Identifying dimensions of perceived patient change: some problems in the use of factor analysis. A study of change in a 'therapeutic community' admission unit is used to illustrate some of the problems associated with the use of factor analysis in evaluating therapeutic outcome. The paper describes the methodological steps involved in an attempt to identify the underlying dimensions of change which reflect how patients see themselves, and are seen by those most closely involved with them. The resulting dimensions, labelled 'pleasantness' and 'illness', are used to illustrate change at three levels: average direction, scatter, and individual cases.


Demoniacal possession and psychoanalytic theory. Psychodynamic explanations of demoniacal possession have been based mainly on Freud's retrospective interpretation of the illness of Christoph Haitzman. Freud applied libido theory to his analysis of the case and subsequently influenced the psychoanalytic theory of paranoid symptom formation and the psychoanalytic approach to the psychoses. Fairbairn, and later Guntrip, applied an object relations theory to the case and conceptualized demoniacal possession in terms of bad internalized objects. They used the example of Haitzman to illustrate an object relations approach to the understanding and treatment of severely disturbed patients. In this paper the psychoanalytic treatment of a case of demoniacal possession is described to indicate the multiple dynamic meanings which possession may have and to demonstrate the necessity for integrating and applying aspects of libido and object relations theories. Current trends in psychoanalysis are towards recognizing the psychotic core which is frequently masked by an apparently psychoneurotic illness so that analysis must reach beyond oedipal conflicts to the primitive internalized object relations. A psychoanalytic study of demoniacal possession contributes much to the understanding of such patients but particularly to the conceptualization of borderline and psychotic states. It is in the treatment of these patients that the role of the psychotherapist is more akin to that of the exorcist.


A first Volunteer Case Aid programme in a mental hospital. Revealing the voluntary helper's role of reality testing between staff and patients. A pilot Voluntary Case Aid programme in an English Mental Hospital using National Health Service resources is described. The selection of ten volunteers and ten patients and the preparation of all personnel was followed by regular contacts for at least a year. Subsequent evaluation suggested high motivation with both Case Aids and patients. Changes towards improved social functioning were evident in most cases by the end of the project. A relationship starting, continuing and finishing is reported in detail and from this study it is clear that the volunteer was a reality tester for staff and patient alike, constructively disturbing and modifying their mutual projections. This was true of most of the Case Aid interventions and the implications are discussed. A method of group supervision for Case Aids is reported.


The status of the anal character: a methodological and empirical reply to Hill. Hill's (1966) critique of the experimental evidence for the anal character is subjected to a critical examination and some of the arguments are shown to be mistaken. In addition new evidence is presented which supports the previous interpretation. It is concluded that there is still factor analytic evidence for the anal character.


Body image, plastic surgery (prosthetic testes) and Kallmann's syndrome. A patient with Kallmann's syndrome received prosthetic testicular implants. In interviews and letters, he demonstrates that delay in electing surgery, and its eventual success, were both related to incorporation of the foreign bodies into the body image.


Assessing the psychological effects of an exercise training programme for patients following myocardial infarction: a pilot study. During a study investigating physiological and other effects of an exercise programme for coronary patients, a questionnaire was administered. Preliminary analysis had suggested some improvement in the patients' morale, but in view of the possible relevance of a number of psychological variables it was decided to carry out further analysis on the available data. The coefficient of discrimination was computed for 32 patients. For 19 patients correlations were computed between scores on subjective fitness, symptoms, anxiety, interest in sex, if at work, age, weight, and workload achieved at a given heart rate. The questionnaire appeared to have satisfactorily high internal and external validity. Patients with a high 'morale' score tended to achieve a greater increase in workload over the course. Although cause and effect cannot be unequivocally assigned, the association is felt to be important, and research is continuing.


Factors associated with low birth weight of infants delivered at term. The singleton births in the 1958 British Perinatal Mortality Survey (Butler and Bonhan, 1963; Butler and Alberman, 1969) were used to study the aetiology of low birth weight of infants delivered at term. After exclusion of macerated stillbirths and infants with lethal congenital defects, 468 infants of 259 days or more gestation and a birth weight which was less than 2500 g were identified. The mothers and their pregnancies were compared with those of the population of 16 994 singleton births. Delivery at term of a low birth weight infant was significantly associated with maternal prepregnant weight, maternal height, maternal smoking, primiparity, maternal employment, low social class, a previous infant of low birth weight, threatened abortion and severe toxaemia. No significant associations were found with illegitimacy, area of residence, previous spontaneous abortion, essential hypertension, mild toxaemia, chronic or acute infections or other conditions. There were significant negative associations with a history of previous large infants and with maternal blood group AB.


Measurement of the fetal heart rate during pregnancy by a new electrocardiographic technique. An external technique for recording the fetal heart rate (FHR) during pregnancy is described. The system is based upon the detection of the fetal electrocardiogram (ECG) from the maternal abdomen and elimination of the maternal QRS complexes by a new system of subtraction. Fetal QRS complexes with an amplitude of 10 microvolts or more were processed to produce heart rate recordings of much higher quality than previously obtained with the phonocardiographic and ultrasonic systems. The proportion of successful recordings was reduced between 27 and 34 weeks of gestation due to the decrease in size of the fetal QRS complex which occurred at this time; this was a major drawback of the ECG system.


Abnormal platelet function in pre-eclampsia. Platelet behaviour was studied in groups of women suffering from mild and severe pre-eclampsia, and compared with normal pregnant and non-pregnant controls. Platelets from women with severe pre-eclampsia were less responsive than normal to a variety of aggregating agents, and this impairment was significant in response to collagen and vasopressin. Women with severe pre-eclampsia had raised plasma adenine nucleotide levels and lowered platelet 5-hydroxytryptamine levels compared with the controls. Platelets from women with mild pre-eclampsia showed only a slight difference from normal. These findings may be the result of platelets having undergone aggregation and disaggregation within the circulation, and suggest that platelets may be involved in the pathogenesis of pre-eclampsia.


Platelet lifespan in normal pregnancy as determined by a nonradioisotopic technique. A nonradioisotopic technique for determination of platelet lifespan was applied to 19 healthy women whose gestational ages were between 32 and 40 weeks. Nine non-pregnant women and five men were also studied. Platelet counts were performed in all subjects. No significant difference was demonstrated between platelet lifespan in pregnant women (9.2 days), in non-pregnant women (9.7 days) and in men (9.2 days). There was no significant different between platelet counts in men and in non-pregnant women, but the mean number of platelets in pregnant women was significantly lower. It is concluded that no significant changes in platelet behaviour appear to occur in normal pregnancy.


Serum lactate dehydrogenase in detecting liver damage associated with pre-eclampsia. Assessment of liver damage by estimation of serum enzymes including lactate dehydrogenase (LDH) and its hepatic fraction, LDH5, was made in 60 normal pregnant women, 20 patients with pre-eclampsia and 10 with eclampsia. Definite evidence of damage was found in patients with pre-eclampsia and eclampsia and was related to the severity of the disease.


Adipose tissue metabolism in pregnancy: the lipolytic effect of human placental lactogen. In vitro adipose tissue lipolysis was investigated in pregnant and non-pregnant women. Basal and hormone-stimulated rates of lipolysis were significantly higher in the pregnant group. Human placental lactogen (HPL) was shown to be lipolytic on adipose tissue from both pregnant and non-pregnant women; the response being more marked in the pregnant group. We postulate that HPL is responsible for this increased lipolytic sensitivity in pregnancy, the result of which is the elevation in levels of plasma free fatty acids in the third trimester.


The effect of indomethacin on spontaneous activity in the isolated human myometrium and on the response to oxytocin and prostaglandin. The spontaneous activity of human myometrial strips obtained during the proliferative and secretory phases of the menstrual cycle and during the early and late stages of pregnancy was studied. Both prostaglandin F2alpha (PGF2alpha) and oxytocin caused contractions which were superimposed on a low level of spontaneous activity but only PGF2alpha was seen to affect high levels of activity. Spontaneous contractions were ultimately abolished by the addition of indomethacin. The strips remained responsive to PGF2alpha but not to oxytocin.


Water relations of the ovarian cycle.

Water relations of the ovarian cycle. This study reports the likely pattern of change in serum osmolality during the normal ovarian cycle, together with that which occurs during ovulation-induction by gonadotrophin. The variation in serum osmolality is attributed to the action of oestrogen on venous distensibility, and the effect of that response on the osmotic equilibrium between intra- and extravascular fluid. Although the action of gonadotrophin is mediated by the products of follicular theca and granulosa cells, the physiological response is ultimately affected by water. Both ovulation and menstruation may therefore depend on achieving a critical water content of follicular fluid and endometrial tissue.


Assessment of ovarian function in perimenopausal women after stopping oral contraceptives. Ovarian function was assessed by measuring plasma gonadotrophins, urinary oestrogen and urinary pregnanediol levels, every week for four weeks, in 34 women, aged 45 to 57 years, after withdrawal of oral contraceptive therapy. A rise in urinary pregnanediol excretion occurred in 14 patients suggesting that ovulation had occurred and that these patients were "premenopausal". Elevated serum gonadotrophin values and persistently low urinary oestrogen and pregnanediol excretion occurred in 13 patients suggesting ovarian failure or a "postmenopausal" state. However, one apparently "postmenopausal" patient later showed hormonal evidence of ovulation, indicating fluctuation in ovarian function. There were 7 patients with "indeterminate" hormone profiles whose ovarian function was difficult to assess. All patients excreting more than 45 nmol of oestrogen a day had a spontaneous period within 45 days of stopping hormone therapy.


Verrucous carcinoma of the vulva.

Verrucous carcinoma of the vulva. A patient with verrucous carcinoma of the vulva is presented. A radical vulvectomy and superficial inguinal lymphadenectomy was done. The short-term result was satisfactory.


Antenatal identification of women at increased risk of being delivered of a low birth weight infant at term. A method is described by which the risk of being delivered of a low birth weight infant at term can be predicted during the third trimester. A composite score is obtained by multiplying together relevant relative risks. The effectiveness of the method is discussed.


Regulation of translation rate during morphogenesis in the fungus Mucor. The present study was undertaken in order to elucidate the molecular mechanisms responsible for regulating changes in the specific rate of protein synthesis during the yeast-to-hyphae morphogenesis in the fungus, Mucor racemosus. The distribution of ribosomes between active polysomes and monosomes and inactive subunits was determined by means of pulse-labeling and density gradient fractionation techniques. The percentage of ribosomes active in protein synthesis was observed to decrease throughout the morphological transition. The rate of amino acid addition to nascent polypeptide chains was calculated and the transit time of messenger RNA translation was measured. The results showed a significant increase in the velocity of ribosome movement along the message which was continuously adjusted throughout hyphal development.


