Yeasts

J Vasc Surg, 1986 Apr, 3(4), 649 - 51
Successful management of Histoplasma capsulatum infection of an abdominal aortic aneurysm; Harris RL et al.; A 65-year-old woman with disseminated histoplasmosis underwent resection of an atherosclerotic abdominal aortic aneurysm . Yeast forms of Histoplasma capsulatum were present in the aneurysm . Surgical resection and revascularization with a Dacron graft followed by systemic amphotericin B therapy and chronic ketoconazole suppressive therapy have resulted in a patient without symptoms 15 months postoperatively . It is important to be aware of the potential for artherosclerotic aortic aneurysm involvement by H . capsulatum.

Appl Environ Microbiol, 1986 Apr, 51(4), 821 - 4
Effect of culture conditions on tremorgen production by some Penicillium species; di Menna ME et al.; Four strains each of seven tremorgenic Penicillium species were grown under various conditions and tested for tremorgen production by intraperitoneal injection of mice and by chemical analysis . Half of the strains had previously been found to be tremorgenic on bioassay after growth on Czapek Dox yeast extract broth or potato-milk-sucrose broth for 3 weeks at 26 degrees C . In the tests reported here nearly all previously nontremorgenic strains were either tremorgenic to mice or produced tremorgens detectable by chemical analysis but did so after longer incubation periods than used in the original screening . Bioassay was not suitable for the estimation of absolute levels but was preferable to chemical analysis when the identity of the tremorgens was not known . Species and strains within species gave different responses to changes in culture medium, incubation temperature, light irradiation, and shaking . Overall, tremorgen production was maximal at 20 or 26 degrees C, increased with time, and was reduced in shaken culture.

Mol Immunol, 1986 Apr, 23(4), 433 - 40
Role of the N-terminal regions of hog C3a, C5a and C5a-desArg in their biological activities; Damerau B et al.; The N-terminal regions of the complement peptides C3a, C5a and C5a-desArg (purified from yeast-activated hog serum) were gradually shortened by incubation with leucine amino peptidase . This treatment led to the following changes in the biological activities of these peptides: the potencies of C5a and C5a-desArg in aggregation of human polymorphonuclear leukocytes and of guinea-pig platelets, and their ability to deactivate these cells were gradually diminished; the chemotactic effect of C5a-desArg on human leukocytes was similarly lowered, while the chemotactic potency of C5a was even increased up to the loss of the first 12 N-terminal amino acids . However, after removal of the whole N-terminal region (i.e . 20 amino acids distal of the first disulfide bridge) the potency of both peptides was decreased to a few percent . In contrast, C3a totally lost its platelet-aggregating as well as deactivating activity already after cleavage of 10-15 N-terminal amino acids by LAP . On leukocytes, on the other hand, C3a retained some activity even after the loss of the whole N-terminal region . These results indicate that the N-terminal regions play an important role for biological activities of the three complement peptides, possibly by stabilizing the optimal conformation of their C-terminal regions which contain the receptor-activating domains.

J Antimicrob Chemother, 1986 Apr, 17 Suppl B, 53 - 7
Effects of pefloxacin on phagocytosis function of rat macrophages and polymorphonuclear leucocytes; Desnottes JF et al.; The influence of pefloxacin on polymorphonuclear leucocytes (PMN) and alveolar macrophages (AM) phagocytosis and microbicidal activity was studied . Phagocytes were collected from Sprague-Dawley rats, mixed with pefloxacin at different concentrations (1, 10, 100 mg/l) and a yeast phagocytosis assay was performed . Three parameters were measured, the phagocytic capacity (number of live and dead yeast/100 cells), the phagocytic activity (number of cells containing 2 or more yeast/100 cells) and the microbicidal activity or % killed (Formula: see text) Both low and high concentrations of pefloxacin increased significantly the phagocytic capacity and activity of AM and PMN . The increase of the AM phagocytic capacity and activity seen with increasing levels of extracellular pefloxacin was not statistically significant . Pefloxacin showed no adverse effect on the microbicidal activity of AM and PMN.

Proc Natl Acad Sci U S A, 1986 Apr, 83(8), 2501 - 5
Identification of the macrophage mannose receptor as a 175-kDa membrane protein; Wileman TE et al.; Mannose-lactoperoxidase, a neoglycoprotein prepared by reaction of lactoperoxidase with cyanomethyl 1-thiomannoside, bound to alveolar macrophages at 4 degrees C (Kd = 5.8 X 10(-8) M) and was rapidly internalized at 37 degrees C (K uptake = 2 X 10(-8) M) . Mannose-lactoperoxidase binding and uptake were blocked by yeast mannan, and mannose-lactoperoxidase inhibited uptake of 125I-labeled mannose-BSA (bovine serum albumin) . Radioiodination of cells with surface-bound mannose-lactoperoxidase was carried out in the presence of glucose and glucose oxidase . A major polypeptide (175 kDa) was radioiodinated by this procedure . Iodination of the 175-kDa polypeptide appeared to be receptor-mediated, since it was blocked by the presence of yeast mannan . Specific iodination was absent from receptor-negative cells . To demonstrate that the 175-kDa species is a ligand-binding protein, cells were iodinated by the standard lactoperoxidase method . Washed cells were then allowed to bind mannose-BSA . Receptor-ligand complexes, prepared by detergent extraction, were passed over anti-BSA IgG affinity columns . Mannose, but not mannose 6-phosphate or galactose, eluted a radioactive protein from the column that migrated with an apparent molecular mass of 175 kDa on NaDodSO4/PAGE . Detergent extracts of crude membranes prepared from macrophage-enriched whole rabbit lung were adsorbed to mannose-Sepharose; the fraction obtained by elution with mannose contained two protein components of 175 and 55 kDa . Subsequent chromatography on N-acetylglucosamine-agarose yielded a single protein of 175 kDa . The 175-kDa polypeptide was shown to bind 125I-labeled mannose-BSA in a precipitation assay . This binding could be blocked with mannan or mannose-BSA . The results indicate that the cell-surface mannose receptor is a 175-kDa protein.

Proc Natl Acad Sci U S A, 1986 Apr, 83(7), 2007 - 11
Molecular cloning of cDNA for the nuclear ribonucleoprotein particle C proteins: a conserved gene family; Nakagawa TY et al.; The C proteins, C1 and C2, are major constituents of the heterogeneous nuclear RNA (hnRNA) ribonucleoprotein (hnRNP) complex in vertebrates . C1 and C2 are antigenically related phosphoproteins that are in contact with hnRNA in intact cells and bind to RNA tightly in vitro . A cDNA clone for the C proteins was isolated by immunological screening of a human lambda gt11 expression vector cDNA library with monoclonal antibodies . The lacZ-cDNA fusion protein is recognized by two different anti-C protein monoclonal antibodies . HeLa cell mRNA that was hybrid-selected with the cDNA clone (1.1 kilobases long) was translated in vitro and yielded both the C1 and C2 proteins (41 and 43 kDa, respectively) . RNA blot analysis showed strong hybridization to two polyadenylylated transcripts, of about 1.4 kb and 1.9 kb, in human cells . Genomic DNA blot analysis showed multiple hybridizing restriction fragments in human and mouse, and homologous DNA sequences are found across eukaryotes from human to yeast . These findings suggest that the sequences encoding the hnRNP C proteins are members of a conserved gene family and they open the way for detailed molecular and genetic studies of these proteins.

Physiologie, 1986 Apr-Jun, 23(2), 139 - 43
Symmetrical-asymmetrical codons and hydrophobic-hydrophilic amino acids; Portelli C et al.; The symmetrical codons present in the first and third positions two purine (P) or two pyrimidine (p) bases (e.g . PNP or pNp), while the asymmetrical codons present in the first and third positions a purine and a pyrimidine base (e.g . PNp or pNP) . The percentages of symmetrical and asymmetrical codons specifying hydrophobic and hydrophilic amino acids are similar in the genes of viruses and vertebrates, which utilise the "universal" genetic code, but they are different for the genes of human mitochondria and yeast mitochondria, which utilize two specific genetic codes.

J Biomol Struct Dyn, 1986 Apr, 3(5), 843 - 57
On loop folding in nucleic acid hairpin-type structures; Haasnoot CA et al.; In a series of studies, combining NMR, optical melting and T-jump experiments, it was found that DNA hairpins display a maximum stability when the loop part of the molecule comprises four or five nucleotide residues . This is in contrast with the current notion based on RNA hairpin studies, from which it had been established that a maximum hairpin stability is obtained for six or seven residues in the loop . Here we present a structural model to rationalize these observations . This model is based on the notion that to a major extent base stacking interactions determine the stability of nucleic acid conformations . The model predicts that loop folding in RNA is characterized by an extension of the base stacking at the 5'-side of the double helix by five or six bases; the remaining gap can then easily be closed by two nucleotides . Conversely, loop folding in DNA is characterized by extending base stacking at the 3'-side of the double helical stem by two or three residues; again bridging of the remaining gap can then be achieved by one or two nucleotides . As an example of loop folding in RNA the anticodon loop of yeast tRNAPhe is discussed . For the DNA hairpin formed by d(ATCCTAT4TAGGAT) it is shown that the loop structure obtained from molecular mechanics calculations obeys the above worded loop folding principles.

J Biol Chem, 1986 Mar 25, 261(9), 4126 - 33
Affinity labeling of nucleotide-binding sites on kinases and dehydrogenases by pyridoxal 5'-diphospho-5'-adenosine; Tamura JK et al.; A new adenine nucleotide analog, {3H}pyridoxal 5'-diphospho-5'-adenosine (PLP-AMP), has been synthesized . The effectiveness of PLP-AMP as an affinity probe has been tested using a number of nucleotide-binding enzymes . In comparison to reaction with pyridoxal 5'-phosphate, PLP-AMP binds more tightly and exhibits greater specificity of labeling for most enzymes tested . PLP-AMP is a very potent inhibitor of yeast alcohol dehydrogenase and rabbit muscle pyruvate kinase, with complete inhibition obtained upon incorporation of 1 mol of reagent/mol of catalytic subunit . The reagent is also a potent inhibitor of yeast hexokinase and phosphoglycerate kinase . When modified in the absence of substrates, these enzymes require 2 mol of reagent/mol of active site for complete inhibition . However, when modified in the presence of sugar substrates, this stoichiometry decreases to 1.1 for the hexokinase-glucose complex and 1.4 for the phosphoglycerate kinase . 3-phosphoglycerate complex . The most potent inhibition by PLP-AMP was observed with rabbit muscle adenylate kinase . Half-maximal inhibition was obtained at a concentration of approximately 1 microM . In contrast to these examples, PLP-AMP, as well as pyridoxal 5'-phosphate, fails to act as a potent or specific inhibitor of beef heart mitochondrial F1-ATP-ase . The high specificity of labeling and the ability of nucleotide substrates to decrease the rate of inactivation of the kinases and dehydrogenase are consistent with the modification of active site residues . The complete reversibility of both modification and inactivation in the absence of reduction by NaBH4 and the absorption spectra of modified enzymes prior to and following reduction indicate reaction with lysyl residues . We conclude that PLP-AMP holds considerable promise as an affinity label for exploring the structure and mechanism of nucleotide-binding enzymes.

Biochem J, 1986 Mar 15, 234(3), 717 - 26
Analysis of Hill slopes predicted by the four-ligand exponential model for a regulatory enzyme; Ainsworth S; The four-ligand exponential model for a regulatory enzyme is described as it is applied to kinetic studies of yeast pyruvate kinase in which the concentrations of four ligands are systematically varied . The Hill slopes predicted by this model are calculated for the two situations in which the fourth ligand is either a substrate or an effector . It is shown that the individual terms that make up the expression for the Hill slope assist the interpretation of the observed behaviour in terms of the constants employed by the model.

Nucleic Acids Res, 1986 Mar 11, 14(5), 2035 - 44
Conservation of the cellular gene encoding the scrapie prion protein; Westaway D et al.; The major protein, PrP 27-30, in purified preparations of hamster scrapie prions is encoded within the genome of the experimental host . DNA sequences related to a PrP cDNA clone can be detected in a wide variety of organisms under relatively stringent conditions where the only signal generated by hamster or mouse DNA corresponds to the PrP gene . Three hosts for scrapie, goat, sheep and rat gave strong hybridization signals . In addition, three invertebrate DNAs reacted with the PrP probe, in the order nematode-Drosophila much greater than yeast . Thus, the sequences detected in goat, sheep, rat, nematode, Drosophila and possibly yeast DNA may arise from authentic PrP genes . This evolutionary conservation is consistent with the notion that PrP proteins participate in essential cellular processes.

Biochim Biophys Acta, 1986 Mar 7, 870(1), 160 - 8
Inhibition of mammalian glyoxalase I (lactoylglutathione lyase) by N-acylated S-blocked glutathione derivatives as a probe for the role of the N-site of glutathione in glyoxalase I mechanism; Al-Timari A et al.; A series of twelve S-blocked and N,S-blocked glutathione derivatives has been studied as inhibitors of glyoxalase I {R)-S-lactoylglutathione methylglyoxal-lyase (isomerising), EC 4.4.1.5) from human erythrocytes . A number of new N,S-blocked glutathiones have been synthesised . Inhibition at pH 7.0, 25 degrees C was linear-competitive in all cases and the Ki values were interpreted in terms of the absence of a specific binding interaction for the N-site of the inhibitor and the absence of coupling between binding processes at N- and S-sites (the regions around the NH2 and HS groups, respectively, of GSH analogues bound to enzyme) . These observations are in strong contrast to previous results with the yeast enzyme . Some Ki values were measured for yeast glyoxalase I . A special binding interaction of the phenyl groups with enzyme from both species was found for glutathione derivatives with N-acyl groups of structure -NH X CO X X X Y X Ph but not for -NH X COPh, where X and Y were variously -CH2-, -NH- and -O- . Studies were made of the range of stability of human erythrocyte glyoxalase I to pH . The pH profiles for the Ki values of S-p-bromobenzyl)glutathione and N-acetyl-S-(p-bromobenzyl)glutathione indicated no pH dependence for the latter and little, if any, for the former inhibitor . The mean Ki over the pH range 5-8.5 for S-(p-bromobenzyl)glutathione was 1.21 +/- 0.37 microM and for N-acetyl-S-(p-bromobenzyl)glutathione in the same pH range, Ki decreased from 1.45 +/- 0.26 microM to 0.88 +/- 0.11 M.

FEBS Lett, 1986 Mar 3, 197(1-2), 134 - 8
Adenylate cyclase activity of NIH 3T3 cells morphologically transformed by ras genes; Levitzki A et al.; The observed homology between G-proteins which regulate adenylate cyclase and ras proteins and the suggested role of ras in the regulation of adenylate cyclase in yeast prompted us to examine the regulation of adenylate cyclase in three cell lines: (i) NIH 3T3 cells, (ii) NIH 3T3 cells transformed by high levels of the normal rasH gene product and (iii) NIH 3T3 cells transformed by a mutated rasH gene product . We found that the regulation of adenylate cyclase by G-proteins is identical in the three cell lines, although the response of the transformed NIH 3T3 cells to agonists is strongly attenuated . Our data suggest that mammalian ras products do not interact directly with adenylate cyclase, although their increased expression may indirectly inhibit the interaction of adenylate cyclase stimulatory receptors with G-proteins.

Acta Physiol Scand, 1986 Mar, 126(3), 413 - 7
Some observations made by intracellular recordings in a primary astrocyte culture and a glioblastoma '138 MG'; Nilsson A et al.; Intracellular recordings were performed in a primary astrocyte culture from rat brain and in a human glioblastoma cell line, 138 MG . The technique proved insufficient to verify the heterogeneous composition of the primary astrocyte culture, since this study shows most cells present in the culture to have similar resting membrane potential, membrane impedance, membrane potential/impedance relationship and K+-sensitivity . With the exception of macrophages, identified by their response to externally applied yeast particles, the results do not allow the identification of different cells that are known to be present . The membrane potential of the primary astrocyte was -68 +/- 14 mV, and the membrane potential of the 138 MG cells -37 +/- 15 mV . The membrane potential of cells in the primary culture have a K+-sensitivity resembling that of astrocytes in situ, whereas the K+-sensitivity of 138 MG more resemble that of a dedifferentiated cell . A reduction of 0.95 mm Ca2+ to o mM depolarizes the astrocyte with 9.6 mV and hyperpolarizes the glioma cell 2.6 mV.

Br J Nutr, 1986 Mar, 55(2), 219 - 25
Nutritional availability to rats of selenium in four seafoods: crab (Callinectes sapidus), oyster (Crassostrea virginica), shrimp (Penaeus duorarum) and Baltic herring (Clupea harengus); Mutanen M et al.; 1 . The present study was conducted to determine the biological availability to rats of the selenium in four high-Se seafoods: crab (Callinectes sapidus), oyster (Crassostrea virginica), shrimp (Penaeus duorarum) and Baltic herring (Clupea harengus) . 2 . Weanling male rats were fed on a Se-deficient Torula yeast diet for 4 weeks followed by either continued depletion or repletion for 4 weeks with 0.05, 0.1 or 0.2 microgram Se as selenite/g, or 0.1 or 0.2 microgram Se as freeze-dried cooked test food/g . Plasma and liver Se levels or glutathione peroxidase (EC 1.11.1.9; GSH-Px) activities were used as indicators of body Se status . 3 . Except for oysters, the biological availability of Se in all these seafoods was close to that of selenite (selenite 100%) when the criterion used was either plasma Se level or plasma GSH-Px activity . 4 . By the criterion of increased liver Se level of restored hepatic GSH-Px activity, only herring-Se had a biological availability comparable to that of selenite-Se under all conditions tested, whereas crab-Se and oyster-Se were distinctly inferior in this regard . 5 . Increasing the amount of crab-Se, oyster-Se or shrimp-Se supplied in the diet from 0.1 to 0.2 microgram/g changed the apparent availability (%) of Se for hepatic GSH-Px restoration from 38 to 78, 22 to 53 and 57 to 90 respectively . 6 . The present study demonstrates that the availability of Se in certain foods is a function of the criterion chosen, the level of Se supplied in the diet, and possibly other unknown interacting dietary factors.

Pharmacol Res Commun, 1986 Mar, 18(3), 241 - 56
Pharmacological activity of FPP028 (2-phenylpyrazolo-4-ethyl-4,7-dihydro {1,5a}pyrimidin-7-one) a new non-steroid anti-inflammatory agent; Pirisino R et al.; The activity of 2-phenylpyrazolo-4-ethyl-4,7-dihydro {1,5a}pyrimidin-7-one (FPP028), a non-acidic, analgesic, antipyretic, and anti-inflammatory compound, was investigated in a number of pharmacological tests performed in rats . The anti-inflammatory properties of FPP028 were evaluated through the carrageenan induced paw edema and the cotton pellet induced granuloma and compared with the activity of indomethacin, phenylbutazone, and isoxicam; as a result, the activity of FPP028 was shown to be similar to that of the latter compounds . To assess the analgesic properties of FPP028 in comparison with indomethacin and phenylbutazone, the Randall and Sellitto and the mouse-writhing tests were used; in both tests, FPP028 demonstrated a significant analgesic activity . FPP028 was shown to possess antipyretic properties in the test of yeast-induced pyrexia . The gastro-erosive activity of phenylbutazone and FPP028 was studied in restraint-stressed rats; in such test the ulcerogenic activity of phenylbutazone appeared to be dose-related; conversely, FPP028 demonstrated a gastro-protective effect since the number of gastric lesions induced either by stress or phenylbutazone treatment was decreased bu FPP028 . Our data show that FPP028 is endowed with most of the pharmacological properties of the classic antiinflammatory drugs . Further studies are however needed to more fully elucidate its mechanism of action because our in-vivo data indicate that FPP028 is not an inhibitor of prostaglandin biosynthesis.

Lab Invest, 1986 Mar, 54(3), 336 - 44
Cell differentiation and cell cycle effects on human promyelocytic leukemia cells induced by 12-O-tetradecanoylphorbol-13-acetate; Yun K et al.; As has been reported, doses of 12-O-tetradecanoylphorbol-13-acetate (TPA) as small as 1 to 100 ng/ml induced human promyelocytic leukemia HL-60 cells to differentiate terminally into macrophage-like cells rather than toward cells of the granulocytic series . This differentiation was accompanied by the appearance of monocyte/macrophage markers and by the disappearance of myeloid markers from the view point of enzyme cytochemistry . Contrasted to untreated HL-60 cells, TPA-treated cells increased in cell size and showed increased phagocytotic activities against opsonized sheep blood red cells and activated yeast . Nitroblue tetrazolium-positive cells increased rapidly after TPA exposure . The alterations of the cell cycle traverse of HL-60 cells by TPA were analyzed by {3H}thymidine autoradiography, flow microfluorimetry, and mitotic cell counting . TPA sequentially caused (a) inhibition of cells to move from G1 to S near G1/S boundary in G1; (b) temporary inhibition in G2; (c) growth arrest of most cells in G1 within 2 to 3 days after TPA exposure.

Semin Respir Infect, 1986 Mar, 1(1), 22 - 8
Immunodiagnosis of blastomycosis; Turner S et al.; The diagnosis of blastomycosis has long been a challenge for the clinician . Primarily a pulmonary disease, blastomycosis is difficult to differentiate from other respiratory infections by clinical symptoms alone . Substantial improvement in the diagnosis of blastomycosis can be attributed to the availability of the purified A antigen of Blastomyces dermatitidis and its homologous antibody and the incorporation of these reagents into diagnostic tests . Reliable procedures for accurately and rapidly immunodiagnosing blastomycosis are available . These methods comprise enzyme immunoassay and immunodiffusion tests for detecting diagnostic serum antibody by B dermatitidis, the exoantigen test for identifying mycelial-form cultures of the etiologic agent, and the fluorescent antibody technique for detecting and identifying yeast-form cells in culture or in tissue . These tests, used alone or in combination, can in many cases provide an early and rapid definitive diagnosis of blastomycosis.

Neuropharmacology, 1986 Mar, 25(3), 309 - 13
Evidence against adenosine 3',5'-monophosphate as a mediator of fever in the brain; Dascombe MJ; The aim of this study was to test the hypothesis that increased concentrations of adenosine 3',5'-monophosphate (cyclic AMP) in the pyrogen-sensitive preoptic/anterior hypothalamic region (PO/AH) mediate fever . Micro-injection of N6-2'-O-dibutyryl adenosine 3',5'-monophosphate (db cyclic AMP) into the preoptic/anterior hypothalamic region in rats produced a dose-dependent fall in body temperature which is inconsistent with the proposal that the nucleotide mediates fever . Hyperthermia was observed in some rats in response to large doses of db cyclic AMP, but this response was associated with convulsions . Endogenous concentrations of cyclic AMP in the preoptic/anterior hypothalamic region, as well as in the cerebral cortex, liver, spleen, thymus, white fat and plasma were unaffected by the febrile response to the subcutaneous injection of yeast in rats . A rise in levels of cyclic AMP was observed in the skeletal muscle of rats treated with yeast . The data presented do not indicate that cyclic AMP is involved in the neuronal events mediating fever in the rat.

Am Rev Respir Dis, 1986 Mar, 133(3), 353 - 6
A genetically engineered, mutant human alpha-1-proteinase inhibitor is more resistant than the normal inhibitor to oxidative inactivation by chemicals, enzymes, cells, and cigarette smoke; Janoff A et al.; Alpha-1-proteinase inhibitors (alpha 1PI) containing methionine (Met-358) or valine (Met----Val-358) at the reactive center were synthesized in and purified to homogeneity from recombinant yeast . The pure proteins were exposed to 1 of 4 different oxidizing systems: N-chlorosuccinimide (chemical oxidation), myeloperoxidase plus peroxide and halide (enzymatic oxidation), activated neutrophils (cellular oxidation), or gas-phase cigarette smoke . The effect of these treatments on the leukocyte elastase inhibitory function of both proteins was then assessed . After brief exposures, substantial inactivation of the normal inhibitor occurred, whereas the mutant inhibitor remained fully active . More prolonged exposures led to complete inactivation of the normal protein and partial inactivation of the mutant inhibitor . These results suggest that the reactive center methionyl residue in alpha 1PI is more rapidly affected by oxidants than are other oxidizable residues in the inhibitor; however, Met-358 is not the only residue in alpha 1PI whose modification can lead to the inactivation of the elastase inhibitory capacity of this protein.

