Monitoring the microtubule nucleation dynamics of sperm centriole after IFV and ICSI in sheep zygotes.

Intracytoplasmic sperm injection (ICSI) is an assisted reproductive (ART) technique that is less efficient in ruminant, comparing to other species. In mammals, the spermatozoa proximal centriole nucleates the microtubule and generates the functional cell centriole of the resulting organism. Our study aimed to investigate eventual difference in the centriole microtubular nucleation in ICSI fertilized oocytes, comparing to control In Vitro Fertilized ones (IVF). In fact, we made the hypothesis that the tail severing step achieved in our ICSI protocol through applying a few piezo pulses, might mechanically damage the proximal centriole. On this basis, Sheep oocytes were in vitro maturated (IVM) for 24 h then were injected by piezo-pulsed spermatozoa, chemically activated by 5 min of incubation with 5 mg/ml ionomycin, washed in H199 for 5 min and cultured in 50 µl drops of Synthetic Oviductal Fluid (SOF) with estrus sheep serum and 16 µM isoproterenol, covered by mineral oil. Fertilization has been arrested around 5h after ICSI, and the presumptive zygotes were processed for immunological detection of tubulin. Zona Pellucida (ZP) was removed with a combined treatment of acid Tyrode and trypsin and zygotes were then fixed with 4% paraformaldehyde (pH7.2) and permeabilized by 0.5% Triton X-100, for 20 min each. Microtubular nucleation was assessed with anti-α-tubulin immunofluorescence under confocal microscopy. No difference was noticed in the dynamics and timing of sperm microtubular aster nucleation, that started around 5h post ICSI (5h30). Therefore, we conclude that abnormal microtubular nucleation by the centriole is not responsible for the low development of ICSI fertilized sheep oocytes

Protocolo para crescimento de isolados bacterianos de nódulos de Acacia mearnsii e eficiência na solubilização de fosfato

Alguns grupos de bactérias rizosféricas apresentam capacidade de realizar a solubilização do fosfato no solo. Entretanto, a pesquisa com bactérias necessita de protocolos para prevenir a presença de fungos contaminantes, durante a fase de avaliação da capacidade de solubilização de fosfato in vitro, afinal o crescimento de fungos pode afetar a eficiência do protocolo adotado. Por esse motivo, o presente trabalho teve como objetivo: 1- estabelecer um protocolo para determinar um meio de cultura e a dosagem de cicloheximida para o crescimento de isolados bacterianos capazes de solubilizar o fosfato e reduzir a contaminação fúngica; e 2 – avaliar a eficiência de diferentes isolados bacterianos na solubilização de fosfato. Foram realizados três ensaios para avaliar a frequência fúngica, grau de contaminação e o índice de solubilização médio (ISM), durante o período de incubação. Os ensaios foram realizados em delineamento de blocos ao acaso. O primeiro em parcelas subdivididas no tempo, com o uso de dez isolados obtidos de nódulos de Acacia mearnsii, submetidos a cinco diferentes dosagens de cicloheximida (0; 25; 50; 75 e 100 mg.L-1). O segundo observando o crescimento dos mesmos isolados em cada meio de cultura e o terceiro para determinar o melhor isolado quanto a solubilização quando comparado aos isolados indicados para a cultura. As avaliações de contaminantes ocorreram diariamente, enquanto que as mensurações do crescimento ocorreram aos 3, 5 e 7 dias de incubação. O cálculo do índice de solubilização médio (ISM) resultou da razão entre o diâmetro do halo e o diâmetro da colônia, os resultados foram submetidos ao teste de Tukey (p < 0,05). As melhores bactérias que promoveram maior solubilização foram CNPF01F, CNPF01G, CNPF10B1. A partir dos resultados, recomenda-se o meio adaptado de Verma et al. (2001) contendo entre 50 e 75mg.L-1 de cicloheximida como protocolo para avaliação de bactérias solubilizadoras de fosfato.