Physical characterization of myosin light chains. This paper reports the results of an investigation into the size and shape of the low molecular weight subunits (light chains) of myosin from several animal species. Hydrodynamic, analytical gel filtration, and fluorescence anisotropy decay measurements indicated that these light chains could be represented by a general ellipsoidal model having a longest axis of about 100 +/- A. Investigation into the stability of the internal structure of the scallop regulatory light chain was carried out by studying the effect of pH, ionic strength, temperature, and guanidine hydrochloride on its circular dichroic spectrum. The nearly complete insensitivity of the circular dichroic spectrum to pH, ionic strength, and temperature variations from 4 to 70 degrees C indicated that this subunit contained regions of very stable structure which probably exist when it is bound to myosin.


Circular dichroism studies on lipid-protein complexes of a hydrophobic myelin protein. Circular dichroism spectra of lipophilin (a hydrophobic protein purified from human central nervous system myelin) were analyzed by the method of Chen et al. (1974) to obtain information on its secondary structure in aqueous and lipid environments. When introduced into phosphatidylcholine vesicles by dialysis from 2-chloroethanol, the protein possessed about 75% alpha helix. A new water-soluble form of lipophilin also containing over 70% alpha helix was obtained by a similar dialysis in the absence of lipid. This product had a higher helical content than two other water-soluble preparations derived by dialysis from phenol-acetic acid-urea. Interaction of all three aqueous forms of the protein with lysolecithin micelles resulted in increases in total helical content or in the average length of helical segments. The amount of beta sheet was at a minimum for lipophilin incorporated into vesicles, where the presence of lipid also provided some protection against thermal denaturation.


Intrinsic fluorescence of a hydrophobic myelin protein and some complexes with phospholipids. The fluorescence characteristics of lipophilin, a proteolipid apoprotein from human myelin, were determined in aqueous and lipid environments. In all cases the tryptophan residues were located in buried hydrophobic sites of uniform, but limited, accessibility to the permeant quenching agent acrylamide; only in the helicogenic solvent 2-chloroethanol were the protein fluorophores exposed to the medium. Quantum yields were dependent on the state of aggregation of the protein in aqueous solution and increased considerably on treatment with lysolecithin micelles, or when the protein was combined with phosphatidylcholine by codialysis from 2-chloroethanol into water. Fluorescence titrations indicated that lipophilin bound to lysolecithin with an association constant greater than 10(6) L/mol. Radiationless singlet excitation energy transfer from tyrosine to tryptophan residues was found to decrease markedly when the protein was combined with lipids. When the protein was introduced into dimyristoylphosphatidylcholine vesicles, the tryptophan fluorescence did not detect any solid-liquid phase change. These results were consistent with strong hydrophobic interactions between lipophilin and phospholipids, which lead to conformational adjustments in the protein, and to establishment of an immobilized layer of boundary lipid in bilayer systems.


Small subunit contacts in ribulose-1,5-bisphosphate carboxylase. The arrangement of subunits of ribulosebisphosphate carboxylase in solution has been studied by exposing the enzyme to the cross-linking agents tetranitromethane, dimethyl suberimidate, and dimethyl adipimidate, and the cleavable cross-linking agent, methyl 4-mercaptobutyrimidate followed by gel electrophoresis in the presence of dodecyl sulfate. All these agents caused the formation of dimers of the enzyme's small subunit, independently of protein concentration. In addition, trimers and tetramers of small subunit were detected in the mercaptobutyrimidate-treated enzyme. The data show that small subunits are closely paired in the native enzyme and may be in layers of four, or a ring of eight.


Biochemistry of the amatoxins: preparation and characterization of a stably iodinated alpha-amanitin. Iodination of alpha-amanitin at the 7-position in the 6-hydroxy-2-sulfoxytryptophan moiety is effected with 1 equiv of iodine monochloride in methanol. The isolated product shows a lambdamax in methanol at 301 nm, compared with 305 nm for the parent alpha-amanitin; in methanolic 0.01 M NaOH the lambdamax are 330 and 332 nm for the product and parent, respectively. Spectrophotometric titration of the phenolic hydroxyl shows a decrease in pKa from 9.72 (alpha-amanitin) to 7.94 (7 iodo-alpha-amanitin). Appropriate spectrophotometric examination therefore distinguishes between parent and product. Proton magnetic resonance shows two aromatic protons (v4H = 7.57; V5H = 6.90 ppm; j4,5 = 9) in the 7-iodo-alpha-amanitin and three aromatic protons (v4H = 7.64; V5H = 6.78; V7H = 6.94 ppm; j4,5 = 9; J5,7 = 2) in alpha amanitin thus establishing the extent and position of iodine substitution. The 7-iodo-alpha-amanitin effectively inhibits RNA polymerase activity with half-maximal inhibition at 2 X 10(-9) M and 10(-4) M for the sea urchin RNA polymerases II and III, respectively. Addition of [125I]-7-iodo-alpha-amanitin (200 Ci/mmol) to crude extracts from sea urchin blastula, MOPC 315 plasmacytoma, and adult Oregon R Drosophila melanogaster followed by resolution on DEAE-Sephadex demonstrates that the radioactive ligand binds stably and specifically with RNA polymerase II in each of these extracts.


Diversity and abundance of polyadenylated RNA from Achlya ambisexualis. The diversity, abundance, and DNA sequence representation of poly(adenylic acid) containing RNA derived from cells of Achlya ambisexualis cultured in defined and undefined media have been determined. The kinetics of hybridization of polyadenylated RNA with complementary DNA were the same for both culture conditions and revealed the presence of three frequency classes containing 29, 220, and 3000 different sequences of an average length of 1150 nucleotides. Complexity estimates derived from experiments in which polyadenylated RNA was hybridized to unique sequence DNA were in good agreement with these results. The kinetics of hybridization of complementary DNA with an excess of nuclear DNA indicate that approximately 10% of the RNA is transcribed from reiterated DNA sequences while the remainder is transcribed from single copy sequences.


T4 polynucleotide ligase catalyzed joining of short synthetic DNA duplexes at base-paired ends. The self-complementary octanucleotide dT-A-G-T-A-C-T-A has been synthesized and its sequence confirmed by two-dimensional fingerprinting. Under conditions used for the T4 polynucleotide ligase reaction, this oligonucleotide forms a dimeric duplex which shows a Tm of 18 degrees C. The optimal rate of joining of the 32P-labeled duplex occurs between 12 and 15 degrees C. The rate is highly concentration dependent, as expected for a bimolecular process. Polyacrylamide gel electrophoretic analysis of this reaction shows the presence of products up to 120 nucleotides in length. In a denaturing gel, each product appears as a double band due to the presence of its 5'-adenylylated activated intermediate. Substrates larger than eight base pairs are utilized more rapidly than the eight base pair duplex, indicating that the T4 ligase has a higher affinity for longer substrates. The low level of nicked intermediates suggests that the joining of both strands requires two steps, the rates of which must be similar.


Nucleotide specificity in microtubule assembly in vitro. A procedure is described for removing most of the GDP bound at the exchangeable GTP binding site (E site) of tubulin. Microtubule protein containing substoichiometric amounts of GDP at the E site is found to polymerize in response to: (a) two nonhydrolyzable ATP analogues, adenylyl imidodiphosphate (AMP-PNP) and adenylyl beta, gamma-methylenediphosphonate (AMP-PCP); and (b) substoichiometric levels of GTP or dGTP. The results are interpreted as suggesting that: (1) when GDP is removed from tubulin, the E site shows broad specificity for nucleoside triphosphates: (2) microtubule assembly can be induced by the binding of substoichiometric amounts of nucleoside triphosphate to the E site.


Action of surface-active substances on biological membranes. II. Hemolytic activity of nonionic surfactants. The hemolytic action of commercially available nonionic surfactants and synthesized polyoxyethylene fatty acids and mercaptans on human erythrocytes was measured. It is shown that the hemolytic power of the detergents depends on the mutual effect of the hydrophobic and hydrophylic fragments of the agent molecule and does not depend on the hydrophile-lipophile balance of the compounds. A graphical image of the structure-activity relationship obtained in the study is similar to the one found in the literature when studying the analgesic effect of imidazoline derivatives on rats in vivo. This fact is discussed on the basis of assumption that the mechanism of both processes in vivo and in vitro is related to influence of the agents on cellular membranes. It is suggested that the role of the polyoxyethylene chain is its effect on the dipole moment and the relative lipophilicity of the compound or in the participation of the fragment in the interaction with the surface components when the agent is sorbed on the membrane. It is concluded that when the correlation between the hydrophile-lipophile balance values and a membrane effect the capacity of the surfactants this indicates that the effect is caused not by destruction of the membrane but by some rearrangement of the membrane structure accompanying the surfactant adsorption.


Isolation and characterization of surface glycoproteins from L-1210, P-388 and HeLa cells. A method is described that permits the rapid extraction of the cell surface glycoproteins of two murine leukemic cells, the P-388 and the L-1210 cells as well as those of the human adenocarcinoma cells, the HeLa cells. Proof of the surface location of these glycoproteins is provided by labeling the intact cells; (a) with 125I by the lactoperoxidase iodination technique; (b) with 3H by the galactose oxidase-reductive tritiation method. Most of these glycoproteins were also shown to incorporate radioactive glucosamine and fucose. By these criteria as well as by the distribution of molecular weights, the surface glycoproteins of the two murine cells are indistinguishable; however, they differ markedly from the surface glycoproteins of HeLa cells. The extracts of the murine cells were shown to contain lectin receptor activity as determined by their ability to inhibit the lectin-induced agglutination of the intact cells.


Cytochrome b-561 in sympathetic nerve terminal vesicles from rat vas deferens. The spectral properties of sympathetic nerve vesicles isolated from the vas deferens of the rat are similar to those of the bovine chromaffin granule membranes and bovine nerve trunk vesicles, indicating the presence of the specific cytochrome b-561. The cytochrome occurs only in the fractions containing nerve vesicles, thus suggesting usefulness as a marker enzyme.


Is the Ca2+-sensitive K+ channel under metabolic control in human red cells? It is widely known that a rise in internal Ca2+ leads to an increased K+ permeability of human red blood cells [1,2,3]. Binding of Ca2+ to some membrane receptors is required for the opening of the K+ channel [4]. This requirement, however, seems to alter after "ageing" red cells in vitro in acid-citrate-dextrose solutions. Thus, the free Ca2+ concentration producing half-maximal effect on K+ permeability ([Ca2+]K+-50) of 4-weeks stored cells is approx. 2.10(-4) M (calculated from ref. 3 using 50% free Ca2+ according to Schatzmann [5]); nearly ten times lower than that reported for fresh cells [6]. This observation suggests the possibility that the K+ channel may become more sensitive to Ca2+ on cold storage. The experiments described below support this idea.


