Notes

Demoralization within the wake up with the COVID-19 crisis: Whereto the long run regarding younger Australians?
A lower level of reactive oxygen species was observed in cells of DMSO-SUC-50FBS when compared to other cryoprotectants. Only cells of DMSO-SUC-50FBS had mitochondrial potential similar to non-cryopreserved cells.

10% DMSO supplemented with 50% FBS and 0.2 M SUC was observed to be the most efficient cryoprotectant for preserving collared peccary somatic cells.
10% DMSO supplemented with 50% FBS and 0.2 M SUC was observed to be the most efficient cryoprotectant for preserving collared peccary somatic cells.
Virus-free sugarcane is difficult to achieve due to the multiple vegetative propagation cycles employed commercially. In vitro culture using small (1 mm) meristematic shoot tips has eliminated viruses but survival is low with small explants.

Droplet-Vitrification (D-V) and V-Cryoplate protocols were investigated for the elimination of Sugarcane mosaic virus (SCMV) from large (c. 3 mm) in vitro-derived shoot tips.

Shoot tips excised from NCo376 and N19 cultivars were exposed to both cryogenic procedures. Virus indexing by RT-qPCR was performed 16 weeks after recovery.

Explants exposed to cryo-treatments that recovered and multiplied was 30-92%, while at least 90% of control explants regenerated. No virus was detected in multiplied shoots from either cultivar after D-V and liquid nitrogen immersion. In NCo376, virus was eliminated after D-V without cooling.

The preliminary findings suggest that cryotherapy and/or osmotherapy are viable options for SCMV removal from infected plants.
The preliminary findings suggest that cryotherapy and/or osmotherapy are viable options for SCMV removal from infected plants.
The cryopreservation process induces osmotic stress, membrane changes and production of reactive oxygen species resulting in damage to the spermatozoa. Together, the presence of oxygen in the extender aggravates the oxidative stress that further reduces the cryosurvival rate of sperm cells.

To study the combined effect of cholesterol loaded cyclodextrin (CLC) and partial deoxygenation on post-thaw semen quality in crossbred bulls.

A total of 18 ejaculates from three crossbred bulls with >3+ mass motility and >70% individual progressive motility were utilized for the study. Each semen sample was divided into four groups Group I (containing extender without partial deoxygenation or CLC addition); Group II (extender containing 3 mg CLC/120X10
spermatozoa); Group III (extender containing 3 mg CLC/120X10
spermatozoa and 4 ppm dissolved oxygen (DO) level); Group IV (extender containing 3 mg CLC/120X10
spermatozoa and 6 ppm DO level). The samples in each group were finally extended to have 80×10
ng post-thaw semen quality in cross-bred bulls.Artificial insemination (AI) with frozen or cooled-stored semen plays a key role in the widespread distribution of germplasm of elite livestock resources and the protection of endangered species. Cryopreservation provides long-term preservation of sperm and also encourages a greater exchange of genetic material between distant populations. However, freezing has some detrimental effects on sperm, including premature induction of acrosome response, reduced sperm motility, reduced viability, and impaired sperm DNA integrity and fertility. The transition of the membrane phase occurs when the sperm cools down, and lipid accumulation damages the micro-domain, thereby impairing membrane functions, leaving a gap between the gel and the liquid membrane region. Coenzyme Q10 (CoQ10) is a vital lipophilic molecule found in all respiratory eukaryotic cells, including spermatozoa. When such a lipophilic antioxidant is added to the sperm, it can directly diffuse into the polyunsaturated lipid chain present in the plasma membrane, thereby affecting the structure and function of the sperm by generating energy and preventing reactive oxygen. Coenzyme Q10 treatment of sperm from various species improves sperm quality during cryopreservation and cooled-stored condition. It is, however, unclear how this antioxidant affects sperm to improve survival during freezing or cooled-stored condition. Thus, this review highlights the potential protective mechanisms of coenzyme Q10 action during the sperm freezing process.
Permeable cryoprotectants (CPAs) are required for successful sperm cryopreservation.

To investigate the efficacy of adding different CPAs to a freezing extender on cat epididymal sperm quality.

Epididymal spermatozoa were suspended in Tris-glucose-citrate-egg yolk extender supplemented either with glycerol, methanol, formamide, ethylene glycol (EG), propylene glycol (PG) and dimethylsulfoxide (DMSO), all at 5% (v/v), and then cryopreserved. Sperm motility, viability, functional membrane integrity, morphology and acrosome integrity were examined at post-thaw.

Glycerol, formamide, EG and DMSO exhibited good, comparable cryoprotective effects, whereas PG showed moderate cryoprotection. Sperm viability in PG was lower than that in glycerol and EG, but was not different in formamide and DMSO. Contrarily, the least efficacy was observed in methanol. Interestingly, the CPA type has no effect on functional membrane integrity and morphology.