Proc Natl Acad Sci U S A, 1986 Mar, 83(5), 1330 - 3
Direct measurements of intramolecular electron transfer rates between cytochrome c and cytochrome c peroxidase: effects of exothermicity and primary sequence on rate; Cheung E et al.; Rapid mixing of ferrocytochrome c peroxidase {cyt c peroxidase(II)} and ferricytochrome c {cyt c(III)} results in the reduction of cyt c(III) by cyt c peroxidase(II) . In 10 mM phosphate, pH 7.0, the rate of decay of cyt c peroxidase(II) and the rate of accumulation of cyt c(II) give equal first-order rate constants: k = 0.23 +/- 0.02 s-1 . Equivalent results are obtained by pulse radiolysis using isopropanol radical as the reducing agent . This rate is independent of the initial cyt c(III):cyt c peroxidase(II) ratios . These results are consistent with unimolecular electron transfer occurring within a cyt c(III)-cyt c peroxidase(II) complex . When cyt c is replaced by porphyrin cyt c (iron-free cyt c), a complex still forms with cyt c peroxidase . On radiolysis, using e-aq as the reducing agent, intracomplex electron transfer occurs from the porphyrin cyt c anion radical to cyt c peroxidase(III) with k = 150 s-1 . This large rate increase with increasing delta G degrees suggests that the barrier for intracomplex electron transfer is large . Finally, we have briefly investigated how the cyt c peroxidase(II)----cyt c(III) rate depends on the primary structure of cyt c(III) . We find the reactivity order to be as follows: yeast (k = 3.4 s-1) greater than horse (k = 0.3 s-1) greater than tuna (k = 0.2 s-1) . These results mirror a report {Ho, P . S., Sutoris, C., Liang, N., Margoliash, E . & Hoffman, B . M . (1985) J . Am . Chem . Soc . 107, 1070-1071} on excited state reactions of the cyt c/cyt c peroxidase couple.

Cancer Res, 1986 Mar, 46(3), 1388 - 94
Expression of tissue transglutaminase in cultured monocytic leukemia (THP-1) cells during differentiation; Mehta K et al.; Retinoic acid (RA) and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) induced differentiation of a human monocytic leukemia cell line, THP-1 . RA- or TPA-treated cells stopped proliferating, became adherent to plastic surfaces, and acquired the ability to phagocytose yeast cells, plain sheep RBCs, and IgG-coated sheep RBCs . The morphological and functional changes, induced by RA or TPA, were associated with a 20-50-fold increase in cellular transglutaminase activity . This increase in enzyme activity was found to be due to the induction of a specific intracellular transglutaminase, tissue transglutaminase . The induction of tissue transglutaminase was a specific response of THP-1 cells to differentiation and was not observed with agents that did not induce their morphological or functional differentiation . Dibutyryl cyclic AMP potentiated the RA-induced expression of tissue transglutaminase . A 15-min exposure to TPA was sufficient to induce differentiation and expression of tissue transglutaminase in THP-1 cells . In contrast, RA required a continuous exposure (48 h) to induce similar changes in morphology or enzyme activity . These results support the view that differentiation of cells of the monocytic lineage is associated with an induction and accumulation of the protein cross-linking enzyme tissue transglutaminase.

Eur J Biochem, 1986 Feb 17, 155(1), 1 - 10
Study of the plasma clearance of antibody--ricin-A-chain immunotoxins . Evidence for specific recognition sites on the A chain that mediate rapid clearance of the immunotoxin; Bourrie BJ et al.; In recent years, antibody--ricin-A-chain immunotoxins have been investigated as anti-neoplastic agents . To achieve in vivo therapy it is necessary that the immunotoxin remains in circulation at a sufficiently high level for a sufficiently long time to allow binding to tumor cells to occur . Therefore, examination of the pharmacology of immunotoxins may elucidate the reasons for the poor in vivo tumoricidal effect of immunotoxin described before . In this study the plasma clearance of antibody--ricin-A-chain immunotoxins, after intravenous injection in animals of different species, has been examined . Sensitive and reproducible techniques were developed to monitor the level of immunotoxin and its constituents in the blood . It is shown that immunotoxins are rapidly eliminated from the bloodstream . Neither the properties of the antibody moiety nor the nature of the linkage binding ricin A-chain to antibody account for the disappearance of immunotoxin from the plasma . On the other hand, we found that the rapid clearance of immunotoxin is due to the mannose residues on the ricin A-chain moiety which are specifically recognized by liver cells . When immunotoxin is administrated together with yeast mannan, which enhances the level of active immunotoxin in circulation by inhibition of liver uptake, the anti-cancer efficacy of immunotoxin in vivo is drastically improved.

J Biol Chem, 1986 Feb 15, 261(5), 2214 - 21
Glycine decarboxylase multienzyme complex . Purification and partial characterization from pea leaf mitochondria; Walker JL et al.; The P, H, and T proteins of the glycine cleavage system have been purified separately from pea leaf mitochondria and demonstrate molecular weights of 98,000, 15,500, and 45,000, respectively, by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The molecular weight of P protein by gel filtration was 210,000, indicating that this enzyme has a native homodimer conformation . Reconstitution assays containing purified P, H, and T proteins and yeast lipoamide dehydrogenase catalyze the oxidation of glycine and demonstrate a strict dependence on pyridoxal phosphate, tetrahydrofolate, NAD+, and dithiothreitol . The released CO2, methylamine-H protein intermediate, and methylenetetrahydrofolate are produced in stoichiometric amounts from glycine during the cleavage reaction . H protein acts as co-substrate with glycine during the decarboxylation reaction, demonstrating an apparent Km value of 2.2 microM . P and H protein alone jointly catalyze the glycine carboxyl-14 CO2 exchange reaction in the presence of pyridoxal phosphate and dithiothreitol . L protein of the glycine cleavage system was immunopurified using monoclonal antibodies . Antigenic and molecular weight similarities of the L protein with the lipoamide dehydrogenase component of the pyruvate dehydrogenase complex were shown suggesting the possibility of common isomers of lipoamide dehydrogenase for the two enzyme complexes.

J Biol Chem, 1986 Feb 5, 261(4), 1941 - 8
hv1 is an evolutionarily conserved H2A variant that is preferentially associated with active genes; Allis CD et al.; Polyclonal antibodies to the Tetrahymena macronuclear-specific histone variant hv1 cross-react with histone-like molecules from yeast, wheat, and mouse . A novel purification scheme has allowed isolation of sufficient hv1 to enable determination of the sequence of 61 amino-terminal residues as well as 27 additional internal residues . These data clearly demonstrate that hv1 shares a number of conserved sequence elements with the H2A family of histones . Comparison of hv1 with H2A.F (= H2A.Z = M1), another evolutionarily conserved H2A variant whose sequence is known, reveals that they share an unblocked amino-terminal alanine (instead of acetylserine) and a distinctive structure in a "variant box" region that distinguishes them from major H2As . In addition, 10 residues have been identified which are identical (or highly similar) in hv1 and H2A.F, but are different from residues conserved in the major H2As . Therefore, in many ways hv1 resembles chick H2A.F more than the major Tetrahymena H2A . The sites of acetylation of hv1 also differ from those of the major Tetrahymena H2As . In spite of their similarities, hv1 and H2A.Z differ significantly in their amino termini, and antibodies against hv1 do not react with H2A.Z . Interestingly, the nucleolar staining pattern reported with anti-hv1 serum is similar to that reported for an antiserum to another H2A variant, mouse testes-enriched H2A.X . Since both H2A.Z and hv1 appear to be enriched in transcriptionally active chromatin, these results suggest that there may be a number of different, functionally distinct, nonallelic variants in the H2A family of histones and that hv1 is a hybrid H2A variant with properties of both vertebrate H2A.Z and H2A.X.

J Med Microbiol, 1986 Feb, 21(1), 7 - 11
Fungicidal activity of murine broncho-alveolar macrophages against Blastomyces dermatitidis; Sugar AM et al.; The fungicidal activity of murine broncho-alveolar macrophages (BAM) against the yeast form of a virulent strain of Blastomyces dermatitidis was studied in the small-volume wells of a Terasaki plate . In a 4-h fungicidal assay, significant killing (16-33%) of the fungus by unstimulated BAM was demonstrated with BAM from normal mice and from mice rendered immune to lethal infection with B . dermatitidis . No significant differences between the activity of BAM from these two sources could be identified . Addition of 10% autologous normal or immune serum did not augment the macrophage fungicidal activity . Simultaneous experiments with peritoneal macrophages (PM) also gave reproducible killing of the yeast form in the wells of the Terasaki plate, but in the larger wells of microtitration plates, PM showed no significant fungicidal activity . On the other hand, BAM had similar fungicidal activity against B . dermatitidis in Terasaki and microtitration-plate wells . The modest fungicidal activity of BAM from immune mice against B . dermatitidis suggests that the resistance of immune mice to respiratory challenge is likely to be based on some augmentation of this first line of defence.

J Cell Sci, 1986 Feb, 80, 123 - 40
Calcium ions initiate the selective depolymerization of the pellicular microtubules in bloodstream forms of Trypanosoma brucei; Dolan MT et al.; Calcium ions (100 microM) were found to initiate the selective and complete depolymerization of the pellicular microtubules of Trypanosoma brucei . The Ca2+-dependent release of tubulin was found to occur without the detectable mediation of calmodulin . The released, depolymerized, pellicular tubulin from T . brucei cross-reacted with a monoclonal antibody raised against yeast tubulin . The pellicular tubulin was found to be composed of two alpha isotypes (apparently equal amounts) and one beta isotype . No other proteins were released from the plasma membrane-microtubule complexes during treatment with Ca2+ . The released pellicular tubulin was capable of reassembly into microtubules with normal ultrastructure . The observations reported here suggest that a special process may be required to accommodate the cleavage furrow during cytokinesis . This process would either be the Ca2+-dependent depolymerization of at least two of the cross-linked pellicular microtubules or the detachment of the cross-bridges between two pairs of pellicular microtubules on opposite sides of the cell.

J Med Vet Mycol, 1986 Feb, 24(1), 41 - 50
Endemic canine and feline histoplasmosis in El Paso, Texas; Kabli S et al.; Seventeen cases of histoplasmosis involving 2 dogs and 15 cats have occurred in the Upper Rio Grande Valley of El Paso since 1978 . The diagnosis, based on clinical signs and radiographic findings, was confirmed by one or more of the following laboratory procedures: demonstration of intracellular Histoplasma capsulatum yeast cells in tissue, positive serology, or isolation of H . capsulatum from various organs of necropsied animals . H . capsulatum was isolated also from a bat cave and soil in the vicinity of some of the houses where the affected animals had resided . Skin-tests of 97 persons for histoplasmosis indicated a 14% positive prevalance in this locale.

Biosci Rep, 1986 Feb, 6(2), 201 - 8
A study of the messenger RNA encoding pyruvate kinase of Neurospora crassa; Devchand M et al.; In Neurospora crassa, there is a single pyruvate kinase (PK) consisting of four identical subunits of approximately 60k daltons . Northern and dot blot hybridization studies, using most of the yeast pyruvate kinase gene as a probe, suggest the presence of two distinct mRNA species for pyruvate kinase, separable on the basis of the length of their polyadenylated tails, by oligo(dT)cellulose chromatography . These messages are present in polysomes, immuno-precipitated by anti-PK antibodies, indicating probable translation in vivo . Fractions containing both messages were translated in vitro in the heterologous systems as well as in a homologous N . crassa lysate, the newly-synthesized PK being detected by immunoadsorption . Protection studies using S1-nuclease suggest no major structural differences in the 5'-untranslated and most of the coding regions of the two messages.

J Biol Chem, 1986 Jan 25, 261(3), 1349 - 54
The binding of the chromosomal protein HMG-2a to DNA regions of reduced stabilities; Brown JW et al.; The interaction of immunopurified high mobility group 2a protein (HMG-2a) with DNA was examined by the nitrocellulose filter binding assay . The relative binding activity of HMG-2a for synthetic polynucleotides was: (dI).(dC) greater than (dA-dT).(dA-dT) greater than (dA).(dT) much greater than (dG).(dC) greater than (dG-dC).(dG-dC) . The protein also exhibited a marked preference for (A + T)-rich restriction fragments derived from rat and Drosophila satellites, yeast centromeres, phage lambda, and the ovalbumin gene and its 5' flanking sequences . These preferential DNA interactions occurred at ionic strengths and temperatures within the physiological range which argue for an in vivo role of DNA stability in dictating the genomic distribution of the large Mr HMG proteins.

Br Med J (Clin Res Ed), 1986 Jan 18, 292(6514), 159 - 61
Antibody responses to recombinant and plasma derived hepatitis B vaccines; Brown SE et al.; The antibody response to hepatitis B surface antigen (anti-HBs) induced in 25 recipients of a recombinant hepatitis B vaccine derived from yeast was compared with that induced in 25 recipients of a vaccine prepared from hepatitis B surface antigen (HBsAg) derived from plasma . Anti-HBs affinity and specificity were compared using assays of antibody affinity with two different antigens, a complex of the major polypeptide of HBsAg (p25; molecular weight 25 000 daltons) covalently linked to its glycosylated form (gp30) prepared from native purified HBsAg, and a cyclical synthetic peptide representing amino acid residues 139-147 of the major polypeptide of HBsAg and known to represent a major part of an a determinant . There was no difference in anti-HBs affinity or molar antigen binding sites of the antibody measured with either antigen between the two groups . All subjects in both groups produced antibody that bound to the gp30/p25 complex antigen, whereas 22 of the recipients of the plasma derived vaccine compared with 24 of those receiving the yeast derived vaccine produced antibodies that bound to the cyclical synthetic peptide 139-147 . These results support the finding of similar levels of anti-HBs, measured by commercial solid phase radioimmunoassay, in the two vaccine groups after three doses of vaccine . These results show no significant difference in the quantity, quality, or specificity of the anti-HBs response induced by the recombinant hepatitis B vaccine and the plasma derived hepatitis B vaccine.

J Biol Chem, 1986 Jan 15, 261(2), 869 - 73
Nuclear genes for cytochrome c oxidase subunits of Neurospora crassa . Isolation and characterization of cDNA clones for subunits IV, V, VI, and possibly VII; Sachs MS et al.; We obtained cDNA clones for cytochrome oxidase subunits IV, V, VI, and possibly VII by constructing a lambda gt11 library of Neurospora crassa cDNA and probing it with antiserum directed against Neurospora cytochrome oxidase holoenzyme . Positive clones were further characterized with antisera directed against individual cytochrome oxidase subunits and subsequently by DNA sequencing . The clones for subunits IV and V encode proteins with regions matching the known N-terminal amino acid sequences of purified Neurospora cytochrome oxidase subunits IV and V, respectively . The sequences of these clones provide the first evidence that cytochrome oxidase subunits IV and V are made as precursors with N-terminal extensions in Neurospora . The N-terminal extensions encoded by these clones share homology, and are rich in arginine, as are signal sequences of other mitochondrially destined proteins . The subunit VI clone codes for the carboxyl terminus of a protein homologous to the carboxy termini of yeast cytochrome oxidase subunit VI and bovine cytochrome oxidase subunit Va . The subunit VII clone contains an open reading frame for a 47-residue protein, the expected size for subunit VII . However, the protein coded by this clone has an unusual amino acid composition . Whether this clone represents an authentic cytochrome oxidase subunit is not established.

Drug Nutr Interact, 1986, 4(3), 309 - 19
Selenium deficiency and detoxication functions in the rat: short-term effects of cadmium; Olsson U; Weanling rats were fed a Torula yeast-based selenium-deficient diet with or without supplementation of sodium selenite (0.2 ppm selenium) in the drinking water . After 5-6 weeks on the diet regimens, the liver glutathione peroxidase activity of the selenium-deficient groups had decreased to about 1% of the supplemented groups, and the rats were then used in experiments . Cadmium-induced effects on the drug-metabolizing system of the liver were measured as the microsomal capacity to perform N- and C-oxygenation of N, N-dimethylaniline . Cadmium in vitro caused a decrease of the cytochrome P-450-dependent C-oxygenation . This effect tended to be more prominent in the selenium-deficient groups . On the other hand, N-oxygenation was increased when cadmium was added in vitro, and no significant difference was found between selenium-deficient and -supplemented groups . However, as was found for the capacity to perform C-oxygenation, there was a tendency for lower N-oxygenation in the selenium-deficient rat . Lipid peroxidation, measured as thiobarbituric acid reactive substances in liver homogenates, was higher in selenium-deficient groups after in vivo treatment or in vitro addition of cadmium, and preincubation or phenobarbital induction enhanced this selenium-dependent difference . Although, the selenium-deficient rat seems more susceptible to cadmium-induced disturbances, 5-6 weeks of selenium deficiency was not enough to cause prominent impairment on the drug-metabolizing system as measured here and with the doses used in the present study.

Biokhimiia, 1986 Jan, 51(1), 59 - 64
{Characteristics of thiamine thiazolone diphosphate-induced inhibition of a pyruvate dehydrogenase complex in vitro and in intact mitochondria}; Iakovleva GM et al.; Thiamine thiazolone diphosphate (TTPP) was capable of penetrating through the mitochondrial membrane and of inhibiting the pyruvate dehydrogenase complex (PDC) in intact mitochondria . TTPP depressed the activity of mammalian PDC in a mixed manner (Ki = 5.10(-8) M) and yeast pyruvate decarboxylase (Ki = 5.10(-6) M) via a competitive mechanism with respect to thiamine diphosphate . It was shown that decarboxylation of pyruvate in intact and disrupted mitochondria of rat liver and brain is less inhibited by TTPP than the overall activity of PDC determined by the formation of acetyl-CoA . It was assumed that TTPP as a transition state analog participates only in oxidative reactions (but not in simple decarboxylation of pyruvate).

Anal Biochem, 1986 Jan, 152(1), 160 - 6
High-performance liquid chromatography of aldehydes and acids formed in monoamine oxidase-catalyzed reactions; Yu PH et al.; A rapid, sensitive, and specific method for the determination of monoamine oxidase (MAO) activities toward different substrates is described . The assay is based on high-performance liquid chromatographic (HPLC) separation and electrochemical detection of the aldehyde or acid products . The aldehyde metabolic intermediates were observed to be quite stable in 0.1 N perchloric acid containing antioxidant and EDTA, and therefore can be used to measure the MAO activity of washed mitochondrial membrane and partially purified or purified MAO . Incomplete conversion of aldehyde to acid was observed when the amine substrates were incubated with the crude enzyme preparations . These aldehydes can be converted to corresponding acids by addition of yeast aldehyde dehydrogenase and beta-NAD and the acid can also be measured by HPLC-electrochemical detection . A deuterium isotope effect in the oxidation of p-{alpha,alpha-2H2}tyramine and {alpha,alpha-2H2}serotonin has been demonstrated by this method.

Arch Biochem Biophys, 1986 Jan, 244(1), 211 - 7
Physiological requirement for biosynthesis of multiple 24 beta-methyl sterols in Gibberella fujikuroi; Nes WD et al.; Studies with Gibberella fujikuroi have been designed to examine the relationship between the biosynthesis and function of fungal sterols . Evidence was obtained through appropriate feeding and trapping experiments for the existence of multiple end products which are produced by separate routes in the later stages of sterol biosynthesis . The three end products, ergosterol (24 beta-methylcholesta-5,7,22E-trien-3 beta-ol), brassicasterol (24 beta-methylcholesta-5,22E-dien-3 beta-ol), and 22(23)-dihydrobrassicasterol (24 beta-methyl-cholesterol), were found to be non-interconvertible during logarithmic phase growth; thus the metabolic route delta 5,7,22-24 beta-CH3----delta 5,22-24 beta-CH3----delta 5-24 beta-CH3 was ruled out . Ergosterol can be further metabolized, viz., to 24 beta-methylcholesta-5,7,9(11),22-tetraen-3 beta-ol, but only as the culture enters into the stationary phase . In the presence of growth inhibitory concentrations of 2,3-iminosqualene, a partial reversal of growth cessation was obtained when all three sterols were concurrently supplied to the medium . Since neither ergosterol nor the other two sterols added individually to the medium was able to overcome the inhibitor's deleterious effect, ergosterol cannot play a dual architectural role (bulk and regulatory) in this fungus as it apparently can do in other fungal systems, i.e., yeast . For G . fujikuroi each sterol end product appears to possess a unique physiological role . Mycelial growth requires more than simply ergosterol.

Toxicol Appl Pharmacol, 1986 Jan, 82(1), 140 - 50
Toxicity of cadmium chloride in vitro: indices of cytotoxicity with the pulmonary alveolar macrophage; Coin PG et al.; Pulmonary alveolar macrophages were isolated from adult, male New Zealand white rabbits by bronchial lavage and exposed to cadmium chloride in vitro . The observed cell sensitivity to this metal was highly dependent upon the incubation conditions used as well as the cytotoxic index selected . An LC50 value, as measured by dye exclusion (erythrosin B), was determined to be 390 microM when these cells were exposed to cadmium in Ham's F12 culture medium for 8 hr at 35 degrees C . The presence of fetal calf serum in the medium (10%; v/v) enhanced this toxicity slightly, LC50 = 235 microM, as did raising the incubation temperature to 37 degrees C, LC50 = 201 microM . No effect on cadmium toxicity was observed when the culture medium was made deficient in Cu, Zn, and Fe, nor was there any effect observed when Hepes buffer was substituted for the bicarbonate/carbon dioxide buffering system . Measurements of cadmium-109 uptake by pulmonary alveolar macrophages were consistent with and could explain, at least in part, the above observations of cytotoxicity . In the standard culture system (an 8-hr exposure period at 35 degrees C in Ham's F12 culture medium plus serum), the appearance in the culture medium of two lysosomal enzyme activities, acid phosphatase and cathepsin D, paralleled cell death . In addition, an EC50 value of 102 microM was found for cadmium when cell respiration (O2 uptake) was measured; an EC50 value of 31 microM was found for cadmium when cell function (engulfment of killed yeast particles) was followed; and scanning electron microscopic studies showed cell membrane changes (loss of fine structure and blebbing) at cadmium concentrations as low as 30 microM . These findings suggest that loss of cell function and/or changes in cell morphology are more sensitive measurements of macrophage exposure to cadmium than is either cell death, lysosomal enzyme release, or cell respiration.

Infect Immun, 1986 Jan, 51(1), 6 - 9
Activation of the alternative complement pathway by Sporothrix schenckii; Scott EN et al.; We studied the activation of complement by Sporothrix schenckii yeast cells . Total complement activity, and the effect of various activators on this activity, were assayed on aliquots of fresh nonimmune human serum with and without prior treatment with chelators . Both total hemolytic complement and C3 were consumed (activated) in serum chelated with magnesium ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, which blocks the classical pathway but leaves the alternative pathway intact . Further, C3 was consumed, but C4 (exclusively a component of the classical pathway) was preserved, in nonchelated serum after challenge by S . schenckii yeast cells . Absorption of serum with S . schenckii yeast cells to deplete antibodies did not alter these results . Furthermore, immunofluorescence studies demonstrated that C3, but not immunoglobulin G, was deposited on yeast cells during incubation with nonimmune serum . These data indicate that S . schenckii yeast cells activate the alternative complement pathway in vitro independently of antibody . These data do not define a role for the alternative pathway in in vivo host defenses against infection with this organism but provide a foundation for studies to evaluate such a role.