Rapid transbilayer movement of phospholipids induced by an asymmetrical perturbation of the bilayer. Phospholipase D is used to convert egg phosphatidylcholine to phosphatidic acid in unilamellar vesicles. The transbilayer distribution of both lipids is determined by 31P NMR using paramagnetic ions. Phosphatidic acid formed in the outer monolayer is translocated to the inner monolayer with a halftime of 30-40 min or less. This is accompanied by an equally fast movement of part of the phosphatidylcholine from the inner to the outer monolayer. During these fast transbilayer movements the barrier properties of the vesicle bilayer are maintained.


Spreading of liposomes at the air/water interface. Two types of film structure are formed when liposomes are spread at the air/water interface. At zero surface pressure, there is a slow transformation of the closed bilayered structure into a lipid monolayer. The internal content of the liposomes is released into the aqueous subphase. In contrast, when multilamellar liposomes are spread against a surface pressure, they retain their internal content at the air/water interface by forming multilayered structures. Among the liposomes which dipped through the interface an important fraction loses its internal content. During the spreading process at zero surface pressure, it seems that the outer layer of the liposome spreads with a better yield as compared with the inner layer. It is possible to use this spreading technique to determine the asymmetrical distribution of lipids across bilayers.


Interferon-mediated effect on ribosomal RNA metabolism. Ribosomal RNA (rRNA) has been shown to be involved in the binding of bacterial messenger RNA (mRNA) and an analogous 18 S rRNA.mRNA complex has been reported in eukaryotic systems. Thus, qualitative changes in host rRNA may be involved in the development of the interferon mediated antiviral state, a process thought to involve the inability of host ribosomes to bind and recognize viral mRNA. Data are reported which suggest that trisomy 21 human fibroblasts respond to human interferon with a marked reduction in cytoplasmic rRNA. [3H]Uridine was used to radioactively label the polysomal RNAs for 24 h beginning 12 h after interferon addition. Subsequent sucrose gradient analysis of the phenol or SDS-extracted RNA revealed that the reduction in radioactive rRNA was nearly complete for the 28 S rRNA. In contrast, considerable residual uridine incorporation was found in the 18 S rRNA species. Corollary data suggesting a net increase in mRNA synthesis and a net decrease in protein synthesis are reported.


Synthesis of ppGpp and chloroplast ribosomal RNA in Chlamydomonas reinhardi. A material identified as guanosine 5',3'-bis-diphosphate (ppGpp) has been detected in extracts of Chlamydomonas reinhardi ac-20 cells grown under mixotrophic conditions or in arg-2 cells deprived of arginine. The material was acid and base labile, susceptible to alkaline phosphatase, resistant to periodate oxidation, had spectral characteristics of a guanine derivative and comigrated on chromatograms with ppGpp from Escherichia coli. In ac-20 ppGpp may be involved in the control of chloroplast ribosomal RNA synthesis. When ac-20 cells were shifted from mixotrophic to autotrophic conditions, the 32Pi labeling of ppGpp, relative to that of GTP, was reduced, while the specific labeling of chloroplast ribosomal RNA was enhanced. Addition of low concentrations of cycloheximide had somewhat similar effects.


Fractionation of aurintricarboxylic acid and effects of its components on nuclear swelling and nucleic acid synthesis. Crude aurintricarboxylic acid synthesized by the conventional method was fractionated into 8 components (A1-A8) by silica gel thin layer chromatography. Some of the components were isolated and their effects were tested on nuclear swelling and on in vitro RNA synthesis and DNA synthesis using isolated polymerases. Only two components with low RF (A7 and A8) induced strong nuclear swelling. The other components were almost inactive. Formaurin-dicarboxylic acid, a contaminating polyanion of crude aurintricarboxylic acid, was synthesized separately. It was as effective as the two active components in nuclear swelling and inhibited RNA and DNA polymerases on naked DNA template. However, it stimulated RNA synthesis on chromatin template probably by dissociating histones from DNA. Unfractionated aurintricarboxylic acid showed the same effects at higher concentrations. A preliminary analysis indicated one of the inactive components (A4) is genuine aurintricarboxylic acid. The results suggest that the observed activity of crude aurintricarboxylic acid is due to some contaminating substances, one of which is formaurindicarboxylic acid.


Bromodeoxyuridine induction of deoxycytidine deaminase activity in a hamster cell line. The Syrian hamster cell line, RPMI 3460, was found to express barely detectable levels of the enzyme deoxycytidine deaminase. In contrast, the cell lines B4 and HAB, which are derived from 3460 cells and have approx. 60 and 100% bromodeoxyuridine substitution in DNA, respectively, show an approx. 50-fold higher enzyme activity. Deoxycytidine deaminase activity can be "induced" in 3460 cells by growth in 10(-5) M bromodeoxyuridine, as well as by the other halogenated pyrimidines, iododeoxyuridine and chlorodeoxy-uridine. The time required for maximal enzyme activity to accrue (approx. 8 days) suggests that new genetic expression is required for enhanced deoxycytidine deaminase activity and inhibition of induction in the presence of Ara. C shows that bromodeoxyuridine must be incorporated into DNA. In addition, the extent of enhanced deoxycytidine deaminase activity is directly related to the level of bromodeoxyuridine substitution in DNA. Another hamster cell line, BHK21/C13, which shows no detectable deoxycytidine deaminase activity, cannot be induced by bromodeoxyuridine. These results are discussed with respect to a mechanism by which bromodeoxyuridine may alter gene expression due to an altered binding of both positive and negative regulatory proteins to DNA.


Effects of inhibitors of S-adenosylmethionine decarboxylase and ornithine decarboxylase on DNA synthesis in rat liver after partial hepatectomy. The effects of inhibitors of polyamine synthesis on DNA synthesis in rat liver regenerating after partial hepatectomy were studied. Neither 1,1'-[(methylethanediylidene)-dinitrilo]-bis-(3-aminoguanidine), a potent irreversible inhibitor of S-adenosylmethionine decarboxylase, nor 1,3-diaminopropane, an indirect inhibitor of ornithine decarboxylase, strongly inhibited [3H]thymidine incorporation into DNA when given as a single injection 1 h after operation and 23 h before DNA synthesis was measured. However, when the two inhibitors were given together, DNA synthesis was completely prevented. The incorporation of [14C]leucine into protein was not affected by this treatment. Combined administration of the inhibitors partially prevented the rise in hepatic spermidine levels normally seen during liver regeneration, but neither drug was effective alone. Hepatic putrescine content measured 12 h after treatment was increased by the combined inhibitors whereas, spermine levels were not significantly changed. By 24 h after operation the effects of the combined inhibitors on spermidine levels had almost worn off but DNA synthesis was greatly inhibited. However, by 40 h after operation the inhibitor treatment had no effect on [3H]thymidine incorporation into DNA. Also treatment with the combined inhibitors abolished DNA synthesis at 24 h after partial hepatectomy only when given more than 6 h before measurement suggesting that the effect was indirect and required time to become apparent. These results are consistent with other recent studies in which prior accumulation of spermidine appeared to be required for normal DNA replication and cell division.


Molecular size and fidelity of DNA polymerase alpha from the regenerating liver of the rat. DNA-dependent DNA polymerase has been extracted from the soluble cytoplasmic fraction of regenerating rat liver and purified using phosphocellulose and DEAE-cellulose chromatography. Glycerol gradient analysis showed that the enzyme was predominantly DNA polymerase alpha, having a sedimentation coefficient of 10.5 S at low ionic strength and of 6--8 S at higher salt concentrations. The fidelity of purified enzyme was assessed using the co-polymer poly(dA-dT).poly(dA-dT) as a template for DNA synthesis. For both the aggregated (10.5 S) and disaggregated (6--8 S) forms, fidelities in the range of 1 wrong base in 100,000--150,000 complementary bases were obtained.


Large-scale purification and subunit structure of DNA-dependent RNA polymerase II from cauliflower inflorescence. DNA-dependent RNA polymerase II (nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) from cauliflower inflorescence (Brassica oleracae, var. botrytis) was highly purified by polyethyleneimine treatment on a large scale. The solubilized enzyme was partially purified by polyethyleneimine fractionation and subjected to chromatography on DEAE-Sephadex and phosphocellulose, and subsequently to sedimentation in a glycerol gradient. The specific activity (231 nmol/mg per 10 min) of this enzyme was comparable to that reported for other purified eukaryotic RNA polymerases. Analysis of the purified RNA polymerase II by polyacrylamide gel electrophoresis under nondenaturing conditions revealed a single band. The subunit composition of the enzyme was analyzed by electrophoresis under denaturing conditions. The RNA polymerase II contained subunits with molecular weights and molar ratios (in parentheses) of 180 000(1), 130 000(2), 48 000(2), 25 000(4), and 19 500(4).


DNA-dependent RNA polymerase III from cauliflower. Characterization and template specificity. Class III DNA-dependent RNA polymerase (EC 2.7.7.6) was highly purified from cauliflower (Brassica oleracea, var. bortytis) by using polyethyleneimine precipitation. The specific activity of the enzyme was comparable to that reported for mammalian enzymes. Glycerol gradient sedimentation analysis indicated that the sedimantation coefficient (23 S) was slightly higher than that of enzyme II from cauliflower. The class III enzyme was inhibited by alpha-amanitin at high concentrations (50% inhibition at 200 microgram/ml). The Km value for nucleoside triphosphate was determined. Template specificities for single synthetic polymers showed that the enzyme read pyrimidine homopolymers as templates and preferred poly(dT) to poly(dC). The enzyme transcribed both strands of homopolymer pairs of poly(dI). poly(dC) and poly(dA).poly(dT). The synthetic polyribonucleotides were not effectively read. Competition experiments with these synthetic polymers indicated that the enzyme had different binding specificities which were not the same as their template specificities. The different binding affinities and template specificites for synthetic templates of the three classes of enzyme suggest that the enzyme can discriminate among different template sequences.


Histone H5 in nucleated erythrocytes of fish as determined by radioimmunoassay. Tissue-specific histone H5 in the nucleated erythrocytes of dogfish, scup, skate, tautog, sea robin and toad fish was studied. The presence of this histone was inferred by its electrophoretic mocility on polyacrylamide gels containing either acid-urea or sodium dodecyl sulfate. By radioimmunoprecipitation assays, cross reaction was observed between fish histones and an anti-H5chicken antibody. The antibody was specific to chicken histone H5; purified chicken histone H1 and calf thymus total histones did not cross react. It is concluded that fish histone H5 shares common antigenic determinants with the chicken H5 histone.


Dose response, wavelength dependence and rate of excision of ultraviolet radiation-induced pyrimidine dimers in mouse skin DNA. The yield of thymine-containing dimers produced in mouse skin DNA in vivo by 290 nm ultraviolet radiation was shown to increase with dose up to around 2000 J/m2 and subsequently at a much slower rate up to 8000 J/m2. The study of wavelength dependence of dimer formation in skin indicated that 290 nm was the most effective wavelength of those investigated, followed by 300, 280 and 260 nm, with 310 nm being by far the least effective. A reduction in the number of dimers present in skin DNA was shown to occur by 24 h post-irradiation in a dose-dependent manner. A significant percentage of the dimers was, however, found to persist in the skin until at least 72 h post-irradiation.


