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

A comparison of small rodent assemblages after a 20 year interval in the Alps

Human-induced environmental alterations in the Alps may importantly affect small mammal species, but evidence in this sense is limited. We live-trapped small rodents in the Central-Eastern Italian Alps in three close-by habitat types (rocky scree, alpine grassland, and heath) at 2100 m a.s.l. during summer-fall, in 1997 and 2016. We compared small rodent assemblages through a Redundancy Detrended Analysis (RDA). In both surveys, we detected two specialist species, i.e., the common vole (Microtus arvalis) and the snow vole (Chionomys nivalis), and, unexpectedly, the forest generalist bank vole (Myodes glareolus). In 1997, grassland was mainly occupied by the common vole, while the bank vole and the snow vole were sympatric in the other habitats. In 2016, the snow vole was detected only in the scree, while other species did not show distribution changes. We discuss a series of hypotheses that might have driven the differences observed across decades, among which is a species-specific response to abiotic and biotic environmental alterations, with the alpine habitat specialist moving out of sub-optimal habitats. We encourage further research on this topic, e.g., via long-term longitudinal studies

Contribution of cryopreservation to the cumulative live birth rate: a large multicentric cycle-based data analysis from the Italian National Registry

Purpose: To estimate the contribution of cryopreservation to the cumulative live birth rate (CLBR) after law modification in Italy in the era of vitrification and freeze-all. Methods: The Italian National Registry performed a cycle-based data collection. Nine Italian IVF clinics were involved incorporating a total of 10,260 fresh cycles performed between January 2015 and April 2016 resulting in 9273 oocyte retrievals and 3266 subsequent warming cycles from the same oocyte retrievals performed up to December 2016. Mean female age was 37 \ub1 4.3\ua0years. Primary outcome measure was CLBR per oocyte retrieval. Confounding factors were tested in multivariate regression analysis, and the relative impact of cryopreservation to the CLBR in different patient categories was calculated. Results: CLBR per oocyte retrieval was 32.6%, 26.5%, 18.7%, 13.0%, and 5.5% for women younger than 36, aged 36\u201339, 40\u201341, and older than 41\ua0years, respectively. The total relative contribution of oocyte/embryo cryopreservation was 40.6% (95% CI 38.41\u201342.75). An association between maternal age, number of oocytes retrieved, fertilization rate, cryopreservation, and cumulative live birth was shown. When adjusted for confounders, a 2.3-fold increase was observed in the chance of live birth when cryopreservation was performed (OR 2.3; 95% CI 1.99\u20132.56). In high responder patients (> 15 oocytes retrieved) where freeze-all was applied in 67.6% of cycles to avoid the risk of hyper stimulation syndrome, the relative contribution of vitrification to the CLBR was 80.6%. Conclusions: Cryopreservation is essential in IVF and should always be available to patients to optimize success rates. Multicentric, cycle-based data analyses are crucial to provide infertile couples, clinicians, and regulatory bodies with accurate information on IVF effectiveness including fresh and cryopreserved cycles

3D Photogrammetry of Bat Skulls: Perspectives for Macro-evolutionary Analyses

Photogrammetry (PH) is relatively cheap, easy to use, flexible and portable but its power and limitations have not been fully explored for studies of small animals. Here we assessed the accuracy of PH for the reconstruction of 3D digital models of bat skulls by evaluating its potential for evolutionary morphology studies at interspecific (19 species) level. Its reliability was assessed against the performance of micro CT scan (µCT) and laser scan techniques (LS). We used 3D geometric morphometrics and comparative methods to quantify the amount of size and shape variation due to the scanning technique and assess the strength of the biological signal in relation to both the technique error and phylogenetic uncertainty. We found only minor variation among techniques. Levels of random error (repeatability and procrustes variance) were similar in all techniques and no systematic error was observed (as evidenced from principal component analysis). Similar levels of phylogenetic signal, allometries and correlations with ecological variables (frequency of maximum energy and bite force) were detected among techniques. Phylogenetic uncertainty interacted with technique error but without affecting the biological conclusions driven by the evolutionary analyses. Our study confirms the accuracy of PH for the reconstruction of challenging specimens. These results encourage the use of PH as a reliable and highly accessible tool for the study of macro evolutionary processes of small mammals