J Hepatol, 1986, 3(2), 190 - 5
Immunogenicity of recombinant hepatitis B vaccine in dialysis patients; Jilg W et al.; Eighty-eight dialysis patients were vaccinated with recombinant hepatitis B vaccine prepared in yeast . Fourty-nine patients were immunized 3 times (months 0, 1, 6) intragluteally with 40 micrograms hepatitis B surface antigen (HBsAg) per dose . Only 32 of them (65.3%) showed anti-HBs concentrations above 10 IU/l with a geometric mean titer (GMT) of 180.7 IU/l after 3 vaccinations, whereas all of the 16 healthy controls, vaccinated 3 times with a 10-micrograms dose of the same vaccine batch, had specific antibodies higher than 10 IU/l (GMT 897.4 IU/l) . Responses of patients were slightly higher than those of dialysis patients vaccinated in an earlier study with plasma-derived vaccine according to the same schedule . Results in 20 patients immunized 6 times intragluteally with 40 micrograms HBsAg/dose in monthly intervals were not better (at month 7, 65% showed anti-HBs concentrations greater than 10 IU/l; GMT = 126.6 IU/l), and 19 patients receiving 6 times 20 micrograms HBsAg monthly showed significantly lower responses (anti-HBs greater than 10 IU/l in 42% of vaccinees, GMT = 89.5 IU/l) . The vaccine was tolerated well; side-effects were slight, and no serious adverse reactions were observed . In conclusion, recombinant hepatitis B vaccine is comparable to plasma-derived vaccine also in the case of dialysis patients; a 6-dose schedule does not seem to have much advantage compared to the conventional 3-dose regimen.

Ann Nutr Metab, 1986, 30(4), 233 - 40
Tolerance of low and high dietary selenium throughout the life span of Syrian hamsters; Birt DF et al.; In lifetime studies on the effects of dietary selenium (Se) levels, Syrian hamsters were fed diets containing low (unsupplemented torula yeast), adequate (0.1 ppm Se supplemented from sodium selenite), or excessive (5 ppm Se supplemented from sodium selenite) levels of Se . A commercial ration was fed to separate groups . Male and female hamsters were assigned to each diet, and blood samples were collected at 54 and 79 weeks of age for determination of Se status . Body weights of male hamsters were generally highest in those fed unsupplemented diets and lowest in those fed 5 ppm Se supplements . Female weights did not differ between the three semipurified diets . Erythrocyte and plasma glutathione peroxidase and blood Se values increased with the increments in dietary Se at the 54- and 79-week measurements . Survival was approximately 40-45% lower in hamsters fed the commercial ration than in those fed semipurified diets, but was not altered by the Se level in the semipurified diet.

J Neurosci Res, 1986, 16(1), 141 - 56
Neuron-specific enolase: complete structure of rat mRNA, multiple transcriptional start sites, and evidence suggesting post-transcriptional control; Forss-Petter S et al.; The protein encoded by a randomly selected rat brain cDNA clone was identified as neuron-specific enolase (NSE; 4.2.1.11; gamma subunit), based on homology to yeast enolase sequences and the presence of the corresponding 2.5-kb mRNA in rat brain but not in liver, kidney, or muscle tissue . The 2,222-nucleotide NSE and mRNA sequence presented identifies a 68-nucleotide 5' noncoding region, a 1,302-nucleotide open reading frame (corresponding to a primary translation product of 434 amino acids), and 852 noncoding 3' bases . Evolutionary implications based on sequence comparisons to yeast enolase and non-neuronal enolase are discussed . Primer extension analysis indicated the presence of several alternative initiation sites for transcription within 60 nucleotides on the NSE gene . The developmental onset of NSE mRNA expression correlates with the appearance of NSE protein; however, the mRNA reaches adult levels by postnatal week 3, whereas the protein continues to accumulate over the next few months, suggesting regulatory mechanisms in addition to transcriptional control.

Mikrobiologiia, 1986 Jan-Feb, 55(1), 120 - 6
{Morphological changes in a chemostat culture of Candida utilis undergoing cyclic changes of pH and pO2}; Lirova SA et al.; The effect of cyclic pH and pO2 changes on the morphology of Candida utilis was studied with time intervals measured in minutes . The pH was varied from 4.5 to 2.6, the pO2 from 70% of the saturation to 0% . The morphology was studied both visually and using the technique of optical-structural computer analysis . The regime with pH changes increased the biomass yield . However, the averaged morphological characteristics showed that the growth was slightly inhibited . Therefore, the yeast population was very heterogeneous, some cells were inhibited while other cells were stimulated, which made the economic coefficient rise . The cells were also inhibited when they were exposed in the conditions of oxygen deficiency for a short period of time . However, the cells could not be inhibited if, at the same time, the pH was extremely low . Changes in the morphology were detected earlier than either the inhibition or stimulation of growth was recorded by measuring the weight of biomass.

J Clin Microbiol, 1986 Jan, 23(1), 163 - 9
Guanine is a growth factor for Legionella species; Pine L et al.; Evaluation of previously described chemically defined media for the growth of Legionella pneumophila showed that these media supported poor growth of several strains of L . pneumophila and did not support growth of certain of the Legionella species described later . Growth was stimulated by the dialysate from yeast extract but not by the nondialyzable fraction . Further investigations indicated that the active factors from the yeast extract dialysate were purine or pyrimidine derivatives, and certain known purines and pyrimidines were found to stimulate growth . Of these, guanine universally stimulated growth of all Legionella strains and was a growth requirement for several of the species tested . A balanced, N-(2-acetamido)-2-aminoethanesulfonic acid-buffered, chemically defined medium having guanine or a purine-pyrimidine mix is presented for the general growth of Legionella species.

Nucleic Acids Symp Ser, 1986, (17), 167 - 70
Enzymatic synthesis of chimeric tRNAs with unusual numbers of base pairs in the anticodon stem; their structure and properties; Nishikawa K; Chimeric tRNATyr molecules have been constructed by enzymatic procedures in vitro from the 5'-half fragment of T . utilis tRNATyr and the 3'-half fragment of yeast tRNATyr, and vice versa . These chimeric tRNAs contain base-mismatching(s) in the anticodon stem and, therefore, have only 3 or 4 base pairs in the stem . Although the Tm of these chimeras are largely decreased, there seems to be no gross difference between the structure of native and chimeric tRNATyrs at physiological temperatures in the presence of 10 mM MgCl2 . Aminoacylation assays also revealed that the tyrosine-acceptance of the chimeras are fully comparable to that of native tRNATyrs . However, the possibility remains that the properties of the chimeras are considerably different from those of native ones at lower Mg++ concentrations.

Acta Paediatr Scand Suppl, 1986, 325, 107 - 11
Experimental research on IGF-1; Skottner A et al.; IGF-1 has been produced by recombinant DNA technology; the host cell is a yeast . Studies in hypophysectomized rats show that IGF-1 has little growth promoting effect unless given by infusion at high doses, and priming with bovine growth hormone did not produce any potentiation . In vitro studies reveal that IGF-1 cross-reacts with the insulin receptor on adipose cells . Immunohistochemical studies have revealed high IGF-1 immunoreactivity in proliferating and differentiating cells in the testes, lymphoid organs and pancreatic islets and particularly in regenerating nerves . Local application of IGF-1 may stimulate regeneration in damaged peripheral nerves . Key words:

Curr Genet, 1986, 10(11), 803 - 10
Plasmids of mitochondrial origin in senescent mycelia of Podospora curvicolla; Bockelmann B et al.; Podospora curvicolla displays symptoms of senescence similar but not quite identical to those reported for Podospora anserina . In Podospora curvicolla single hyphae may escape from death leading to a new growth front and consequently to a mode of growth characterized by alternating phases of growth and non-growth . Restriction analyses and hybridization experiments have revealed that the Podospora curvicolla type of senescence is correlated with plasmids originating from amplification of a single distinct region of the mitochondrial DNA containing the 1rRNA gene . In the yeast transformation system sequences of this region may function as autonomously replicating sequences (ARS) . Plasmids (pl1, pl2 and pl3) isolated from different, independently aged mycelia are largely homologous to each other but differ in their excision/junction sites and have different sizes: 10.85 kb (p11), 9.01 kb (pl2) and 10.50 kb (pl3) . The sequence of the most frequently occurring plasmid in ageing strains of Podospora anserina is absent in Podospora curvicolla either as free plasmid DNA or as an integrated part of the mtDNA . Possibly there is a correlation between the absence of this particular sequence in Podospora curvicolla and the type of senescence displayed in this organism.

Curr Genet, 1986, 10(10), 755 - 60
Characterization of Phycomyces blakesleeanus mutants temperature-sensitive for heat-shock induced germination; Micol JL et al.; Sixty-nine mutants of the fungus Phycomyces blakesleeanus, temperature-sensitive for heat-shock induced germination, have been characterized . All of them show a low viability at 26 degrees C and normal viability at 16 degrees C . Eleven mutants recover the wild type phenotype if yeast extract is added to the minimal medium; the mutant phenotype of eight of these mutants is also suppressed by the addition of putrescine or other polyamines . The majority of the mutants are affected very early in germination . Spontaneous, heat-shock and acetate induced germination are not equally impaired by some of the mutations, so specific and independent steps seem to be involved in part of the activation mechanism of germination.

Curr Genet, 1986, 10(6), 459 - 62
Identification of an autonomously replicating sequence near a histone gene of Physarum polycephalum; Wilhelm ML et al.; Fragments of DNA which function as autonomous replication sequences in yeast were cloned from Physarum polycephalum . The ars activity is located in a 1.2 kbp fragment extending 1.5 kbp to 2.7 kbp upstream of the 5' end of a histone H4 gene . Our recent finding that a replication origin is located at a distance less than 3 kbp of this histone gene suggests that the ars element identified coincides with a specialized replication origin and can be used to direct chromosome replication in Physarum polycephalum.

Princess Takamatsu Symp, 1986, 17, 23 - 30
Two new retroviral onc genes, sea and jun; Bos TJ et al.; Avian leukemia virus S13 induces erythroblastosis, granulocytic leukemia, fibrosarcoma, anemia, and endothelial neoplasia . It transforms chick embryo fibroblasts and primitive erythroid cells in culture and is defective in replication . Its onc gene, sea, is expressed as transformation specific env-sea fusion glycoprotein of 155 kDa . Gp155 is proteolytically processed into gp85env and gp70env-sea . The latter shows tyrosine specific protein kinase activity . Avian sarcoma virus 17 induces fibrosarcoma and transforms chick embryo fibroblasts in culture . Its cell derived onc gene, jun, is not related to known onc genes and appears to be expressed as a gag-jun fusion protein of 55 kDa . The amino acid sequence of jun shows homology in its C-terminal region to the C-terminal DNA binding region of the yeast regulatory protein GCN4, suggesting that the jun protein may bind to DNA.

Curr Genet, 1986, 10(10), 767 - 75
Behaviour of recombinant plasmids in Aspergillus nidulans: structure and stability; Barnes DE et al.; A pyrG- Aspergillus strain was transformed with plasmid pDJB-1, derived from pBR325 by insertion of the Neurospora crassa pyr4 gene (orotidine 5'-phosphate carboxylase), giving mitotically unstable transformants . Aspergillus DNA which acted as an "autonomously replicating sequence" (ARS) in yeast was inserted into pDJB-1 and the resulting construct, pDJB12.1, gave mitotically stable transformants when introduced into Aspergillus . Transformants obtained with pDJB-1 and pDJB12.1 gave few pyr- progeny in crosses to a pyrG+ strain . Southern hybridisation analysis of pyr+ transformants obtained with pDJB-1 revealed restriction fragments expected for integrated plasmid but transformants obtained with pDJB12-1 showed only bands derived from free plasmid . pDJB-1 and derivatives of pDJB12.1 could be recovered from transformants . These derivatives could not be explained by straightforward excision of integrated pDJB12.1 sequences but could result from recombination between plasmid molecules . Hybridisation of undigested transformant DNAs showed that the transforming DNA was present in a high molecular weight form . These results suggest: (1) pDJB12.1 derivatives and possibly pDJB-1 can replicate autonomously in Aspergillus; (2) A . nidulans DNA acting as an ARS in yeast enhances replication and/or segregation of transforming plasmids in Aspergillus; and (3) recombinant plasmids may undergo rearrangements when introduced into Aspergillus.

Acta Paediatr Scand Suppl, 1986, 325, 85 - 9
Current research on recombinant human growth hormone and the related growth factors, IGF-1 and GRF; Fryklund L; A brief introduction is provided to the physiological actions of somatostatin, somatomedins and growth hormone releasing factor (GRF) in relation to human growth hormone (hGH) . The development of recombinant DNA techniques allowed the biosynthesis of somatrem (methionyl hGH), and as the methods were refined, authentic recombinant hGH (methionyl-free) has been produced . rhGH is currently undergoing clinical trials in several countries . Recombinant IGF-1 has also been developed by similar methods, utilizing a yeast as the host cell . In contrast, GRF is too small to be biosynthesized, but can be produced by classical peptide synthesis . Only the N-terminal sequence is required for biological activity, and may be 29, 40 or 44 residues long . Research into the biological effects of both IGF-1 and GRF is underway.

Immunol Res, 1986, 5(2), 129 - 38
Phagocytosis of unopsonized zymosan particles by trypsin-sensitive and beta-glucan-inhibitable receptors on bone marrow-derived murine macrophages; Kadish JL et al.; Murine bone marrow cells, plated at 4 X 10(4) cells/well and cultured in 50% fibroblast CM, yielded pure populations of large, individual, adherent cells that were phagocytic and morphologically indistinguishable from macrophages . Adherent macrophages appeared in small numbers with 24 h of culture, increased to maximal cell numbers within 10 days of culture, and remained at these cell densities for at least 11 weeks in culture . The capacities of adherent macrophages to ingest unopsonized zymosan particles and EsIgG, at inputs of 1.25 X 10(7) targets, were expressed by 7 and 40% of the cells derived from 24-hour cultures, respectively, were increased at nearly identical rates to comparable maximal levels within 10-14 days of culture and were exhibited by essentially all adherent cells derived from 2-11-week cultures . The percentage of adherent macrophages from twelve 3-6-week cultures ingesting greater than or equal to 1, greater than or equal to 6 and greater than or equal to 10 zymosan particles was 89 +/- 5, 47 +/- 11 and 14 +/- 9% (mean +/- SD, n = 12), respectively, and the percentage ingesting greater than or equal to 1, greater than or equal 6 and greater than or equal to 10 EsIgG was 86 +/- 5, 49 +/- 10 and 14 +/- 8%, respectively . Incubation of adherent macrophages with mannan-free ss-glucan particles at inputs of 5 X 10(5)-5 X 10(7)/ml initiated a phagocytic response comparable to that obtained with the same doses of zymosan particles which contained mannan and beta-glucan . Preincubation of adherent macrophages with 100 micrograms/ml of a fully soluble beta-glucan, laminarin, and solubilized barley beta-glucan reduced subsequent macrophage phagocytosis of greater than or equal to 6 zymosan particles by 53 and 40%, respectively . In contrast, yeast alpha-mannan was less than 1% as active, and 10 mg/ml reduced the number of adherent macrophages ingesting greater than or equal to 1 zymosan particles by 64% . At concentrations as high as 2 mg/ml, laminarin and barley beta-glucan had no effect on Fc receptor-mediated ingestion of EsIgG, and mannan at 20 mg/ml also failed to inhibit EsIgG ingestion . Pretreatment of adherent macrophages with 20 micrograms/ml of trypsin reduced the number of cells ingesting greater than or equal to 1 zymosan particles from 89 to 10% and those ingesting greater than or equal to 6 zymosan from 43 to 0%, whereas pretreatment with as much as 100 micrograms/ml of trypsin failed to decrease macrophage ingestion of EsIgG.(ABSTRACT TRUNCATED AT 400 WORDS)

Ann Clin Res, 1986, 18(1), 13 - 7
Selenium in food and nutrition in Finland . An overview on research and action; Koivistoinen P et al.; For geochemical reasons Finland is a low-selenium area . In the 1960's several diseases associated with serious Se deficiency were observed in domestic animals . Selenium medication of animals and selenium supplementation of animal feeds from 1969 effectively eliminated these diseases . An extensive study of the trace element content of foods consumed in Finland in the 1970's demonstrated that the dietary intake of selenium was exceptionally low (25 micrograms/day/10 MJ) during the years when domestic grains were used . A study carried out in 1981 showed that supplementation of healthy middle-aged men with high selenium wheat or yeast or selenate double the glutathione peroxidase activity in platelets . Prospective epidemiological studies based on cohorts that were followed in the 1970's suggested that low selenium (less than 45 ng/ml serum) might be a risk factor for cardiovascular diseases and cancer . Technologies to increase the selenium content of foods and feeds were developed and an official decision was reached to add, starting in 1984, sodium selenate to the main fertilizers to increase the selenium content of domestic grain to about 100 micrograms/kg . This measure will increase the average selenium intake above 50 micrograms/d even in the years when grain with a high selenium content is not imported.

Basic Life Sci, 1986, 37, 207 - 15
Recombinant DNA approaches to feline leukemia virus immunization; Luciw P et al.; We have utilized 2 recombinant DNA strategies for immunization against FeLV in cats: (a) modified live virus was attenuated by mutation and recombination, and (b) an immunogen, consisting of subunit envelope protein, was prepared in genetically engineered yeast . Results indicated that the genetically manipulated live virus preparations were not protective against FeLV challenge because they were either not attenuated in virulence or were not sufficiently antigenic . Immunization with yeast-synthesized FeLV envelope protein followed by the modified live virus gave protective immunity in cats under experimental conditions . Future immunization attempts will concentrate on enhancing the immunogenic potency of the yeast- synthesized FeLV envelope protein.

Microbiol Immunol, 1986, 30(1), 1 - 11
Active substance of Histoplasma capsulatum that inhibits blastogenic response of lymphocytes; Yamamoto Y et al.; A substance inhibiting blast transformation of murine spleen lymphocytes stimulated with various mitogens, such as LPS, PHA, and PWM, was obtained from yeast-form cells of Histoplasma capsulatum . This active substance was partially purified from the cell-free extract by DEAE-Sepharose CL-6B column chromatography . As a result of this partial purification, the inhibitory activity was 1.26 micrograms/ml in terms of ID50 . Materials from H . capsulatum also inhibited blast transformation of murine spleen lymphocytes stimulated with the antigen PPD as well as mitogens LPS, PHA, and PWM . However, the con A-induced proliferative response was only slightly affected . A similar result was observed for the MLR . These inhibitory activities were abolished by heating at 70 C for 30 min . These results suggest that the heat-labile active substance produced by H . capsulatum might directly affect the lymphocytes, leading to inhibition of their blast transformation.

Arch Dermatol Res, 1986, 279(1), 59 - 65
On the interaction between anthralin and mitochondria: a revision; Fuchs J et al.; Anthralin is an inhibitor of oxidative phosphorylation at concentrations found in vivo . ADP-stimulated oxygen consumption is diminished . Consequently, the rate of ATP synthesis is reduced and mitochondrial ATP content declines . Neither the isolated ATPase (F1F0-ATPase), nor the mitochondrial membrane-bound ATPase are influenced by the drug . Respiration under resting conditions is not affected . The experimental data unequivocally indicate that anthralin is not an uncoupler of oxidative phosphorylation, as previously stated . Furthermore, the interpretation that respiratory deficiency induced in yeast strains by anthralin is a consequence of petite mutations has to be reconsidered . Under in vivo conditions, anthralin inhibits respiration per se . Our experiments, including the electron spin resonance spectroscopy, reveal that anthralin alters mitochondrial membrane structure and function simultaneously . A redox or free-radical mediated step may be involved . In consequence, inhibition of ATP production occurs which may become the limiting factor for increased cellular metabolism in psoriasis.

Mol Biochem Parasitol, 1986 Jan, 18(1), 103 - 12
Cysteine proteinase of Entamoeba histolytica . I . Partial purification and action on different enzymes; Scholze H et al.; A method for a 50-60-fold purification of a cysteine proteinase from trophozoites of Entamoeba histolytica using 35-80% ammonium sulphate fractionation, gel chromatography on Sephadex G-75, and preparative isoelectric focusing is described . The enzyme was examined for its proteolytic potencies towards native enzyme substrates . The amebic proteinase directly inactivates aldolase and glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle as well as glucose-6-phosphate dehydrogenase from yeast . The inactivation of citrate synthase from porcine heart proceeds rather slowly, whereas malate dehydrogenase from porcine heart is not affected by the amebic proteinase under the condition used . With the exception of aldolase all inactivated enzyme substrates have been cleaved by limited proteolyses yielding major cleavage products . The inactivation of aldolase probably functions by the release of a small segment from a terminus being essential for aldolase activity.

Curr Genet, 1986, 10(8), 625 - 30
Characterization of inverted repeats from plasmid-like DNAs and the maize mitochondrial genome; Braun CJ et al.; Integrated inverted repeat (IR) sequences similar to those of the S plasmids have been isolated from the genomes of the normal and S type male-sterile cytoplasms of maize mitochondria . The nucleotide sequences of both the IRs and their flanking regions have distinguished and characterized several different types of repeats . The repeats may be involved in the recombinational process that occurs continuously in the mitochondrial genome . One cloned fragment, derived from a fertile revertant and containing sequences similar to S-2, does not appear to act as a typical transposable element during reversion . Several of the flanking regions examined contain a small repeat of 34 base pairs, in which a nonanucleotide segment is found with similarity to the yeast mitochondrial promoter.

Curr Genet, 1986, 10(7), 515 - 25
Analysis of the mitochondrial and nuclear genomes of two basidiomycetes, Coprinus cinereus and Coprinus stercorarius; Weber CA et al.; The mitochondrial and nuclear genomes of Coprinus stercorarius and C . cinereus were compared to assess their evolutionary relatedness and to characterize at the molecular level changes that have occurred since they diverged from a common ancestor . The mitochondrial genome of C . stercorarius (91.1 kb) is approximately twice as large as that of C . cinereus (43.3 kb) . The pattern of restriction enzyme recognition sites shows both genomes to be circular, but reveals no clear homologies; furthermore, the order of structural genes is different in each species . The C . stercorarius mitochondrial genome contains a region homologous to a probe derived from the yeast mitochondrial var1 gene, whereas its nuclear genome does not . By contrast, the C . cinereus nuclear, but not mitochondrial, genome contains a region homologous to the var1 probe . Only a small fraction of either the nuclear or mitochondrial genomes, perhaps corresponding to the coding sequences, is capable of forming duplexes in interspecies solution reassociations, as measured by binding to hydroxylapatite . Those sequences capable of reassociating were found to have approximately 15% divergence for the mitochondrial genomes and 7%-15% divergence for the nuclear genomes, depending on the conditions of reassociation.

Cancer Surv, 1986, 5(1), 81 - 91
Biochemical and cellular determinants of bleomycin cytotoxicity; Sikic BI; Bleomycin is a mixture of cytotoxic glycopeptides which function as mininucleases, binding to DNA and producing single and double strand breaks by the formation of an activated oxygen complex . Bleomycin is an effective agent against a few human cancers, notably lymphomas, testicular and ovarian germ cell cancers and certain squamous carcinomas . Most human cancers are resistant to bleomycin a priori, however, and those which are initially sensitive frequently develop resistance to the drug during therapy . Several potential modes of resistance to bleomycin have been identified in cell culture and animal tumour models, although their relative importance in determining the responsiveness of human cancers to the drug is not well understood . Bleomycin is selectively toxic to cells in the M and G2 phases of the cell cycle, and generally more effective against actively dividing rather than resting cells . Thus, the cytokinetic state of the tumour cell population is an important determinant of drug activity . Oxygen is an essential substrate for bleomycin's action, with the degree of cytotoxicity directly related to ambient oxygen . Both acutely and chronically hypoxic cells form a substantial fraction of the cell population of many tumours, and may serve as a reservoir of cells resistant to bleomycin on this epigenetic basis . Metabolic inactivation of bleomycin is a mechanism of resistance to the drug in some cells and may influence toxicity in normal tissues . Bleomycin hydrolase activity is low in lungs and skin, the two major sites of normal tissue toxicities, and levels of this enzyme have been elevated in some but not all tumour cell lines selected for resistance to bleomycin . The capacity to repair or withstand single and double strand DNA breaks may also be an important determinant of resistance to the drug . Most yeast and mammalian cell mutants, which are hypersensitive to ionizing radiation because of defects in DNA repair, are also more sensitive to bleomycin than wild-type cells . A number of agents which interact with membranes or inhibit DNA repair, such as ethanol, lidocaine, verapamil and caffeine, have been reported to sensitize cells to bleomycin in vitro.