Molecular orbital studies on the structure of nucleoside analogs. I. Conformation of 8-azapurine nucleosides. PCILO (perturbative configuration interaction using localized orbitals) computations have been carried out for the conformational properties of 8-azapurine nucleosides. The results indicate an anti conformation for Xcn and a gg conformation for phiC(4')-C(5') for C(2')-endo 8-aza analogs compared to the syn and gg conformation for the corresponding purine nucleosides. For C(3')-endo sugar puckering, both molecules prefer the syn conformation due to intramolecular hydrogen bonding between O(5')-H of the sugar and N(3) of the base, the preference being more profound in 8-aza analogs. The crystallographic conformation 8-azaadenosine has been attributed to crystal forces. The available NMR data on 8-azapurine nucleosides are in agreement with the PCILO predictions.


Differential inhibitory effect of 5-fluorouridine on RNA labelling in dipteran salivary gland cells. In Chironomus salivary glands, 5-fluorouridine inhibits labelling of ribosomal RNA. The drug does not prevent labelling of other types of RNA- like heterodisperse messenger-like RNA, 75-S RNA of Balbiani ring origin and 4-S RNA, which furthermore are exported to cytoplasm. The potential use of this drug for the study of RNA metabolism in insect salivary glands can be compared with the use of low doses of actinomycin D in cultured mammalian cells.


Interaction of normal and unusually modified microbial DNA with cultured mammalian cells. Breakdown and reincorporation vs. uptake of polymerized DNA. The uptake of radioactively labeled bacterial and phage DNA and the incorporation of acid-soluble DNAase I digests of these DNAs by cultures of human foreskin and 3T3 cells were studied. The presence of large amounts of unusually modified pyrimidine residues in donor phage DNAs allowed radioactive donor DNA in the nuclei of DNA-treated cells to be distinguished from host DNA labeled with breakdown products derived from donor DNA. This distinction could be made because it was found that radioactively labeled 5-methylcytosine residues in predigested XP-12 DNA and glucosylated 5-hydroxymethylcytosine residues in predigested T4 DNA could not be incorporated in an unaltered form into animal cell DNA. The results obtained from the study of uptake of these DNAs suggest that approx. 4--40 ng of phage DNA per 10(6) cells was transported to the nuclei of DEAE-dextran-pretreated cells during 3 days of incubation in medium after treatment with the DNA. However, interpretation of the results is complicated by the finding of considerable amounts of donor DNA binding to and persisting at the cell surface, which might attach to nuclei during subcellular fractionation.


Characterization of unstable poly (A)-RNA in Caulobacter crescentus. A significant amount of poly(A)-RNA in Caulobacter crescentus is located on polysomes and the size distribution of this polysomal poly(A)-RNA is small compared to the total pulse-labeled RNA in these cells. These observations suggest that the poly (A)-RNA represents a subset of small messenger RNA molecules. Poly (A)-RNA, and presumably the poly (A) portion of these molecules is extremely unstable: as assayed by binding to oligo (dT)-cellulose the poly (A)-RNA turns over with a chemical half-life of 15--20 s compared to a half-life of approx. 2 min for total cellular messenger RNA. The presence of adenosine in hydrolysates of poly(A) tracts showed that these sequences are located at the 3'-OH end of the RNA. The ratios of AMP/adenosine in the samples confirmed that the length of the A-tracts is approx. 13-17 nucleotides (Ohta, N., Sanders, M. and Newton, A. (1975) Proc. Natl. Acad. Sci. U.S.A. 72, 2343--2346)


The relationship between hnRNA+-poly(A) and mRNA+-poly (A) in non-dividing human lymphocytes. Evidence for distinct synthetic pathways for mRNA precursor- and nonprecursor-hnRNA. The processing of hnRNA+-poly(A) to mRNA+-poly(A) has been studied in resting lymphocytes from human peripheral blood. In pulse-chase experiments, two types of hnRNA+-poly(A) have been distinguished: the first is labeled predominantly with exogenous radioactive precursors supplied during the pulse, and the second incorporates primarily scavenged labeled precursors made available during a chase incubation. When the disappearance of both types of hnRNA+-poly(A) was quantitatively compared with the appearance of stable and labile mRNA+-poly(A), only 10% of the anticipated cytoplasmic material was actually obtained. Statistically, 90% of the poly(A)-bearing hnRNA molecules processed were degraded. The two types of hnRNA+-poly(A) were found to be functionally different. Pulse-labeled material was processed to poly(A)-bearing mRNA; "chase-labeled" molecules did not leave the nucleus and never served as precursors for cytoplasmic mRNA. The data fit a model in which there are distinct pathways for mRNA precursor- and nonprecursor-hnRNA+-poly(A).


The putative 15 S precursor of globin mRNA contains a poly (A) sequence. [3H] Uridine or [3H] adenosine pulse-labelled nuclear RNA was isolated from chicken immature red blood cells and separated on denaturing formamide sucrose gradients. RNA of each gradient fraction was hybridized with unlabelled globin DNA complementary to mRNA (cDNA) and subsequently digested by RNAase A and RNAase T1. The experiments revealed two RNA species with globin coding sequences sedimenting 9 S and approx. 15 S, the latter probably representing a precursor of 9 S globin mRNA. A poly (A) sequence was demonstrated in this RNA by two different approaches. Nuclear RNA pulse-labelled with [3H] uridine was fractionated by chromatography on poly (U)-Sepharose. Part of the 15 S precursor was found in the poly(A)-containing RNA. In the second approach 15 S RNA pulse-labelled with [3H]adenosine was hybridized with globin cDNA, incubated with RNAase A and RNAase T1 and subjected to chromatography on hydroxyapatite. The hybrids were isolated and after separation of the strands degraded with DNAase I, RNAase A and RNAase T1. By this procedure poly(A) sequences of approximately 100 nucleotides could be isolated from the 15 S RNA with globin coding sequences. The poly(A) sequence was completely degraded by RNAase T2.


Comparison of the structural characteristics of the 4-aminobutyrate:2-oxoglutarate transaminases from rat and human brain, and of their affinities for certain inhibitors. 4-Aminobutyrate:2-oxoglutarate (4-aminobutyrate:2-oxoglutarate amino-transferase, EC 2.6.1.19) from human brain has been purified 2500-fold with respect to the initial homogenate. The enzyme, which appears to be pure by polyacrylamide gel electrophoresis, N-terminal analysis and immunodiffusion, was compared to rat brain 4-aminobutyrate transaminase, purified to the same extent in an earlier study [15]. The two enzymes, which have approximately the same molecular weight, show large differences in their tryptic fingerprints and in the peptides produced by cyanogen bromide cleavage. The Km values (limit) for 4-aminobutyrate are different, the human enzyme having four times greater affinity for this substrate. A series of branched-chain fatty acids (including n-dipropylacetate), which are structural analogues of 4-aminobutyrate and inhibit rat brain 4-aminobutyrate transaminase, are less powerful inhibitors of the human enzyme.


The binding of lithium and of anionic metabolites to phosphoglucomutase. Intercept inhibition of rabbit-muscle phosphoglucomutase (alpha-D-glucose-1,6-bisphosphate: alpha-D-glucose-1-phosphate phosphotransferase, EC 2.7.5.1) produced by several nucleotide diphosphates and compounds related to coenzyme A was re-examined in order to re-evaluate an earlier suggestion that this enzyme has an allosteric regulatory site. However, in all cases intercept inhibition constants were much larger than those previously reported, and in all but two cases were too large to assess in the assay system, i.e., were greater than 10 mM. Most of the intercept inhibition previously observed apparently was caused by the use of the Li+ salts of inhibitors. Thus, Li+ binds competitively with the natural activator, Mg2+, and in the presence of glucose phosphates binds almost as well as Mg2+: Kd approximately 10 micrometer. The observation that glucose phosphates bind to the Li+ complex of phosphoglucomutase some 900 times more tenaciously than to the corresponding Mg2+ complex could provide a partial rationale for the lack of reactivity of the Le+ form of the enzyme. Attempts to verify the dimeric structure of phosphoglucomutase that was previously reported also produced negative results.


Acid glycohydrolase in Chinese hamster with spontaneous diabetes II. N-acetyl-beta-D-hexosaminidase in plasma and tissues. Excessively high activity of N-acetyl-beta-D-hexosaminidase (2-acetamido-2-deoxy-beta-D-glucoside acetamidodeoxy-glucohydrolase, EC 3.2.1.30) was found in the plasma of hereditary diabetic XA line animals, which however showed similar activity of this enzyme in both 12 00 X g supernatant and precipitate fractions of kidney homogenates as the nondiabetic M line animals. 0.1% Triton X-100 extracts of kidney, spleen, hind leg muscle, cheek pouch and spinal cord of XA and M line animals also showed similar activities of this enzyme whereas the XA animal liver extracts showed significantly higher activity than the M extracts. On a Sepharose CL-6B column, plasma N-acetyl-beta-D-hexosaminidase was eluted as two major peaks at 0 and 0.05 M NaCl (isozyme B1 and B2). Both isozymes showed pH optima between 3.5 and 4.0 and the same Michaelis constants for p-nitrophenyl-N-acetyl-beta-D-glucosaminide at pH 4.5, i.e. 0.18 mM. Isozymes from XA and M animals showed identical properties. N-acetyl-beta-D-hexosaminidase in the liver extracts was separated into 3 isozymes, A, B1 and B2, by successive column chromatography runs on Sepharose 6B and DEAE-Sepharose CL-6B. At 49 degrees C, isozyme B1 showed thermostability whereas B2 and A lost 20% and 76% of their activities after 30 min incubation, pH optima for A, B1 and B2 were 4.0--4.5, 3.5 and 3.5--4.0 respectively. The Km values for p-nitrophenyl-N-acetyl-beta-D-glucosaminide were 0.48 mM for A and 0.19 for B1 and B2. The XA animal liver extracts showed higher activities in all three isozymes than the M animal livers. Identical results, however, were obtained for liver isozymes from M and XA animals with regard to thermostability, pH vs. activity, elution profile on ion exchange column and affinity to p-nitrophenyl-N-acetyl-beta-D-glucosaminide.


Properties of a beta-D-mannosidase from Aspergillus niger. The beta-D-mannosidase (beta-D-mannoside mannohydrolase, EC 3.2.1.25) from culture filtrate of Aspergillus niger has been purified in large amounts by fractionation with (NH4)2SO4 and DEAE-cellulose chromatography. The removal of traces of alpha-D-galactosidase was performed on a Sepharose-epsilon-aminocaproyl-galactosylamine column. The final enzyme preparation (specific activity 188 units) has no other glycosidase activity and is judged homogeneous. The enzyme has a molecular weight of 130 000 +/- 5000 and an isoelectric point of 4.7. The amino acid composition of the enzyme is characterized by high proportion of acidic amino acids and no cysteine residues and a single chain structure of the enzyme is suggested. The enzyme shows maximum activity on p-nitrophenyl-beta-D-mannopyrano-side at pH 3.5 and at 55 degrees C. The presence of 80% of beta-sheet structure in the protein and 20.8% of monosaccharides (Gal : 1.3; Man : 7; GlcNAc : 1) could explain this relative high heat stability (up to 2 h at 55 degrees C). Enzyme activity is inhibited by mannose (Ki = 7.85 mM) and the specificity is examined.


