In vitro cultivation of mouse embryos during the initial stages of organogenesis

Optimal conditions for in vitro cultivation of mouse embryos were sought. The embryos at 8--9.5 days of development were explanted into the culture in rotating tubes. A number of nutritional media were studied: the human umbilical blood serum; the rat blood serum, as well as its mixture (1 : 1) with Eagle's medium. It was demonstrated that the optimal medium for cultivation of 8-day-old embryos was the mixture of the rat blood serum with Eagle's medium, and for 8.5--9.5-day-old embryos--the rat blood serum, as well. Comparing in vitro and in vivo embryonic development, it was evident that in the cultivated embryos the parameters characterizing their growth decreased significantly. Possible causes of this phenomenon are discussed.     

The clinical significance of MDR-1 gene expression in the hemopoietic cells of patients with acute leukemias in different phases of the disease

AIM: Analysis of cytostatic therapy effects on expression of gene MDR-1 in hemopoietic cells of patients with acute leukemia (AL) in complete clinicohematological remission (CCHR). MATERIALS AND METHODS: The study included 48 AL patients. 27 of them were untreated, 25 were resistant to or had recurrent AL. 4 patients were followed up. Bone marrow mononuclear fraction was investigated. Expression of MDR-1 gene in the cells was evaluated at hybridization. RESULTS: High expression of MDR-1 gene occurred in leukemic blast cells either upon achievement of CCHR or at least 6 months after its onset. When using schemes of chemotherapy containing two potential inductors of gene MDR-1, expression of this gene was registered significantly more frequently than in using schemes based on one inducing drug (p < 0.05). Frequency of occurrence of enhanced expression of gene MDR-1 in leukemic blasts significantly correlated with frequency of CCHR (p < 0.05). Correlation between occurrence of the gene's expression in normal hemopoietic cells in CCHR and occurrence of early AL recurrences was not found. CONCLUSION: The findings may facilitate design of new AL treatment programs.     

Identification and characterization of differentiating soluble antigens of sheep and goat poxviruses

Soluble antigens of sheep and goat poxviruses (SPV, GPV) were isolated and purified from scab suspensions prepared from lesions of experimentally infected homologous hosts. The soluble antigens were then subjected to sequential ammonium sulphate precipitation. All the obtained fractions reacted in counter immunoelectrophoresis (CIE) with both the antisera against SPV and GPV except the fraction obtained at 30% saturation level (30% SSPV), which did not react with antiserum against GPV. This differentiating soluble SPV antigen was found to consist of 210 K proteins in exclusion chromatography. The 210 K proteins contained 3 polypeptides of 100, 35 and 17 K in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate (SDS-PAGE). The study thus gave an evidence that the SPV-specific proteins are of a higher molecular mass nature.     

Visualization of vortex shedding at the proximal side-to-end artery-graft anastomosis

The motion of guanine particles was recorded by video to visualize transitional flow phenomena in models of a proximal side-to-end anastomosis. Close examination of successive video fields revealed that above a critical Reynolds number, particles were periodically shed into the graft from a vortex situated near the anastomosis heel, and this disturbed the flow patterns in the graft causing vortex shedding to occur near to the toe of the anastomosis. The images clearly demonstrated that periodic flow structures propagated distally along the graft for over 15 tube diameters from the proximal anastomosis. The frequency of the vortex shedding was found to increase with Reynolds number. Under pulsatile conditions, the primary vortex at the heel of the anastomosis became unstable during the deceleration phase of the flow cycle and particles were shed downstream into the graft. Although it was possible briefly to observe the characteristic banded structure in the bypass graft, the flow patterns were highly three-dimensional and were quickly broken up by the accelerating flow. Dynamic flow visualization using guanine particles was found to be a complementary technique to particle tracer flow visualization and was highly effective in identifying transitional flow phenomena and the mass transport mechanisms associated with them.     