Dev Biol Stand, 1986, 63, 141 - 6
A brief overview of the new vaccines against hepatitis B virus infection: immunogenic gene products and peptide analogues of antigenic epitopes; Vyas GN; Immunologically recognizable antigens encoded by hepatitis B virus (HBV) DNA include: (i) the surface antigen (HBsAg), (ii) the pre-S antigen, (iii) the X antigen (HBxAg) and (iv) the core/e antigens (HBcAg/HBeAg) . Each one of these antigens or their combination may be prepared for the next generation of vaccines against HBV infection . The synthesis of HBsAg by expression of recombinant DNA in the yeast system has now reached the stage of clinical trials and will certainly provide the first alternative to the currently licensed plasma-derived HBsAg vaccine . The recombinant vaccinia containing the gene encoding HBsAg and transfected cell-lines expressing HBsAg/pre-S gene products are also contenders for alternative vaccines . Synthetic peptide analogues produced by organic synthesis provide experimental immunogens whose potential for the third generation of vaccines appears promising . Among the following group of immunogenic synthetic peptide analogues (PA), PA (139-147) is more promising, because human antibodies in HBsAg-vaccinated individuals predominantly react with this amino acid sequence: HBsAg sequence PA 110-137, 117-137, 125-137, 134-146, 139-147, 138-149 . The N-terminal 22 amino acid sequence of the 55 residues preceding the HBsAg sequence, Pre-S (120-145), with the property of binding polymerized albumin . PA HBxAg sequence (100-115, 144-154) . PA'HBcAg sequence (73-84).

J Biol Chem, 1985 Dec 25, 260(30), 16156 - 61
The Euglena gracilis chloroplast ribulose-1,5-bisphosphate carboxylase gene . I . Complete DNA sequence and analysis of the nine intervening sequences; Gingrich JC et al.; The nucleotide sequence of 6225 base pairs (bp) of Euglena gracilis chloroplast DNA including the complete DNA sequence of the chloroplast-encoded ribulose-1,5-bisphosphate carboxylase large subunit gene along with the flanking DNA sequences is presented . The gene is greater than 5.5 kilobase pairs in length and is organized as 10 exons coding for 475 amino acids, separated by 9 introns . The exons range in size from 45 to 438 bp, while the introns range in size from 382 to 568 bp . The introns have highly conserved boundary sequences with the consensus, 5'-N GTGTGGATTT...(intron)...TTAATTTTAT N-3' . The introns are 82-85 mol% AT, with a pronounced T greater than A greater than G greater than C base bias in the RNA-like strand . They do not appear to encode any polypeptides . In addition, the introns have a conserved sequence 30-50 bp from their 3'-ends with the consensus, 5'-TACAGTTTGAAAATGA-3' . The 5'-TACA sequence bears some homology to the 5'-end of the TACTAACA sequence found in a similar location in yeast nuclear mRNA introns . The conserved sequences of the Euglena rbcL introns may be indicative of a splicing mechanism similar to that of eucaryotic nuclear mRNA introns and group II mitochondrial introns.

N Engl J Med, 1985 Dec 19, 313(25), 1586 - 90
The prospects for and pathways toward a vaccine for AIDS; Francis DP et al.; PIP: This article reviews prospects for a vaccine for acquired immunodeficiency syndrome (AIDS), considered the only certain barrier to further spread of the disease . The development of an effective vaccine against another retrovirus, feline leukemia virus, suggests the possibility that a retrovirus vaccine for humans might work . However, vaccine production depends on the ability to manufacture large quantities of a safe antigen that stimulates protective immunity when it is injected into humans . There are also many nonscientific problems that must be solved, including the reluctance of the private sector to invest in such a project . If it is found that the genetic variation of human T-lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV) is limited and predictable, vaccine-induced antibodies directed toward its structural proteins, especially envelope proteins, could protect against subsequent infection . Because retroviruses have a predilection for recombination and have been associated with malignant diseases, a live attenuated retrovirus vaccine is unlikely . A killed-virus vaccine should be more acceptable, but there are problems with selecting a cell line for producing the virus, choosing methods for purifying the viral antigens, and freeing them from viral nucleic acid . Cells derived from human tumors may be necessary to support the large-scale production of immunogenic antigens . Perhaps the best option at present is recombinant DNA technology . The advantage of antigen-production systems is that the infective virus cannot be produced because only a small portion of the viral genome is incorporated . Decisions must be made about which recombinant system to use (bacterial, yeast, or mammalian cells) and which antigen or antigens should be incorporated into the vaccine . Once expressed envelope proteins are available in adequate quantity and purity for testing, an evaluation pathway that will document safety and efficacy while minimizing the time for approval needs to be established .

Arch Tierernahr, 1985 Dec, 35(12), 847 - 53
{Initial research on the use of mixed corn grain and stalk silage in broiler fattening}; Jeroch H et al.; The suitability of a corn and cob maize silage (CCM) as a component rich in energy in the fattening ration was investigated in a growth experiment with male broilers . The CCM silage with a dry matter content of 60.2% contained the following substances determining its value (per 1 kg dry matter of the feed): 102 g crude protein, 61 g crude fibre and 682 energetic feed units for chickens (EFUc) . CCM silage was either fed as a component of a feed mixture consisting of 70.5% CCM silage and 29.5% protein concentrate (all values of original substance) or both components of the ration were fed to broilers separately . The protein concentrate was composed as follows (values per kg): 650 g soybean oilmeal, 110 g fishmeal, 85 g tankage from rendering plants, 55 g torula yeast, 70 g mineral mixture and 30 g of a mixture of biologically active substances . The broilers of the control group received commercial fattening feed for broilers with coarse maize meal as cereal component . The feed mixture with CCM silage and separate CCM silage were accepted readily . When CCM silage and protein concentrate were offered alternatively, the same amount of CCM silage was consumed but 8% less protein concentrate . The broilers fed with CCM silage plus protein concentrate reached 96-97% of the live weight of the control animals (1.952 g/animal, 49th day), which received conventional feed, although their net energy intake was 10% lower . This proves a more favourable net energy expenditure for the test groups fed with CCM silage.

Arch Biol Med Exp (Santiago), 1985 Dec, 18(3-4), 253 - 9
Search for compartments of glucose metabolism in the microinjected frog oocyte; Ureta T et al.; Microinjection of frog oocytes allows the modification of intracellular levels of substrates, intermediates, cofactors and enzymes . Use of labeled glucose at specific positions has led us to conclude that oocytes utilize glucose mainly for glycogen synthesis and to a lesser extent for the pentose-P pathway . Glycolysis, glycogenolysis and gluconeogenesis are not operative in these cells . The subject of compartmentation of glucose utilization has been addressed in this paper . First, we show that microinjection of glucose results in a 30-fold increase of carbon incorporation into glycogen when compared to oocytes incubated at saturating glucose concentrations . On the other hand, carbon incorporation into CO2, remains at about the same levels in both conditions Second, microinjection of NADP+ increases CO2 release and inhibits glycogen synthesis from glucose . Third, co-injection of unlabeled intermediates affects differentially glycogen synthesis and CO2 production from labeled glucose . Finally, microinjection of pure yeast hexokinase stimulates markedly 14CO2 release and inhibits glycogen synthesis . We conclude that two separate pools of glucose-6-P exists in oocytes: one pool is committed to the pathway of glycogen synthesis while a second pool serves as substrate for the operation of the pentose-P pathway.

Mol Cell Biol, 1985 Dec, 5(12), 3560 - 76
Characterization of the multigene family encoding the mouse S16 ribosomal protein: strategy for distinguishing an expressed gene from its processed pseudogene counterparts by an analysis of total genomic DNA; Wagner M et al.; Two genes from the family encoding mouse ribosomal protein S16 were cloned, sequenced, and analyzed . One gene was found to be a processed pseudogene, i.e., a nonfunctional gene presumably derived from an mRNA intermediate . The other S16 gene contained introns and had exonic sequences identical to those of a cloned S16 cDNA . The expression of this gene was demonstrated by Northern blot analysis of nuclear poly(A)+ RNA with cDNA and unique sequence intron probes . Each S16 intron contains a well-preserved remnant of the TACTAAC motif, which is ubiquitous in yeast introns and known to play a critical role in intron splicing . A sequence comparison with two other mouse ribosomal protein genes analyzed in our laboratory, L30 and L32, revealed common structural features which might be involved in the control and coordination of ribosomal protein gene expression . These include the lack of a canonical TATA box in the -20 to -30 region and a remarkably similar 12-nucleotide pyrimidine sequence (CTTCCYTYYTC) that spans the cap site and is flanked by C + G-rich sequences . The nature of the other members of the S16 family was evaluated by three types of experiment: a DNase I sensitivity analysis to measure the extent of chromatin condensation; an analysis of the thermal stability of cDNA-gene hybrids to estimate the extent of divergence of each gene sequence from that of the expressed gene; and a restriction fragment analysis which distinguishes intron-containing genes from intronless processed genes . The results of these analyses show that all genes except the expressed S16 gene are in a condensed chromatin configuration associated with transcriptional quiescence; that most of the genes within the S16 family have sequences greater than 7% divergent from the expressed S16 gene; and that at least 7 of the 10 S16 genes lack introns . We conclude that the ribosomal protein S16 multigene family contains one expressed intron-containing gene and nine inactive pseudogenes, most or all of which are of the processed type.

Mech Ageing Dev, 1985 Dec, 33(1), 103 - 13
Growth rate and life span in Drosophila V . The effect of prolongation of the period of growth on the total duration of life (J.H . Northrop, 1917)--revisited; Economos AC et al.; Sixty-eight years ago Northrop observed a constant life span in Drosophila after a progressive increase of the duration of development of the flies achieved by using a yeastless nutrient medium to which he added yeast with a progressively increasing delay . This evidence against the more recent concept of an increased life span following an experimentally decreased developmental rate has generally been ignored due, presumably, to the imprecise methodology employed by Northrop at a time that Drosophila research was just commencing . We describe here a study that aimed at re-examining and extending Northrop's work by developing the flies in either a yeastless or a lightly yeasted medium . While in a yeastless medium development of flies was virtually arrested until yeast was added, in the yeasted medium a slow growth of the larvae was possible before yeast was added . With another method, larval growth rate was reduced over the entire developmental period by adding a relatively low amount of yeast in four portions and with various delays between portions (the first portion being added without delay) . Our study confirmed the "Northrop-effect", i.e . the absence of an effect on life span from increased duration of development by a virtual arrest of growth of the larvae for a number of days . Further, it showed that manipulation of growth rate by portioning the yeast amount did not unequivocally support the concept that a lower growth rate leads to an increased life span.

J Am Mosq Control Assoc, 1985 Dec, 1(4), 506 - 10
Feeding rate of larval Aedes vexans stimulated by food substances; Aly C; Feeding rates of fourth instar larvae of Aedes vexans were compared by counting substrate filled gut segments after exposure to food or inert particles . Food particles (wheat flour, fishmeal or yeast) were ingested approximately 3 times faster than inert particles (kaolin, pumice or synthetic cellulose) . Aqueous fishmeal extract accelerated ingestion of inert particles to the level of ingestion of food particles, demonstrating gustatory stimulation of larvae . Absolute amounts of ingested materials were calculated on the basis of dry weights of guts completely filled with test substrates . At 21 degrees C, 46-74 micrograms of food particles, and 15-33 micrograms of inert particles were ingested per larva during 10 min of exposure . In the subsequent time span, ingestion rates of food particles decreased continuously with satiation of larvae.

Arch Biol Med Exp (Santiago), 1985 Dec, 18(3-4), 317 - 23
Pyruvate kinase: studies on affinity labeling and active-site structure using the rabbit muscle enzyme; Eyzaguirre J et al.; Important advances have been made in recent years in the study of the structure of pyruvate kinase: the amino acid sequence of the enzymes from chicken muscle and yeast have been established and the three-dimensional structure of the cat muscle enzyme has been determined at 0.26 nm resolution . Work in our laboratory has shown that dialdehyde-ADP (oADP) can be used as an affinity label of rabbit muscle pyruvate kinase: if the enzyme is incubated with cold oADP in the presence of high ADP concentrations, dialyzed and then incubated with 14C-oADP, the enzyme inactivates and one mole of radioactive oADP incorporates per mole of enzyme subunit . A labeled peptide with a molecular weight of about 5900 has been purified from a tryptic digest of the modified enzyme . The first 26 residues of the peptide have been sequenced and this sequence is identical to a region in the chicken muscle enzyme and a peptide isolated from the bovine muscle enzyme specifically labeled with trinitrobenzenesulfonate . High homology is also found with a region of the yeast enzyme . All this suggests that the isolated peptide is part of the active site; the modified amino acid, probably a lysine, seems to be located in one of the alfa helices of domain A of the enzyme, according to the x-ray data.

An Acad Bras Cienc, 1985 Dec, 57(4), 497 - 506
Kinetic study of hemoglobin Porto Alegre {beta 9(A6)Ser----Cys} disulfide polymer reduction; Tondo CV et al.; The cleavage of HbPA disulfide polymer by GSH and its indirect cleavage by yeast glutathione reductase, via reduced glutathione is obtained . Decreasing the initial proportion of GSH in the hemolysate increases the formation of HbPA disulfide polymer . In the experimental conditions used, yeast glutathione reductase is unable to perform the direct cleavage of the mixed disulfide of HbPA and GSH, using it as substrate . The reduction of HbPA polymer to tetramers by DTE is analyzed by a pseudo-first-order kinetic and two rate constants are obtained . That of 265 X 10(-3) min-1 should be concerned with one disulfide of the closed ring and one of the open ring structure of dodecamer, while that of 38 X 10(-3) min-1 is related to disulfide reduction of the octamer . The enthalpy of activation values of 8.0 kcal.mol-1 an 17.4 kcal.mol-1 obtained, from the Arrhenius plot, for the "fast" and "slow" rate disulfide reduction, respectively, are indicative that a strong conformational strain of S--S bonds in the closed ring structure is maintained . The entropy of activation values of 24 e.u . and 52 e.u . are found for the activation of disulfides from dodecamers and octamers, respectively.

Arch Biol Med Exp (Santiago), 1985 Dec, 18(3-4), 331 - 58
{Concurrence of multiple and integrated mechanisms in the modulation of enzyme activities: significance for the regulation of metabolic fluxes}; Niemeyer H et al.; The activity of some enzymes in a given metabolic pathway is modulated through multiple mechanisms, which operate in a simultaneous and coherent way to produce either stimulation or inhibition . The operation of these mechanisms is illustrated with several enzymes involved in glucose metabolism, by choosing examples from the presentations at the Symposium . Thus the reciprocal interactions of the regulatory mechanisms acting upon hexokinase D ('glucokinase'), phosphofructokinase, fructose 1,6-bisphosphatase and pyruvate kinase were discussed, as well as their relationships with the induction of enzyme conformational changes . In addition, the effects of covalent interconversions on glutamine synthetase activity were briefly analyzed . An outstanding feature exhibited by all these enzymes is the display of a great number of elasticity coefficients, which are differential quotients measuring the dependence of enzymatic activity on each variable that modulates it . A general assumption is that these enzymes make an important contribution to the control of the metabolic flux in which they participate . The flux control, however, appears to be shared in different degrees by all the components of the system, and may be quantified through the differential quotient denominated control coefficient . Some of the problems that emerge in any attempt to estimate these coefficients in the living cells are discussed . The problems derive partly from the complex subcellular structure, the formation of functional compartments resulting from reversible association of the enzymes, one to another and to different cellular components, and the actual state of cell water . These problems make that the results obtained with purified and highly diluted enzymes in most enzymological studies should not be extrapolated directly to what happens in vivo, without a careful evaluation of each particular case . The regulatory role of enzyme activity of fructose 2,6-bisphosphate and its eventual participation as an intermediary in the hormonal control of glycolytic and gluconeogenic fluxes are emphasized . The regulation of yeast fructose 1,6-bisphosphate activity is discussed in relation to the eventual role of allosteric modulators and covalents interconversions as signals for the initiation of intracellular degradation of the enzyme during catabolic inactivation.

J Clin Invest, 1985 Dec, 76(6), 2368 - 76
Interaction of human monocytes, macrophages, and polymorphonuclear leukocytes with zymosan in vitro . Role of type 3 complement receptors and macrophage-derived complement; Ezekowitz RA et al.; Macrophages take up zymosan in the absence of exogenous complement via receptors for iC3b (type 3 complement receptors) acting with or without lectin-like receptors for mannosyl-fucosyl-terminated glycoconjugates . We previously provided evidence that macrophages themselves secrete complement-alternative pathway components able to opsonize zymosan locally (Ezekowitz et al., J . Exp . Med . 1984 . 159:244-260) . We show here that covalently bound C3 cleavage products C3b and iC3b can be eluted from zymosan particles cultivated with 36-h adherent human monocytes in the absence of serum . The ligand binding site of type 3 complement receptors is involved in macrophage-zymosan interactions as shown by inhibition studies of zymosan binding and uptake with Fab fragments of anti-C3 antibodies and monoclonal antireceptor antibodies M01 and OKM10 . In contrast, antibody IB4, which binds to a receptor epitope distinct from the binding site, does not inhibit zymosan uptake . Selective modulation of macrophage receptors onto anticomplement receptor antibody and mannose-rich yeast mannan, respectively, confirms that the complement and lectin-like receptors are distinct . Human polymorphonuclear leukocytes, which express receptors for complement, but are not known to secrete complement proteins, bind and ingest only exogenously opsonized zymosan . Unopsonized zymosan is a poor trigger of respiratory burst activity in neutrophils or 7-d adherent human macrophages, but induces cell aggregation and secretion of large amounts of superoxide anion when these cells are co-cultivated in serum-free medium and challenged with zymosan . Our studies indicate that complement and/or other products synthesized by macrophages at extravascular sites could play an important role in opsonization and lysis of pathogens able to activate the alternative pathway and mediate macrophage-neutrophil collaboration in first-line host defence.

J Mol Biol, 1985 Nov 20, 186(2), 321 - 35
Organization of the actin multigene family of Dictyostelium discoideum and analysis of variability in the protein coding regions; Romans P et al.; There are 17 to 20 actin genes in the genome of the cellular slime mold Dictyostelium discoideum . Genomic clones of 15 of the genes have been isolated . Extensive nucleotide sequence within the protein-coding regions has been determined, including the complete nucleotide sequence of four genes representing the three distinct evolutionary groups of Dictyostelium actin genes . All are similar to mammalian cytoplasmic actins at diagnostic amino acid positions, and there is generally less variability among Dictyostelium actin genes than among Drosophila actin genes . Two genes, Actins 3-sub 1 and 3-sub 2 differ substantially from all the rest in terms of replacement amino acid substitutions and probably encode actin-related proteins rather than bona fide actins . Each contains several amino acid substitutions that should alter the secondary structure of the resulting proteins, and Actin 3-sub 2 encodes four additional amino acids at the C terminus . This gene is as divergent from other Dictyostelium actin genes as is the yeast or a soybean actin gene . At present, evidence suggests that all 15 genes examined are expressed, except the previously identified Actin 2-sub 2 . We suggest that Dictyostelium might maintain a high number of functional actin genes for the purpose of regulating the level of actin synthesis within narrow limits, rather than because most genes perform different functions.

Experientia, 1985 Nov 15, 41(11), 1421 - 2
Lowering of liver acetaldehyde but not ethanol concentrations by pretreatment with taurine in ethanol-loaded rats; Watanabe A et al.; A rise in blood and liver acetaldehyde concentrations following ethanol loading (1.5 g/kg b.wt) was significantly reduced when rats were pretreated orally with taurine (0.5 g/kg), a potent in vitro activator of yeast aldehyde dehydrogenase . This taurine pretreatment produced a 4-fold increase in liver taurine content.

Arch Biochem Biophys, 1985 Nov 15, 243(1), 46 - 61
Purification and properties of rabbit liver cathepsin M and cathepsin B; Erickson-Viitanen S et al.; Cathepsins M and B from rabbit liver lysosomes were separated by chromatography on Ultrogel AcA34 at low ionic strength and purified to homogeneity, and their catalytic and molecular properties were compared . Cathepsin M was relatively inactive with synthetic peptide substrates . Thus, it hydrolyzed benzoyl arginine naphthylamide at only one-fifth the rate observed with cathepsin B, and no activity was detected with Gly-Phe naphthylamide which is a relatively good substrate for cathepsin B . On the other hand, cathepsin M exhibited a preference for protein substrates . It was more active than cathepsin B in catalyzing the inactivation of the following enzymes: rabbit muscle or liver fructose-1,6-bisphosphate aldolases, rabbit liver fructose-1,6-bisphosphatase and pyruvate kinase, yeast glucose-6-phosphate dehydrogenase, and rabbit muscle glyceraldehyde-3-phosphate dehydrogenase . With glucagon as substrate, both enzymes showed similar peptidyl dipeptidase activities with some minor differences in peptide bond specificity . Cathepsins M and B are similar in size, with apparent molecular weights of 30,200 for cathepsin M and 28,800 for cathepsin B, and in amino acid composition and carbohydrate content . Each contains approximately 2-3 equivalents/mol glucosamine, 3 equivalents/mol mannose, and no fucose or galactosamine . They also show similar microheterogeneity in sodium dodecylsulfate-gel electrophoresis and isoelectric focusing; this microheterogeneity is probably related to differences in glycosylation . Extensive homology in primary structure for the two proteins was indicated by the similar patterns of peptides formed on digestion with trypsin.

J Biol Chem, 1985 Nov 15, 260(26), 14173 - 9
Fructose-1,6-bisphosphatase from rat liver . A comparison of the kinetics of the unphosphorylated enzyme and the enzyme phosphorylated by cyclic AMP-dependent protein kinase; Ekdahl KN et al.; A purification procedure for rat hepatic fructose-1,6-bisphosphatase, described earlier, has been improved, resulting in an enzyme preparation with a neutral pH optimum and with both phosphorylatable serine residues present . The subunit Mr was 40,000 . Phosphorylation in vitro with cyclic AMP-dependent protein kinase resulted in the incorporation of 1.4 mol of phosphate/mol of subunit and led to an almost 2-fold decrease in apparent Km for fructose-1,6-bisphosphate . In contrast to yeast fructose-1,6-bisphosphatase, fructose-2,6-bisphosphate had no effect on the rate of phosphorylation or dephosphorylation of the intact enzyme . The effects of the composition of the assay medium, with regard to buffering substance and Mg2+ concentration, on the apparent Km values of phosphorylated and unphosphorylated enzyme were investigated . The kinetics of phosphorylated and unphosphorylated fructose-1,6-bisphosphatase were studied with special reference to the inhibitory effects of adenine nucleotides and fructose-2,6-bisphosphate . Unphosphorylated fructose-1,6-bisphosphatase was more susceptible to inhibition by both AMP and fructose 2,6-bisphosphate than phosphorylated enzyme, at high and low substrate concentrations . Both ATP and ADP had a similar effect on the two enzyme forms, ADP being the more potent inhibitor . Finally, the combined effect of several inhibitors at physiological concentrations was studied . Under conditions resembling the gluconeogenic state, phosphorylated fructose-1,6-bisphosphatase was found to have twice the activity of the unphosphorylated enzyme.

Eur J Biochem, 1985 Nov 4, 152(3), 625 - 31
Ribosomes are stalled during in vitro translation of alfalfa mosaic virus RNA 1; Lindhout P et al.; In the presence of plant tRNAs the full-length translation product of alfalfa mosaic virus RNA 1 is produced in rabbit reticulocytes only at low mRNA concentration . At higher mRNA concentration translation is restricted to the 5' half of RNA 1 . At high mRNA concentration the full-length product can be formed when additional plant tRNA and glutamine are supplied to the translation mixture . In contrast, in the presence of yeast or calf liver tRNA the translation pattern of alfalfa mosaic virus RNA 1 always results in the synthesis of the full-length product . Pulse-chase experiments in the presence of plant tRNAs show that the ribosomes pause at several positions in the 5' half of RNA 1 . The pausing time is different at the different 'halting places' . Protein synthesis is resumed upon addition of glutamine, even when the addition is delayed for more than 3 h after the start of protein synthesis . Only one tRNA species, purified from wheat germ or tobacco, could promote full-length translation of RNA 1 . This tRNA can be charged with glutamine . Analysis of the position of glutamine codons on RNA 1 shows a correlation between the positions of the CAA codons and the halting places of the ribosomes . The CAA codon (for any other codon) on its own cannot be responsible for the pausing of the ribosomes, since a variety of RNAs, known to contain all sense codons, are translated efficiently in rabbit reticulocyte lysates in the presence of plant tRNAs . Apparently other elements can restrict decoding of normal codons during protein chain elongation.