Purification and properties of alpha-l-arabinofuranosidase from plant Scopolia japonica calluses. alpha-L-Arabinofuranosidase (alpha-L-arabinofuranoside arabinofuranohydrolase, EC 3.2.1.55) from the culture medium of Scopolia japonica calluses was partially purified. Various properties of the enzyme were studied and the effects of lactones on the activity were determined.


Isorenin, pseudorenin, cathepsin D and renin. A comparative enzymatic study of angiotensin-forming enzymes. 1. Renin was purified 30 000-fold from rat kidneys by chromatography on DEAE-cellulose and SP-Sephadex, and by affinity chromatography on pepstatinyl-Sepharose. 2. The enzymatic properties of isorenin from rat brain, pseudorenin from hog spleen, cathepsin D from bovine spleen, and renin from rat kidneys were compared: Isorenin, pseudorenin and cathepsin D generate angiotensin from tetradecapeptide renin substrate with pH optima around 4.9, renin at 6.0. With sheep angiotensinogen as substrate, isorenin,  pseudorenin and cathepsin D have similar pH profiles (pH optima at 3.9 and 5.5), in contrast to renin (pH optimum at 6.8). 3. The angiotensin-formation from tetradecapeptide by isorenin, pseudorenin and cathepsin D was inhibited by albumin, alpha-and beta-globulins. These 3 enzymes have acid protease activity at pH 3.2 with hemoglobin as the substrate. Renin is not inhibited by proteins and has no acid protease activity. 4. Renin generates angiotensin I from various angiotensinogens at least 100 000 times faster than isorenin, pseudorenin or cathepsin D, and 3000 000 times faster than isorenin when compared at pH 7.2 with rat angiotensinogen as substrate. 5. The 3 'non-renin' enzymes exhibit a high sensitivity to inhibition by pepstatin (Ki less than 5.10(-10) M), in contrast to renin (Ki approximately 6-10(-7) M), at pH 5.5. 6. It is concluded from the data that isorenin from rat brain and pseudorenin from hog spleen are closely related to, or identical with cathepsin D.


Ethane and ethylene formation by mitochondria as indication of aerobic lipid degradation in response to wounding of plant tissue. During aerobic incubation of potato slices or potato mitochondria at acidic pH, ethane and ethylene in a ratio of approx. 50 : 1 are generated from an endogenous substrate. Both ethane and ethylene production are stimulated by the addition of alpha-linolenic acid. Ethane formation from linolenic acid is a radical mechanism dependent on oxygen and is not significantly influenced by mitochondrial electron transport. Ethane production may represent a sensitive marker for membrane damage.


The effect in vitro of ionizing irradiation and small rises in temperature on the uptake and release of labelled lipids by the human erythrocyte membrane. 1. The effect of X-irradiation (50 000 rad) and an increase in temperature from 37 to 42 degrees C on the synthesis, uptake and release of labelled lipids by erythrocytes was studied in plasma incubations in vitro. 2. Both irradiation and a rise in temperature resulted in an inhanced synthesis of [32P]phosphatidic acid in the erythrocytes. 3. The uptake by the erythrocytes of 14C- and 3H-labelled cholesterol, [14C, 32P]phosphatidylethanolamine and [14C, 32P]phosphatidylcholine from plasma lipoproteins was increased by a rise in temperature but not by irradiation. These labelled lipids were apparently taken up in the ratio in which they were found in plasma. They were not released from the erythrocytes in the same manner.


A new chromatographic approach to the resolution of individual gangliosides. Ganglioside mapping. 1. Anion-exchange column chromatographies on DEAE-Sephadex, DEAE-Sepharose and QAE-Sephadex were tested for fractionation of ganglioside-molecular species. DEAE-Sepharose gave the best resolution, with good separation of mono-, di-, tri- and even tetrasialogangliosides. Even minor gangliosides could be resolved and detected by silica gel thin-layer chromatography of successive fractions of effluent from a DEAE-Sepharose column. In this two-step chromatographic system, the first step of elution from the column depends on differences in anionic charge and the second step of development on a silica gel plate depends on differences in polarity. With this ganglioside-mapping technique, at least 25 unidentified gangliosides were separated from bovine and human brains in addition to the well-known compounds, G7, GM3, GM2, GM1, GM1 (GlycNeu), GD2, GD3, GD1a, GD1a-GAN, GD1a(AcNeu, GlycNeu), GD1b, GT1a, GT1b and GQ. 2. The procedure was used to compare the gangliosides in human (3, 5 and 35 years old), bovine, cat, rat, rabbit, chicken and dog brains. The ganglioside profiles of human, cat, rat, rabbit and dog brains only differed in minor components. However, the gangliosides in chicken brain were unexpectedly complex, at least 30 minor gangliosides, including 15 monosialogangliosides being recognized. Gangliosides containing N-glycolylneuraminic acid (GDIa and GM1 type) were only found in bovine brain. The concentrations of tri- and tetrasialogangliosides in human brain were found to increase during maturation. 3. The long chain bases of each ganglioside fraction, in which the content of sialic acid was confirmed by measuring the ratio of sialic acid to stearic acid, were also analyzed as their aldehydes. The ratios of C-20 to C-18 sphingosine increased in the series from the mono- to tetrasialoganglioside fraction (0.216-1.777) in all animal brains tested.


An anti-A1 lectin in the seeds of Amphicarpaea bracteata. An anti-A1 lectin has been isolated from the extract of Amphicarpaea bracteata seeds by affinity chromatography on Epoxy-activated Sepharose 6B coupled to N-acetyl-D-galactosamine. The yield of the purified lectin was 86 microgram/g of seeds. The purified lectin shows one main band on electrophoresis in sodium dodecyl sulfate-polyacrylamide. The amino acid and neutral sugar composition indicate that this lectin is an acidic glycoprotein with a neutral sugar content of approx. 2%. The composition of the lectin is different from that of the Dolichos biflorus lectin but the two lectins have some common characteristics. The most powerful inhibitors of the agglutination of A1 red blood cells by the A. bracteata lectin is N-acetyl-D-galactosamine. Much weaker inhibitors of the agglutination are alpha-lactose, D-fucose, and five other sugars.


Isolation and chemical characterization of two distinct "link proteins" from bovine nasal cartilage proteoglycan complex. Bovine nasal cartilage proteoglycan aggregates have been dissociated and separated by dissociative density gradient centrifugation into proteoglycan sub-units and "link fraction". The latter contained mainly the two "link proteins" as shown by analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The two "link proteins" were then separated by preparative gel electrophoresis under dissociative conditions. Molecular weight and amino acid composition of both proteins are presented.


The isolation of "link proteins" from bovine nasal cartilage. Methods of isolating the water insoluble 'link proteins' from preparations of bovine nasal cartilage proteoglycan aggregates have been investigated. Upon chromatography on Sepharose 4B in 0.1% sodium dodecyl sulfate, the 'link proteins' are found in a discrete, included peak, whereas the bulk of the proteoglycan emerges at the void volume. Some low molecular weight proteoglycan is associated with the 'link proteins'. An alternative procedure, i.e. chromatography on DEAE-cellulose in sodium dodecyl sulfate, has also been examined. In 0.1% sodium dodecyl sulfate, proteoglycan monomer is not absorbed by the column, whereas 'link proteins' are. Subsequently, the 'link proteins' with a minor fraction of the proteoglycan are eluted with 1.0% sodium dodecyl sulfate. Both procedures serve to separate the 'link proteins' from the bulk of proteoglycan present in an aggregate preparation, but additional steps are necessary to achieve homogeneity. Thus a 'link protein' preparation, fractionated from proteoglycan aggregate by equilibrium density gradient centrifugation under dissociative conditions, can be finally purified by chromatography on DEAE-cellulose in sodium dodecyl sulfate.


Proteolytic modification of pig and rat liver pyruvate kinase type L including phosphorylatable site. The phosphorylated or phosphate-accepting site of pyruvate kinase from pig and rat liver was removed without inactivation by incubation with subtilisin. At different time intervals the subtilisin was inactivated with phenylmethylsulfonyl fluoride and the amount of remaining phosphorylatable or phosphorylated sites of pyruvate kinase estimated by incubation with an excess of [32P]-ATP and protein kinase. It was found that to get the same rate of modification the subtilisin concentration required to modify unphosphorylated pyruvate kinase was approximately ten times higher than that used for removal of the phosphorylated site of phosphorylated site of phosphorylated enzyme. It was shown that the proteolytically-modified pyruvate kinase had an increased apparent Km for phosphoenolpyruvate without a change in V, when compared to unmodified unphosphorylated and phosphorylated pyruvate kinase. The removal of the phosphorylated site was not associated with loss of the allosteric sites for ATP and Fru-1,6-P2. The possibility that phosphorylation of the pyruvate kinase increases its degradation rate in vivo is briefly discussed.


Molecular weights of wheat gamma2-, beta6-, alpha7-, alpha8- and alpha9-gliadins. The molecular weights of wheat gamma2-, beta6-, alpha7-, alpha8- and alpha9-gliadins were calculated with the aid of a computer technique from sedimentation equilibrium data obtained in an ultracentrifuge equipped with photoelectric scanner. The dissociative solvents, all at pH 3.1 by addition of HCl, included 3 M urea, 0.15 M KCl; 8 M urea, 0.15 M KCl and 6 M guanidine-HCl. The minimum molecular weights for gamma2-, alpha7- and alpha9-gliadins, obtained in 6 M guanidine-HCl, were 34 600, 30 400 and 30 900, respectively. The beta6- and alpha8-gliadins gave minimum molecular weights of 33 000 and 36 900, respectively, in 3 M urea, 0.15 M KCl.


Aggregates of human erythrocyte membrane sialoglycoproteins in the presence of deoxycholate and dodecyl sulfate. Gel electrophoresis in the presence of deoxycholate of human erythrocyte membranes solubilized with deoxycholate resolves four glycoprotein zones. Electrophoresis in dodecyl sulfate in a second dimension reveals several components, three of which migrate in the region of PAS-2. One of the zones in deoxycholate gel electrophoresis contains component PAS-3, and this glycoprotein seems to exist as a monomer in deoxycholate, but aggregates partially upon addition of dodecyl sulfate. The major sialoglycoprotein migrates as a diffuse zone in dodecyl sulfate. The major sialoglycoprotein migrates as a diffuse zone in deoxycholate gel electrophoresis, indicating association and dissociation during the electrophoresis. The use of deoxycholate followed by dodecyl sulfate in two-dimentional electrophoresis gave high resolution of membrane proteins and can be used for detection of complexes in one of the detergents.


