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

First case of traumatic myiasis caused by Calliphora vicina in a crested porcupine Hystrix cristata L. in Italy

Calliphora vicina (Diptera: Calliphoridae) it is a facultative ectoparasite responsible for traumatic myiasis in humans and warmblooded vertebrates in the world. In this work one case of traumatic myasis caused by C. vicina (Diptera Calliphoirdae) is reported for the first time in a vulnerable crested porcupine (Hystrix cristata Linnaeus, 1758). A total of 30 larvae located in the posterior-dorsal area of the animal were removed from inside the lesion and either preserved in ethanol or reared to the adult stage. This report shows the great ability of C. vicina to use many organic matter for the food source's offspring, including many live vertebrates

Retreating to nature : rethinking 'therapeutic landscapes'

There is a long history of removing oneself from ‘society’ in order to recuperate or repair. This paper considers a yoga and massage retreat in Southern Spain, and what opportunities this retreat experience might offer for recuperation and the creation of healthy bodies. The paper positions ‘nature’ as an active participant, and as ‘enrolled’ in the experiences of the retreat as a ‘therapeutic landscape’, and questions how and what particular aspects of yoga practice (in intimate relation with place) give rise to therapeutic experiences

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

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

Valor nutritivo de pastagens de capim-elefante manejadas sob sistema convencional e agroecológico.

O capim-elefante é utilizado, na sua grande maioria, em sistemas convencionais de produção animal. O objetivo deste trabalho foi comparar o valor nutritivo do capimelefante em sistemas de manejo agroecológico e convencional, quanto a proteína bruta (PB), fibra em detergente neutro (FDN) e digestibilidade in vitro da matéria seca (DIVMS). Foram usados quatro piquetes, com 0,12ha cada um. No sistema convencional, o capim-elefante foi estabelecido singularmente. No sistema agroecológico, o plantio foi feito em linhas afastadas de 3m. Nas entrelinhas, estabeleceu-se a aveia e o azevém no período hibernal, enquanto que no período estival permitiu-se o desenvolvimento de espécies espontâneas. A adubação foi feita com fertilizantes orgânicos (150kg ha-1 de N). No sistema agroecológico, foram conduzidos sete pastejos, de 24/04/2004 a 05/05/2005. Na pastagem convencional, usouse a mesma quantidade de N (uréia), sendo conduzidos quatro ciclos de pastejo, de 06/10/2004 a 05/05/2005. Para ambos os sistemas foram utilizadas vacas da raça Holandês, recebendo complementação alimentar de 3,5kg dia-1 de concentrado com 20% de proteína bruta, constituindo-se nos animais experimentais. Nas avaliações, considerou-se a massa de forragem inicial com base na matéria seca (MS), os componentes botânicos da pastagem e estruturais do capimelefante. As análises de qualidade foram feitas em amostras de pastejo simulado. O delineamento experimental foi o inteiramente casualizado com dois tratamentos, convencional e agroecológico, duas repetições (piquetes) e em parcelas incompletas subdivididas no tempo (pastejos). Houve interação (P<0,05) entre tratamentos e pastejos em todas as variáveis. Na pastagem agroecológica, o modelo que melhor se ajustou foi o cúbico para todas as variáveis, em função do tempo de pastejo. Na pastagem convencional, a PB e a DIVMS ajustaram-se melhor ao modelo linear, com taxa positiva de crescimento, sendo observado comportamento inverso para FDN, com o decorrer dos pastejos. Tanto na pastagem convencional quanto na agroecológica encontraram-se associações negativas entre lâmina foliar do capim-elefante com PB e DIVMS e positiva com FDN. Ambos os sistemas apresentaram teores qualitativos elevados das pastagens, considerando-se a adubação, o manejo e o tempo de utilização

Oocyte and ovarian tissue cryopreservation in European countries : statutory background, practice, storage and use

STUDY QUESTION: What is known in Europe about the practice of oocyte cryopreservation (OoC), in terms of current statutory background, funding conditions, indications (medical and ‘non-medical’) and specific number of cycles? SUMMARY ANSWER: Laws and conditions for OoC vary in Europe, with just over half the responding countries providing this for medical reasons with state funding, and none providing funding for ‘non-medical’ OoC. WHAT IS ALREADY KNOWN: The practice of OoC is a well-established and increasing practice in some European countries, but data gathering on storage is not homogeneous, and still sparse for use. Ovarian tissue cryopreservation (OtC) is only practiced and registered in a few countries. STUDY DESIGN, SIZE, AND DURATION: A transversal collaborative survey on OoC and OtC, was designed, based on a country questionnaire containing information on statutory or professional background and practice, as well as available data on ovarian cell and tissue collection, storage and use. It was performed between January and September 2015. PARTICIPANTS/MATERIALS, SETTING AND METHODS: All ESHRE European IVF Monitoring (EIM) consortium national coordinators were contacted, as well as members of the ESHRE committee of national representatives, and sent a questionnaire. The form included national policy and practice details, whether through current existing law or code of practice, criteria for freezing (age, health status), availability of funding and the presence of a specific register. The questionnaire also included data on both the number of OoC cycles and cryopreserved oocytes per year between 2010 and 2014, specifically for egg donation, fertility preservation for medical disease, ‘other medical’ reasons as part of an ART cycle, as well as for ‘non-medical reasons’ or age-related fertility decline. Another question concerning data on freezing and use of ovarian tissue over 5 years was added and sent after receiving the initial questionnaire. MAIN RESULTS AND THE ROLE OF CHANCE: Out of 34 EIM members, we received answers regarding OoC regulations and funding conditions from 27, whilst 17 countries had recorded data for OoC, and 12 for OtC. The specific statutory framework for OoC and OtC varies from absent to a strict frame. A total of 34 705 OoC cycles were reported during the 5-year-period, with a continuous increase. However, the accurate description of numbers was concentrated on the year 2013 because it was the most complete. In 2013, a total of 9126 aspirations involving OoC were reported from 16 countries. Among the 8885 oocyte aspirations with fully available data, the majority or 5323 cycles (59.9%) was performed for egg donation, resulting in the highest yield per cycle, with an average of 10.4 oocytes frozen per cycle. OoC indication was ‘serious disease’ such as cancer in 10.9% of cycles, other medical indications as ‘part of an ART cycle’ in 16.1%, and a non-medical reason in 13.1%. With regard to the use of OoC, the number of specifically recorded frozen oocyte replacement (FOR) cycles performed in 2013 for all medical reasons was 14 times higher than the FOR for non-medical reasons, using, respectively, 8.0 and 8.4 oocytes per cycle. Finally, 12 countries recorded storage following OtC and only 7 recorded the number of grafted frozen/thawed tissues. LIMITATIONS, REASONS FOR CAUTION: Not all countries have data regarding OoC collection, and some data came from voluntary collaborating centres, rather than a national authority or register. Furthermore, the data related to use of OoC were not included for two major players in the field, Italy and Spain, where numbers were conflated for medical and non-medical reasons. Finally, the number of cycles started with no retrieval is not available. Data are even sparser for OtC. WIDER IMPLICATIONS OF THE FINDINGS: There is a need for ART authorities and professional bodies to record precise data for practice and use of OoC (and OtC), according to indications and usage, in order to reliably inform all stakeholders including women about the efficiency of both methods. Furthermore, professional societies should establish professional standards for access to and use of OoC and OtC, and give appropriate guidance to all involved. STUDY FUNDING/COMPETING INTEREST(S): The study was supported by ESHRE. There are no conflicts of interest.peer-reviewe

Is "option B+" also being adopted in pregnant women in high-income countries? Temporal trends from a national study in Italy

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

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