile-up. In this work, counting errors associated with neglecting cascade effects are calculated for measurements in live-time and real-time mode.

alpha-Particle and gamma-ray spectrometry of a plutonium solution for impurity determination.

A highly enriched (240)Pu solution was measured by alpha-particle and gamma-ray spectrometry to determine other radionuclides present in the material as impurities. Low activities of (238)Pu, (241)Am, (243)Cm and (244)Cm were determined by measuring thin sources, made from the original solution, in a high-resolution alpha-particle spectrometer. The sources were prepared by evaporating the plutonium solution on quartz plates in a vacuum chamber. From the ingrowth of (241)Am in the original solution, the amount of (241)Pu could be calculated. After radiochemical separation of (241)Am, the plutonium was measured by high-efficiency alpha-particle spectrometry to determine the amount of (238)Pu. The enriched (240)Pu material was also measured by high-resolution gamma-ray spectrometry, using two different HPGe detectors to determine the impurities of (239)Pu and (241)Am. The preparation of the sources and the measurement methods are described and discussed. The measured impurities, given in % of the (240)Pu activity, are compared with the values on the certificate.

Linearity check of an ionisation chamber through 99 mTc half-life measurements.

The half-life of 99 mTc was measured at the JRC using the ionisation chamber 'IC1' (type Centronic IG12). The result, T1/2(99 mTc) = 6.00660 (18) h, is in good agreement with literature data. One experiment was performed in IC1's default set-up with the ionisation current being integrated over an air capacitor and read out as a voltage increase over time. This ensured excellent linearity and precision throughout the dynamic range, but the maximum current was limited to 2 nA. In a second test, the current was directly read out with a Keithley 6517 A electrometer. Applying correction factors for the automatic range switching of the electrometer, an acceptable linearity was demonstrated over a range of 12 half-life periods starting at 20 nA. Range switching and autocorrelation of the current readout increase the systematic and random error propagation factors. Piecewise fitting of the decay curve over periods of 6 h yields the same 99 mTc half-life value within 0.04% (0.0025 h) standard deviation over an activity range spanning at least 10 half-life periods (3 orders of magnitude).

Deconvolution of 238,239,240Pu conversion electron spectra measured with a silicon drift detector.

Internal conversion electron (ICE) spectra of thin 238,239,240Pu sources, measured with a windowless Peltier-cooled silicon drift detector (SDD), were deconvoluted and relative ICE intensities were derived from the fitted peak areas. Corrections were made for energy dependence of the full-energy-peak counting efficiency, based on Monte Carlo simulations. A good agreement was found with the theoretically expected internal conversion coefficient (ICC) values calculated from the BrIcc database.

Is decay constant?

Some authors have raised doubt about the invariability of decay constants, which would invalidate the exponential-decay law and the foundation on which the common measurement system for radioactivity is based. Claims were made about a new interaction - the fifth force - by which neutrinos could affect decay constants, thus predicting changes in decay rates in correlation with the variations of the solar neutrino flux. Their argument is based on the observation of permille-sized annual modulations in particular decay rate measurements, as well as transient oscillations at frequencies near 11 year-1 and 12.7 year-1 which they speculatively associate with dynamics of the solar interior. In this work, 12 data sets of precise long-term decay rate measurements have been investigated for the presence of systematic modulations at frequencies between 0.08 and 20 year-1. Besides small annual effects, no common oscillations could be observed among α, ß-, ß+ or EC decaying nuclides. The amplitudes of fitted oscillations to residuals from exponential decay do not exceed 3 times their standard uncertainty, which varies from 0.00023 % to 0.023 %. This contradicts the assertion that 'neutrino-induced' beta decay provides information about the deep solar interior.

Measurement of absolute γ-ray emission probabilities in the decay of 235U.

Accurate measurements were performed of the photon emission probabilities following the α decay of 235U to 231Th. Sources of highly enriched 235U were characterised in terms of isotopic composition by mass spectrometry and their activities were standardised by means of alpha-particle counting at a low defined solid angle. The standardised sources were subsequently measured by high-resolution γ-ray spectrometry with calibrated high-purity germanium detectors to determine the photon emission probabilities. Four laboratories participated in this work and reported emission probabilities for 33 γ-ray lines. Most of them agree with previously published evaluated data. In addition, new values are proposed for γ-lines which have been measured only once in the past.

The solid angle subtended by a circular detector for a linear source.

An exact solution is presented for the solid angle subtended by a circular detector for a linear source. It is based on Conway's equation for a non-coaxial point source, involving an integral of Bessel functions [Conway, J.T., 2006. Generalizations of Ruby's formula for the geometric efficiency of a parallel-disk source and detector system. Nucl. Instrum. Methods A 562, 146-153]. Some numerical examples are calculated and compared with the results obtained with a recently published closed formula (Galiano, E., Pagnutti, C., 2006. An analytical solution for the solid angle subtended by a circular detector for a symmetrically positioned linear source. Appl. Radiat. Isot. 64, 603-607), which is incorrect but can be improved by applying a correction factor.