Do our methods lead to insomniacs' madness?: Daytime testing after laboratory and home-based polysomnographic studies

Complaints of daytime dysfunction are common among chronic insomniacs, but laboratory comparisons of insomniacs and age-matched and gender-matched normal controls have generally failed to document these complaints. However, a few studies, which allowed subjects to sleep in their homes on the nights before daytime testing, have shown some relative diurnal deficits among insomniacs. The current study compared the effects of nocturnal laboratory and home polysomnogram (PSG) studies on subsequent daytime test results among older insomniacs and normal sleepers. Insomniacs (n = 32) and normal sleepers (n = 32) were randomly assigned to first undergo three nights of nocturnal PSG monitoring either in the sleep laboratory (16 insomniacs, 16 normal sleepers) or in their homes (16 insomniacs, 16 normal sleepers). Following the third night of PSG monitoring, subjects spent 1 day in the sleep laboratory, where they completed a four-trial multiple sleep latency test along with four trials of a computer-administered performance test battery. Results showed that insomniacs, as a group, were slightly, albeit consistently, sleepier than were normal sleepers following nights of home sleep monitoring, but a reverse of this trend was found among subjects who underwent nocturnal laboratory PSG before daytime testing. Furthermore, normal sleepers showed faster reaction times on a signal detection task than did insomniacs within the subgroup who underwent home PSGs prior to such testing. However, within the subgroup that underwent nocturnal laboratory PSGs, insomniacs' signal detection reaction times were significantly faster than those shown by normal sleepers. Results provide some support for the speculation that the nocturnal PSG monitoring site, used as a precursor to daytime testing, may systematically affect daytime comparisons between insomniacs and matched controls. Moreover, these results suggest that the use of home-based nocturnal PSG monitoring prior to daytime testing may provide an enhanced understanding of insomniacs' diurnal complaints.     

Lipid peroxidation, arachidonic acid and products of the lipoxygenase pathway in ischaemic preconditioning of rat heart

Progenitor and pluripotent stem cells reside within connective tissue compartments. They are also present in granulation tissue. This study examined the effects of treating these two cell populations with eight bioactive factors. Cells were assayed for DNA content as a measure of proliferation and for tissue-specific phenotypic markers as measures of lineage progression and lineage commitment. Platelet-derived endothelial growth factor and insulin-like growth factor-II did not induce proliferation in either population. However, dexamethasone, insulin, insulin-like growth factor-I, muscle morphogenetic protein, platelet-derived growth factor-AA, and platelet-derived growth factor-BB stimulated proliferation in one or both cell populations. Platelet-derived growth factor-BB was the most potent stimulator of proliferation in either population. Phenotypic expression markers were induced in the progenitor cells by insulin, insulin-like growth factor-I, insulin-like growth factor-II, dexamethasone, and muscle morphogenetic protein. However, only dexamethasone and muscle morphogenetic protein induced phenotypic expression markers in the pluripotent cells. Platelet-derived endothelial cell growth factor, platelet-derived growth factor-AA, and platelet-derived growth factor-BB did not induce phenotypic expression markers in progenitor or pluripotent cells. This study suggests the potential for using progenitor and pluripotent cells as an in vitro model to ascertain the effects of various bioactive factors on stem cells potentially involved in tissue maintenance and repair.     

Effect of hydroxylamine hydrochloride on the bacterium Shigella dysenteriae 1 and antigens isolated from them

A study was made into the functional status of hypophysis, adrenal cortex, ovaries, and thyroid gland in patients with kraurosis and leukoplakia vulvae. The above patients demonstrated profound disturbances in the system hypophysis-ovaries, that attest to the need for applying corrective therapies to treat the disturbed hormonal homeostasis.     

Effect of testosterone on long-term organ cultures of canine prostate

Organ cultures of rodent and human prostate glands have shown marked differences in their morphological response to testosterone. In this study, explants from 19 canine prostate glands were cultivated for a minimum of 9 days in Trowell's T-8 medium. Groups of explants were exposed to media containing from 0.05 to 100 mum testosterone. While the higher testosterone levels (50 and 100 mum) markedly decreased explant viability, explants cultivated at lower levels (0.05 to 5 mum) appeared similar to control explants in testosterone-free Trowell's T-8 medium. Atmospheric mixtures containing either 95% or 50% oxygen were equally effective. Shortly after the cultures were initiated, large amounts of secretory product were liberated into the lumen. After 9 or more days in vitro, glandular epithelium appeared cuboidal and never revealed the acid phosphatase-rich secretory granules seen in the preculture control. However, the epithelium exhibited an increase in alkaline phosphatase and lipid content following cultivation.