Preparation and Electrochemical Performance of Three-Dimensional Vertically Aligned Graphene by Unidirectional Freezing Method

Three-dimensional vertically aligned graphene (3DVAG) was prepared by a unidirectional freezing method, and its electrochemical performances were evaluated as electrode materials for zinc−ion hybrid supercapacitors (ZHSCs). The prepared 3DVAG has a vertically ordered channel structure with a diameter of about 20−30 μm and a length stretching about hundreds of microns. Compared with the random structure of reduced graphene oxide (3DrGO), the vertical structure of 3DVAG in a three−electrode system showed higher specific capacitance, faster ion diffusion, and better rate performance. The specific capacitance of 3DVAG reached 66.6 F·g−1 and the rate performance reached 92.2%. The constructed 3DVAG zinc−ion hybrid supercapacitor also showed excellent electrochemical performance. It showed good capacitance retention up to 94.6% after 3000 cycles at the current density of 2 A·g−1.

The effect of different vitrification protocols on cell survival in human ovarian tissue: a pilot study

Background Vitrification has superseded the slow freezing method for cryopreservation of oocytes, embryos, and sperm, but there are as yet no standard protocols for its use in ovarian tissue cryopreservation (OTC). Published protocols diverge mainly with regard to the extent of supplementation of dimethyl sulfoxide (DMSO) to the vitrification medium, and to the use of an open or closed vitrification system. We investigated the viability of cells after vitrification/warming, using ovarian tissue of transgender patients, by means of Fluorescence Activated Cells Sorting (FACS), and histomorphological analyses using a DMSO-containing (P1) and a DMSO-free protocol (P2) in an open or closed vitrification setting. Results Twelve ovarian samples were donated from female-to-male transgender patients: 6 were vitrified according to protocol 1, the other 6 according to protocol 2. The amount of viable cells was 90.1% (P1) and 88.4% (P2) before vitrification. After vitrification and subsequent warming, viable cells were reduced to 82.9% (P1, p = 0.093) and 72.4% (P2, p = 0.019). When comparing the closed and the open systems, the decline in cell viability from pre- to post-vitrification was significant only for the latter (p = 0.037). Histological examination reveals no significant differences with respect to degenerated follicles before or after vitrification. Conclusion These results led us to conclude that a protocol containing DMSO results in a higher viability of ovarian cells than a protocol that uses ethylene glycol as cryoprotective agent in vitrification. The use of an open vitrification system led to significant decline in the rate of viable cells. Trial registration NCT03649087, retrospectively registered 28.08.2018.

Preservation of Biologically Active Compounds and Nutritional Potential of Quick-Frozen Berry Fruits of the Genus Rubus

Cryoprotective freezing methods are increasingly being developed and used as an effective means of protecting valuable bioactive compounds in processed berry fruits. The quick-freezing method allows the bioactive compounds in the plant material to be preserved over a longer period of time, thus providing a high-quality product with significant antioxidant capacity. The aim of this study was to determine the effects of the quick-freezing method on physico-chemical properties and bioactive compounds content of fruits in three soft fruit species: tayberry, raspberry, and blackberry, and to evaluate the stability of specific phytochemicals during the three-month storage period. The freezing method had a significant effect on the physicochemical properties with a significantly less drip loss observed after thawing in fruit frozen by quick-freezing (at −34 °C for 25 min) compared to fruit frozen classically (−18 °C to 24 h). The color of quick-frozen fruits also changed significantly less compared to fresh fruits. Of the bioactive compounds analyzed, it should be noted that there was a significantly lower loss of ascorbic acid recorded during quick-freezing. On average, the quick-frozen fruits contained 28% more ascorbic acid than the classical frozen fruits. In general, the quick-freezing procedure contributed to a better preservation of total polyphenolic compounds and anthocyanins, and thus berry fruits also showed higher values of antioxidant capacity during quick freezing than during the classical procedure. During the storage period of three months, a decrease in the content of all the bioactive compounds studied was observed, although it should be emphasized that this loss during storage was not as pronounced in fruits frozen by the quick-freezing method as in classically frozen fruits. It can be concluded that the quick-freezing contributes significantly to the preservation of valuable bioactive compounds of berries and that this processing method can be considered important for maintaining the nutritional properties of berry fruits.

Initial Characterization of Male Southern Stingray (Hypanus americanus) Reproductive Parameters and Preliminary Investigation of Sperm Cryopreservation

