Impact of Dorsal Root Ganglia Cryoextract on Histological Steatures in Dices and Contractility of Uterus in Differently Aged Rats

To date, the number of women giving birth to their first child in late reproductive age is increasing around the world. This stipulates a need in designing the new approaches to restore the uterine contractile activity. In this paper, the histological features of uterus and its contractile activity have been experimentally studied in differently aged rats after the dorsal root ganglia cryoextract (DRGCE) administration. The cryoextract was derived from dorsal root ganglia of neonatal piglets by three-fold freezing down to –196°C in saline, followed by thawing at room temperature, homogenization and centrifugation. Here, we used the female rats of reproductive age (RA, 6-month-old) and those of late reproductive age (LRA, 14-month-old). Animals of both age groups received intraperitoneally either DRGCE (experimental groups) or saline (control groups) for 9 days by 0.2 ml. To days 28–29 after administration beginning the animals were sacrificed and the uterine fragments were taken for histological examination and study of oxytocin (OT)-induced uterine contractile activity (UCA). The strength of OT-induced uterine isometric contraction was found to decrease by 28.6% in LRA rats vs. the RA ones. The DRGCE administration to LRA rats increased the UCA indices, namely the contractile frequency, contractile amplitude and strength of isometric contraction augmented by 25, 9.8 and 30% respectively, as compared with the group of the same age without DRGCE introduction. This effect was observed on the background normal uterine histological structure and unchanged myometrial thickness.

Thermoplastic Analysis of Cluster Crystallization of Cryoprotective Solutions

For the first time the possibility of using thermomechanical analysis to construct the fragments of state diagrams of cryoprotective solutions in the zone of glass transition temperatures has been considered. A method for studying cluster crystallization of cryoprotective solutions based on thermomechanical curves has been developed. The parameters of thermomechanical curves of frozen aqueous solutions of dimethyl sulfoxide (DMSO), polyethylene oxide 1500 (PEO-1500), glycerol were analyzed and the relationship between these parameters and the cluster crystallization kinetics for these solutions was established. On the basis of experimentally obtained thermomechanical curves for the frozen solutions of DMSO and PEO-1500 the possibility of formation of clusters of two types has been shown: on the basis of ice and cryoprotective substance microcrystals. Additional experimental data were obtained to construct a complete state diagrams of cryoprotective solutions, which include the existing cluster phase areas.

Theoretical Estimation of Optimal Linear Cooling Rate for PK-15 Cell Suspension

Preservation of cells during crystallization of the cell suspension is influenced by two types of damaging factors. The first type of cryoinjury occurs during the crystallization of the extracellular environment and is caused by dehydration of cells, increasing the concentration and ionic strength of extracellular and intracellular solutions. As the cooling rate rises, the damage rate of the first type decreases as a result of the reduced time of action of damaging factors. The second type of cryoinjury is intracellular crystallization, the probability of which enhances at high cooling rates, is considered the most destructive to cells. The optimal linear cooling rate for PK-15 cells is determined using a physico-mathematical model, which describes the probability of cryoinjury of cells in the linear freezing mode and is based on the two-factor theory of cryoinjury, thermodynamic theory of homogeneous crystallization and general theory of activation-type processes. The findings have shown that within the range of cooling rates < 0.5 °C/min the cryoinjury of PK-15 cells occurs mainly due to the effects of the solution, and at cooling rates > 2.5 °C/min this was mainly resulted from an intracellular crystallization. The dependence of the percentage of damaged cells on the cooling rate has a relatively wide minimum within the range of cooling rates of 0.5 °C/min… 2.5 °C/min.