A Season of American Football Is Not Associated with Changes in Plasma Tau

American football athletes are routinely exposed to sub-concussive impacts over the course of the season. This study sought to examine the effect of a season of American football on plasma tau, a potential marker of axonal damage. Nineteen National Collegiate Athletic Association (NCAA) football athletes underwent serial blood sampling over the course of the 2014–2015 season at those times in which the number and magnitude of head impacts likely changed. Non-contact sport controls (NCAA men's swim athletes; n = 19) provided a single plasma sample for comparison. No significant differences were observed between control swim athletes and football athletes following a period of non-contact (p = 0.569) or a period of contact (p = 0.076). Football athletes categorized as starters (n = 11) had higher tau concentrations than non-starters (n = 8) following a period of non-contact (p = 0.039) and contact (p = 0.036), but not higher than swimmers (p = 1.000 and p = 1.000, respectively). No difference was noted over the course of the season in football athletes, irrespective of starter status. Despite routine head impacts common to the sport of American football, no changes were observed over the course of the season in football athletes, irrespective of starter status. Further, no difference was observed between football athletes and non-contact control swim athletes following a period of non-contact or contact. These data suggest that plasma tau is not sensitive enough to detect damage associated with repetitive sub-concussive impacts sustained by collegiate–level football athletes

The plants, rituals and spells that 'cured' helminthiasis in Sicily

<p>Abstract</p> <p>Background</p> <p>The author reports on the plants, rituals and spells used against worms and the so-called <it>scantu </it>(fright) in some areas of Sicily. The work is based on ethnobotanical research carried out, prevalently, between 2002-2006, in some areas of Eastern, South-Eastern, North-Central and South-Central Sicily.</p> <p>Methods</p> <p>This research is based on dialogue. Senior 'healers' were contacted; furthermore, doctors, teachers, farmers and in general 'experts' with herbs and 'magic' rituals. Information was collected about the way the plants of folk medicine are prepared. The interviewees were also invited to recite prayers and spells against helminthiasis.</p> <p>Results</p> <p>The author has highlighted the importance of how, in some parts of Sicily, some ailments like helminthiasis and other correlated pathologies like <it>scantu </it>are 'treated' and, especially within the rural social classes, by folk medicine remedies, herbal practises, particular prayers, rituals and spells.</p> <p>Conclusion</p> <p>As regards health/illness, it should be noted that in the last ten years conventional medicine has provided very satisfactory results even resolving potentially mortal pathologies. However, in certain social classes, there is no real collaboration between conventional and folk medicine; so for some senior citizens, the 'healer' with his rituals and empirical and magical herbs is still the person to turn to for the 'cure' of particular ailments. Interest in these practises from ancestral heritage in an advanced country like Italy, is only relevant if the aim is to recoup a cultural identity which is already in decline.</p> <p>It is significant to report a piece: on 14 October 2007 the news on a well-known national Italian TV channel reported an interview with a 94 year-old man from Arbatax (Sardinia) referred to as a 'healer' because both his townspeople and others from all over the world go to him for his cures. He is not paid except in kind and has been known to cure St. Anthony's fire, burns, scalding and marine fungal infections, by smearing his saliva over the infected part and reciting 'special words'.</p

Towards accurate and precise T1 and extracellular volume mapping in the myocardium: a guide to current pitfalls and their solutions

Mapping of the longitudinal relaxation time (T1) and extracellular volume (ECV) offers a means of identifying pathological changes in myocardial tissue, including diffuse changes that may be invisible to existing T1-weighted methods. This technique has recently shown strong clinical utility for pathologies such as Anderson- Fabry disease and amyloidosis and has generated clinical interest as a possible means of detecting small changes in diffuse fibrosis; however, scatter in T1 and ECV estimates offers challenges for detecting these changes, and bias limits comparisons between sites and vendors. There are several technical and physiological pitfalls that influence the accuracy (bias) and precision (repeatability) of T1 and ECV mapping methods. The goal of this review is to describe the most significant of these, and detail current solutions, in order to aid scientists and clinicians to maximise the utility of T1 mapping in their clinical or research setting. A detailed summary of technical and physiological factors, issues relating to contrast agents, and specific disease-related issues is provided, along with some considerations on the future directions of the field. Towards accurate and precise T1 and extracellular volume mapping in the myocardium: a guide to current pitfalls and their solutions. Available from: https://www.researchgate.net/publication/317548806_Towards_accurate_and_precise_T1_and_extracellular_volume_mapping_in_the_myocardium_a_guide_to_current_pitfalls_and_their_solutions [accessed Jun 13, 2017]

