Erlandsenmcintyre4909
Sperm cryopreservation is presently used for conservation of male gametes in assisted reproduction technologies (ART). Despite the benefits of sperm banking, freeze-thawing process is injurious to sperm integrity due to induced oxidative stress by cold stress. Oxidative stress reduces sperm motility, viability and DNA integrity.
To investigate the effect of alpha lipoic acid (ALA) on human sperm function during the freeze-thawing process.
Thirty semen samples were collected and different concentrations (0, 0.05, 0.1, 0.2, 0.4, 0.8, and 8mM) of ALA were added to a sperm freeze medium and its effects on sperm motility, DNA damage, and lipid peroxidation of frozen-thawed spermatozoa were assessed.
The addition of 0.2 mM ALA to the sperm freeze medium resulted in significant improvement in percentage of sperm motility, less DNA damage and decreased lipid peroxidation during freeze-thawing process (p<0.05).
ALA improves the cryo-protective capacity of sperm freeze medium used for human semen by protecting the sperm from ROS attack induced by the freezing-thawing process. We suggest that sperm freeze medium supplemented with 0.2 mM ALA would be beneficial for the cryopreservation of male gametes in ART.
ALA improves the cryo-protective capacity of sperm freeze medium used for human semen by protecting the sperm from ROS attack induced by the freezing-thawing process. We suggest that sperm freeze medium supplemented with 0.2 mM ALA would be beneficial for the cryopreservation of male gametes in ART.
Adipose-derived mesenchymal stem cells (ADSCs) have emerged as a promising modality for cellular therapy. However, techniques of ADSC cryopreservation, which can facilitate their clinical application, haven't been established yet.
To determine optimal conditions for ADSC cryopreservation.
We used three cryoprotectants [serum containing 10% dimethyl sulfoxide; CP-1
(5% dimethyl sulfoxide, serum-free); Stem-CellBanker
(dimethyl sulfoxide and serum-free)], two storage temperatures (-80°C, -150°C) and two cell densities (1 × 10
, 7 × 10
cells/mL). Storage was up to 18 months using cryovials. We didn't use a rate-controlled freezer or liquid nitrogen storage.
We found that CP-1
was a suitable cryoprotectant. Storage at -150°C and higher cell density (7×10
cells/mL) kept the best viability of ADSCs, but storage at -80°C and a lower cell density (1×10
cells/mL) is acceptable for up to 9 months. We also confirmed large quantities of ADSCs, stored with CP-1 in a cryobag, were still viable after -150°C cryopreservation for 24 months.
We have developed a safe, cost-effective way to cryopreserve ADSCs that could be used in the clinical setting.
We have developed a safe, cost-effective way to cryopreserve ADSCs that could be used in the clinical setting.
Canine embryo cryopreservation and subsequent transfer are relevant in the use of reproductive technologies.
The purpose of this study is the identification and quantification of the gene expression BAX and Bcl2, AQP3, Na+/K+ ATPase alpha-1 and beta-1 and LIFr in canine embryos obtained in vivo and after freezing.
For the collection of embryos, the bitches were identified at pro-estrous until the detection of 80-90% superficial cells. After that, they were artificially inseminated with fresh semen. The embryos were collected after ovariohysterectomy. RNA was extracted and amplified, and embryos were randomly distributed into fresh (Fr) and frozen/thawed (Ft) groups.
Eighteen blastocysts were collected from three bitches. Genes BAX, AQP3 and LIFr did not differ among the studied groups.
We suggest, through these results, that the genes BAX, Bcl2, AQP3, Na + / K + ATPase alpha-1 and beta-1 and LIFr were expressed in canine blastocysts collected in vivo and after slow freezing cryopreservation.
We suggest, through these results, that the genes BAX, Bcl2, AQP3, Na + / K + ATPase alpha-1 and beta-1 and LIFr were expressed in canine blastocysts collected in vivo and after slow freezing cryopreservation.
The gray catfish known as Surubim-do-Paraíba (Steindachneridion parahybae), which is endemic to the Paraíba do Sul river basin, is on the red list of Brazilian fauna threatened with extinction and the cryopreservation of germ cells of this fish is needed in support of conservation.
We aimed to assess the effect of storage temperature on S. parahybae mature and immature oocytes.
Two trials were carried out. Trial I (TI.1-3) used 30 mature oocytes (diameter >1.8 mm) placed in cryoprotectant solutions and submitted to three different techniques. Trial II (TII.1-3) used 30 immature oocytes (diameter <1.6 mm) placed in cryoprotectant solutions and submitted to three storage temperatures (i.e., TII.1 at room temperature for 120 min; TII.2 in the freezer for 120 min; TII.3 in liquid nitrogen for 24 hours.
The mature oocytes were sensitive to every protocol used, including at room temperature. In contrast, the immature oocytes had increased sensitivity according to the temperature reduction to which they were submitted, with the treatment in liquid nitrogen causing greater damage.