Binding of calcium by parvalbumin fragments. Parvalbumin fragments from carp pI 4.47 parvalbumin corresponding to its residues 1--75 and 76--108 bind Ca2+ with affinities corresponding to Kd 0.9 . 10(-4) M and Kd 3 . 10(-3) M, respectively.


A methodological problem in thermoanalytic studies arising from admixed fluoride in the samples. At a temperature just above the collagen denaturation point the aluminium walls of the pans used in microcalorimetric measurements react exothermically with NaF in test solutions. This formation of the fluor-aluminium complex cryolite is assumed to be catalyzed by the denaturing collagen.


The effect of ammonium nitrate on the synthesis of nitrogenase and the concentration of leghemoglobin in pea root nodules induced by Rhizobium leguminosarum. The effects of NH4NO3 on the development of root nodules of Pisum sativum after infection with Rhizobium leguminosarum (strain PRE) and on the nitrogenase activity of the bacteroids in the nodule tissue were studied. The addition of NH4NO3 decreased the nitrogenase activity measured on intact nodules. This reduction of nitrogen fixation did not result from a reduced number of bacteroids or a decreased amount of bacteroid proteins per gram of nodule. The synthesis of nitrogenase, measured as the relative amount of incorporation of [35S]sulfate into the components I and II of nitrogenase was similarly not affected. The addition of NH4NO3 decreased the amount of leghemoglobin in the nodules and there was a quantitative correlation between the leghemoglobin content and the nitrogen-fixing capacity of the nodules. The conclusion is that the decrease of nitrogen-fixing capacity is caused by a decrease of the leghemoglobin content of the root nodules and not by repression of the nitrogenase synthesis.


The effect of phenobarbitone on cytoplasmic NADP-linked dehydrogenase activities in rat liver. Phenobarbitone administered in drinking water (0.5 g/l) or by daily intraperitoneal injection (100 mg/kg body weight) consistently caused an elevation of hepatic NADP-linked malic enzyme in rats maintained on a pellet diet. Three to four days appeared to be required for maximum response. The effect was also observed in animals maintained on a protein rich diet, in which the basic hepatic malic enzyme activity was low, but not in animals maintained on a sucrose rich diet, in which the basic enzyme activity was almost twice normal. Methyl cholanthrene, administered by daily intraperitoneal injection (40 mg/kg body weight), resulted in elevated hepatic levels not only of malic enzyme but also of the pentose phosphate pathway dehydrogenases. The timing of the "starve-refeed" response of the hepatic NADP-linked dehydrogenases in phenobarbitone-treated rats was similar to that in controls, and similar maximum enzyme activities were reached. The role of cytoplasmic NADP-linked dehydrogenases in the provision of reducing equivalent is discussed, particularly in relation to hepatic microsomal drug metabolism.


Emission Mössbauer studies of some organocobalamins. Subtle changes in the Mössbauer parameters are observed while going from methyl- to ethyl- to adenosylcobalamin, and also when the "base" is detached from the cobalt. The observation of these changes demonstrates that the Co-C bond, among others, remains intact after the Auger event, accompanying the electron-capture decay of the cobalt-57. The differences between ethylcobalamin and the other two organocobalamins in the magnitude of the quadrupole splittings have been interpreted on the basis of the sigma-donating tendency of the organic moiety and the Co-C bond length. The latter is presumably determined by the steric hindrance offered to the group in approaching the cobalt atom. The ethyl- and adenosylcobalamins in their "base-off" form exhibit a larger quadrupole splitting than the corresponding "base-on" form. In the "bas-off" form, the cobalt atom is perhaps raised above the mean plane of the four equatorial nitrogen atoms of the corrin ring, which may result in the diminution of the delocalization of the 3dpi electron density. The higher population of dpi orbitals and the enhanced metallic character of the dz2, resulting from shrinkage of the Co-C bond length, enhances the magnitude of the quadrupole splitting.


Enzymatic sulfation of steroids. II. The sulfation of corticosterone by the glucocorticoid sulfotransferases of rat liver cytosol. A radioisotopic assay for the cytoplasmic corticosterone sulfotransferase activity of rat liver was developed. The steroid inhibits the enzyme reaction. For reliable results, a complex assay method, using three different corticosterone concentrations, each studied with several different amounts of enzyme, was necessary. This "mosaic" assay compensates for observed biological, gonadal and seasonal enzyme fluctuations. Cytosols from female rats contain 6--9-times the enzyme activity found in males. The sulfation product with both sexes is corticosterone-21-sulfate. The effects of castration and of androgen administration on hepatic cortisol and corticosterone sulfation were compared in female rats. Ovariectomy resulted in 20--32% and 25--35% decreases of hepatic corticosterone and cortisol sulfotransferase activity, respectively. Androgen administration caused 37--55% and 40--60% decreases of sulfation of the two steroids. The data suggest the equivalence of hepatic cortisol and corticosterone sulfotransferases. Fractionation of cytosols from female rats, on DEAE-Sephadex A-50 columns, resolved three peaks of corticosterone sulfotransferase activity which eluted concurrently with the hepatic cortisol sulfotransferases I, II and III. They appear to be the same enzymes. Cytosol from males contained cortisosterone sulfotransferase activity due mostly to sulfotransferase III. Sulfotransferases I and II appear to have higher turnover numbers for hepatic cortisol than for corticosterone. The reverse is true for sulfotransferase III.


Cell disease: desialylation of beta-hexosaminidase and its effect on uptake by fibroblasts. The pinocytosis by fibroblasts of beta-hexosaminidase (EC 3.2.1.30) excreted by cultured skin fibroblasts from a patient with I-cell disease was not enhanced by neuraminidase treatment of the enzyme. The uptake of sialic acid-rich normal plasma beta-hexosaminidase was minimal and neuraminidase treatment did not appreciably enhance uptake. In contrast, sialic acid-rich normal seminal fluid beta-hexosaminidase was readily pinocytosed regardless of neuraminidase treatment. Thus the presence of sialic acid on beta-hexosaminidase does not influence uptake and a neuraminidase deficiency in I-cell disease may not be directly responsible for excessive extracellular enzyme.


Interrelation of aerobic glycolysis and lipogenesis in isolated perfused liver of well-fed rats. The rates of glycolysis and lipogenesis in isolated perfused liver of well-fed rats were studied. When liver was allowed to synthesize [14C]glycogen prior to perfusion, no more than 9% of the degraded [14C]glycogen was recovered in lactate and 6% in lipid. Addition of glucose, fructose and sorbitol enhanced concomitantly the formation of lactate and pyruvate and the rate of release of triglyceride and free fatty acid. Glucose was less efficient than fructose or sorbitol. The incorporation of 14C from these 14C-labelled substrates into lactate, pyruvate and lipids confirmed their role as carbon sources. Incorporation of 14C into the glycerol moiety of neutral lipid exceeded that found in the fatty acids, suggesting that these substrates contributed largely to the esterification of fatty acids. The total rate of de novo fatty acid synthesis was correlated with the formation of lactate and pyruvate. It is concluded that increased rates of aerobic glycolysis are related to increased rates of lipogenesis.


The adenylate cyclase system of planaria Polycelis tenuis: activation by serotonin and guanine nucleotides. The particulate fraction prepared after homogenization of planaria Polycelis tenuis in a buffer containing 3 mM EDTA and 15 mM 2-mercaptoethanol possesses an adenylate cyclase activity which was enhanced two-fold by serotonin and 20-fold by the nucleotide analog guanosine 5'-(beta-gamma-imino)triphosphate, Gpp(NH)p; when present together, the two activators exhibited a marked synergistic effect. The effect of serotonin was dose dependent, with a KA of 2 micrometer and a Hill coefficient of 0.4. In the presence of 10 micrometer Gpp(NH)p, these values became 45 nM and 1.5, respectively. The effect of serotonin was due to an increase in the maximal velocity of the enzyme and was specifically inhibited by methiotepin. The effect of methiotepin was half-maximal at 0.2 micrometer in the absence of Gpp(NH)p and at 5.0 micrometer in its presence. Planaria thus appear to be the lowest organisms in which guanine nucleotides are active upon adenylate cyclase. As serotonin is normally present in planaria, it is postulated that a serotonin-dependent regulation of adenylate cyclase activity plays a physiological role in this species.


Effects of a polyoxyethylene detergent (Brij 58) on platelet aggregation, release and clotting activity. Low concentrations of a polyoxyethylene detergent, Brij 58, inhibited the secondary phase of platelet aggregation induced by ADP in human citrated platelet-rich plasma but had no effect on primary aggregation. Thrombin-induced aggregation of washed human platelets suspended in Tyrode's buffer was inhibited after incubation of cells with 4.10(-6) M detergent. Efflux of [14C]serotonin, 45Ca2+ and labile aorta contracting substance (thromboxane A2) and development of prothrombin-converting activity (platelet factor 3) were abolished concomitantly. Aggregation of washed platelets either by sodium arachidonate or by collagen was also inhibited by the same concentration of Brij 58 which inhibited thrombin aggregation. This concentration did not itself produce any release of a cytoplasmic marker, lactate dehydrogenase, from platelets. Higher concentrations of Brij 58, exceeding 4.10(-5) M, lysed the cells liberating lactate dehydrogenase, serotonin and Ca2+. When albumin was included as a platelet stabilizer in the suspending medium the concentration of detergent required for the inhibitory effects was increased ten-fold. This could be attributed to competitive binding of the detergent to albumin, demonstrated with [14C]acetylated Brij 58. A variety of other polyoxyethylene detergents, at concentrations from 8.10(-4) to 5.10(-3) M, also inhibited platelet aggregation induced by thrombin. It is concluded that low concentrations of Brij 58 stabilize the platelets against the action of aggregating agents, while higher concentrations produce membrane destabilization and cell lysis.


[Dark transition of 1st level reduction products of porphyrins to 2d level]. Transition of the products of the first restoration of porphyrines to the second one in the darkness and practically in the absence of a reducer and oxygen has been studied. Its cause is the disproportioning of dyhydroform into tetrahydro- and the initial pigment. Regeneration of porphyrines under these conditions is also shown. The rate of both reactions significantly increases in the light.


[Kinetic model of a single muscle contraction]. A model of single muscle contraction is proposed. The kinetics of two reactions have been used: interaction between the excitation mediator and the muscle membrane receptor, and the reaction of enzymatic mediator splitting. The muscle contraction parameters have been correlated with the concentration of the mediator-receptor complex.