Different populations and sources of human mesenchymal stem cells (MSC): A comparison of adult and neonatal tissue-derived MSC

The mesenchymal stroma harbors an important population of cells that possess stem cell-like characteristics including self renewal and differentiation capacities and can be derived from a variety of different sources. These multipotent mesenchymal stem cells (MSC) can be found in nearly all tissues and are mostly located in perivascular niches. MSC have migratory abilities and can secrete protective factors and act as a primary matrix for tissue regeneration during inflammation, tissue injuries and certain cancers

Reducing the time of spermatozoa/oocyte interaction improves embryonic development in sheep.

In vitro fertilization (IVF) is a well-established Assisted Reproductive Technique (ART) routinely applied in human infertility treatment and in large animals, both to boost reproductive performances of selected genotypes, and also for basic research. Despite its value, IVF has seen very little progress in the last two decades, and relies to established paradigms, like overnight sperm-egg co-incubation. However, the long exposure to the relatively high spermatozoa concentration in a dish increases the risk of polyspermy and could be detrimental for early embryonic development. In our work we have identified the time window within which the fertilization occurs in order to refine the procedure and reduce sperm-egg co-incubation, comparing embryo development after short (sIVF) and overnight spermatozoa/oocyte co-incubation (o/nIVF). A total of 144 in vitro matured sheep oocytes were co-incubated with spermatozoa in IVF medium [synthetic oviductal fluid (SOF) with 20% oestrus sheep serum and 16µM isoproterenol]. Then, small batches of oocytes were collected every 30 minutes to check for the presence of a fertilizing spermatozoon. To assess that, cumulus cells were removed and presumptive fertilized oocytes were fixed and stained with propidium iodide for nuclei and Pisum sativum agglutinin for zona pellucida (ZP) detection respectively. Pronuclear formation (PN) and embryo development were evaluated after 16 hours (PN), 24 hours (2 cells) and 7 days of culture (blastocyst). We found that spermatozoa engaged with the ZP not earlier than 90 minutes and penetrated the oocytes within 3-4 hours after IVF. A trend for a lower polyspermic fertilization (>2PN) was detected in sIVF (7.7%) vs o/nIVF (18.4%,). Likewise, cleavage and blastocyst rate were higher after sIVF compared to o/n-IVF (2-cells: 38.6% vs 23.8%; blastocyst: 18.1% vs 9.5%; sIVF vs o/nIVF). This work demonstrates that 4 hours spermatozoa/oocyte interaction are sufficient to achieve fertilization, reduce polyspermy and improve early embryonic development in sheep

A new intraoral device to facilitate preparation of the guide plane for removable dental prostheses

The design of the ParalAB device facilitates transfer of the path of insertion from the diagnostic cast to the mouth quickly and efficiently

Exploring Algorithmic Fairness in Deep Speaker Verification

To allow individuals to complete voice-based tasks (e.g., send messages or make payments), modern automated systems are required to match the speaker’s voice to a unique digital identity representation for verification. Despite the increasing accuracy achieved so far, it still remains under-explored how the decisions made by such systems may be influenced by the inherent characteristics of the individual under consideration. In this paper, we investigate how state-of-the-art speaker verification models are susceptible to unfairness towards legally-protected classes of individuals, characterized by a common sensitive attribute (i.e., gender, age, language). To this end, we first arranged a voice dataset, with the aim of including and identifying various demographic classes. Then, we conducted a performance analysis at different levels, from equal error rates to verification score distributions. Experiments show that individuals belonging to certain demographic groups systematically experience higher error rates, highlighting the need of fairer speaker recognition models and, by extension, of proper evaluation frameworks