The immature stages exhibit more promising results, encouraging further studies using the combination of different CPSs, mainly penetrating ones, in oocyte cryopreservation protocols.
The immature stages exhibit more promising results, encouraging further studies using the combination of different CPSs, mainly penetrating ones, in oocyte cryopreservation protocols.
Although potato as a crop is commercially grown from seed tubers, plants grown from tissue culture plantlets are often used in physiological studies including freezing tolerance determination.
This study aimed to examine the effects of the source of plants on freezing tolerance of potato plants at young developmental stages.
We compared freezing tolerance and contents of soluble proteins and sugars of Solanum tuberosum plants derived from tissue culture with those derived from tubers before and after cold acclimation.
Tuber-derived plants showed significantly higher freezing tolerance than tissue-culture-derived plants after cold acclimation, although non-acclimated plants did not show any marked differences. Soluble protein contents were higher in tuber-derived plants regardless of cold acclimation. Sucrose content increased to a higher level in tuber-derived plants after cold acclimation.
These results suggest that source of plant tissue can have a significant effect on the response of young potato plants to freezing stress and that the use of tissue culture plants in freezing tolerance studies may not accurately reflect the frost tolerance of commercially grown plants.
These results suggest that source of plant tissue can have a significant effect on the response of young potato plants to freezing stress and that the use of tissue culture plants in freezing tolerance studies may not accurately reflect the frost tolerance of commercially grown plants.Congenital heart disease (CHD) is the most common birth defect. Fetal screening ultrasound provides five views of the heart that together can detect 90% of complex CHD, but in practice, sensitivity is as low as 30%. Here, using 107,823 images from 1,326 retrospective echocardiograms and screening ultrasounds from 18- to 24-week fetuses, we trained an ensemble of neural networks to identify recommended cardiac views and distinguish between normal hearts and complex CHD. We also used segmentation models to calculate standard fetal cardiothoracic measurements. In an internal test set of 4,108 fetal surveys (0.9% CHD, >4.4 million images), the model achieved an area under the curve (AUC) of 0.99, 95% sensitivity (95% confidence interval (CI), 84-99%), 96% specificity (95% CI, 95-97%) and 100% negative predictive value in distinguishing normal from abnormal hearts. Model sensitivity was comparable to that of clinicians and remained robust on outside-hospital and lower-quality images. The model's decisions were based on clinically relevant features. Cardiac measurements correlated with reported measures for normal and abnormal hearts. Applied to guideline-recommended imaging, ensemble learning models could significantly improve detection of fetal CHD, a critical and global diagnostic challenge.Machine learning techniques have great potential to improve medical diagnostics, offering ways to improve accuracy, reproducibility and speed, and to ease workloads for clinicians. In the field of histopathology, deep learning algorithms have been developed that perform similarly to trained pathologists for tasks such as tumor detection and grading. However, despite these promising results, very few algorithms have reached clinical implementation, challenging the balance between hope and hype for these new techniques. This Review provides an overview of the current state of the field, as well as describing the challenges that still need to be addressed before artificial intelligence in histopathology can achieve clinical value.3D correlative microscopy methods have revolutionized biomedical research, allowing the acquisition of multidimensional information to gain an in-depth understanding of biological systems. With the advent of relevant cryo-preservation methods, correlative imaging of cryogenically preserved samples has led to nanometer resolution imaging (2-50 nm) under harsh imaging regimes such as electron and soft X-ray tomography. These methods have now been combined with conventional and super-resolution fluorescence imaging at cryogenic temperatures to augment information content from a given sample, resulting in the immediate requirement for protocols that facilitate hassle-free, unambiguous cross-correlation between microscopes. We present here sample preparation strategies and a direct comparison of different working fiducialization regimes that facilitate 3D correlation of cryo-structured illumination microscopy and cryo-soft X-ray tomography. Our protocol has been tested at two synchrotron beamlines (B24 at Diamond Light Source in the UK and BL09 Mistral at ALBA in Spain) and has led to the development of a decision aid that facilitates experimental design with the strategic use of markers based on project requirements. This protocol takes between 1.5 h and 3.5 d to complete, depending on the cell populations used (adherent cells may require several days to grow on sample carriers).Existing protocols for full-length single-cell RNA sequencing produce libraries of high complexity (thousands of distinct genes) with outstanding sensitivity and specificity of transcript quantification. These full-length libraries have the advantage of allowing probing of transcript isoforms, are informative regarding single-nucleotide polymorphisms and allow assembly of the VDJ region of the T- and B-cell-receptor sequences. Since full-length protocols are mostly plate-based at present, they are also suited to profiling cell types where cell numbers are limiting, such as rare cell types during development. A disadvantage of these methods has been the scalability and cost of the experiments, which has limited their popularity as compared with droplet-based and nanowell approaches. VTX-27 cell line Here, we describe an automated protocol for full-length single-cell RNA sequencing, including both an in-house automated Smart-seq2 protocol and a commercial kit-based workflow. The protocols take 3-5 d to complete, depending on the number of plates processed in a batch.