[Quantum-mechanical model of the genesis of the early receptor potential in the retina]. An account is given of a possible mechanism of the initiation of the retina early receptor potential due to quantization of the electron energetic spectrum in its fine biolayers. A connection is established between the fine structure of lamellar systems of biological origin and function. The mechanism suggested can be used when studying photoelectric processes in chlorophyll layers, in beta-carotin films for the search of means to increase the photopotential value in the films of photosynthetic pigments, when explaining the receptors adaptation property.


[Propagation of perturbations in active media with autooscillatory properties]. Propagation of an activity impulse in the medium where autooscillatory relaxation biochemical reaction takes place is examined theoretically and with the help of the computer experiment. Propagation velocities of the front and shape of the impulse are found. The results are compared with axiomatic theory data on spontaneous activity perturbated media.


[Selective activation of fast and slow electrogenesis of cardiac membranes in the excitable period of the cardiac cycle]. Two types of electrogenic alterations of cardiac cell membranes during vulnerability are shown. The first one: selective activation only of slow response close to the source of miocardium stimulation and a simultaneous appearance of an extrasystolic impulse of great latency (100 ms and more) far from the source of stimulation. Extrasystolic latency is considered as the time of slow response propagation. Slow propagation is due to the small size of reentry during vulnerability. The second one: selective activation only of fast Na channels leads to generation of a very short response (10-20 ms duration)--cardiac spike. Appearance of such a spike is also favourable for reentry during vulnerability.


[Effect of the curvature of the dorsal hippocampus on the shape of the electric field which it generates]. A mathematical model is presented of spatial configuration of the field generated by hippocampus surface. Calculations have shown that the potential induced on the cortex surface is almost 5-fold smaller than that under the hippocampus surface. Theoretical results well agree with the experimental data and allow a conclusion to be inferred that passive penetration of fields generated by subcortical structures does not exceed 20% of the total level of spontaneous EEG.


[Quaternary structure of butyrylcholinesterase from horse blood serum]. The molecular weight of butyrylcholinesterase from horse blood serum was determined using the method of analytical ultracentrifugation. The molecular weight of the enzyme subunit was also determined by its mobility in gel containing sodium dodecyl sulphate. The number of active sites was determined by titrating butyrylcholinesterase with a specific inhibitor, namely di-isopropyl phosphorofluoridate. A conclusion is made that butyrylcholinesterase is a tetramer with one active site per unit.


[Participation of singlet and triplet states of tryptophanyls in protein inactivation]. Using energy transfer from 4-aminobenzene-sulfonamide triplet level to tryptophanyl one of the main energy contributions to trypsin and chymotrypsin inactivation at the temperature of liquid N2 has been shown to make singlet excited state of their tryptophanyls.


[Origin of the electroencephalogram].

[Origin of the electroencephalogram]. An attempt is made to estimate the contribution of the activity of each cortical layer of rabbit's brain and subcortical structure in EEG formation. It has been shown that the main generator layers of spontaneous EEG are the I layer--the layer of apical dendrites and the V--the layer of pyramidal neuron somas. To perform a quantitative estimation of passive penetration of the electric fields from deep-seated structures, chiefly from the hippocampus, biopotentials studied were recorded from I, V cortex layers and hippocampus simultaneously, the cortex being disengaged by the method of propagating depression. Significant suppression of surface-activity of the motor cortex was achieved with its preservation in the hippocampus.


[Spectral characteristics of immobilized peroxidase]. Visible spectra of peroxidase adsorbed on insoluble supports were studied. It has been shown that immobilization leads to essential conformational changes of the enzyme which affect its bond with imidasol.


[Adjustment of the cat visual cortex receptive fields to certain spatial frequency bands within definite ranges of movement frequencies of stimulating gratings]. Reactions of simple and composite receptive fields (RF) of the 17th and 18th fields of cat's visual cortex to grating with various spatial frequency moving through the field with different velocities were studied. It has been shown that for each RF the velocity range is revealed within which spatial-frequency selectivity of RF exists. The narrow-band tuning for high frequency RF exists in the range of low velocities, and for low frequency ones in the range of higher velocities.


[Application of a graphic method for analyzing the kinetics of multienzyme reactions]. A new graphic method is proposed to solve kinetic equations for polyenzymic reactions. Each graph apex is corresponded by the transmitting function deduced from kinetic equations by means of Laplas transformation. Application of this procedure allows to simplify the solution of kinetic equations and its analysis. The procedure suggested makes it possible to use the methods of automatic control when solving theoretical problems of enzymology.


[Denaturation temperature of collagen as a linear function of its amino acid composition]. TD of collagen have been constructed as linear function of fractions of each from 18 amino acids using experimental data for 29 proteins. For coefficient estimation the least squire method have been used. As a result the TD of any collagen may be calculated by using of the function obtained. 14 amino acids strongly influence on TD. The suggestion arises that poly(Gly-Ile-Lys) may produce collagen like structure.


[Movement of dissolved oxygen in a constant magnetic field]. Some regularities of the movement of liquid-solved oxygen in the constant magnetic field have been studied. Redistribution and specific dynamics of the changes of pO2 in a vessel with a physiological solution under the effect of the magnetic field are shown. The data obtained make us to believe that when interpreting the results of biological experiments applying magnets movement of oxygen in the magnetic field should be taken into account as well as possible change of metabolic processes in this case.


[Photochemical and photoelectronic properties of components of the photosynthetic apparatus. III. Heat-stimulated conductivity in the lamellar system chlorophyll a--n-chloranil]. Traps with the bedding depth of 0.1 ev in the layers of chlorophyll alpha treated with the electron acceptor n-chloranil were discovered by the method of thermostimulating conductivity. It is suggested that the traps catch mobile vacancies, which occur due to the decomposition of the excited states of the pigment on electron-acceptor centres of the dissociation.


[Stabilization of bilayer lipid membranes with polymeric resins]. Bilayer lipid membranes on the bases of monoglyceride of oleic acid with the inclusion of polysterene are described. The inclusion of the polymer allows to obtain high stable membranes without sufficient changes in the nature of their conductivity.


[Effect of chlorpromazine on the relation between temperature and the binding of ANS by human erythrocytes]. It has been shown that under the effect of chlorpromazin (concentration to 2.10(-5) M) breaks observed on Arrhenius graphs of the rate constant of ANS binding with erythrocytes (k) at 28 and 36 degrees C are shifted to the region of low temperatures approximately to a similar interval (14-15 degrees C). The value k measured at 22 degrees C is not changed within the pH range of 5-7. It is concluded that breaks characterised the initial and final stages of temperature transition initiated in the zone of acid phospholipid of membranes.


[Cooperative nature of the denaturation process in tissues and cell nuclei]. The heat absorbance curves for liver tissue, liver tissue of embrione and for splin of white rats reveal two peaks corresponding to melting processes of protein and DNA. There are three peaks for cell nucleous: the first (corresponding lower temperature) responsible for membrane melting process. Two other peaks coinside with nucleoprotein denaturation process.


[Electric conductivity of tissue measured in the physiological potential gradient]. An ability to measure the electric conductivity of tissue in physiological potential gradient is proposed. It has been shown that the resistance of plant leaf during the development of the injured potential is constant.


[Picosecond flash photolysis of Halobacterium halobium bacteriorhodopsin at room and low temperatures]. The primary stage of photoexcitation of bacteriorhodopsin from Halobacterium halobium upon the action of ultrashort (tau equal to 25 ps) laser impulse of 530 nm wavelength and of energy 2.5.10(-3) J has been studied. The primary photoproduct with a maximum of 630 nm is shown to occur in the differential spectrum in a time less than 25 ps both at room temperature (+20 degrees C) and at a low temperature (-150 degrees C).


[Composition and structure of phospholipids in hydrocarbon yeasts. I. Phosphatidylcholine and phosphatidylethanolamine]. From fooder yeast grown on purified liquid n-parafins two fractions--phosphatydylcholin and phosphatydyletanolamin--have been isolated. Structure, composition and properties of these compounds have been studied. It is established that double bonds in fatty acids ends are located near their polar group.


[Role of non-histone proteins in chromatin solubility]. Treatment of chromatin gel with low ionic strength solution of tRNA has produced the dioxyribonucleoprotein (dnptRNA) in which only part of non-histone proteins was removed without loss of any major histone fraction. The solubility of DNP in the presence of 0.15 M NaCl and 1 to 5 mM MgCl2 was considerably higher than that of initial untreated chromatin. It has been assumed that the solubility of chromatin depended primarily on some non-histone proteins and not on H1 histone.


[Photochemiluminescence of tryptophan-containing peptides and proteins during photooxidation. VII. Mechanism of sensitization by eosin of the photochemiluminescence of human serum albumin solutions]. Dependence of photochemoluminescence (PCL) intensity of serum albumin solutions with eosine initiated under light illumination with the wave length of 546 nm on oxygen concentration in solution has been studied. The luminescence intensity was shown to increase gradually with the increase of oxygen concentration. The relationship obtained is described by an empyric formula. Experimental data on the change of PCL intensity dependent on protein and dye concentrations have been obtained. A three-dimensional diagram of such relationship is plotted: this surface has a complex profile. An uncomplicated relationship between PCL intensity and protein concentration was analysed on the strength of the data obtained. The experimental, rated and literature data allowed a quantitative description of sensitized PCL of serum albumin solutions with eosine.


[Possible use of ultraviolet fluorescence for detecting protein conformational transitions in intact tissues]. Spectra of fluorescence (SF) excited by the light with the wave length of 260 nm were measured with the aim of revealing the character of structural reconstructions in water-dificient plant cells. Hypocotyls of Cucumis sativus L. seedlings during soil draught served as the objects of study. With an increase of water deficiency the SF maximum is shifted to 5-8 nm to the long wave region. In wilted plant tissues free triptophane was accumulated whose SF maximum was shifted to the long wave region as compared to that of triptophane bound in proteins. The data obtained point to a limited usage of UV-fluorescence in detecting structural transitions of proteins in intact cells and tissues. In cases when the content of free triptophane is changed it is difficult to make certain conclusions about the character of conformational changes of protein macromolecules.


[Contribution of individual structural components to stabilization of the secondary structure of double-helical polynucleotides]. Contributions of individual structural components of the double-helical polynucleotide to the stabilization of its secondary structure have been studied. The energy of intramolecular interactions was calculated by the method of atom-atom potentials. Sections of the energy function were constructed according to the parameters determining mutual location of base pairs with the optimal conformation of ribose-phosphate backbone in the points close to A- and B-forms of DNA. A complicated nature of the contribution of different structural components of the polynucleotide to its stabilization was revealed by means of different parameters. Relationships between the change of conformational parameters corresponding to optimal values of energy for A- and B-families of nucleic acids were found.


