Cryoprotectant-free vitrification of human spermatozoa in new artificial seminal fluid

Vitrification is a new method that has been recently introduced in Assisted Reproduction Technique programs. The aim of this study was to design a new medium similar to normal human seminal fluid (SF), formulation artificial seminal fluid (ASF), and to compare the cryoprotective potency of this medium with SF and human tubal fluid (HTF) medium. Thirty normal ejaculates were processed with the swim-up technique and sperm suspensions were divided into four aliquots: (i) fresh sample (control); (ii) vitrification in HTF medium supplemented with 5 mg/mL human serum albumin and 0.25 mol sucrose (Vit HTF); (iii) vitrification with patients' SF (Vit SF); and (iv) vitrification in ASF (Vit ASF). After warming, sperm parameters of motility, viability, and morphology were analyzed using WHO criteria. Also, sperm pellets were fixed in 2.5% glutaraldehyde and processed for scanning electron microscopy and transmission electron microscopy observations. The results showed that progressive motility (46.09 ± 10.33 vs. 36.80 ± 13.75), grade A motility (36.59 ± 11.40 vs. 16.41 ± 11.24), and normal morphology (18.74 ± 8.35 vs. 11.85 ± 5.84) and viability (68.22 ± 10.83 vs. 60.86 ± 11.72) of spermatozoa were significantly higher in Vit ASF than in Vit HTF. All parameters were better in Vit ASF than in Vit SF, but only viability was significantly different (p = 0.006). After cryopreservation, deep invagination in cytoplasm and mechanically weak point sites and folded tail were commonly observed. But, this phenomenon was more significant in Vit HTF and Vit SF than in ASF (p < 0.05). In transmission electron microscopy evaluation, acrosome damage, plasma membrane loss, chromatin vacuolation, and disruption of mitochondria arrangement and structures were observed in all vitrified groups. Adherence of several tail sections together was also seen in all cryo groups. But this was seen more in Vit HTF and Vit SF than in ASF (p < 0.05). In conclusion, vitrification of human spermatozoa with ASF can effectively preserve the quality of sperm motility in comparison with Vit HTF. © 2016 American Society of Andrology and European Academy of Andrology

Ultrastructure of cytoplasmic fragments in human cleavage stage embryos

Purpose: The goal of this study was to evaluate the ultrastructure of cytoplasmic fragments along with the effect of cytoplasmic fragment and perivitelline space coarse granulation removal (cosmetic microsurgery) from embryos before embryo transfer on ART outcomes. Methods: One hundred and fifty intracytoplasmic sperm injection cycles with male factor infertility were included in this prospective study. Patients were divided into three groups of case (n = 50), sham (n = 50), and control (n = 50). Embryos with 10–50 % fragmentation were included in this study. Cosmetic microsurgery and zona assisted hatching were only performed in case and sham groups respectively. Extracted fragments were evaluated ultrastructurally by transmission electron microscopy (TEM). Rates of clinical pregnancy, live birth, miscarriage, multiple pregnancies, and congenital anomaly in the three groups were also compared. Results: Micrographs from TEM showed that mitochondria were the most abundant structures found in the fragments along with mitochondria-vesicle complexes, Golgi apparatus, primary lysosomes, and vacuoles. There were no significant differences in demographic characteristics, laboratory and clinical data, or embryo morphological features between the groups. The rate of clinical pregnancy in control, sham, and case groups had no significant differences (24, 18, and 18 %, respectively). The rates of live birth, miscarriage, multiple pregnancy, and congenital anomaly were also similar between the different groups. Conclusions: Our data demonstrated that cosmetic microsurgery on preimplantation embryos had no beneficial effect on ART outcomes in unselected groups of patients. As mitochondria are the most abundant organelles found in cytoplasmic fragments, fragment removal should be performed with more caution in embryos with moderate fragmentation

Fine morphological assessment of quality of human mature oocytes after slow freezing or vitrification with a closed device: a comparative analysis