Conservation status of Eurasian Otter Lutra lutra in Italy

The southern Italian otter population is considered an ESU, as it is completely isolated from other European populations. The two former disjointed nuclei of this core population are now a unique metapopulationb. Expansion at the edge of this range is proceeding inconstantly with presence/absence varying year by year, especially in Abruzzo and Apulian rivers (Foro, Pescara, Alto Sangro, Candelaro) (Fig. 1). However, despite not all grid cells were resampled over time and sampling designs were not homogeneous and consistent along time and space, all trends are positive (Fig. 2). As to northern Italy, the otter expansion in Austria and Slovenia promoted the return of the species on the eastern Alpine chain since 2011, where its rapid expansion is giving hope for the establishment of a viable population in the short-medium term. Also, a new nucleus on the western part of the Alps was detected in 2020 on the Roya catchmenti across the French border. The Italian Alpine range is also hosting a isolated B-line population in the Ticino river, a population derived from reintroduction of individuals crossbred with L. l. barang. However, a recent study revealed this population is likely unviable and no otter signs have been recorded in the area since 2018 (Fig. 1)

Pregnancy outcomes and cytomegalovirus DNAaemia in HIV infected pregnant women with CMV

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Rate , correlates and outcomes of repeat pregnancy in HIV-infected women

Objectives: The aim of the study was to assess the rate, determinants, and outcomes of repeat pregnancies in women with HIV infection. Methods: Data from a national study of pregnant women with HIV infection were used. Main outcomes were preterm delivery, low birth weight, CD4 cell count and HIV plasma viral load. Results: The rate of repeat pregnancy among 3007 women was 16.2%. Women with a repeat pregnancy were on average younger than those with a single pregnancy (median age 30 vs. 33 years, respectively), more recently diagnosed with HIV infection (median time since diagnosis 25 vs. 51 months, respectively), and more frequently of foreign origin [odds ratio (OR) 1.36; 95% confidence interval (CI) 1.10–1.68], diagnosed with HIV infection in the current pregnancy (OR: 1.69; 95% CI: 1.35–2.11), and at their first pregnancy (OR: 1.33; 95% CI: 1.06–1.66). In women with sequential pregnancies, compared with the first pregnancy, several outcomes showed a significant improvement in the second pregnancy, with a higher rate of antiretroviral treatment at conception (39.0 vs. 65.4%, respectively), better median maternal weight at the start of pregnancy (60 vs. 61 kg, respectively), a higher rate of end-of-pregnancy undetectable HIV RNA (60.7 vs. 71.6%, respectively), a higher median birth weight (2815 vs. 2885 g, respectively), lower rates of preterm delivery (23.0 vs. 17.7%, respectively) and of low birth weight (23.4 vs. 15.4%, respectively), and a higher median CD4 cell count (+47 cells/μL), with almost no clinical progression to Centers for Disease Control and Prevention stage C (CDC-C) HIV disease (0.3%). The second pregnancy was significantly more likely to end in voluntary termination than the first pregnancy (11.4 vs. 6.1%, respectively). Conclusions: Younger and foreign women were more likely to have a repeat pregnancy; in women with sequential pregnancies, the second pregnancy was characterized by a significant improvement in several outcomes, suggesting that women with HIV infection who desire multiple children may proceed safely and confidently with subsequent pregnancies

Amniocentesis and chorionic villus sampling in HIV-infected pregnant women: a multicentre case series.

Objectives: To assess in pregnant women with HIV the rates of amniocentesis and chorionic villus sampling (CVS), and the outcomes associated with such procedures. Design: Observational study. Data from the Italian National Program on Surveillance on Antiretroviral Treatment in Pregnancy were used. Setting: University and hospital clinics. Population: Pregnant women with HIV. Methods: Temporal trends were analysed by analysis of variance and by the Chi-square test for trend. Quantitative variables were compared by Student's t-test and categorical data by the Chi-square test, with odds ratios and 95% confidence intervals calculated. Main outcome measures: Rate of invasive testing, intrauterine death, HIV transmission. Results: Between 2001 and 2015, among 2065 pregnancies in women with HIV, 113 (5.5%) had invasive tests performed. The procedures were conducted under antiretroviral treatment in 99 cases (87.6%), with a significant increase over time in the proportion of tests performed under highly active antiretroviral therapy (HAART) (100% in 2011–2015). Three intrauterine deaths were observed (2.6%), and 14 pregnancies were terminated because of fetal anomalies. Among 96 live newborns, eight had no information available on HIV status. Among the remaining 88 cases with either amniocentesis (n = 75), CVS (n = 12), or both (n = 1), two HIV transmissions occurred (2.3%). No HIV transmission occurred among the women who were on HAART at the time of invasive testing, and none after 2005. Conclusions: The findings reinforce the assumption that invasive prenatal testing does not increase the risk of HIV vertical transmission among pregnant women under suppressive antiretroviral treatment. Tweetable abstract: No HIV transmission occurred among women who underwent amniocentesis or CVS under effective anti-HIV regimens. © 2016 Royal College of Obstetricians and Gynaecologist