Vopr Virusol, 1985 Nov-Dec, 30(6), 697 - 700
{Comparative study of the antiviral activity of natural double-stranded RNAs in experimental tick-borne encephalitis}; Knoroz MIu et al.; A comparative study of the antiviral effect of interferon inducers from the group of natural double-stranded RNAs (yeast plasmid dsRNA, phage phi 6 and phage f2 dsRNAs was carried out on the model of experimental tick-borne encephalitis . The possibility of inducing up to 60% protection against 10 LD50 of TBE virus by prophylactic inoculation of interferon inducers alone was demonstrated . The therapeutic effect was observed only with yeast dsRNA (30% protection when the inducer was administered 4 hours after virus infection of mice) . The prophylactic effect of inoculation of interferon inducers (yeast dsRNA and f2 dsRNA) to immune mice correlated with the protective effect of inducers alone . At the same time, the therapeutic effect of inoculation of yeast dsRNA to immune animals (4 hours after TBE virus, 40% protection) was more marked . In the therapeutic use of f2 dsRNA for pre-immunized animals a synergetic effect of the preparation was observed (56.7% protection in 4 hours and 26.7% protection 24 hours postinfection).

Mech Ageing Dev, 1985 Nov, 32(2-3), 193 - 204
Growth rate and life spain in Drosophila . IV . Role of cell size and cell number in the biphasic relationship between life span and growth rate; Economos AC et al.; The patterns of variation of wing cell size and number were studied under developmental conditions leading to a biphasic relationship between life span and growth rate while duration of development remained constant (development on an agar-only medium with a varying added yeast amount, constant temperature (25 degrees C) and constant larval density) . Across the yeast range, a 125% increase of body weight was accompanied by a roughly 30% increase in the wing linear dimensions, wing cell size and wing cell number while estimated duration of cell division and its reciprocal mitotic division rate remained constant . Furthermore, cell size (but not cell number) varied with growth rate in a similar biphasic pattern to that observed for life span . Finally, from a simultaneous examination of the covariation patterns of life span, growth rate, cell size and cell number with decreasing yeast amount, it became apparent that there was a "critical" yeast amount, approximately 125 mg/120 eggs, below which: (a) cell number abruptly started to decrease linearly from a roughly constant value; (b) the rate of the slow decrease of cell size now tripled and that of growth rate increased even more; and (c) life span which, in the upper yeast range, increased slowly with decreasing yeast, apparently reached a maximum at the critical yeast level and decreased three times faster below that level . These data taken together suggest that: (i) the decrease of all parameters (including life span) below the critical yeast level results from a presumably suboptimal or disturbed development because of and in proportion to the lack of nutrients and (ii) the increase of life span with decreasing yeast amount above the critical yeast level has not been definitely explained but some possibilities are suggested such as changes in subcellular organelle numbers, size and/or functional properties, or other changes due to a phenomenon equivalent to food restriction in rats, probably without changes in overall metabolic rate of the flies.

Am J Clin Nutr, 1985 Nov, 42(5), 829 - 35
Selenium status of exclusively breast-fed infants as influenced by maternal organic or inorganic selenium supplementation; Kumpulainen J et al.; A longitudinal dietary Se supplementation study on lactating mothers was performed to determine the possibilities of improving the Se status of exclusively breast-fed infants . A total of 200 mothers randomized into three groups received either no Se supplements, 100 micrograms of selenite, or 100 micrograms of yeast-Se daily . Maternal and infant serum Se concentrations showed a linear correlation during exclusive breast-feeding . Yeast-Se in the dose administered was safe and more effective than selenite in increasing the Se concentrations of maternal serum and milk, and infant serum . The mean estimated daily Se intakes of the infants were 7.7 +/- 2.2, 8.9 +/- 2.2, and 11.5 +/- 4 micrograms, in the control, selenite, and yeast-Se groups respectively . Though the infant Se intakes of the unsupplemented and selenite-supplemented mothers were below the lower limit of the safe and adequate range as set by the US National Research Council, their serum Se concentrations increased steadily over the 6-mo study period . As maternal serum Se also increased by over 50% during the same period the results suggest that a maternal daily intake of 50-75 micrograms is adequate during lactation.

J Immunol, 1985 Nov, 135(5), 3388 - 93
Properties of glycans that activate the human alternative complement pathway and interact with the human monocyte beta-glucan receptor; Czop JK et al.; The glycans used in an earlier study to define the ligand specificity of the human monocyte phagocytic receptor for unopsonized particulate activators were assessed for their capacities to activate the proteins of the human alternative complement pathway . Normal human serum was preincubated with glycans under conditions of chelation to prevent activation of the classical complement pathway, and the activation-depletion of the alternative complement pathway was determined by the subsequent capacity of the serum to lyse rabbit erythrocytes (Er) . When serum was preincubated at a 1/2 dilution in 8 mM EGTA/2 mM Mg with increasing numbers of yeast glucan or zymosan particles, and was evaluated at final serum dilutions of 1/8, its capacity to lyse Er was found to be reduced by 50% with 1.9 X 10(6)/ml yeast glucan particles and 1.4 X 10(6)/ml zymosan particles . At 2 mg/ml of serum diluted 1/2 in 8 mM EGTA/2 mM Mg, nonturbid preparations of mannan, laminarin, or pyrogen-free inulin and turbid suspensions of cellulose, Sephadex, agarose, or purified inulin failed to activate the alternative complement pathway . In contrast, activation-depletion of the alternative pathway was induced by turbid preparations of crude inulin, nigeran, pachyman, barley beta-glucan, and pustulan, which at 700 micrograms/ml, 500 micrograms/ml, 350 micrograms/ml, 60 micrograms/ml, and 27 micrograms/ml, respectively, effected 50% reductions in the subsequent lysis of Er . After centrifugation of 2 mg/ml suspensions of barley beta-glucan at 1100 X G for 5 min and at 15,000 X G for 15 min, the supernatants contained 90 to 92% and 65% of the barley beta-glucan, respectively, as determined by the anthrone method . On a weight basis, the 1100 X G supernatant exhibited the same capacity to activate the alternative pathway as the corresponding original suspension, whereas the 15,000 X G supernatants had less than 3% of the original anti-complementary activity . Preincubation of adherent human monocytes with increasing concentrations of barley beta-glucan suspensions, 100,000 X G supernatants containing 64% of the original beta-glucan, and laminarin all decreased subsequent ingestion of 1.25 X 10(6) zymosan particles in a dose-related fashion . The numbers of monocytes from three different donors phagocytosing zymosan were reduced by 50% after pretreatment with 30 to 65 micrograms/ml, 25 to 48 micrograms/ml, and 12 to 15 micrograms/ml of barley beta-glucan suspensions, 100,000 X G supernatants of barley beta-glucan, and laminarin, respectively, even though the latter two preparations were fully soluble and had no capacity to activate the alternative pathway.(ABSTRACT TRUNCATED AT 400 WORDS)

J Lab Clin Med, 1985 Nov, 106(5), 573 - 7
Limiting role of 6-phosphogluconolactonase in erythrocyte hexose monophosphate pathway metabolism; Beutler E et al.; The natural product of the glucose-6-phosphate dehydrogenase reaction is 6-phosphoglucono-delta-lactone, which must be hydrolyzed to 6-phosphogluconic acid before it can be further metabolized by 6-phosphogluconate dehydrogenase . Because this lactone is very unstable, it has been uncertain whether the enzyme that hydrolyzes it, 6-phosphogluconolactonase, is required for functioning of the hexose monophosphate pathway . We have purified glucose-6-phosphate dehydrogenase, 6-phosphogluconolactonase, and 6-phosphogluconate dehydrogenase from human erythrocytes to the point where each enzyme is essentially free of each of the other activities . We constructed an artificial hexose monophosphate pathway from these enzymes, providing as substrate 14C-labeled glucose-6-phosphate either directly or by continual generation from 14C-glucose by yeast hexokinase and adenosine triphosphate . The oxidation of 6-phosphogluconic acid was estimated by measuring the CO2 formed . In the absence of a reduced nicotinamide-adenine dinucleotide phosphate (NADPH)-oxidizing system, such as oxidized glutathione (GSSG)-glutathione reductase or phenazine methosulfate, little CO2 was formed, and the presence of 6-phosphogluconolactonase did not affect the amount that was produced . When the hexose monophosphate pathway was stimulated by providing an NADPH-oxidizing system, CO2 was produced two and a half to five times as fast in the presence of 6-phosphogluconolactonase as in its absence . These studies suggest that 6-phosphogluconolactonase is required for the functioning of the hexose monophosphate pathway when the rate of oxidation of NADPH is accelerated.

Allergol Immunopathol (Madr), 1985 Nov-Dec, 13(6), 463 - 9
Mononuclear phagocytic cells in peritoneal exudates of guinea pigs: a comparison of inducing agents; Ganguly R et al.; Peritoneal macrophages are used for immunologic assessment of the lymphokine, migration inhibitory factor (MIF) . For this purpose, these cells are induced intraperitoneally in animals with inflammatory agents such as mineral oil (MO) and peptone water (PW) . Purpose of the present study was two-fold: To assess the changes in biologic properties of guinea pig macrophages induced peritoneally with MO or PW as compared to control cells after administration of normal saline (NS); and to examine the suitability of induced macrophages to respond to MIF in vitro . Peritoneal exudate cells were harvested from guinea pigs 7 days following intraperitoneal injection of MO, PW or NS . They were enumerated in hemocytometers, their differential counts determined by Wright's stain and morphologic characteristics assessed by scanning electron microscopy . Functional activation of peritoneal macrophages was determined by lysosomal enzyme functions, as well as by yeast phagocytosis in vitro . Random cell migration and responses to migration inhibitory factor (MIF) were determined in Mackaness chambers . Mineral oil injection resulted in significantly higher yield of peritoneal macrophages . Greater than 70% of peritoneal exudate cells were macrophages in all three groups . Spread out structures and ruffled borders were seen in electron micrographs of MO induced cells . Such structures were less evident in PW induced cells and were absent in controls . Acid phosphatase (ACP) and beta-N-acetylglucosaminidase (B-NAG) activities as well as yeast phagocytosis significantly increased in MO and PW induced cells . Random migration and responses to MIF in Mackaness chambers remained comparable in the three experimental groups.

Clin Exp Immunol, 1985 Nov, 62(2), 270 - 7
Binding of C3 fragments to the Trypanosoma cruzi surface in the absence of specific antibodies and without activation of the complement cascade; Krettli AU et al.; Bloodstream trypomastigotes (BTry) of Trypanosoma cruzi were found to bind to their surface membrane in-vitro fragments of C3, namely C3b and/or C3bi . Different reagents specific for C3 were used: anti-C3c serum and anti-C3 monoclonal antibodies; bovine conglutinine and yeast particles . The BTry, isolated from acutely infected mice and maintained on a complement-free culture medium (TC-199), were strongly agglutinated by these reagents or formed clumps with the yeast only after being previously treated with fresh or heat inactivated normal human serum . It is suggested that the binding of C3b fragments to the parasite membrane may influence the extent or the nature of the early events during T . cruzi infection allowing the BTry to escape complement destruction by the alternative complement pathway.

Arch Int Pharmacodyn Ther, 1985 Nov, 278(1), 23 - 44
Pharmacological effects of SCH 30497--a novel analgesic substance; Chipkin RE et al.; SCH 30497 (2-{3-(1,2,3,6-tetrahydro-4-2(methylphenyl)-pyridin-1-yl) -propyl}-1,2,4-triazolo{4,3-a} pyridin-3-(2H)-one) was tested for analgesic effects in the rat, mouse and squirrel monkey . SCH 30497 showed dose-related analgesic effects in the rat yeast-paw test; at peak times the ED7sec (95% confidence limits) in mg/kg via the oral, subcutaneous, intramuscular and intravenous routes of administration were as follows, respectively: 4.7 (2.1-9.8), 5.7 (3.4-9.6), 6.4 (4.1-9.8) and 1.4 (0.6-3.2) . SCH 30497 was also analgesic in the acetic acid-induced writhing test in mice (ED50 = 1.9 mg/kg p.o.) and the squirrel monkey shock titration test (ED50 = 14.5 mg/kg p.o.) . It was inactive in the mouse (100 mg/kg p.o.) and rat (40 mg/kg p.o.) tail-flick tests . Thus, SCH 30497 was efficacious versus chemical, mechanical and electrical nociceptive stimuli . Naloxone antagonism of the analgesic effects of SCH 30497 was species specific with significant inhibition observed only in the rat and not in the mouse or monkey . SCH 30497 did not produce Straub tail or hyperactivity in mice . Twice daily dosing at 30 mg/kg p.o . to rats for 5 days failed to produce tolerance; in separate experiments, daily injections for 10 days at 20 or 100 mg/kg p.o . failed to induce signs of dependence following naloxone challenge . SCH 30497-induced analgesia was not attenuated in rats previously made tolerant to narcotics by implantation of a morphine pellet . SCH 30497 showed a weak ability to displace 3H-Met5-enkephalin from its binding sites on rat brain membranes (IC50 = 48 microM) . SCH 30497 (100 microM) did not affect prostaglandin synthesis in vitro . In vivo, the drug did not have anti-inflammatory or ulcerogenic effects up to 80 mg/kg p.o . Acute behavioral, neurological and autonomic side effects were primarily depressant in rodents and occurred at doses greater than 15 times those that were analgesically relevant . Moderate doses in the monkey (2.5 times the ED50) and high doses in mice produced convulsions . It is hypothesized that SCH 30497-like drugs represent a new class of analgesics based on this unique pharmacological spectrum of activity.

Biochem Biophys Res Commun, 1985 Oct 30, 132(2), 780 - 6
The mitogenic/oncogenic p21 Ki-RAS protein stimulates adenylate cyclase activity early in the G1 phase of NRK rat kidney cells; Franks DJ et al.; tsK-NRK rat cells infected with a temperature-sensitive mutant of the Kirsten murine sarcoma virus were arrested in the G0/G1 phase of their cell cycle by incubation in serum-deficient medium at a temperature (41 degrees C) which inactivates the virus' abnormally thermolabile mitogenic/oncogenic 21 kDa (p 21) RAS protein product . Reactivating the viral RAS protein by lowering the temperature to a permissive 36 degrees C rapidly (within 1 hour) stimulated adenylate cyclase, sensitized the enzyme to stimulation by GTP and forskolin and caused the tsK-NRK cells to transit G1 and start replicating their DNA about 10 hours later . The 41 degrees C----36 degrees C shift did not affect adenylate cyclase or stimulate G1 transit in uninfected NRK cells . Thus, an oncogenic viral RAS protein was able to stimulate adenylate cyclase and G1 transit in a mammalian cell just as other RAS proteins appear to do in yeast cells.

J Biol Chem, 1985 Oct 15, 260(23), 12769 - 72
Structural organization of ribosomal RNAs from Novikoff hepatoma . I . Characterization of fragmentation products from 40 S subunit; Choi YC; The internal topography of rRNAs in situ was probed with RNase T1 . Isolated polysomes were treated with RNase T1 to examine the unprotected regions of rRNAs contained in polysomes . The 40 S subunit yielded 23 cleavage products of 18 S rRNA, of which the fragments from three domains were isolated, characterized, and compared with the secondary structure of 18 S rRNA proposed by Chan et al . (Chan, Y.-L., Gutell, R., Noller, H.F., and Wool, I.G . (1984) J . Biol . Chem . 259, 224-230) . There were two consistent fragments (1-71, 1-74) derived from the 5' domain, where two alternative sites were cleaved at a loop, indicating conformational flexibility of 40 S subunit . There was a fragment (1760-1874) consisting of the 3'-end portion derived from the 3'-minor domain where a single site was cleaved at a loop . These patterns of cleavages at the single-stranded regions are similar to those of bacterial 16 S rRNA . In contrast to the phylogenetic similarity of cleavages between 16 S rRNA and 18 S rRNA (Stiegler, P., Carbon, P., Zucker, M., Ebel, J . P., and Ehresmann, C . (1981) Nucleic Acids Res . 9, 2153-2172), a difference was found in one fragment (777-840) derived from the unassigned long insert of the central domain . Based on the determination of its cleavage sites, a secondary structure model is proposed, which conserves a phylogenetic consistency among yeast, Xenopus, rat, and rabbit.

Nature, 1985 Oct 10-16, 317(6037), 551 - 5
Site-directed mutagenesis shows that tyrosine 248 of carboxypeptidase A does not play a crucial role in catalysis; Gardell SJ et al.; The residue Tyr 248 of carboxypeptidase A (CPA) is thought to play a role in catalysis by contributing a proton to the incipient amine anion generated during cleavage of peptide substrates . To test this hypothesis we have modified the rat CPA cDNA by site-directed mutagenesis so that the codon for Tyr 248 is replaced by that for Phe . Here, we report the expression of the cDNAs for proCPA and its Tyr-to-Phe variant in yeast via the alpha-factor system . Following zymogen activation by trypsin, wild-type CPA (CPA-WT) and variant CPA (CPA-Phe 248) were purified to homogeneity and characterized enzymatically . CPA-Phe 248 displays essentially undiminished values for the catalytic constant (kcat) towards various peptide and ester substrates . However, the Michaelis constants (Km values) of peptide substrates and the inhibition constant (Ki) of the potato carboxypeptidase inhibitor are increased 6-fold and 70-fold, respectively . These data suggest that the phenolic hydroxyl of Tyr 248 does not act as the requisite general acid catalyst but participates in ligand binding.

J Biol Chem, 1985 Oct 5, 260(22), 12142 - 7
Effect of divalent metal ions and pH upon the binding reactivity of human serum amyloid P component, a C-reactive protein homologue, for zymosan . Preferential reactivity in the presence of copper and acidic pH; Potempa LA et al.; The serum amyloid P component (SAP) has been found to associate in vitro with a variety of polysaccharide and proteinaceous ligands including the yeast cell wall polysaccharide preparation, zymosan, in the presence of calcium at neutral pH . In the present study, we have investigated the role of copper and zinc and other divalent cations and acidic pH on the binding of SAP to zymosan . We report that binding occurs not only in the presence of calcium, but in the presence of copper, zinc, and cadmium as well . No binding occurs in the absence of added metal, or in the presence of barium, cobalt, magnesium, manganese, or nickel . 125I-SAP binding in the presence of metals is inhibited by presaturating the zymosan surface with unlabeled SAP . Whereas calcium-mediated binding decreases by more than 50% as the pH is lowered to 5, copper-mediated binding increases substantially at the more acidic pH values while zinc-mediated binding is essentially unchanged . These data indicate that, in addition to calcium at neutral pH, copper (and zinc) at neutral and particularly acidic pH values mediates SAP binding to polysaccharide ligands . This suggests that SAP may well be considered a copper- as well as a calcium-dependent protein under certain conditions and that this reactivity is favored under those conditions of lowered pH which may result from metabolic processes occurring at local sites of inflammation.

Mycopathologia, 1985 Oct, 92(1), 37 - 43
Association of anurans with pathogenic fungi; Mok WY et al.; In a study of 450 Amazonian anurans, we isolated yeasts and yeast-like fungi from 54 animals (Bufo granulosus, B . marinus, Dendrophrynyscus sp., Hyla geographica, H . lanciformes, Ololygon rubra, Adenomera hylaedactyla, Eleutherodactylus fenestratus, Leptodactylus fuscus, L . ocellatus, L . pentadactylus) . The internal organs of these animals did not show any macroscopic anomaly nor histopathology . We recovered 105 fungal isolates from the anuran liver, lung, kidney, spleen, heart and gonad . The isolates were made up of 30 fungal species, 9 of which (48 isolates, 46%) were fungi with known pathogenic potentials, namely: Candida guilliermondii, C . parapsilosis, C . tropicalis, C . glabrata, Geotrichum candidum, Aureobasidium pullulans, Wangiella dermatitidis, Trichosporon cutaneum and Exophiala werneckii . Eleven animals harbored identical fungi in more than one of their internal organs; seven animals had more than one fungal species colonizing a single organ . Our findings indicated probable natural subclinical infections of candidiasis, geotrichosis or phaeohyphomycosis, and also symbiotic presence of non-pathogenic fungi among neotropical anurans.

EMBO J, 1985 Oct, 4(10), 2617 - 26
(AT)n is an interspersed repeat in the Xenopus genome; Greaves DR et al.; We have observed (AT)34 and (AT)23 tracts close to the coding sequences of the Xenopus laevis tadpole alpha T1 and adult beta 1 globin genes, respectively . We show that (AT)n sequences are found as interspersed repeats within the Xenopus globin and histone gene loci . Using (AT)n co-polymer in filter hybridisation experiments we estimate that there are 10(4) (AT)n tracts per haploid Xenopus genome . Hybridisation to genomic blots of DNA from yeast, slime mold, trypanosome, fruit fly, salmon, chicken, rat, human, crab and Xenopus species shows that strictly alternating AT of sufficient length to hybridise appears to be most abundant in Xenopus and crab genomes . We show that the specificity of the co-polymer probe for strictly alternating AT is, however, dependent on the length of the probe . Hybridisation experiments using (TG)n copolymer suggest that this highly conserved repeat is found as clustered repeats in the Xenopus genome in contrast to other eukaryotic genomes so far studied.

Infect Immun, 1985 Oct, 50(1), 183 - 9
Evolution of inflammatory response and cellular immune responses in a murine model of disseminated blastomycosis; Deepe GS Jr et al.; A reproducible model of disseminated blastomycosis was established in C57BL/6 mice by intravenous injection of 10(6) yeast-phase Blastomyces dermatiditis organisms . The infection progressed over 5 weeks to involve lungs, brains, superficial fascia, livers, and spleens of mice . By week 5, there was a greater number of organisms in lungs and brains than in livers and spleens . The tissue response in lungs, brains, and livers progressed from acute neutrophilic invasion before week 1 to pyogranuloma formation by week 5 . Lymph nodes and spleens were remarkably spared . By week 5, infected mice became anergic to intradermal challenge with both specific Blastomyces antigen and a nonspecific antigen (sheep erythrocytes) . At this time, the response to concanavalin A or phytohemagglutinin by splenocytes was markedly less than that of normal controls . Likewise, the plaque-forming cell response to sheep erythrocytes by splenocytes from infected mice was diminished . In coculture studies, splenocytes from 5-week-infected mice reduced the plaque-forming cell response by normal splenocytes . The development of this murine model should prove useful for elucidating the perturbations of immunoregulation associated with disseminated blastomycosis.

Chemioterapia, 1985 Oct, 4(5), 406 - 12
In vivo tests for antimycotic drugs; Gargani G et al.; Authors emphasize the pathogenetic importance of fungous dimorphism and its consequence in the usefulness of antimycotic drugs . For the study of drugs' activity on C . albicans, two models for the evaluation on yeast or mycelial form were developed . The first model is of candida growth on an animal cell culture, where a good mycelial form develops during the first 24 hours: authors demonstrated a synergism of the amphotericin B-rifampin combination, which is less evident in the traditional in vitro tests, and some activity of tolcyclate . The second model is an intracutaneous injection in the guinea pig of C . albicans, which quickly develops in the mycelial form, infiltrating the skin layers . Some interesting differences were recorded, comparing amphotericin B and ketoconazole . Moreover the authors used a therapy test in mice, based mainly on examination of the kidney.

Sabouraudia, 1985 Oct, 23(5), 335 - 8
Lack of antibody induction by Histolyn-CYL, a new skin-testing reagent for histoplasmosis; Scalarone GM et al.; Histolyn-CYL, a yeast phase skin-test reagent, was administered to 85 histoplasmin-sensitive subjects living in the United States and South America . In a multicenter study, sera were obtained at the time the skin tests were read and again 3 weeks later . In no instance did the skin test induce significant complement-fixing antibody changes or antibodies detected by immuno-diffusion.