[Ion permeability of bilayer membranes formed from synthetic phospholipids in the phase transition region]. Ion permeability of black lipid membranes formed from synthetic phospholipids has been studied. The resistance of BLM formed from phosphatidylcholine, tiophosphatidylcholine, threealkylphosphate and threealkyltiophosphate was 10(7)--10(8) Ohm.cm2. It was shown that the membrane potential of the 10--30 mV arised in KCl gradient indicating the preference cation conductance in synthetic lipid membranes. A sharp decrease of the membrane conductance near to the phase transition temperature was discovered. The change of conductance by phase transition temperature was sensitive to chemical nature of the polar head of phospholipids used.


[Intermediate reduction products of porphyrins and their structure]. It was shown on the basis of NMR-H spectra that the addition of proton to dianions of Zn-octaethylchlorine, Zn-octaethylporphin, and Zn-monoasaethioporphyrin results in the formation of products with the structure of alpha-dihydroflorine, alpha-florine, and gamma-florine respectively. This indicates that electron density greatly increases when the anions are formed in the centres which had a decreased density in the initial molecule.


[Interaction of a spin-labeled analog of ATP with myosin]. By means of spin labeled analogs of ATP we have shown that conformational changes in myosin molecule induced by variation of temperature take place in the region of the active centre. In case of Mg-ATP and unmodified myosin conformation of the active centre changes monotonously with the change in temperature but after the modification of S1 thiol groups by N-ethylmaleimide on the temperature dependence curve of rotational mobility of the spin label a discontinuous is observed at 14-16 degrees C. It is also observed in case of K+-EDTA-ATP, or Ca2+-ATP and unmodified myosin. It is shown that the chemical analogs of Mg2+-paramagnetic ion Mn2+ are directly connected with the myosin active centre in the presence of ATP(ADP), i. e. a triple complex enzyme-bivalent cation-substrate is formed.


[Calculation of parameters of ion transfer through bilayer membranes on the basis of steady-state measurement data]. The multi-site model of ion transfer through thin membranes is used to derive the steady-state current-voltage characteristics. It is shown that experimental data are in a good agreement with the theory predictions.


Substitution reactions of a water-soluble metalloporphyrin with azide and 1,1,3,3-tetramethyl-2-thiourea. The substitution reactions of tetrakis-(4-N-methylpyridyl)porphinecobalt (III) (CoIIITMpyP) with azide and with 1,1,3,3-tetramethyl-2-thiourea (TMTU) have been studied as a function of pH at 25 degrees and an ionic strength of 0.5 M. The mechanistic pathway proposed for thiocyanate [1] and pyridine [2] is applicable to these ligands as well once allowance is made for two attacking forms of azide, N3- and HN3. A TMTU axial substituent has about the same influence on the rate of further ligand substitution as does SCN- and a much larger influence than does azide. Similar behavior between bound SCN- and bound TMTU is also shown in electron-transfer reactions with Ru(NH3)62+. Whereas both sulfur-containing ligands enhance the rate relative to the diaquo complex, the azide complex undergoes reduction an order of magnitude more slowly than does the diaquo complex.


MCD spectra of iron-sulfur complexes with or without inorganic sulfur. The magnetic circular dichroism spectra were observed for various iron-sulfur complexes with and without inorganic sulfur as models for rubredoxin and 2-Fe ferredoxin. The MCD band shapes ascribed the bands around 390 and 490 nm to Faraday A terms for mononuclear iron sulfur complexes. These bands are probably assigned to the charge-transfer transitions from the thiol sulfur orbital to the iron t2 and e 3d-orbitals, respectively. The MCD magnitudes decreased by more than one-half for binuclear iron-sulfur complexes with inorganic sulfur in comparison with those for the mononuclear complexes. The low MCD magnitude as well as the possible core symmetry as low as D2d attributed the MCD bands to Faraday B terms. Incorporation of inorganic sulfur produced new MCD bands, some of which can be assigned to the charge-transfer transitions from the inorganic sulfur orbital to the iron t2 and e 3d-orbitals. Among complexes studied here, the bis(o-xylyldithiolato) ferrate(III) monoanion gave the MCD spectrum which resembles that of a rubredoxin. This implies that the MCD spectroscopy also assessed complex as a good rubredoxin model. However the binuclear complex bis[o-xylyldithiolato-micron2-sulfidoferrate(III)] dianion failed to offer the MCD spectrum similar to that of the spinach ferredoxin.


[Interaction of paramagnetic probes with submitochondrial particle membranes]. Redox-transformations of some paramagnetic probes differing in chemical structure have been studied during their interaction with the membranes of submitochondrial particles. Dependence of the probe restoration kinetics on the conditions of functioning of the electron-transport chain, presence of oxygen, respiration substrate, electron transport inhibitors (antimycin, alpha tenoiltriftoracetate, potassium cyanide) in the reaction mixture is shown. Water-soluble as well as strongly hydrophobic spin probes are restored when the respiration substrate (succinate) is present; they interact with the lipid-soluble carrier ubiquinone.


Lanthanide octaethylprophyrins: preparation, association, and interaction with axial ligands. The synthesis of virtually all the lanthanide octaethylporphyrin complexes have been achieved by heating appropriate anhydrous lanthanide halide and octaethylporphyrin in imidazole melt at 210 degrees C for two hours. The lighter lanthanide porphyrin complexes are very susceptible to hydrolysis, the middle lanthanide porphyrin complexes are moderately stable, and the heavier lanthanide porphyrin complexes are relatively more stable to hydrolysis. Two out of four lanthanide porphyrin complexes studied in detail, namely ytterbium and lutetium octaethylporphyrins, aggregate in benzene and the Soret bands in their absorption spectra are about 6 nm shifted to higher energies upon a hundred-fold increase in their concentrations. The aggregations of these lanthanide porphyrin complexes in non-coordinating solvents have been further verified by 1H NMR spectral studies. This spectral behavior can be interpreted qualitatively in terms of the model of the molecular exciton interactions with stacking of at least two prophyrins. A dimeric structure of these lanthanide porphyrin complexes has been proposed on the basis of geometrical considerations. On the contrary, the europium and gadolinium octaethylporphyrins associate very weakly in benzene in the concentration range studied. All four lanthanide porphyrin complexes interact with pyridine and piperidine, and the Soret bands in their absorption spectra are about 8 nm shifted to low energies as compared with their values in pure benzene.


[Thermal transitions in erythrocyte membranes revealed by their permeability to ANS]. A number of breaks were recorded on the curve of Arrhenius relationship of the rate constant of the dye 1-anilino-8-naphthalenesulphonate sodium salt (ANS) input into human erythrocytes of 20, 28, 36, 42 and 46 degrees C. Variations in the values of activation energies within the temperature range of 28-36 degrees and 42-46 degrees C obtained in various blood samples allow to consider these temperatures as those at which structural changes of the membranes take place. The values of activation energy of the process for temperature "conformers" of the erythrocyte membrane are 12(10-20 degrees C), 26.5 (20-28 degrees C), 34.2(36-42 degrees C) and 47 kcal/mol (t is greater than 46 degrees C). Within the temperature range of 28-36 degrees and 42-46 degrees C an irreversible decrease of permeability to ANS of the erythrocyte ghost after their incubation for 10 min at increased temperatures were observed. Thus the temperature regions of the change in erythrocyte permeability correspond to those at which the resealing of ghost takes place. The break in Arrhenius graph at 20 degrees C seems to characterize a highly cooperative "point" transition. The lipid nature of the initiator of structural transition within 28-36 degrees C is proved by a sharp increase of the permeability of liposomes prepared from erythrocyte membrane lipids to ANS at 28 degrees C. The nature of the initiators of two other thermal transitions is discussed.


[Dipole cooperative model of ion channels in excitable biomembranes. I. Gating currents]. The experimental data on gating currents are analyzed in terms of dipole cooperative model of ionic channel. The dependence of displaced charge on potential is calculated. A comparison between the theory and experiment allows to establish the cooperative part of the charged gating subunits energy. It is shown that kinetics of gating particles displacement is nearly exponential. The theoretical dependence of characteristic times on the potential during testing pulse of voltage agrees qualitatively with the experimental curves.


Interaction of the anti-tumour drug cis-Pt(en)Cl2 with Cytidilyl-3'--5'-guanosine. Chemical shifts occurring in 1H nuclear magnetic resonance and Sephadex G 25 chromatography are used to follow the reaction of the dinucleotide C3'p5'G (cytidilyl-3'phosphate5'-guanosine) in aqueous solution and to separate its products. Binding of the platinum occurs in first place at the cytosine, then at the guanine base. Two major complexes are produced, Pt:CpG and (Pt:CpG)2, which account for respectively for an internal and an external cross-linking effect.


[Mechanism of oxidative phosphorylation. V. Conditions contributing ot control of brain mitochondria respiration by adenosine diphosphate]. By changing the quantities of rat brain homogenate or mitochondria, adding succinic acid and ADP a situation was created in polarographic cell leading to the initiation of Lardy and Chance control upon respiration. It is concluded that this control is characteristic of in vitro conditions, but not only of functional properties of mitochondrial preparations. For studying oxidative phosphorylation the disappearance of nonorganic phosphate must be estimated.


[Conformational fluctuations in chloroplasts]. Characteristic features of conformational changes in chloroplasts have been studied by microfilm technique at BMI-13 installation. The oscillatory character of changes in chloroplast cross-section area in time, determined earlier by the tele-lazer microscopy technique, has been confirmed by our data. When chloroplasts are simultaneously seen in the microscope it is possible to observe their synchronous oscillation. The stationarity check of conformational changes shows that is a non-stationary process in the general case. So it is concluded that the standard technique of analysing accidental non-stationary processes can be used for study of conformational changes in chloroplasts provided that the corresponding parts of their kinetics are selected or trends are eliminated depending on the type of non-stationarity.


[Spectral transformations of photoproducts in Halobacterium halobium cells]. Long-living spectral products observed earlier in purple membranes (PM) are discovered in the cells of Halobacterium halobium (Hh) under the action of yellow light. Blue spectral region of the light has been found to be optimal for PM arising in the cells cultivated under light. On the basis of the changes discovered in the optical spectra of Hh we may suppose that PM arising in the cells, is the dark processes, occur due to a hypothetic enzyme, which appears under light. The rate of PM arising is maximal during the periods of thermogenesis maximum.


Effects of certain antitumor platinum compounds on kidney esterases. In a study of the biochemical mechanism of renal toxicity of certain antitumor platinum compounds, particularly cis-dichlorodiammineplatinum(II) (NSC-119875), qualitative and quantitative studies of the soluble nonspecific kidney esterases were carried out using (C57BL/L X DBA/2) mice. There was a major suppression of the testosterone-dependent esterases of treated male mice; these levels dropped to levels below those found in untreated females within 72 h after certain of the drugs were administered. This effect appeared to be in inverse relationship to the numerical value of the LD50 values of the compounds investigated.


[On the localization of ribosomal proteinase]. The localization of proteolytic activity in rat liver ribosomes was studied. 40S and 60S subparticles were obtained upon dissociation of polyribosomes by puromycin. It was shown that the proteolytic activity is associated with small subparticles or with proteins removed from the latter.