Mathematical analysis of two-phase mass transfer in a batch reactor for the chemical transformation of a steroid

A reactor is described for the conversion of the slightly water-soluble steroid testosterone (T) to 4-androstene-3, 17-dione (4-AD) by enzyme in the presence of excess cofactor. Since the enzyme is subject to substrate inhibition, reaction rates are strong functions of aqueous substrate concentration. High concentrations of the substrate, testosterone, per unit reactor volume are maintained within poly(dimethylsiloxane) beads that are suspended in the aqueous enzyme solution. Mass transfer (controlled by bead size, polymer to water volume ratio, enzyme loading) is used to control the degree and rate of conversion. The reactor dynamics are predicted over a wide range of reaction conditions. The product steroid is recovered in the polymeric beads from the enzyme solution.     

Generation of a transmembrane gradient of Na+ in Methanosarcina barkeri

A transmembrane Na+ gradient was generated by Methanosarcina barkeri during methanogenesis. The intracellular Na+ concentration amounted to approximately one fifth of the extracellular one. A secondary Na+/H+ antiport system was shown to be responsible for Na+ extrusion. This system could be inhibited by amiloride. In the presence of amiloride the delta pH across the cytoplasmic membrane increased and a transmembrane Na+ gradient could neither be generated nor maintained. The possible role of Na+ in the oxidation of methanol to the level of formaldehyde is discussed.     

Common misinterpretations of the “linear,no-threshold” relationship used in radiation protection

Summary Absorbed doseD is shown to be a composite variable, the product of the fraction of cells hit (I _H ) and the mean dose (hit size)z to those cells.D is suitable for use with high level exposure (HLE) to radiation and its resulting acute organ effects because, sinceI _H = 1.0, it approximates closely enough the mean energy density in the cell as well as in the organ. However, with low level exposure (LLE) to radiation and its consequent probability of cancer induction from a single cell, stochastic delivery of energy to cells results in a wide distribution of hit sizesz, and the expected mean value,z, is constant with exposure. Thus, with LLE, onlyI _H varies withD so that the apparent proportionality between dose and the fraction of cells transformed is misleading. This proportionality therefore does not mean that any (cell) dose, no matter how small, can be lethal. Rather, it means that, in the exposure of a population of individual organisms consisting of the constituent relevant cells, there is a small probability of particle-cell interactions which transfer energy. The probability of a cell transforming and initiating a cancer can only be greater than zero if the hit size (dose) to the cell is large enough. Otherwise stated, if the dose is defined at the proper level of biological organization, namely, the cell and not the organ, only a large dosez to that cell is effective.Dedicated to Prof. L.E. Feinendegen on the occasion of his 60th birthday     

Synthesis and physical characterization of DNA fragments containing N4-methylcytosine and 5-methylcytosine.

The synthesis of N4-methyl-2'-deoxycytidine and its fully protected mononucleotide, suitable for the oligonucleotide synthesis by phosphotriester method is described. A set of octanucleotides - d(CGCGCGCG), d(CG5mCGCGCG), d(CG4mCGCGCG) and dodecanucleotides - d(GGACCCGGGTCC), d(GGA5mCCCGGGTCC), d(GGA4mCCCGGGTCC) has been synthesized in a solution. Physical characterization of the oligonucleotide duplexes by means of UV and CD spectrometry provides the evidence that 4mC similarly to 5mC favours the B--greater than Z transition, although both of these methylated cytosines inhibit the B--greater than A conformational change. N4-Methylcytosine in contrast to 5-methylcytosine reduces the DNA double helix thermal stability.     

A novel method of protein analysis for prediction of biological function: application to tumor toxins

Informational spectra method was applied to the analysis of lymphotoxin and tumor necrosis factor. The correlation between the information contained in primary structure of these tumor toxins and some oncogen transforming proteins was established. This correlation implies the possibility of a competitive action between these two groups of proteins. "Hot spots" positions in the primary structure of lymphotoxin and tumor necrosis factor for the functional "up" and "down" mutations were predicted.     

An antioxidant prevents and reverses calcium-induced uncoupling of rat liver mitochondria

Accumulation of Ca2+ by rat liver mitochondria in the presence of inorganic phosphate results in spontaneous activation of respiration accompanied by a progressive loss of the accumulated cation. The lipid peroxidation inhibitor, ionol, completely prevents and reverses the Ca2+/phosphate-induced loss of accumulated Ca2+ and restores the respiration to state 4 level without having any effect on the rate of Ca2+ accumulation and respiration in the presence of an uncoupler. No correlation between the ionol-dependent loss of Ca2+ and the formation of malonic dialdehyde in mitochondria was found. The measurements of delta psi across the inner mitochondrial membrane during a progressive loss of Ca2+ suggest that the Ca2+/phosphate-induced "uncoupling" is mainly due to the appearance of electrogenic fluxes (but not Ca2+ cycling) which is under control of some products of initial steps of lipid peroxidation.     