Sabouraudia, 1985 Oct, 23(5), 377 - 87
In-vitro killing of Candida species by murine immunoeffectors and its relationship to the experimental pathogenicity; Vecchiarelli A et al.; Killing of yeast cells of several species of Candida by murine phagocytic cells was assessed in vitro by a radiolabel release microassay and measurement of colony forming units . The most effective candidacidal phagocytes, i.e . polymorphonuclear and bone marrow cells, were able to kill equally well cells of any species or isolate tested, given sufficient time (4 h) and an appropriate effector: target ratio . However, C . guilliermondii, C . krusei and C . parapsilosis were killed by polymorphonuclear and bone marrow cells much more promptly (1 h) and at a significantly lower effector:target ratio than C . albicans, C . tropicalis and C . viswanathii . Moreover, there were immune effectors such as peritoneal resident macrophages and, mostly, spleen cells which were practically ineffective against C . albicans and C . tropicalis but showed significant activity against C . guilliermondii, C . krusei and C . parapsilosis, even in mice immuno-depressed with cyclophosphamide . Three isolates of C . albicans, differing in the capacity to form germ tubes, also differed in mouse virulence: the germ-tube forming isolate was the most virulent . However, they showed an identical pattern of susceptibility to killing by mouse immunoeffectors, suggesting that virulence is probably not due to the resistance of hyphal cell to phagocytosis.

J Clin Pathol, 1985 Oct, 38(10), 1175 - 8
Defective activation of neutrophils after splenectomy; Foster PN et al.; Neutrophil chemotaxis and phagocytosis in the presence of serum from 20 patients who had undergone splenectomy and from 15 healthy volunteers was studied . The mean distance migrated by normal neutrophils in the presence of serum from the patients after splenectomy was significantly less than that when normal serum was used (p less than 0.005) . The percentage of neutrophils phagocytosing a yeast was also significantly reduced in the presence of serum from patients after splenectomy (p less than 0.02) . In addition, when neutrophils from these patients were studied both chemotaxis and phagocytosis were enhanced in normal compared with autologous serum (p less than 0.05).

EMBO J, 1985 Oct, 4(10), 2695 - 703
T4 polynucleotide kinase; cloning of the gene (pseT) and amplification of its product; Midgley CA et al.; The T4 gene (pseT) for polynucleotide kinase (pnk) has been cloned in lambda . Induction of a lambda E-W-S-cI857 prophage in which the pseT gene can be transcribed from the late lambda promoter, PR1, leads to greater than 100-fold amplification of pnk activity; pnk comprises approximately 7% of the total soluble cell protein . The purified enzyme, as expected, is both a 5'-kinase and a 3'-phosphatase . The amino acid sequence deduced from an open reading frame identified as the pseT gene contains a sequence which corresponds particularly well with that part of the adenine nucleotide binding site of adenylate kinase shown to form a flexible loop . A deletion mutant that lacks 5'-kinase activity, and possibly also 3'-phosphatase activity, has lost two amino acids from within the proposed loop structure . A second region of the pnk sequence shares homology with phosphoglycerate kinase, yeast inorganic pyrophosphatase and histone 2b from various organisms.

Am J Surg Pathol, 1985 Oct, 9(10), 744 - 51
Pneumocystis carinii pneumonitis . The nature and diagnostic significance of the methenamine silver-positive "intracystic bodies"; Watts JC et al.; The cyst forms of Pneumocystis carinii in specimens stained with methenamine silver contain single or paired discrete foci of enhanced staining that measure 1-2 micron in maximum size . The nature of these foci and their location within the cysts have been disputed . We demonstrate by electron microscopy of silver-stained sections that the darkly stained foci correspond to a focal thickening of the cyst wall and are unrelated to sporozoites and other intracystic organelles . The morphology of these structures by light microscopy is characteristic, and their recognition is helpful in identifying P . carinii cysts and in differentiating them from yeast-form fungi and argyrophilic tissue elements in histologic sections and cytology specimens.

Med J Aust, 1985 Sep 16, 143(6), 234 - 5
An elimination diet for chronic urticaria of childhood; Kemp AS et al.; Twenty-three children with chronic urticaria were treated with an elimination diet for two weeks . Eighteen completed the period of dietary elimination; in seven of the 18 children there was a marked remission of the urticaria during the second week of the diet . The administration of challenge capsules provoked an exacerbation of urticaria in five of the 14 (36%) children given aspirin . The incidence of reactions to tartrazine, sodium benzoate and yeast (7%) was not significantly different from those to the lactose placebo (9%) . In selected cases, elimination diets with controlled reintroduction of foods have a role in the management of chronic urticaria in childhood.

Biochem Biophys Res Commun, 1985 Sep 16, 131(2), 557 - 63
Protection of phenylalanine ammonia-lyase from proteolytic attack; Gilbert HJ et al.; Phenylalanine ammonia-lyase contained within permeabilized cells of Rhodosporidium toruloides was protected from proteolytic attack by trypsin, chymotrypsin and duodenal juice . The inactivation by the proteases was biphasic . The enzyme contained within the yeast cells had a similar Km for phenylalanine and Ki for cinnamic acid to the protein in free solution . Phenylalanine ammonia-lyase present in the yeast depleted duodenal juice of free phenylalanine, while the enzyme in free solution did not . The possibility of using permeabilized cells of R . toruloides as a vehicle for protecting orally ingested therapeutic enzymes from proteolytic inactivation is discussed.

Biochem J, 1985 Sep 15, 230(3), 771 - 6
Essentiality of the active-site arginine residue for the normal catalytic activity of Cu,Zn superoxide dismutase; Borders CL Jr et al.; Chemical modification of bovine and yeast Cu,Zn superoxide dismutases with phenylglyoxal diminishes the catalytic activities by greater than or equal to 98%, and treatment of these enzymes with butanedione plus borate leads to greater than or equal to 96% inactivation . The activity loss is accompanied by the modification of less than two arginine residues per subunit with no concomitant loss of Cu or Zn . The phenylglyoxal-modified enzymes require at least a 20-fold greater concentration of cyanide for 50% inhibition than do the corresponding native enzymes . Polyacrylamide-gel electrophoresis and activity staining of the phenylglyoxal-inactivated enzymes demonstrate that the residual activity is largely associated with modified forms that bear lower net positive charge than the native superoxide dismutases.

J Med Virol, 1985 Sep, 17(1), 57 - 62
Safety and immunogenicity of a recombinant hepatitis B vaccine; Dandolos E et al.; A hepatitis B vaccine produced in yeast by recombinant DNA technology was evaluated using 5-micrograms and 10-micrograms doses in a randomized trial lasting 7 months in 110 male armed forces recruits aged 17-19 years . Results were compared to those of an identical trial of a plasma-derived vaccine . No allergic reactions were observed, and the rate of mild side effects was similar to the plasma-derived vaccine . Seroconversion rates in the first month were 60% (33/55) and 67% (37/55) with the 5-micrograms and 10-micrograms doses of the recombinant vaccine, respectively . All participants seroconverted by 3 months, and none lost antibody . These results are very similar to those for plasma-derived vaccine . Comparison of titres of antibody to hepatitis B surface antigen (anti-HBs) showed a slightly higher level with the 10-micrograms than with the 5-micrograms dose of the recombinant vaccine . Geometric mean titres of anti-HBs after the booster dose were similar in the 5-micrograms and 10-micrograms dose recombinant vaccine groups (2,620 and 2,748 IU/l, respectively) and in the 5-micrograms plasma-derived vaccine group (3,591 IU/l) but significantly higher (9,227 IU/l) with the 10-micrograms dose of the plasma-derived vaccine . These results confirm the safety and immunogenicity of the recombinant vaccine, although further study is needed on the duration of immunity.

J Clin Microbiol, 1985 Sep, 22(3), 422 - 4
Isolation of Legionella pneumophila from blood with the BACTEC system: a prospective study yielding positive results; Rihs JD et al.; In a prospective study of 16 patients with Legionnaires disease confirmed by cultural isolation of Legionella pneumophila from the respiratory tract, 38% (6 of 16) had positive blood cultures . Daily subcultures were made onto buffered charcoal-yeast extract plates from 6B aerobic and 7C anaerobic BACTEC blood culture bottles (Johnston Laboratories, Inc., Towson, Md.) . Isolation of L . pneumophila was achieved from both aerobic and anaerobic bottles . L . pneumophila growth indices failed to exceed the BACTEC threshold limits; thus, the organism would have been overlooked despite its presence in the blood culture bottles . Bacteremic patients had statistically significant higher quantities of L . pneumophila isolated from sputum and visualized on direct fluorescent antibody stains . Thus, the potential exists for improved diagnosis of Legionella infection by a relatively noninvasive procedure (blood culture) with an instrument already in use in many hospital laboratories.

J Cell Physiol, 1985 Sep, 124(3), 545 - 53
Opposing effects of dexamethasone on the clonal growth of granulocyte and macrophage progenitor cells and on the phagocytic capability of mononuclear phagocytes at different stages of differentiation; Shezen E et al.; Dexamethasone, a synthetic glucocorticosteroid, was shown to modulate the colony-stimulating factor-dependent clonal growth of myeloid progenitor cells in semisolid agar cultures, enhancing the formation of granulocyte colonies (50-100%) and suppressing the formation of macrophage colonies (75-97%) . Modulation of the pattern of myeloid colony formation by dexamethasone (12-125 nM) was brought about when the steroid was administered to 6-day cultures at the time of culture initiation and up to 72 hr later . Dexamethasone inhibited myeloid cell proliferation when administered to 5-day liquid cultures at culture initiation and up to 96 hr later . Dexamethasone (12-250 nM) also enhanced the phagocytic activity of bone marrow-derived mononuclear phagocytes toward heat-killed (HK) yeast cells (up to 100%) and IgG-coated sheep red blood cells (up to 60%) . Enhancement of the phagocytic capability depended critically on the stage in culture at which dexamethasone was administered . Exposure to dexamethasone for 28 hr up to 96 hr of 96-hr cultures of bone marrow cells did not lead to a modulation of phagocytic activity of the developing mononuclear phagocytes . The presence of dexamethasone during the critical period of 96 hr to 120 hr after culture initiation led to an enhanced phagocytic capability, which was statistically significant already 12 hr after the administration of the glucocorticoid . Dexamethasone induced an enhanced phagocytic activity when administered at any time after culture initiation provided that it was in culture during this critical period . When added at 120 hr of culture, dexamethasone no longer enhanced the phagocytic capability of mononuclear phagocytes and when added later than 156 hr of culture suppressed it . Dexamethasone also suppressed (up to 68%) the phagocytic capability of resident and elicited peritoneal macrophages . The results suggest that glucocorticoids shift the balance of granulocyte vs . macrophage formation at early stages of precursor cell differentiation . Reduction in mononuclear phagocyte growth and enhancement of its phagocytic capability might reflect accelerated differentiation/maturation steps . The inhibitory effect of dexamethasone on macrophage formation and on the phagocytic capability of mature mononuclear phagocytes and peritoneal macrophages might be a relevant aspect of the in vivo immune suppression encountered after glucocorticoid administration.

Exp Cell Res, 1985 Sep, 160(1), 95 - 105
Receptor-mediated uptake of horseradish peroxidase in innervated and denervated skeletal muscle; Tagerud S et al.; The in vitro uptake of {3H}inulin and horseradish peroxidase (HRP) has been studied in innervated and 6 days denervated extensor digitorum longus muscle of the mouse . Both markers were taken up at a higher rate in denervated muscle . The increase in uptake after denervation was, however, larger for HRP than for {3H}inulin . After 2 h incubation at 37 degrees C, pH 7.3, in the presence of equimolar concentrations of HRP and {3H}inulin (approx . 2.1 microM), the uptake of HRP was approx . 8 times as great as the uptake of {3H}inulin in the same innervated muscles . In denervated muscle the HRP uptake was approx . 19 times as great as the {3H}inulin uptake in the same muscles . Various possible explanations of these differences in uptake have been considered and tested experimentally . {3H}Inulin uptake in skeletal muscle has previously been shown to obey bulk kinetics . The present investigation shows the HRP uptake to obey saturation kinetics . The HRP uptake shows dependency on divalent cations and is reduced if incubation is carried out at pH 6.4 . The uptake of HRP, when used at a low, non-saturating concentration (10 micrograms/ml approx . 0.25 microM), is inhibited greater than or equal to 60% by yeast mannan (0.1 mg/ml), ribonuclease B (0.1 mg/ml, approx . 7.4 microM), mannose (30 mM), monodansylcadaverine (1 mM), chloroquine (100 microM), trifluoperazine (25 microM) or maleic acid (2 mM) . It is concluded that HRP is taken up in innervated and denervated skeletal muscle by a process of receptor-mediated endocytosis and that this uptake is under neurotrophic control.

Am J Clin Pathol, 1985 Sep, 84(3), 323 - 7
Progressive disseminated penicilliosis caused by Penicillium marneffei . Report of eight cases and differentiation of the causative organism from Histoplasma capsulatum; Deng ZL et al.; Eight patients with fatal penicilliosis caused by Penicillium marneffei are reported . All were natives of southern rural Guangxi, and none had a predisposing illness or evidence of altered immunity . The distinctive features of P . marneffei include proliferation of yeast-like cells within histiocytes, followed by the development of focal necrosis and, eventually, large abscesses . Outside the histiocytes, the fungi elongate, become slightly curved, and form septa . In vitro, P . marneffei produces a red pigment which diffuses into the culture medium . The differentiation between P . marneffei and Histoplasma capsulatum is described, and possible reservoirs for P . marneffei are discussed.

J Nutr, 1985 Sep, 115(9), 1196 - 202
Effect of dietary selenium deficiency on incidence and size of 1,2-dimethylhydrazine-induced colon tumors in rats; Pence BC et al.; Torula yeast diets deficient (less than or equal to 0.02 ppm) in selenium (Se) and a control diet supplemented with sodium selenite (0.1 ppm Se), were fed to 52 male Sprague-Dawley rats per diet for 23 wk following weaning . 1,2-dimethylhydrazine (DMH) was administered (20 mg/kg body weight) in 20 weekly i.p . injections after 3 wk of feeding . After 23 wk, 67% of the rats fed the Se-deficient diet had intestinal adenocarcinomas versus 63% on the 0.1 ppm Se diets . Diet also had no effect on the number or size of tumors per tumor-bearing animal or on the location of the tumors within the colon . The effects of Se deficiency on the hematological variables of white blood cell count, hematocrit, serum urea nitrogen and cholesterol concentrations were examined . Serum urea nitrogen levels were lower in Se-deficient DMH-treated rats (9.6 +/- 0.7 vs . 13.7 +/- 0.9 mg/dl, P less than 0.01) and serum cholesterol was higher in Se-deficient DMH-treated animals (69.0 +/- 5.5 vs . 50.7 +/- 3.9 mg/dl, P less than 0.05) . Body weights of the Se-depleted group were lower in the DMH-treated animals (P less than 0.01) although food consumption did not differ . Controls without DMH did not show differences due to Se status for any variable examined . Se deficiency appears to affect DMH toxicity but nutritional adequacy (0.1 ppm Se) does not inhibit tumor development . The results of this study do not support the belief that Se deficiency enhances colon carcinogenesis.

Br J Nutr, 1985 Sep, 54(2), 367 - 72
Low-iodine diet for the production of severe I deficiency in marmosets (Callithrix jacchus jacchus); Mano MT et al.; A low-iodine diet consisting of maize, peas (Pisum sativum), torula yeast, meat meal, maize oil and added vitamins, minerals and amino acids was given to eight pairs of adult, common cotton-eared marmosets (Callithrix jacchus jacchus) . Eight control pairs were given the same diet to which potassium iodate was added . Both groups also received low-I apple and deionized water . The diet provided adequate nutrition, as confirmed by the maintenance of body-weight and good health . In the I-deficient marmosets the concentration of plasma thyroxine was decreased from 140.1 nmol/l to 22.4 nmol/l and thyroid-stimulating hormone increased significantly from 1.8 ng/ml to 9.0 ng/ml compared with control marmosets, thereby indicating severe I deficiency . Compared with newborn offspring from control marmosets, the thyroid glands from the I-deficient offspring showed an increase in weight, a decrease in I content and, on histological examination, hyperplasia, hypertrophy and a total absence of colloid material in the follicles.

Tsitologiia, 1985 Sep, 27(9), 1055 - 8
{Structural and functional analysis of the respiratory burst in the phagocytosing macrophage}; Khramtsov AV et al.; The interrelation between structural changes and oxygen consumption by the phagocyting macrophage was studied . The mean number of phagocyted particles was estimated by the method of stereological transformation . It is found that the uptake of yeast particles and CN- -nonsensitive oxygen consumption is related to the concentration of yeast cells in the incubation medium . A positive correlation was established between the oxygen consumption and the mean number of phagocyted particles . The results obtained may suggest that the "respiration burst" takes place in the contact area of the macrophage and the phagocyted material, and its extent probably depends on the surface of that contact area.

J Ethnopharmacol, 1985 Sep, 14(1), 45 - 51
Antipyretic studies on some indigenous Pakistani medicinal plants; Khattak SG et al.; Significant oral antipyretic activity in rabbits was exhibited by hexane-, chloroform- and water-soluble extracts of Artemisia absinthium, Viola odorata, Melia azadirachta and Fumaria parviflora comparable in potency aspirin . Pyresis was induced by subcutaneous yeast injections . Antipyretic activity was more prominent in the hexane-soluble portions of these plants . Insignificant to no antipyretic effects were produced by extracts of Butea frondosa, Berberis lycium and Sisymbrium irio . No obvious toxic effects were noted for any of the plant extracts up to doses of 1.6 g/kg.

Agents Actions, 1985 Sep, 16(6), 542 - 7
Dexamethasone fails to produce antipyretic and analgesic actions in experimental animals; Nakamura H et al.; In order to explore the role of phospholipase A2 inhibition in the mechanisms of the action of glucocorticoids, it was investigated whether the steroid exhibits the analgesic and antipyretic actions as well as cyclo-oxygenase inhibitors such as indomethacin or not . Dexamethasone has been reported to produce the anti-inflammatory action with a lag time of at least 1 h at doses of up to 0.1 mg/kg in mice and rats . However, dexamethasone when given 4 h beforehand had no significant analgesic activity even at doses of 1 and 10 mg/kg i.v . in the acetic acid writhing test in rats . In mice, the significant reduction in writhes counts was seen when dexamethasone (1 and 10 mg/kg i.v.) was given 15 min or 4 h before phenylquinone injection; i.e . the activity had not the lag time . On the other hand, dexamethasone showed a strong antipyretic activity against both the fevers caused by LPS and yeast in rats . In the yeast-febrile rats, the antipyretic activity had a lag time of about 1 h, and was dose-related at doses as low as 0.03 to 0.3 mg/kg i.v.; the steroid markedly reduced the increased PGE2 content in the cerebrospinal fluid . The antipyretic activity after local injection into the cerebroventricle or the yeast pouch was stronger than that after systemic injection into the tail vein, although so large a difference in the activity between the dosage routes was not seen, suggesting that the site of the antipyretic action is in both the brain and periphery . The antipyretic activity of dexamethasone (10 mg/kg i.v.) was not seen in rabbits with fever caused by LPS.(ABSTRACT TRUNCATED AT 250 WORDS)

Biochem J, 1985 Sep 1, 230(2), 535 - 42
The phosphonomethyl analogue of 3-phosphoglycerate is a potent competitive inhibitor of phosphoglycerate mutases; McAleese SM et al.; The phosphonomethyl analogue of 3-phosphoglycerate (2-hydroxy-4-phosphonobutanoate) is a potent competitive inhibitor of cofactor-dependent phosphoglycerate mutase from yeast and of cofactor-independent phosphoglycerate mutase from wheat germ . For the yeast enzyme Ki is 1.3 mM (Km for substrate is 0.71 mM); for the wheatgerm enzyme Ki is 18 mM (Km for substrate is 0.86 mM) . This analogue should be a useful tool for n.m.r . spectroscopic studies on the mechanism of action of the two mutases . The arsonomethyl analogue of 3-phosphoglycerate (4-arsono-2-hydroxybutanoate) was a relatively poor inhibitor.

Proc Natl Acad Sci U S A, 1985 Sep, 82(17), 5588 - 92
High-affinity-receptor-mediated uptake and degradation of glucose-modified proteins: a potential mechanism for the removal of senescent macromolecules; Vlassara H et al.; Proteins that have been modified by long-term exposure to glucose accumulate advanced glycosylation end products (AGE) as a function of protein age . In these studies, we have characterized the interaction of AGE-protein with mouse peritoneal macrophages, using AGE-modified bovine serum albumin (AGE-BSA, prepared by incubation with glucose) as a probe . AGE-BSA was specifically bound to cells at 4 degrees C and was taken up and degraded at 37 degrees C; these processes were concentration dependent and saturable . Competition experiments with AGE-BSA, BSA incubated with phosphate-buffered saline rather than glucose, and yeast mannan demonstrated that macrophages specifically recognize AGE on proteins by a receptor that is completely distinct from the mannose/fucose receptor . Scatchard analysis of AGE-BSA binding data indicated that there are approximately 1.06 X 10(5) receptors per macrophage, with an affinity constant of 1.75 X 10(-11) M . Specific binding of AGE-BSA to the macrophage receptor was competitively inhibited by BSA that had been chemically coupled to a synthetic analogue of the specific AGE, 2-(2-furoyl)-4(5)-(2-furanyl)-1H-imidazole (FFI-BSA) . FFI-BSA was also taken up by macrophages in a concentration-dependent, saturable manner . Prior incubation of macrophages with AGE-BSA failed to influence the subsequent uptake and degradation of added AGE-BSA . Thus, the AGE receptor does not appear to be down-regulated by exposure to AGE-proteins . Results from these studies suggest that AGE could act in vivo as a specific signal for recognition and degradation of senescent macromolecules . Incomplete removal of AGE-proteins by macrophages may ultimately give rise to some of the physiologic changes that occur with normal aging.

Trop Med Parasitol, 1985 Sep, 36(3), 163 - 70
The induction of functional mononuclear and multinuclear macrophages in murine brugian filariasis: morphological and immunological properties; Mackenzie CD et al.; This present study describes the ontogeny and morphology of mononuclear and multinuclear macrophages in murine Brugia pahangi infections . Intraperitoneal infection with L 3 resulted in the development of adult worms and the production of microfilariae in many mice; however, all worms were usually removed by 100 days post infection (p.i.) . The infection induced predominantly mononuclear and multinuclear macrophage responses in the peritoneal cavity, with maximum numbers of peritoneal cells being achieved by 8-12 weeks p.i . and at this time some 20% of the macrophages were multinucleated . Lymphocyte numbers were also increased during infection . The reactions around the filariae involved macrophages (both single and multinucleated), eosinophils, fibroblasts and the laying down of collagen . Most worms were involved in these cellular reactions by 5-6 weeks p.i., with the multinuclear macrophages generally seen lying close to the parasites . All stages of parasites developed vacuolar internal changes early in their degeneration with calcification being a later change . The destruction and removal of the sheath was not apparently a prerequisite for degeneration of the body of microfilariae . Rosetting techniques showed that 75% of peritoneal cells (macrophages) in infected animals possessed Fc receptors compared with 32% of those in uninfected mice, and for C3 receptors the respective results were 35% and 5% . The multinuclear cells possessed both types of receptors at levels similar to those of mononuclear macrophages . Likewise both cell types were found to readily phagocytose yeast and latex particles, as well as staining positively for non-specific esterases.(ABSTRACT TRUNCATED AT 250 WORDS)

Arch Biochem Biophys, 1985 Sep, 241(2), 348 - 55
Mechanistic investigation on the temperature dependence and inhibition of corn root plasma membrane ATPase; Tu SI et al.; The kinetics of corn root plasma membrane-catalyzed Mg-ATP hydrolysis may be satisfactorily described by a simple Michaelis-Menten scheme . It was found that the Km of the process was relatively insensitive to changes in temperature . This property allowed us to conveniently estimate the activation energy of the enzyme turnover process as approximately 14 kcal mol-1 in the temperature range of 10 to 45 degrees C . The enzyme activity was inhibited by the presence of diethystilbestrol (DES), miconazole, vanadate, and dicyclohexylcarbodiimide (DCCD) . The inhibition caused by DES and miconazole was strictly uncompetitive and inhibition by vanadate was noncompetitive . The inhibition by DCCD showed a substrate concentration dependence, i.e., competitive at high and uncompetitive at low concentrations of Mg-ATP . The 1/V vs {I} plots suggested that there were different but unique binding sites for DES, vanadate, and miconazole . However, the modification of the plasma membrane by DCCD exhibited interaction with multiple sites . Unlike yeast plasma membrane ATPase, the enzyme of corn root cells was not affected by the treatment with N-ethylmaleimide . Although the enzyme activity was regulated by ADP, a product of the reaction, the presence of inorganic phosphate showed no inhibition to the hydrolysis of Mg-ATP.

