Functionally important lysine residues in inorganic pyrophosphatase from E. coli. II. Isolation and characteristics of modified tryptic peptide and analysis of the functional role of lysine residues controlling enzyme activity

Inactivation of inorganic pyrophosphatase from E. coli by pyridoxal-5'-phosphate is due to the modification of a lysine residue located in the tryptic peptide with the Asp-Leu-Pro-Glu N-terminal sequence. In course of the enzymatic process this lysine-residue appears to be in the protonated state and either operators as the proton donor for the product of the enzymatic reaction or is involved in stabilization of the transition state.     

Temperature regulation in a burrow-nesting tropical seabird,the Wedge-tailed Shearwater (Puffinus pacificus)

Summary At low air temperatures (2.3–13.9°C), Wedge-tailed Shearwaters (Puffinus pacificus) shivered and their oxygen consumption increased to as much as 283% of the mean value (0.77 ml O₂/g·h) within the thermoneutral zone of air temperature (23–34°C). The minimal thermal conductance of the tissues and plumage was similar to the value predicted from the body mass (320.5 g). The oxygen consumption of the birds within their thermoneutral zone was lower than predictions based on body mass. At elevated air temperatures, the shearwaters panted at respiratory frequencies as high as 260 respirations/min; maximal respiratory frequencies were not invoked until the birds had become hyperthermic. During exposure to a hot environment, the oxygen consumption of the birds increased and in most instances the shearwaters were not able to lose heat equivalent to their concurrent metabolic heat production.Symbols and abbreviations BMR basal metabolic rate - C _total total thermal conductance - f respiratory frequency - TEWL total evaporative water loss - T _st stomach temperature - T _re rectal temperature     

Purification of acetylcholinesterase from horse erythrocytes using affinity chromatography

A commercial preparation of water-soluble acetylcholinesterase from horse red cells has been purified to a specific activity of 2380 U/mg of protein (a 1660-fold purification) by a twofold affinity chromatography on the known sorbent of Sepharose-p-[NH-(CH2)5-C(O)NH(CH2)5C(O)NH-]-C6H4-N+(CH3)3 X Br- at pH 7.5. A selective elution of the enzyme was carried out from 10 mM of the phosphate buffer solution which contains 0.2% of triton X-100. Subsequent desorption of the enzyme proceeded with 5 mM of phenyltrimethylammonium bromide introduced into the buffer. Such effective preparations of acetylcholinesterase have not been previously produced. Effectiveness of the affinity sorbents considerably depends on the nature of the ligand which is covalent-linked with a Sepharose matrix and on the length of the attachment spacer arm ("insert") between them. A reversible inhibitory effect of certain ligands (tetramethylammonium, phenyltrimethylammonium) and their derivatives on acetylcholinesterase is estimated in comparison.     

Endocrine function of the heart. Structure and biological properties of peptides secreted by the heart atrium

Apart from the generally known functions, the heart has also an endocrine function. Atrial cardiocytes, being typical secretory cells, release peptide hormones into the blood stream: atrial natriuretic peptide containing 28 amino acids and cardiodilatin. The structure of atrial peptides was determined. It was shown that both peptides were derived from their common precursor, a protein containing 151 amino acids. The presence of specific receptors is demonstrated on plasmatic membranes of cells of kidney epithelium, arterial smooth muscle, arterial endothelium, kidney cortex and hypophysis. The interaction of atrial peptides with these receptors activates the guanylate cyclase system. The biological action of atrial peptides manifests itself in the quick, massive and instantaneous increase of diuresis and electrolyte excretion, elevated clearance of creatinine, decrease of kidney vascular resistance, intensification of glomerular filtration, inhibition of stimulated secretion of aldosterone, relaxation of blood vessels, elimination of arterial and intestinal spasm induced by various endogenous and exogenous vasoconstrictors and in correction of kidney hypertension. Various radioimmunoassays for the presence of atrial peptides in human plasma were developed; it was shown that in patients with congestive heart failure the content of atrial peptides is increased.     

5-Amino-4-imidazolecarboxamide is a mutagen in E. coli

5-Amino-4-imidazolecarboxamide (5A4IC), the base moiety of a common intermediate in de novo purine biosynthesis, was found to be mutagenic in E. coli. Using a series of mutants in the tryptophan synthetase A gene, 5A4IC was observed to cause transition and transversion mutations at similar levels. At 400 micrograms/ml in the growth medium, it stimulates the base substitution GC----AT 4.8-fold; AT----GC 20-fold; AT----CG (2 sites) an average of 6.0-fold; AT----TA 7.8-fold; and GC----CG 6.1-fold. The transversion GC----TA was not tested. In contrast to the base, 5-amino-4-imidazolecarboxamide riboside is not mutagenic at a similar molar concentration.     

Dynamics of immunoglobulins and immunocompetent lymphocytes in guinea pigs with experimental ascariasis

The paper presents data on variations in the level of serous immunoglobulins (Ig M and Ig G classes), immunocompetent T- and B-lymphocytes in lymphoid organs (spleen, mesenteric, portal and mediastinal lymph nodes) and specific antiparasitic antibodies in guinea pigs during the dynamics of experimental ascariasis.     

Method for histological preparation of bone sections containing titanium implants

A thin sectioning technique involving hand grinding has been developed to produce 20-40-microns-thick sections of bone-titanium implant sites. Components include: 1) surface staining of sections prior to mounting on slides so bone labels (oxytetracycline-HCl and 2,4-bis(N,N-dicarbomethyl)aminomethylfluorescein (DCAF] can be seen in sections viewed with transmitted light, 2) a pneumatic sample press for bonding sections to slides with a thin, uniform glue line and without trapped air bubbles, and 3) bonding methyl methacrylate embedded sections to clear acrylic slides with methyl methacrylate monomer to provide enhanced bond strength and grinding properties compared to those obtainable with glass slides. Sample cracking and distortion is minimized and the tissue-implant interface can be kept intact. The expense of start-up equipment for this technique is minimal.     

Effect of the cytoplasm of irradiated cells on chromatin degradation in mouse thymocytes and hepatocytes

The addition of a cytoplasmic fraction, isolated from cells 3h after irradiation of mice, to exposed or intact thymocyte nuclei causes a 2- or 3-fold acceleration of chromatin degradation in the nuclei incubated in conditions optimum for activity of Ca2+,Mg2+-dependent endonuclease to be manifest. In contrast to thymocytes, no chromatin degradation products are found in liver cells of irradiated mice. The cytoplasmic fraction isolated from hepatocytes of irradiated animals fails to activate chromatin degradation in thymocyte nuclei.     

Catecholamine content and acetylcholinesterase activity in the brain of rats exposed to lasers

The catecholamine content was quantitated and acetylcholinesterase activity determined in rat brain locally exposed to laser radiation. It was shown that the adrenaline/noradrenaline ratio changed in the tissues under study and the cholinergic system was involved in the abscopal effect of laser radiation.