This study investigated the reproductive biology and sperm cryopreservation of ex situ southern stingrays (Hypanus americanus) by semen collection and characterization and the development and validation of an enzyme-linked immunoassay for plasma total testosterone. Semen was collected in March and June using a manual massage technique, and the ejaculates were assessed for volume, pH, osmolarity, motility, status (0–5 scale: 0 = no forward progression, 5 = rapid linear progression) and total sperm count. Semen was extended in Hank’s elasmobranch ringer solution containing 10% DMSO, 10% glycerol or 5% glycerol with 5% N-methylformamide and cryopreserved using a conventional freezing method (~−50 °C/min) or a modified slow freezing method (~−3 °C/min). Body condition was scored from 1–5 and was noted to be low in March (1.93 ± 0.07) due to feeding practices and increased by June (2.93 ± 0.05) after dietary corrections were made. A concomitant increase (p < 0.05) in plasma total testosterone concentration and sperm motility was noted between March (8.0 ± 7.2 ng/mL, 5.71 ± 2.77%) and June (97.3 ± 11.3 ng/mL, 51.4 ± 14.3%). Samples cryopreserved using a modified slow freeze method (~−3 °C/min) had higher post-thaw motility and plasma membrane integrity than conventionally cryopreserved samples. Data indicate that southern stingray sperm morphometrics adheres to those of other elasmobranch species and that a slow cooling rate may be an avenue of research to improve southern stingray sperm survival during cryopreservation.

Direct freezing of whole blood enables analysis of leucocyte markers by flow cytometry: a proof-of-concept study

Aim: A new one-step flow cytometry procedure has been recently demonstrated for identifying subjects with infections, but only for fresh whole blood samples. The goal of this study was to assess its applicability on frozen samples, by proposing a new method to perform the sample freezing directly and easily. Methods: Fresh blood was tested, then frozen either directly or with dimethylsulfoxide and serum. Common markers of white blood cells as well as infection-related biomarkers were tested. Results: All percentages of leucocyte subsets and levels of infection-related biomarkers were significantly correlated between frozen and fresh samples. Conclusion: The direct freezing method enables an accurate assessment of common cellular sub-populations and of levels of important infectious biomarkers via flow cytometry.

Effects of Different Freezing Methods on Water Distribution, Microstructure and Protein Properties of Cuttlefish during the Frozen Storage

To study the effect of different freezing methods on the quality changes of cuttlefish during the frozen storage of cuttlefish, fresh cuttlefish was treated with six freezing methods (refrigerator direct-freezing, saline solution impregnation freezing, flat freezing, tunnel type continuous freezing, air-blast freezing and liquid nitrogen freezing) and then stored at −18 °C for 90 days. The time to pass the maximum ice crystal generation zone for the above six freezing methods in this experiment was 165.5, 67.5, 34.5, 21.8, 20.4 and 1.5 min, respectively. In this study, water retention (thawing loss rate, centrifugal loss rate, and cooking loss), pH, malondialdehyde content, TVB-N value, and sulfhydryl content were measured to evaluate the quality after thawing. Protein secondary structure was measured by attenuated total reflection infrared spectroscopy (ATR-FTIR), water migration was determined by low-field NMR, and muscle microstructure was observed by scanning electron microscopy. The results showed that among the six freezing methods, liquid nitrogen freezing took the shortest time to pass through the maximum ice crystal generation zone. And it had the highest water retention, the lowest TVB-N content, the highest sulfhydryl content and the least irregular curling of protein secondary structure after 90 days of frozen storage. However, liquid nitrogen freezing can cause cracks and breakage in cuttlefish due to cryogenic fracture caused by ultra-low temperature, which affects its sensory evaluation. Although the freezing speed of flat freezing is faster than refrigerator direct-freezing and saline solution impregnation freezing, the muscle is extruded and deformed during the freezing process, and the damage is more serious, and the frozen storage quality is the worst. The comprehensive analysis results showed that the freezing speed of air- blast freezing was faster and the quality of cuttlefish in the freezing process was better, which was the more recommended freezing method, and this study provided some theoretical basis for the selection of freezing method in the actual production of cuttlefish.

The Effect of Different Vitrification Protocols on Cell Survival in Human Ovarian Tissue: A Pilot Study

BackgroundVitrification has superseded the slow freezing method for cryopreservation of oocytes, embryos, and sperm, but there are yet no standard protocols for its use in ovarian tissue cryopreservation (OTC). Published protocols diverse mainly in the supplementation of dimethyl sulfoxide (DMSO) to the vitrification medium and the use of an open or closed vitrification system. We investigated the vitality of ovarian tissue of transgender patients by Fluorescence Activated Cells Sorting (FACS) and histomorphological analyses using a DMSO-containing (P1) and a DMSO-free protocol (P2) in an open or closed vitrification setting. ResultsTwelve ovarian samples were donated from female-to-male transgender patients: 6 were vitrified according to protocol 1, the other 6 according to protocol 2. The amount of vital cells was 90.1% (P1) and 88.4% (P2) before vitrification. After vitrification and subsequent warming, vital cells were reduced to 82.9% (P1, p=0.093) and 72.4% (P2, p=0.019). When comparing the closed and the open systems, the decline in cell vitality from pre- to post-vitrification was significant only for the latter (p= 0.037). Histological examination reveals no significant differences with respect to degenerated follicles before or after vitrification. ConclusionThese results lend support to the hypothesis that a protocol containing DMSO results in a higher vitality of ovarian cells than a protocol that uses ethylene glycol as cryoprotective agent in vitrification. The use of an open vitrification system led to significant decline in the rate of vital cells. Trial registration: NCT03649087, retrospectively registered 28.08.2018