Nucleic Acids Res, 1985 Aug 26, 13(16), 5857 - 67
Isolation and sequence analysis of a cDNA encoding the ATP/ADP translocator of Zea mays L; Baker A et al.; A cDNA complementary to the mRNA for the ATP/ADP translocator of maize (Zea mays L.) has been identified by virtue of hybridisation with the homologous gene from yeast . The cloned cDNA has been shown by DNA sequence analysis to contain an open reading frame of 954bp., which encodes a polypeptide of molecular weight 40,519 . This polypeptide exhibits a high degree of homology to the translocator polypeptides of beef heart and Neurospora crassa mitochondria.

J Biol Chem, 1985 Aug 15, 260(17), 9509 - 12
Covalent modification of phenylalanyl-tRNA synthetase with phenylalanine during the amino acid activation reaction catalyzed by the enzyme; Rapaport E et al.; Yeast phenylalanyl-tRNA synthetase (PRS) is shown to undergo autoaminoacylation with phenylalanine under in vitro amino acid activation conditions . Phenylalanyl adenylate enzyme complex yields a covalent phenylalanyl isopeptide exclusively with the beta subunit of the alpha 2 beta 2 enzyme . Contrary to previously reported cases of autoaminoacylation of aspartyl-tRNA synthetase and tryptophanyl-tRNA synthetase, the autoaminoacylation of PRS occurs under a specific set of conditions and results in the identification of only one labeled tryptic peptide on two types of high pressure liquid chromatography columns . The ability of PRS to undergo this covalent modification directly correlates with its ability to catalyze the synthesis of diadenosine 5',5"'-P1,P4-tetraphosphate from enzyme-bound phenylalanyl adenylate . Both reactions require the presence of low levels of zinc or cadmium and are inhibited by tRNAPhe or by low levels of low molecular weight thiols . Since diadenosine 5',5"'-P1,P4-tetraphosphate synthesis is known to be catalyzed in vivo in response to oxidation stress, it is also likely that the autoaminoacylation of phenylalanyl-tRNA synthetase may occur in vivo under a similar set of conditions . These reactions are thus not simply the result of accumulation of phenylalanyl adenylate and probably reflect conformational changes in the protein which are brought about by its interaction with zinc or cadmium.

J Biol Chem, 1985 Aug 15, 260(17), 9559 - 66
Primary structure of copper-zinc superoxide dismutase from Neurospora crassa; Lerch K et al.; The complete amino acid sequence of copper-zinc superoxide dismutase from Neurospora crassa is reported . The subunit consists of 153 amino acids and has a Mr of 15,850 . The primary structure was determined by automated and manual sequence analysis of peptides obtained by digestions of the carboxymethylated and aminoethylated enzyme with trypsin and thermolysin . The protein is devoid of tryptophan and methionine and displays a free amino terminus . Comparison of the amino acid sequence with those from human erythrocyte, bovine erythrocyte, horse liver, swordfish liver, and yeast copper-zinc superoxide dismutases reveals a high degree of sequence homology among the six enzymes . Most prominently, the regions containing the amino acid residues participating in the metal-binding and the half-cystine residues forming the intramolecular disulfide bridge are highly conserved . The invariant amino acids Pro 74 and Asp 76 of the four vertebrate and yeast superoxide dismutases were found to be substituted by arginine and alanine, respectively, in the Neurospora enzyme . These radical substitutions occurring in the zinc ligand region, known to form a characteristic loop structure in bovine erythrocyte copper-zinc superoxide dismutase (Tainer, J . A., Getzoff, E . D., Beem, K . M., Richardson, J . S., and Richardson, D . C . (1982) J . Mol . Biol . 160, 181-217), however, do not affect the catalytic properties of the Neurospora enzyme.

Eur J Biochem, 1985 Aug 15, 151(1), 153 - 65
The structure of the double-stranded RNA pentamer 5'(CACAG) . 5'(CUGUG) determined by nuclear Overhauser enhancement measurements: interproton distance determination and structure refinement on the basis of X-ray coordinates; Clore GM et al.; The structure in solution of the duplex RNA pentamer 5'(CACAG) . 5'(CUGUG), comprising the stem of the T psi C loop of yeast tRNAPhe, has been investigated by means of one- and two-dimensional nuclear Overhauser enhancement measurements . All non-exchangeable base and sugar proton resonances with the exception of the H5'/H5" sugar resonances are assigned in a sequential manner . From the relative intensities of the cross-peaks obtained in the pure-phase absorption two-dimensional nuclear Overhauser enhancement spectra at several mixing times, it is deduced that the RNA pentamer adopts an A-type conformation in solution . Cross-relaxation rates and interproton distances are determined from the time dependence of the nuclear Overhauser effects, principally by one-dimensional measurements . The structure of the RNA pentamer is then refined by restrained least-squares minimization on the basis of both distance and planarity restraints using fibre diffraction data as an initial model . The refined structure of the RNA pentamer is of the A type but exhibits local structural variations in glycosidic bond and backbone torsion angles as well as in propeller twist, base roll and base tilt angles.

Am J Ophthalmol, 1985 Aug 15, 100(2), 252 - 8
Subretinal neovascularization after experimental ocular histoplasmosis in a subhuman primate; Jester JV et al.; Subretinal neovascularization was demonstrated in a subhuman primate eye (Macaca speciosa) one year after the internal carotid injection of yeast-phase Histoplasma capsulatum . In this animal model of ocular histoplasmosis, initial injection of viable H . capsulatum results in the development of self-limiting acute multifocal choroiditis . Acute lesions resolve within six months, forming chorioretinal scars that are clinically similar to human histo spots . Detailed ultrastructural study of a peripapillary scar 30 months after the injection showed the presence of subretinal neovascularization located between Bruch's membrane and degenerated retinal inner segments . These vessels appeared to be continuous, had tight junctional complexes, and a well-developed basal lamina with occasional pericytes . Failure of fluorescein angiography to demonstrate the presence of this neovascular net may be explained by the presence of tight junctions within the subretinal capillaries.

Antibiot Med Biotekhnol, 1985 Aug, 30(8), 595 - 8
{Comparative evaluation of the effect of immunostimulants on the dehydrogenase activity of the mouse thymus}; Gapochko KG et al.; Comparative biochemical estimation of the influence of yeast RNA, prodigiosan and levamisole on the activity of lactate dehydrogenase, malate dehydrogenase and glutamate dehydrogenase in the thymus of mice was carried out . It was shown that the immunostimulants induced different qualitative and quantitative changes in the activity of the enzymes . Administration of yeast RNA resulted in activation of lactate dehydrogenase, malate dehydrogenase and glutamate dehydrogenase . Prodigiosan increased the activity of lactate dehydrogenase and glutamate dehydrogenase but not that of malate dehydrogenase . The action of levamisole was characterized by long-term activation of glutamate dehydrogenase . The possible mechanisms of the influence of these drugs on the metabolic activity of the thymus are discussed.

Parasitology, 1985 Aug, 91 ( Pt 1), 1 - 7
A rapid chromatographic method for elimination of fungal contamination in in vitro cultures of Leishmania spp; Okot-Kotber BM; It was demonstrated that antimycotic agents are unsuitable for elimination of mycotic contamination in in vitro cultures of Leishmania spp . Ion-exchange chromatography was designed to separate the protozoan parasites from the contaminating yeast or fungal cells based on their surface net charges . The method was found to be efficient and rapid, and would also reduce bacterial contamination in cultures which have been exposed to both contaminants . Viability of the cells has been shown to be unaltered by this treatment as demonstrated by their ability to multiply rapidly in culture medium, as was seen in the controls.

Am J Obstet Gynecol, 1985 Aug 1, 152(7 Pt 2), 959 - 61
Treatment of recurrent vaginal candidiasis; Forssman L et al.; In recurrent vulvovaginal candidiasis, predisposing factors should be eliminated wherever possible . Reduction of the gastrointestinal yeast flora by oral antimycotic treatment does not prevent recurrences . Perianal application of antifungal cream might be an alternative in preventing reinfection from the rectum . Venereal spread of yeasts does not seem to be an important factor, and partner treatment does not significantly alter treatment results . Traditionally, prolonged treatment periods are recommended to prevent recurrences . Patient compliance may then become a factor limiting efficacy . One-dose treatment has been shown to give results that compare favorably with traditional treatment schedules . Results in recurrent cases were comparable with those in primary infections . Thus one-dose treatment could be an acceptable alternative to longer treatment periods in schedules for treatment of recurrent cases . Intermittent prophylactic one-dose application appears to be a promising method of reducing recurrences.

J Biomol Struct Dyn, 1985 Aug, 3(1), 127 - 44
Backbone conformation in nucleic acids: an analysis of local helicity through heminucleotide scheme and a proposal for a unified conformational plot; Malathi R et al.; A relationship has been established to express the local helicity of a polynucleotide backbone directly in terms of the virtual bonds spanning the conformationally equivalent heminucleotide repeats, with a view to provide a better understanding of the cumulative effects of all the chemical bond rotational variations on local helicity . Using this, an analysis made with a few oligodeoxynucleotide crystal structures clearly brings forth that it is the concerted movements manifested in the near neighbour correlations between the pair of chemical bonds C4'-C5' and P-O5' and C4'-C3' and P-O3' of the 5' and 3' heminucleotides respectively that are primarily responsible for the observed non-uniform helical twists both in A and B type helical backbones . That these need not be restricted to oligodeoxynucleotides but may be a feature of oligoribonucleotides backbone also is shown from an analysis of helical segments of yeast tRNA(Phe) . A proposal of a unified or a grand two dimensional conformational plot which would help visualise succinctly the overall effect of the variations in all the repeating six chemical bonds of a polynucleotide backbone is made . Apart from considerable simplification, the plot affords identification on it regions characteristic of helical, and loop and bend conformations of nucleic acid backbone chain.

Proc Natl Acad Sci U S A, 1985 Aug, 82(15), 4992 - 4
Sea urchin metallothionein sequence: key to an evolutionary diversity; Nemer M et al.; The metallothioneins (MTs) constitute a diverse family of proteins, which are enriched in cysteines and bind heavy metals . The amino acid sequence of sea urchin MT has been obtained from its mRNA sequence and compared with MT sequences of various sources . A largely conserved sequence of 10 amino acids, the "central segment," is located near the center of the MT molecules of Neurospora, yeast, and Drosophila and the center of putative domains in mammalian and sea urchin MTs . The sea urchin carboxyl-terminal-half MT resembles the mammalian 9-cysteine amino-terminal MT domain I, both in the presence of this central segment and in the relative placement of cysteine residues . Conversely, the sea urchin amino-terminal-half MT, containing 11 cysteines, resembles the mammalian carboxyl-terminal MT domain II in its exclusive enrichment in vicinal cysteines . The reversed order of these sea urchin and mammalian MT halves appears to be just one aspect of a diversity based on the elaboration of structures containing the central segment . Still another variation in this diversity is the duplication of the central segment, apparent in Drosophila and crab MTs.

Mol Cell Biol, 1985 Aug, 5(8), 1993 - 6
Nuclear polyadenylate-binding protein; Sachs AB et al.; Polyadenylate-binding activity can be detected in eluates from sodium dodecyl sulfate gels by a nitrocellulose filter-binding assay . Nuclear extracts from rat liver show a single peak of binding activity at 50 to 55 kilodaltons; cytoplasmic extracts show a single peak at 70 to 80 kilodaltons, corresponding to a 75-kilodalton protein previously described . Similar results are obtained with yeast and mouse fibroblasts, indicating a high degree of conservation of both nuclear and cytoplasmic polyadenylate-binding proteins . The activity from rat liver nuclei has been purified 125-fold on the basis of specific binding to polyadenylate and shows two main bands in sodium dodecyl sulfate gels at 53 and 55 kilodaltons.

Cutis, 1985 Aug, 36(2), 129 - 30
Erythema annulare centrifugum: an unusual case due to hydroxychloroquine sulfate; Hudson LD; Erythema annulare centrifugum presents as a cutaneous hypersensitivity to diverse causes including fungal and yeast infections, parasitic infestations, drugs, and, rarely, occult malignancies . A prolonged case of erythema annulare centrifugum secondary to the use of hydroxychloroquine sulfate is presented . The prolonged time needed for clearing after discontinuation of the medication is thought to result from the melanocyte-binding characteristics of the drug.

Exp Hematol, 1985 Aug, 13(7), 652 - 7
Hematopoiesis on cellulose ester membranes (CEM) . VIII . Studies of the hematopoietic microenvironment developing on intraperitoneally implanted CEM in S1/S1d and S1+/S1+ mice; Knospe WH et al.; Cellulose ester membranes (CEM) were implanted intraperitoneally into S1/S1d and S1+/ S1+ mice . These CEM rapidly became coated with peritoneal cells capable of supporting primarily granulocytic colony development after seven days . S1/S1d-coated CEM showed a diminished capacity to support colony development compared with S1+/ S1+ CEM, perhaps reflecting the defect in the hematopoietic microenvironment of these mice . Marrow cells from S1+/S1+ and S1/S1d mice generated similar numbers of colonies on S1+/S1+ CEM . When CEM were transferred from a primary to a secondary host there was a tendency to remodel the CEM toward the characteristics of the secondary host . Peritoneal cells coating CEM from S1/S1d mice had less phagocytosis of yeast particles than peritoneal cells from S1+/S1+ mice . The cell coat on the membranes from S1/S1d mice was fewer cell layers in thickness than those on membranes coated in S1+/ S1+ mice.

Ophthalmology, 1985 Aug, 92(8), 1159 - 64
Disseminated bilateral chorioretinitis due to Histoplasma capsulatum in a patient with the acquired immunodeficiency syndrome; Macher A et al.; A 31-year-old white male homosexual was healthy until March 1984, when he developed Pneumocystis carinii pneumonia, which resolved with treatment . In April 1984 he developed fever, followed by hepatosplenomegaly, headaches, blurred vision, pancytopenia and pulmonary infiltrates . On June 11, intracytoplasmic yeast were noted within leukocytes on a peripheral blood smear, and amphotericin B was started . The patient developed progressive respiratory and renal insufficiency and died on June 13, 1984 . Autopsy histopathology demonstrated disseminated histoplasmosis and Histoplasma capsulatum was cultured from numerous tissues . Ocular histopathologic examination using special fungal stains and electron microscopy revealed numerous budding yeasts characteristic of Histoplasma capsulatum in the choroid, retina and central retinal vein . Their identification as H . capsulatum was confirmed by immunofluorescent staining.

Arch Microbiol, 1985 Aug, 142(3), 242 - 7
Detection of an antigenic cell wall layer in Histoplasma capsulatum . An immunoelectron microscopic study; Edwards MR et al.; Histoplasma capsulatum yeast cells have been studied by immunoelectron microscopy using rabbit polyclonal antisera and a biotin-avidin-peroxidase detection system . An antigenic surface layer has been visualized in the cell wall of immunostained organisms . This layer was not seen in samples prepared by standard electron microscopic methods or in negative controls used with the immunocytochemical technique . Without immunostaining the cell wall of Histoplasma appeared almost transparent . In contrast, after immunoperoxidase staining the cell wall was conspicuous, bounded by the darkly stained outer layer . This electron dense layer, appeared to be a reservoir of surface antigens that were recognized by anti-Histoplasma antibodies.

Biochem Biophys Res Commun, 1985 Jul 31, 130(2), 533 - 9
Is hydroxyl radical generated by the Fenton reaction in vivo?
Bilinski T, Krawiec Z, Liczmanski A, Litwinska J.
Yeast mutants deficient in activities of cytosolic superoxide dismutase and catalase A and T were exposed to four different kinds of oxygen stress . The response of the cells contradicts suggestions, that hydroxyl radical is formed in vivo through the Fenton reaction . The results suggest that superoxide radicals are directly responsible for cytotoxic effects of oxygen.

Cancer, 1985 Jul 15, 56(2), 318 - 20
Candida rugosa in immunocompromised infection . Case reports, drug susceptibility, and review of the literature; Sugar AM et al.; Candida rugosa was isolated from two patients . One patient had acute leukemia and developed invasive disease due to this yeast on two occasions while granulocytopenic . Her infection was eventually cured after treatment with amphotericin B . In another immunocompromised patient, the yeast was isolated from the sputum in the presence of a pulmonary infiltrate, but there was no other evidence for a pathogenic role . Antifungal susceptibility testing of the first patient's isolate and three environmental isolates showed all four to be susceptible to amphotericin B, miconazole, and flucytosine, and only the patient isolate was resistant to ketoconazole . These results suggest possibilities for therapy in future encounters . It appears that C . rugosa, a common pathogen in cattle, can be pathogenic in humans under the appropriate circumstances.

J Biol Chem, 1985 Jul 15, 260(14), 8483 - 91
8-Azidoflavins as photoaffinity labels for flavoproteins; Fitzpatrick PF et al.; 8-Azidoflavins have been synthesized and their potential as photoaffinity labels for flavoproteins has been explored . They are very photolabile, and in aqueous media they react with solvent to yield 8-aminoflavins and 8-hydroxlaminoflavins as the main products . They fulfill the criteria expected of a good photoaffinity label, since they bind stoichiometrically at the flavin-binding site of flavoproteins, thus minimizing problems of nonspecific labeling . Second, they absorb strongly in the visible, so that the reactive nitrene can be generated without short wavelength light, minimizing the possibility of light-induced damage of the protein . Third, in the absence of light, 8-N3-flavins are stable, permitting a study of their binding to apoproteins . 8-Azidoflavins have been bound to hen egg white riboflavin-binding protein, Megasphera elsdenii flavodoxin, yeast Old Yellow Enzyme, Aspergillus niger, glucose oxidase, and pig kidney D-amino acid oxidase, and the effect of exposure to visible light has been determined . Only small extents of covalent attachment of the flavin to the protein were found with flavodoxin, D-amino acid oxidase, and Old Yellow Enzyme; much more extensive labeling was obtained with glucose oxidase and riboflavin-binding protein . In addition to their photoreactivity, 8-azidoflavins have been found to be converted to 8-aminoflavins by reaction with sulfite or upon reduction . Similar reactions occur with 8-hydroxylamino-, 8-(O-methyl)hydroxylamino-, and 8-hydrazinoflavins, which serve as models for possible flavin-protein covalent linkages which could be formed in the photolabeling procedure . Some of the properties of these flavins, which were obtained by reaction of 8-F-flavin with the corresponding nucleophiles, are also described.

Nucleic Acids Res, 1985 Jul 11, 13(13), 4971 - 81
Intron splicing: a conserved internal signal in introns of Drosophila pre-mRNAs; Keller EB et al.; The introns of Drosophila pre-mRNAs have been analysed for conserved internal sequence elements near the 3' intron boundary similar to the T-A-C-T-A-A-C in yeast introns and the C/T-T-A/G-A-C/T in introns of other organisms . Such conserved internal elements are the 3' splice signals recognized in intron splicing . In the lariat splicing mechanism, the G at the 5' end of an intron joins covalently to the last A of a 3' splice signal to form a branch point in a splicing intermediate . Analysis of 39 published sequences of Drosophila introns reveals that potential 3' splice signals with the consensus C/T-T-A/G-A-C/T are present in 18 cases . In 17 of the remaining cases signals are present which vary from this consensus just in the middle or last position . In Drosophila introns the 3' splice signal is usually located in a discrete region between 18 and 35 nucleotides upstream from the 3' splice point . We note that the Drosophila small nuclear U2-RNA has sequences complementary to C-T-G-A-T, one variant of the signal, and to C-A-G, one variant of the 3' terminus of an intron . We also note that the absence of any A-G between -3 and -19 from the 3' splice point may be an essential feature of a strong 3' boundary.

Biochimie, 1985 Jul-Aug, 67(7-8), 811 - 7
Solvent distribution in crystals of B- and Z-oligomers; Westhof E et al.; Two crystal structures of deoxyoligomers were refined to high resolution using the programs NUCLIN and NUCLSQ developed for refining the structure of yeast aspartic acid tRNA . The B form oligomer is the dodecamer d(5'OH-C-G-C-G-A-A-T-T-C-G-C-G-3'OH) solved by Dickerson and coworkers in 1981; the data used for the refinement are those deposited in the Brookhaven Data Bank . The crystal structure of the Z form hexamer d(5'OH-5BrC-G-5BrC-G-5BrC-G-3'OH) was resolved in Strasbourg . During the refinement each compound exhibits a different behaviour . Over sixty water molecules were located for each oligomer . The distribution of water molecules is typical of each helical form . Probably due to disorder, the hydration of the B form is not extensive . The minor groove of the B form presents the "spine of hydration" much discussed by Dickerson and coworkers . The Z form hexamer presents its most extended hydration network in the deep cavernous groove corresponding to the minor groove . Intermolecular contacts occur in both oligomers in the minor groove: in the B form through twisted guanine-guanine hydrogen bonding, and in the Z form through base-base stacking and the water network . The role and importance of solvent structure for DNA structure is discussed in the light of the results presented.

Ann Inst Pasteur Microbiol, 1985 Jul-Aug, 136B(1), 9 - 27
Ultrastructural study of the interaction of the fungi Sporothrix schenckii and Ceratocystis stenoceras with bone-marrow-derived murine macrophages; Ryter A et al.; Bone-marrow-derived macrophages of C57BL/6 mice cultivated in vitro were infected with the yeast form of Sporothrix schenckii or Ceratocystis stenoceras . Observations made in light and electron microscopy showed that part of the S . schenckii-containing phagosomes rapidly fused with lysosomes and fungal cells were digested . Surviving fungal cells elongated very rapidly and were liberated into the culture medium after 48 h upon macrophage lysis . The cells of the non-pathogenic isolate C . stenoceras did not elongate and were almost all digested after 8 days, while macrophages were unaltered . Staining for acid phosphatase showed that this enzymatic activity increased soon after ingestion with both isolates . However, this increase was less pronounced with S . schenckii than with the non-pathogenic isolate . The search for a toxic substance produced by S . schenckii and responsible for the low content in acid phosphatase and subsequent macrophage lysis remained negative . It is thus probable that both phenomena essentially resulted from fungus filamentation which led to a dramatic distortion of phagosomes and host cells.

Biochem J, 1985 Jul 1, 229(1), 213 - 9
Iodinated fibroblast beta-glucuronidase as a ligand for receptor-mediated endocytosis; Dean MF et al.; Antibodies raised to human placental beta-glucuronidase were shown to cross-react with the beta-glucuronidase secreted by mouse 3T3 fibroblasts, but did not react with other lysosomal enzymes . The beta-glucuronidase secreted by 3T3 cells was purified 15000-fold by chromatography on an affinity column made from this antibody and resolved into a single component, of Mr 68000, by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis . Iodinated samples of purified enzyme were taken up into mouse peritoneal macrophages by receptor-mediated endocytosis at a rate similar to that calculated previously for unlabelled enzyme, and uptake was competitively inhibited by yeast mannan . Binding of beta-glucuronidase to macrophages was saturable, with a Kd of 7 X 10(-9)l/mol, an affinity comparable with that calculated for the binding of mannosylated bovine serum albumin (Kd 1.3 X 10(-9)l/mol), a ligand specific for mannose receptors . Four times as many molecules of mannosylated albumin (12000) as of beta-glucuronidase (3000), however, bound to each cell . This purification and iodination procedure did not therefore have any adverse effect on the uptake properties of secreted beta-glucuronidase, and provides a ligand with which to investigate binding and specific endocytosis into a range of different types of cell.