Gel Chromatographic Examination of Serum of Rats and Hamsters Under Artificial and Natural Hibernation

In this study, molecular composition of hamster and rat blood was studied by gel permeation chromatography under natural (NH) and artificial hibernation (AH). The control group was represented by 5 fractions of molecules in hamsters and 7 in rats. The areas under peaks of the fractions similar in molecular weight in hamsters and rats were larger in rats. NH was characterized by appearance of new fractions (1,350, 2,350, and 6,350 Da) and an increase in areas under peaks of the control fractions (1,140 and 1,980 Da). Artificial hibernation in rats led to an increase in areas under peaks of 1,140 Da fraction, a decrease in that of 530 Da, and disappearance of 1290 Da, as well as the appearance of new fractions (650, 830, 950, 2350, and 5110 Da). Two hrs of later AH, the areas under peaks of 1,140 and 1,520 Da fractions were greater and that of 530 Da was lesser; 650, 2,350 and 5,110 Da fractions disappeared, 1,290 Da reappeared and new fraction of 4,030 Da appeared. New fractions of 5,820 and 6,530 Da were found 24 hrs later. In hamsters under AH, the areas under peaks of 1,140, 1,600, and 3,330 Da increased; as well as a new peak in 5,280 Da appeared, both in the control rats and those in 24 hrs after AH.

Cultural Properties of Cryopreserved Thymic Multipotent Stromal Cells and Fetal Skin and Muscle-Derived Cells

The paper provides a comparison of properties of cryopreserved fetal murine multipotent stromal cells (MSCs) of skin-muscular origin and those derived from adult thymus in culture in vitro. Fetal MSCs showed a 30% higher number of average population doublings within 24 hrs, and 41% lower average population doubling time. It was found that the fetal MSCs of the 4th passage had a 39% higher clonogenic activity than the adult thymus-derived ones. Fetal MSCs and those derived from adult thymus differentiated in osteogenic and adipogenic lineages with equal efficiency in special culture media. Fetal and thymus-derived MSCs were characterized by almost the same high ability of contact interaction with thymocytes, and the fibroblast-lymphocyte rosette (FLR) formation. They were far less active in FLR formation with lymph node cells. This indicated the presence of membrane affinity for immature lymphoid cells in both MSC subpopulations. The results showed the fetal MSCs to be significantly different from the adult thymus-derived MSCs by more active kinetics of growth and clonogenic potential. However, both cell subpopulations had virtually the same ability for linear differentiation and showed high activity during contact with immature lymphoid cells. Linear differentiation and the ability to interact with lymphocytes were found to be quite stable properties of MSCs, but a proliferative activity and in vitro colony formation distinguished significantly in different types of MSCs. This can be taken into account when choosing the cells for therapy, research and results assessment.

Role of Liquid-Liquid Phase Transitions in Mechanism of Erythrocyte Protection During Cooling with CRIHBT-115 Cryopreservative Agent

The presence in the system of critical liquid-liquid phase transition (PT) by the mechanism, resulting in formation of dispersion system, namely high-concentrated emulsion, has been established here during cooling when using polarized light microscopy and fixation of critical opalescence phenomenon in erythrocyte concentrate with glycerol-containing cryopreservative agent, designed at the Central Research Institute of Haematology and Blood Transfusion (Russia) (CRIHBT-115 ). The studied cryobiological system displayed no signs of crystallization. A phase behaviour of cryopreservative and supernatant has been studied during cooling-warming cycle. Changes in the volume of cryopreservative and erythrocyte concentrate were comparatively and qualitatively evaluated during cooling. The mechanism of protective action of cryopreservation solution has been determined. The similarity between physical and chemical processes during cooling-warming of erythrocyte cytoplasm and garlic meristem cells (germinal plant tissue) when entering cold anabiosis has been established.

Placental Cryoextract and Renin-Angiotensin-Aldosterone System Blockade Mitigate Renal Failure in Rats

Here, we have studied the impact of administration of rat placental cryoextract (PCE), drug blockade of the renin-angiotensin-aldosterone system (RAAS) with enalapril and spironolactone and their combination on the rat kidney tissue structure and excretory function at different stages of chronic renal failure (CRF) development using the glycerol model. In 3 weeks after glycerol introduction, the animals from all the groups showed low values of glomerular filtration rate, impaired blood flow in renal cortex, tubular epithelial dystrophy, inflammation and edema of interstitium, indicating the onset of CRF development. Tubulo-interstitial nephritis and nephrosclerosis were dominated in untreated rats 16 weeks later. The use of RAAS drug blockade, as well as a comprehensive therapy with RAAS blockers and placental cryoextract stopped the inflammatory processes in renal tissue, restored blood circulation and normalized excretory function, which persisted for up to 16 weeks of observation.