BACKGROUND: Human mature oocytes are very susceptible to cryodamage. Several reports demonstrated that vitrification might preserve oocyte better than slow freezing. However, this is still controversial. Thus, larger clinical, biological and experimental trials to confirm this concept are necessary. The aim of the study was to evaluate and compare fine morphological features in human mature oocytes cryopreserved with either slow freezing or vitrification. METHODS: We used 47 supernumerary human mature (metaphase II) oocytes donated by consenting patients, aged 27-32 years, enrolled in an IVF program. Thirtyfive oocytes were cryopreserved using slow freezing with 1.5 M propanediol +0.2 M sucrose concentration (20 oocytes) or a closed vitrification system (CryoTip Irvine Scientific CA) (15 oocytes). Twelve fresh oocytes were used as controls. All samples were prepared for light and transmission electron microscopy evaluation. RESULTS: Control, slow frozen/thawed and vitrified/warmed oocytes (CO, SFO and VO, respectively) were rounded, 90-100 mum in diameter, with normal ooplasm showing uniform distribution of organelles. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates and small mitochondria-vesicle (MV) complexes were the most numerous structures found in all CO, SFO and VO cultured for 3-4 hours. M-SER aggregates decreased, and large MV complexes increased in those SFO and VO maintained in culture for a prolonged period of time (8-9 hours). A slight to moderate vacuolization was present in the cytoplasm of SFO. Only a slight vacuolization was present in VO, whereas vacuoles were almost completely absent in CO. Amount and density of cortical granules (CG) appeared abnormally reduced in SFO and VO, irrespective of the protocol applied. CONCLUSIONS: Even though, both slow freezing and vitrification ensured a good overall preservation of the oocyte, we found that: 1) prolonged culture activates an intracellular membrane "recycling" that causes the abnormal transformation of the membranes of the small MV complexes and of SER into larger rounded vesicles; 2) vacuolization appears as a recurrent form of cell damage during slow freezing and, at a lesser extent, during vitrification using a closed device; 3) premature CG exocytosis was present in both SFO and VO and may cause zona pellucida hardenin

A meteo-hydrological prediction system based on a multi-model approach for precipitation forecasting

International audienceThe precipitation forecasted by a numerical weather prediction model, even at high resolution, suffers from errors which can be considerable at the scales of interest for hydrological purposes. In the present study, a fraction of the uncertainty related to meteorological prediction is taken into account by implementing a multi-model forecasting approach, aimed at providing multiple precipitation scenarios driving the same hydrological model. Therefore, the estimation of that uncertainty associated with the quantitative precipitation forecast (QPF), conveyed by the multi-model ensemble, can be exploited by the hydrological model, propagating the error into the hydrological forecast. The proposed meteo-hydrological forecasting system is implemented and tested in a real-time configuration for several episodes of intense precipitation affecting the Reno river basin, a medium-sized basin located in northern Italy (Apennines). These episodes are associated with flood events of different intensity and are representative of different meteorological configurations responsible for severe weather affecting northern Apennines. The simulation results show that the coupled system is promising in the prediction of discharge peaks (both in terms of amount and timing) for warning purposes. The ensemble hydrological forecasts provide a range of possible flood scenarios that proved to be useful for the support of civil protection authorities in their decision

Simulation of a flash-flood event over the Adriatic Sea with a high-resolution atmosphere–ocean–wave coupled system

On the morning of September 26, 2007, a heavy precipitation event (HPE) affected the Venice lagoon and the neighbouring coastal zone of the Adriatic Sea, with 6-h accumulated rainfall summing up to about 360 mm in the area between the Venetian mainland, Padua and Chioggia. The event was triggered and maintained by the uplift over a convergence line between northeasterly flow from the Alps and southeasterly winds from the Adriatic Sea. Hindcast modelling experiments, using standalone atmospheric models, failed to capture the spatial distribution, maximum intensity and timing of the HPE. Here we analyze the event by means of an atmosphere-wave-ocean coupled numerical approach. The combined use of convection permitting models with grid spacing of 1 km, high-resolution sea surface temperature (SST) fields, and the consistent treatment of marine boundary layer fluxes in all the numerical model components are crucial to provide a realistic simulation of the event. Inaccurate representations of the SST affect the wind magnitude and, through this, the intensity, location and time evolution of the convergence zone, thus affecting the HPE prediction. © 2021, The Author(s)