Using Tris-glucose-citrate-egg yolk extender, formamide, EG and DMSO could substitute glycerol as permeable CPAs for cat epididymal sperm cryopreservation.
Using Tris-glucose-citrate-egg yolk extender, formamide, EG and DMSO could substitute glycerol as permeable CPAs for cat epididymal sperm cryopreservation.
Human induced pluripotent stem cells (hiPSCs) are valuable resources for cell therapy and drug discovery. Cryopreservation is a key technique used to realize these applications, which require a large number of cells. However, standard protocols for the preservation of the adherent cells involve multiple complicated steps, which can lead to technical difficulties.

To develop a more efficient method for cryopreservation of adherent cells using culture dishes.

Ice-seeding treatment was employed to avoid intercellular freezing, and rapid warming used to improve cell viability.

The immediate survival rate after thawing was 48%. The recovery period of cells cryopreserved by the dish culture method was shortened upon subsequent passage culture, and the time for re-cultured cells to reach the appropriate confluency was reduced by two days.

The hiPSCs can be successfully cryopreserved in culture dishes with improved viability and faster recovery. The optimization of the ice-seeding temperature and cooling rate increased the survival rate.
The hiPSCs can be successfully cryopreserved in culture dishes with improved viability and faster recovery. The optimization of the ice-seeding temperature and cooling rate increased the survival rate.
Plate cryolipolysis is a method of applying cooling without a vacuum system, which can be used in regions with less possibility of skin suction or fibrosis.

To investigate the effects of cryolipolysis with the use of plate-shaped applicators (CrioPlaceTM) for localized fat treatment.

The sample consisted of men aged 20 to 45 years with complaints of localized adiposity in the abdominal region and flanks. Two plates were positioned in the flank and abdomen regions, respectively. They received two 60-min applications in the temperature of -2°C. The anthropometric, thermographic and ultrasound assessments were performed, and a satisfaction questionnaire was applied after treatment. The re-evaluations occurred 30 and 60 days after the first intervention.

A reduction in adiposity was observed in flank region plicometry (p<0.05) and abdominal and flank ultrasound (p < 0.05). About 66.7% of the volunteers reported less water retention, about 41.7% reported that their clothes were looser, and 100% reported overall satisfaction. Fifty percent rated the treatment as excellent and 58.3% felt improvement in overall aesthetics.

The CrioPlaceTM method was effective in reducing localized adiposity, with clinical satisfaction of measurement reduction, both in plicometry and ultrasound analyses, with highlights to the flank region results.
The CrioPlaceTM method was effective in reducing localized adiposity, with clinical satisfaction of measurement reduction, both in plicometry and ultrasound analyses, with highlights to the flank region results.
Tiger beetles are a widely distributed group including species that may be exposed to sub-freezing temperature overwinter. Despite being well studied, little is known about tiger beetle cold tolerance.

We investigated seasonal changes in cold hardiness of two northerly distributed tiger beetle species (Cicindela repanda and Cicindela limbalis).

We monitored the supercooling point (SCP), glycerol concentration, and hemolymph osmolality of adult tiger beetles during a 3.5-month acclimation to winter.

SCP decreased during winter acclimation for C. repanda, but not for C. limbalis. Both species modestly increased glycerol concentration, and C. ALK inhibitor repanda increased hemolymph osmolality by 38%.

This initial investigation into the cold-hardiness of adult tiger beetles suggests that they are capable of lowering their SCP as winter approaches, which may help them survive sub-freezing winter temperatures. Further assessment of their chill and freeze tolerance and of their overwintering conditions in the field is needed to better understand their winter physiology.
This initial investigation into the cold-hardiness of adult tiger beetles suggests that they are capable of lowering their SCP as winter approaches, which may help them survive sub-freezing winter temperatures. Further assessment of their chill and freeze tolerance and of their overwintering conditions in the field is needed to better understand their winter physiology.
The redistribution of basic ions between the cell cytoplasm and its surrounding medium due to osmotic action affects transmembrane potential and plasma membrane integrity at all stages of low temperature preservation.

To develop a physical-mathematical model describing the redistribution of osmotically active solutes between the cell and its hypertonic solutions of penetrating cryoprotectants that enables the calculation of kinetic changes in cell volume, cryoprotectant and ion concentrations, as well as the cell transmembrane potential during cell equilibration with cryoprotectant solutions.

The study has modeled the mass transfer process of mouse oocytes upon exposure to 1.5 M DMSO and 1,2-Propanediol (1,2-PD) solutions.

Equations for changes of the normalized volume as well as intracellular concentrations of DMSO, 1,2-PD, potassium, sodium, chlorine and transmembrane potential have been obtained in a dimensionless form. The membrane permeability coefficients for DMSO and 1,2-propanediol have been determined and compared with the data of Paynter et al (5).

The study shows that the incorporation of transmembrane ion movement and electrical potential change in the mathematical model leads to lower values of mouse oocyte membrane permeability coefficients for water and cryoprotectants in comparison with data determined by the traditional model.
The study shows that the incorporation of transmembrane ion movement and electrical potential change in the mathematical model leads to lower values of mouse oocyte membrane permeability coefficients for water and cryoprotectants in comparison with data determined by the traditional model.
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