Role of protein kinase C in regulating smooth muscle electrical and contractile activity: the effect of phorbol ester

The effects of protein kinase C activation by 12-O-tetra-decanoyl-phorbol-13-acetate (TPA) on the functions of guinea-pig smooth muscle taenia coli have been studied, using double-sucrose-gap method. A 15-20-min treatment of the muscle with 2 X 10(-8) M TPA caused a progressing inhibition of spontaneous electrical activity and mechanical tension, suppression of post-hyperpolarizing electrical and contractile "off-responses", a decrease in the number of action potentials during superthreshold membrane depolarization, depression of electrical and mechanical responses induced by acetylcholine, histamine, bradykinin mediators. The treatment of pre-depolarized (140 mM kappa+) muscle with 2.10(-8) TPA has led to a considerable reduction in contractile responses induced by the above mediators. The results obtained indicate that protein kinase C is capable of regulating both voltage-sensitive and receptor-operated ionic channels in smooth muscle cells.     

Effect of dibunol and isoptin on the creatine kinase and myoglobin content of the blood serum in dogs undergoing postischemic coronary reperfusion

The effect of an antioxidant dibunol and calcium antagonist verapamil on postperfusion release of myoglobin (Mb) and MB-creatine kinase (MB-CK) has been assessed in 30 dogs with experimental coronary occlusive myocardial infarction. It has been shown that reperfusion after 3-hour ischemia does not only accelerate the release of intracellular proteins, but also leads to pronounced myoglobinemia and blood enzymes. In postischemic blood flow recovery with combined dibunol and verapamil preliminary injections, an almost threefold decrease in MB-CK and Mb blood content, as compared to "reperfusion" indexes, was observed by the 10th minute of reperfusion.     

The dosage thresholds for disordered physical work capacity in mice and rats following irradiation

Physical efficiency of mice and rats at early times following irradiation with doses of 35 to 200 Gy was estimated by the muscle endurance and motor coordination indices. A threshold nature of the effects was exhibited at high doses. For instance, as large as 40 Gy was the dose threshold at which stability in the accomplishment of the known operations was impaired; at 70-100 Gy, the accomplishment decreased down to 50% of the initial level, by the 2nd hour after irradiation, with the subsequent partial restoration; 200 Gy was the threshold dose at which the disturbances were irreversible. These values were also similar to those of the threshold doses at which clinical picture of the affection was changed.     

New lines of spontaneously transformed cells obtained from "precrisis" cultures of embryonic rat cells

A new approach to selection of lines of spontaneously transformed cells from the rat embryo "precrisis" cultures is described and their phenotypes at the initial and advanced stages during a long-term cultivation are characterized. The new selective system, referred to as 2T7, differs from the well known 3T3, 2T6 and 3T12 systems (Todaro, Green, 1963; Aaronson, Todaro, 1968). It is based on the maintenance of cultures under maximum cell densities. Such an approach facilitated and accelerated the start of the "crisis" stage (up to 3-8 passages) with the following gradual death of almost the whole normal senescent cell population, the colony formation resulting from the proliferation of single clonogenic cells. The frequency of clonogenic cells was about 6 x 10(-6). Six lines of spontaneously transformed cells from embryos of noninbred white rats (LRec-1--LRec-6) and one line from the Wistar embryos (LRec-7) were established. All the lines are characterized as diploid or near-tetraploid, with 1-4 different marker chromosomes formed from chromosome 7, as was reported elsewhere (Artsybasheva et al., 1988). The values of saturation densities and the time of population doubling for all the 7 lines differed from those for the rat embryo primary cultures cells. LRec-1--LRec-6 cells were unable to form the colonies in soft agar, while LRec-7 cells were able to grow in agar. The lines LRec became oncogenic for 1-2 day old rats after different periods of cultivation in vitro--from 3 to 7 months. The line LRec-7 Wistar appeared to be highly oncogenic from the very beginning after its selection. The histological analysis revealed that the LRec-1 tumors could be classified as polymorphocellular sarcoma. Up to 20 passages the LRec-1 line had numerous clonogenic cells (50-60%) in sparse cultures independently on the serum content in the media. By a 3-step selection of LRec-1 cells, on cultivation in media with lower serum contents (1-0.1-0%), a semisuspension of LRec-1sf subline (serum free) was established. This line was highly oncogenic for 1-2 day old rats, was easily cryopreserved and proliferated in the serum-free media for unlimited time, forming small colonies in agar. Thus, the new approach allows to establish with high effectiveness spontaneous lines of rat embryo cells with differently transformed phenotypes, i.e. preneoplastic and oncogenic ones.(ABSTRACT TRUNCATED AT 400 WORDS)