Growth factor-binding compounds and methods of use

Growth factor binding compounds having a plurality of acyclic isophthalic acid groups attached to a non-peptide organic scaffold and pharmaceutical compositions of the same are disclosed. Methods of administering and using the growth factor binding compounds or the growth factor binding compositions are also taught. These novel growth factor binding compounds are useful for treating angiogenesis, excessive cellular proliferation, tumor growth, and a combination thereof as well as inhibiting growth factor binding to cells and phosphorylation

Radiogenomics in clear cell renal cell carcinoma: correlations between advanced CT imaging (texture analysis) and microRNAs expression

Purpose: A relevant challenge for the improvement of clear cell renal cell carcinoma management could derive from the identification of novel molecular biomarkers that could greatly improve the diagnosis, prognosis, and treatment choice of these neoplasms. In this study, we investigate whether quantitative parameters obtained from computed tomography texture analysis may correlate with the expression of selected oncogenic microRNAs. Methods: In a retrospective single-center study, multiphasic computed tomography examination (with arterial, portal, and urographic phases) was performed on 20 patients with clear cell renal cell carcinoma and computed tomography texture analysis parameters such as entropy, kurtosis, skewness, mean, and standard deviation of pixel distribution were measured using multiple filter settings. These quantitative data were correlated with the expression of selected microRNAs (miR-21-5p, miR-210-3p, miR-185-5p, miR-221-3p, miR-145-5p). Both the evaluations (microRNAs and computed tomography texture analysis) were performed on matched tumor and normal corticomedullar tissues of the same patients cohort. Results: In this pilot study, we evidenced that computed tomography texture analysis has robust parameters (eg, entropy, mean, standard deviation) to distinguish normal from pathological tissues. Moreover, a higher coefficient of determination between entropy and miR-21-5p expression was evidenced in tumor versus normal tissue. Interestingly, entropy and miR-21-5p show promising correlation in clear cell renal cell carcinoma opening to a radiogenomic strategy to improve clear cell renal cell carcinoma management. Conclusion: In this pilot study, a promising correlation between microRNAs and computed tomography texture analysis has been found in clear cell renal cell carcinoma. A clear cell renal cell carcinoma can benefit from noninvasive evaluation of texture parameters in adjunction to biopsy results. In particular, a promising correlation between entropy and miR-21-5p was found

High resolution simulations of a flash flood near Venice.

Abstract. During the MAP D-PHASE (Mesoscale Alpine Programme, Demonstration of Probabilistic Hydrological and Atmospheric Simulation of flood Events in the Alpine region) Operational Period (DOP, 1 June–30 November 2007) the most intense precipitation event observed south of the Alps occurred over the Venice Lagoon. In the early morning of 26 September 2007, a mesoscale convective system formed in an area of convergence between a south-easterly low level jet flowing along the Adriatic Sea and a north-easterly barrier-type wind south of the Alps, and was responsible for precipitation exceeding 320 mm in less than 12 h, 240 mm of which in only 3 h. The forecast rainfall fields, provided by several convection resolving models operated daily for the D-PHASE project, have been compared. An analysis of different aspects of the event, such as the relevant mechanisms leading to the flood, the main characteristics of the MCS, and an estimation of the predictability of the episode, has been performed using a number of high resolution, convection resolving models (MOLOCH, WRF and MM5). Strong sensitivity to initial and boundary conditions and to model parameterization schemes has been found. Although low predictability is expected due to the convective nature of rainfall, the forecasts made more than 24 h in advance indicate that the larger scale environment driving the dynamics of this event played an important role in favouring the achievement of a relatively good accuracy in the precipitation forecasts