Med Radiol (Mosk), 1985 Jul, 30(7), 41 - 6
{Mathematical model of the combined effects of ionizing radiation and hyperthermia on mammalian cells}; Komarov VP et al.; A previously proposed mathematical model for the depiction of the effects of combined ionizing radiation and hyperthermia on yeast cells of different species was applied to the description of experimental data on mammalian cells . The model suggests that the synergistic effect of combined ionizing radiation and hyperthermia is caused by additional lethal damages arising from the interaction of "sublesions" induced by both agents . These "sublesions" are not lethal after the action of these agents, each taken alone . It has been shown that the thermal enhancement ratios are strongly determined by the ratios of radiation and hyperthermia-induced lethal damages . The model predicts that the maximal synergistic effect after a separate action of these modalities will be achieved if the former agent induces a greater part of lethal damages as compared to the latter one . It has been shown that the model depicts quantitatively the synergism of the simultaneous action of the agents used for Chinese hamster cells and predicts the maximal value of the synergistic effect, conditions under which it can be achieved and the dependence of the synergistic effect on the do se rate.

Biochem Pharmacol, 1985 Jul 1, 34(13), 2287 - 90
Selenium and drug metabolism--III . Relation of glutathione-peroxidase and other hepatic enzyme modulations to dietary supplements; Reiter R et al.; Male mice were fed a torula yeast-based diet containing different amounts of added selenium for a period of 4 months . Liver glutathione peroxidase activity assayed with H2O2 showed a logarithmic dependence on dietary selenium with a saturation plateau above 2 ppm Se and an extrapolated zero of 0.02 ppm Se . In contrast, liver selenium content and GSH-Peroxidase activity showed a linear correlation . Glutathione peroxidase activity became undetectable at a liver Se content of about 90 ng Se/g liver wet wt . Thus, about 10% of liver selenium is not related to GSH-Px activity . Five dietary groups were supplemented, respectively, with 0, 0.05, 0.5, 5.0 and 10 ppm Se in the form of Na2SeO3 . Some changes in drug metabolism enzymes were observed with the high Se diets . An increase occurred in Non-Se-GSH activity as well as in ethacrynic acid-assayed GSH transferase, these are interpreted as early signs of Se toxicity . The diet containing 0.01 ppm Se with no supplementary Se produced the multiple hepatic enzyme modulations which were previously reported . The animals raised on this very low Se diet had normal hepatic contents of glutathione, alpha-tocopherol, calcium, magnesium, iron, zinc, copper and manganese compared to controls supplemented with 0.5 ppm Se . However, significant changes in the microsomal fatty acid pattern were observed while the total phospholipid content as well as membrane fluidity showed no differences between the two dietary groups.

Biochim Biophys Acta, 1985 Jul 1, 829(3), 354 - 7
Anomeric specificity of mannose phosphorylation by hexokinase; Giroix MH et al.; In rat parotid or pancreatic islet homogenates incubated at 7 degrees C, hexokinase displayed a greater affinity for but a lower maximal velocity with the alpha-anomer, as distinct from beta-anomer, of D-mannose . The anomeric specificity of mammalian hexokinase was similar in the case of D-mannose and D-glucose, but represented a mirror image of that of yeast hexokinase.

Biochem J, 1985 Jul 1, 229(1), 167 - 71
Inactivation of rabbit muscle phosphoglycerate mutase by limited proteolysis with thermolysin; Price NC et al.; Rabbit muscle phosphoglycerate mutase is inactivated by proteolysis with thermolysin . Inactivation is correlated with the breakage of one (or a few) bond(s) near one end of the polypeptide chain . There is no change in the overall conformation, quaternary structure or binding to Cibacron Blue on proteolysis . The possible analogy with the existence of a flexible tail in the yeast enzyme is discussed.

J Mol Biol, 1985 Jun 25, 183(4), 633 - 8
A plasmodial alpha-tubulin cDNA from Physarum polycephalum . Nucleotide sequence and comparative analysis; Krammer G et al.; As the first step towards correlating structure and function of tubulin in the slime mold Physarum polycephalum we have elucidated the nucleotide sequence of a cDNA that appears to code for all but the last 25 to 30 C-terminal amino acids of a plasmodial alpha-tubulin . Differences in amino acid sequence from those of other alpha-tubulins are distributed fairly evenly throughout the sequence, although a relatively extensive conserved region is found in position 396 to 426 near the C terminus . A small region in position 298 to 307 contains a cluster of amino acid residues unique to Physarum alpha-tubulin . The sequence is 70% homologous to two yeast alpha-tubulins and about 83% homologous to five animal alpha-tubulins . A comparison of the homologies of all the known alpha-tubulins indicates that a large decrease in the accepted point mutation rate has occurred during the evolution of the metazoa, suggesting a major functional specialization of microtubules.

Nucleic Acids Res, 1985 Jun 25, 13(12), 4411 - 6
Sequence and codon recognition of bean mitochondria and chloroplast tRNAsTrp: evidence for a high degree of homology; Marechal L et al.; Bean mitochondria and chloroplast tRNAsTrp, purified by RPC-5 chromatography and two-dimensional gel electrophoresis, have been sequenced using post-labeling techniques . The high degree of sequence homology between bean mitochondria and chloroplast tRNAsTrp shows that these two tRNAs are coded for by closely related genes which have probably evolved from a common ancestor gene . The anticodon of bean mitochondria tRNATrp is CmCA, which can recognize UGG (the codon for tryptophan in the universal code) and is complementary neither to UGA (which codes for tryptophan in mammalian and yeast mitochondria) nor to CGG (which could be a tryptophan codeword in plant mitochondria).

Nucleic Acids Res, 1985 Jun 25, 13(12), 4311 - 32
Structure of a Neurospora RNA polymerase I promoter defined by transcription in vitro with homologous extracts; Tyler BM et al.; A Neurospora in vitro transcription system has been developed which specifically and efficiently initiates transcription of a cloned Neurospora crassa ribosomal RNA gene by RNA polymerase I . The initiation site of transcription (both in vitro and in vivo) appears to be located about 850 bp from the 5' end of mature 17S rRNA . However, the primary rRNA transcripts are normally cleaved very rapidly at a site 120-125 nt from the 5' end in vitro and in vivo . The nucleotide sequence surrounding the initiation site has been determined . The region from -16 to +9 exhibits partial homology to the corresponding sequences from a wide variety of organisms including yeast, but the most striking similarity is to the initiation region from Dictyostelium discoideum which displays 73% homology to the Neurospora sequence from -23 to +47 . The Neurospora sequences from -96 to +97 have been shown to be sufficient for transcription . This region contains two sequences displaying 8/9 bp matches to elements of the 5S rDNA promoter.

Life Sci, 1985 Jun 3, 36(22), 2153 - 61
Degradation and covalent cross-linking of glutathione reductase by hemin; Aft RL et al.; Hemin (ferriprotoporphyrin IX-chloride) can mediate the covalent cross-linking and degradation of yeast glutathione reductase . This reaction requires both NADPH and oxygen suggesting the involvement of a reduced oxygen species in the cross-linking and degradation process . During the course of the reaction the enzymatic activity of glutathione reductase is rapidly destroyed . Implications of these findings for a regulatory role of hemin in cell biology are discussed.

J Pharmacobiodyn, 1985 Jun, 8(6), 432 - 9
Effects of buthiobate, a fungicide, on cytochrome P-450 of rat liver microsomes; Yoshida Y et al.; Effects of buthiobate (S-butyl S'-p-tert-butylbenzyl N-3-pyridyldithiocarbonimidate), a fungicide which inhibits lanosterol 14 alpha-demethylation catalyzed by a cytochrome P-450 (P-450 14DM) of yeast, on cytochrome P-450 of rat liver microsomes were studied . Buthiobate bound to limited forms of cytochrome P-450 in the microsomes with three different Kd values, 0.38, 1.56 and 13.6 microM . Buthiobate inhibited lanosterol 14 alpha-demethylase activity of the microsomes with a 50%-inhibition concentration of 0.4 microM . This concentration was comparable to the lowest Kd of buthiobate for the microsomal cytochrome P-450 and also to the 50%-inhibition concentration of the inhibitor for lanosterol 14 alpha-demethylation by yeast P-450 14DM . Buthiobate partially inhibited 7-ethoxycoumalin O-deethylase activity of the microsomes but inhibited neither benzphetamine N-demethylation nor p-nitroanisole O-demethylation . These observations suggest that cytochrome P-450s catalyzing drug oxidations are rather insensitive to buthiobate . These observations indicate that buthiobate is an unique inhibitor for hepatic microsomal cytochrome P-450 system, which inhibits cholesterol biosynthesis effectively but causes a little effect on drug oxidations.

Sabouraudia, 1985 Jun, 23(3), 179 - 88
Phaeoannellomyces and the Phaeococcomycetaceae, new dematiaceous blastomycete taxa; McGinnis MR et al.; Phaeoannellomyces McGinnis et Schell gen . nov., which is based upon P . elegans McGinnis et Schell sp . nov., is proposed for dematiaceous yeasts which produce percurrently proliferating conidiogenous cells . Cladosporium werneckii Horta is transferred to Phaeoannellomyces as P . werneckii (Horta) McGinnis et Schell comb . nov . because the most stable and distinctive synanamorph produced by this fungus consists of annellidic yeast cells . The Phaeococcomycetaceae McGinnis et Schell fam . nov . is proposed in the class Blastomycetes, division Fungi Imperfecti for the dematiaceous yeast genera Phaeoannellomyces and Phaeococcomyces de Hoog.

Mycopathologia, 1985 Jun, 90(3), 181 - 6
Caffeine degradation and increased ochratoxin A production by toxigenic strains of Aspergillus ochraceus isolated from green coffee beans; Tsubouchi H et al.; The growth and ochratoxin A production of Aspergillus ochraceus strains S-235-100 and IFM 0458, which were isolated from green coffee beans and glutinous rice, respectively, were examined in yeast extract-sucrose (YES) medium containing 0.1 to 1.0% caffeine . The mycelial growth and ochratoxin A formation of strain IFM 0458 was inhibited by caffeine at concentrations over 0.1%, and ochratoxin A was not produced at caffeine levels of 0.5% and 1.0% . Contrary to this, A . ochraceus strain S-235-100 produced a larger amount of ochratoxin A in the presence of 0.5% and 1.0% caffeine when grown on YES medium, reaching a maximum after 15 to 20 days of incubation . The formation of ochratoxin A by nine additional strains of A . ochraceus, three strains of A . elegans and one strain of A . sclerotiorum isolated from green coffee beans was determined on rice and ground green coffee media . A significant degree of degradation of caffeine in the green coffee medium was demonstrated with cultures of nine A . ochraceus isolates from green coffee beans . Most of these isolates showed the potential to grow on moist green coffee beans and to produce a significant amount of ochratoxins.

Cutis, 1985 Jun, 35(6), 536 - 8
Malassezia folliculitis in immunocompromised patients; Yohn JJ et al.; Four cases of Malassezia folliculitis in immuno-compromised patients with leukemia, papillary adenocarcinoma of the lung, and chronic renal failure are reported . This condition manifests with multiple bland asymptomatic follicular papules of the trunk and arms . Biopsy specimens show dilated follicles containing unipolar budding yeast forms . Malassezia is a common infection that must be differentiated from the cutaneous manifestations of systemic candidiasis.

J Am Acad Dermatol, 1985 Jun, 12(6), 1007 - 12
Fixed cutaneous sporotrichosis: unusual histopathology following intralesional corticosteroid administration; Bickley LK et al.; The fixed cutaneous type of sporotrichosis is difficult to diagnose because clinical lesions are variable in appearance and the cells of Sporothrix schenckii are usually scarce in skin biopsy specimens . We have described two patients with lesions of fixed cutaneous sporotrichosis that resembled other inflammatory skin conditions and were treated with intralesional corticosteroids . Subsequent skin biopsies from these lesions demonstrated an unusually large number of yeast cells.

Eur J Epidemiol, 1985 Jun, 1(2), 84 - 9
Studies on histoplasmosis farciminosi (epizootic lymphangitis) in Egypt . Isolation of Histoplasma farciminosum from cases of histoplasmosis farciminosi in horses and its morphological characteristics; Selim SA et al.; Isolation of Histoplasma farciminosum from five horses, showing typical signs of histoplasmosis farciminosi (epizootic lymphangitis) was successfully attempted . The mycelial form of H . farciminosum was isolated on Sabouraud dextrose agar enriched with 2.5% glycerol, brain heart infusion (BHI) agar enriched with 10% horse blood and PPLO dextrose glycerol agar . The last medium proved to be the most effective, both for primary isolation and subculturing of the fungus . It was found that on primary isolation, the lag phase of the mycelial form of the fungus was relatively long, involving 4-8 weeks at 25 degrees C . Colonies of the mycelial form of H . farciminosum appeared on subculture as a yellowish, light brown to deep brown, convoluted, waxy, cauliflower-like growth tending to form scant aerial growth . Conversion of the mycelial form to the yeast form of H . farciminosum was successful by subculturing either on BHI agar with 5% blood or on Pine's medium and incubating at 35-37 degrees C . Complete conversion to the yeast form was achieved only after 4-5 repeated serial transfers onto fresh media every 8 days . The yeast colonies were flat, raised, slightly or deeply wrinkled, white to light gray to grayish brown, and were pasty in consistency.

J Am Mosq Control Assoc, 1985 Jun, 1(2), 186 - 90
Laboratory colonization of Mansonia uniformis, Ma . indiana and Ma . bonneae in Malaysia; Chiang GL et al.; Methods are described for the laboratory colonization of Mansonia uniformis, Ma . indiana and Ma . bonneae in Malaysia . Gravid females oviposited in 500 ml beakers with a layer of water covered with small leaves of Salvinia . Newly hatched larvae were set up in a basal medium of guinea pig dung and water or liver powder, yeast powder and water . Larvae attached to aquatic plants or 'Keaykolour' ruffia snow white paper . The cultures with paper gave better yields than those with plants . Production of Ma . uniformis was higher than the other two species . Twelve generations of Ma . uniformis and 11 generations of Ma . indiana and Ma . bonneae were monitored in the laboratory.

Proc Natl Acad Sci U S A, 1985 Jun, 82(11), 3635 - 9
Synthesis of small nuclear ribonucleoprotein particles by the malarial parasite Plasmodium falciparum; Francoeur AM et al.; Sera from patients with autoimmune diseases have been used to identify small nuclear ribonucleoprotein particles (snRNPs) present in higher eukaryotic cells and also in dinoflagellates . Previously these sera have not detected crossreactive snRNP protein antigens of other lower eukaryotes such as yeast, Tetrahymena, or Dictyostelium . We report that anti-Sm, anti-U1-RNP, and anti-La/SS-B human antisera react with specific snRNP protein antigens synthesized by the protozoan Plasmodium falciparum, the human malarial parasite . These results suggest that the structure and antigenicity (and thus probably the function) of snRNPs have been widely conserved in eukaryote evolution.

Arch Int Physiol Biochim, 1985 Jun, 93(2), 107 - 12
Effects of amino-acid mixtures on food utilization and growth in Tenebrio molitor L; Davis GR; Ten holidic diets, varying in amino-acid concentration or composition, were fed to larvae of Tenebrio molitor for four weeks at 27 +/- 0.25 degrees C and 65 +/- 5% r.h . Effects of diet on growth, food utilization and energy utilization were recorded for individual larvae . Differences in gains in fresh weight or in dry matter among larvae fed diets containing 0% to 5% of the amino-acid mixture were not demonstrated . However, larvae fed 10% or 20% of this mixture gained more than the former, but less than larvae fed a diet of ground whole wheat and brewer's yeast (9:1, w/w) . When the amino-acid mixture was supplemented with alanine, aspartic acid, and serine, or with these three and asparagine, and was fed to larvae at the 10% level, growth was slower than with the unsupplemented mixture . Supplementation of the amino-acid mixture with the first three amino acids did not reduce larval growth when fed at the 20% level . Amounts of food and of energy utilized were positively correlated with larval fresh-weight and dry-matter gains . Energy utilization was negatively correlated with dietary amino-acid level . Food and energy utilization and larval growth were influenced by dietary amino acids, either metabolically or through phagostimulation.

Life Sci, 1985 May 27, 36(21), 2017 - 23
Strychnine antagonizes vaginal stimulation-produced analgesia at the spinal cord; Roberts LA et al.; Vaginal-cervical mechanostimulation (VS) suppresses vocalization and withdrawal responses to noxious stimulation . To determine whether the inhibitory neurotransmitter, glycine, contributes to the action of VS, strychnine, a specific glycine receptor antagonist was administered perispinally via intrathecal catheter in dosages of 1,5,25 and 100 micrograms . Prior to strychnine administration, VS (400 g force) elevated thresholds to elicit vocalization in response to graded intensities of tail shock, and blocked vocalization elicited by stimulation of a skin area, previously sensitized by intradermal injection of a 20% yeast solution . After strychnine administration the analgesic effects of VS were significantly attenuated . These findings suggest that the analgesic action of VS is partially mediated by glycine at the spinal level.

Nucleic Acids Res, 1985 May 24, 13(10), 3667 - 83
Mechanism of codon recognition by transfer RNA studied with oligonucleotides larger than triplets; Labuda D et al.; The binding of yeast tRNAPhe to UUCA, UUCC, UUCCC, UUCUUCU, U4, U5, U6 and U7 was analysed by fluorescence temperature jump and equilibrium sedimentation measurements . In all cases the two observed relaxation processes can be assigned to alpha) an intramolecular conformation change of the anticodon loop and beta) preferential binding of the oligonucleotides to one of the anticodon conformations . The anticodon loop transition is associated with inner sphere complexation of Mg2+ and proceeds with rate constants of about 10(3) s-1 . The rate constants of oligonucleotide binding are between 4 and 10 X 10(6) M-1s-1 and reflect an increase of the association rate with the number of binding sites compensated to some degree by electrostatic repulsion in the preequilibrium complex . Neither temperature jump nor equilibrium sedimentation experiments provided evidence for UUCA or UUCC induced tRNA dimerisation, although UUC binding leads to strong tRNA dimerisation under equivalent conditions . The results obtained for the longer oligonucleotides are similar . In the case of UUCUUCU with its two potential binding sites for tRNAPhe there was no evidence for the formation of 'ternary' complexes . Apparently tRNAPhe binds preferentially to the second UUC of this 'messenger' and forms additional contacts with residues on either side of the codon . Some evidence for the formation of ternary complexes is obtained for U6 and U7, although the extent of this reaction remains very small . Our results demonstrate that the mode of tRNA binding to a codon is strongly influenced by residues next to the codon . The formation of cooperative contacts between tRNA molecules at adjacent codons apparently requires support by a catalyst adjusting an appropriate conformation of messenger and tRNA molecules.

Nature, 1985 May 23-29, 315(6017), 347 - 50
Size variation in chromosomes from independent cultured isolates of Plasmodium falciparum; Kemp DJ et al.; The complexity of the life cycle of the protozoan malaria parasite Plasmodium falciparum has hindered genetic analysis; even the number of chromosomes in P . falciparum is uncertain . The blood stages of rodent malaria parasites are haploid and hybridization with cloned complementary DNAs similarly suggests a haploid genome in P . falciparum blood stages (ref . 4 and our unpublished results) . A novel approach to karyoptic and linkage analysis in P . falciparum has been provided recently by the technique of pulsed-field gradient (PFG) gel electrophoresis, which allows the fractionation of DNA molecules of 30-3,000 kilobases (kb), a range including the sizes of intact chromosomal DNA molecules from eukaryotes such as yeast and trypanosomatids . We describe here the fractionation by PFG electrophoresis of chromosomal DNA molecules from P . falciparum into at least seven discrete species which vary in size by up to 20% between different isolates . Several genes for P . faciparum antigens which contain repetitive sequences are located on different chromosomes . Surprisingly, two of the chromosomes seem to contain the same sequences.

Experientia, 1985 May 15, 41(5), 622 - 3
Modulation of protein kinases and phosphoprotein phosphatases by a small acidic protein from bovine brains; Kuo WN et al.; A small, acidic and heat-stable protein was purified from bovine brains by column chromatography on DEAE-cellulose, Bio-Gel HTP, Affi-Gel phenothiazine and Sephadex G-75 . This protein stimulates megamodulin-dependent protein kinase I from brains and phosphoprotein phosphatases from either brain or yeast . However, it inhibits cyclic AMP-dependent protein kinases from skeletal muscle.

Nature, 1985 May 30-Jun 5, 315(6018), 430 - 2
A role for branchpoints in splicing in vivo; Rautmann G et al.; The nucleotides immediately surrounding intron/exon junctions of genes transcribed by RNA polymerase B can be derived from 'consensus' sequences for donor and acceptor splice sites by only a few base changes . Studies in vivo have underlined the importance of these junction nucleotides for splicing . In higher eukaryotes, no evidence has been found for specific internal intron sequences involved in splicing . However, the recent discovery that, in vitro, introns are excised in a lariat form where the 5' end of the intron is joined via a 2'-5'-phosphodiester linkage to an A residue (branchpoint acceptor) close to the 3' end of the intron, suggests that internal intron sequences may nonetheless be important for splicing . Indeed, in yeast nuclear genes, the internal sequence 5'-TACTAAC-3' (or close homologue) is essential for splicing in vivo . A proposed consensus sequence for branchpoints in mammalian introns is 5'-CT(A/G)A(C/T)-3' . This sequence resembles the essential yeast internal sequence . Are branchpoints involved in the splicing of introns of higher eukaryotes in vivo? We show here that a branchpoint sequence from a human globin gene (5'-CTGACTCTCTCTG-3') greatly enhances the efficiency of splicing of a 'synthetic' intron in HeLa cells . A mutated branchpoint sequence, 5'-CTCCTCTCTCTG-3', in which the branchpoint acceptor nucleotide A has been deleted and the neighbouring purine G mutated to a C, does not exhibit this enhancing capability . We conclude that branchpoints have an important function in the splicing process in vivo.

Poult Sci, 1985 May, 64(5), 955 - 62
Liver response to diet and estrogen in white Leghorn and Rhode Island Red chickens; Takahashi K et al.; Experiments were conducted to compare the response of Rhode Island Reds (RIR) and White Leghorns (WL) to estradiol (E2) administration and diets of different composition . In growing chickens injected with E2 for 7 days, RIR accumulated significantly more hepatic lipid and had significantly less plasma lipid than WL . Both RIR and WL, injected for 4 days with E2, had lower hepatic lipids when fed a diet containing fish meal, alfalfa meal, and torula yeast (FAY) than when fed a corn-soybean meal diet (CS) . Plasma glutamic oxaloacetic transaminase activity was significantly lower for RIR . Feeding a FAY diet to laying hens significantly reduced hepatic lipids in WL but not in RIR . Implantation of tubes containing E2 in RIR-laying hens resulted in a significant body weight loss and 25% mortality, while a similar treatment in WL-laying hens caused no body weight loss or mortality . Injecting both RIR and WL-laying hens with E2 caused a marked body weight loss and high mortality in both breeds although WL were less severely affected . The results show that breeds of chickens respond differently to both E2 administration and changes in diet composition.

Poult Sci, 1985 May, 64(5), 947 - 54
Dietary effects on content of hepatic lipid, plasma minerals, and tissue ascorbic acid in hens and estrogenized chicks; Brenes A et al.; The purpose of this study was to determine if plasma mineral levels and plasma or hepatic ascorbic acid would be affected by the composition of the diet fed to chickens with higher circulating estrogen levels . Three-week-old broiler chicks were implanted with estradiol dipropionate to give estimated release rates of 0, 3.2, and 8.0 micrograms/bird/day . The chicks were fed a corn-soy diet (CS) or a diet containing 5% fish meal, 5% alfalfa meal and 10% torula yeast (FAY) . The FAY diet resulted in significantly lower hepatic lipid and plasma iron, copper, manganese, and zinc in chicks administered the highest level of estrogen . In two experiments laying hens were fed either CS or diets containing fish meal (FM), alfalfa meal (AM), distillers dried grains with solubles (DDGS) or wheat-soy (WS) for 4 weeks . In the first experiment, liver lipid content was not significantly affected by diet composition, but plasma iron was significantly reduced by the AM, DDGS, and WS diets . No significant effects on copper, manganese, and zinc in plasma were observed among the dietary treatments . In the second experiment, relative liver weight was significantly reduced with the AM, DDGS, and WS diets and liver lipid by the AM diet . Plasma iron levels were significantly reduced by feeding all diets compared with the CS diet, but no significant differences in total iron binding capacity were observed . Plasma and hepatic ascorbic acid were significantly increased by the AM, DDGS, and WS diets, but no significant differences in hepatic ascorbic acid were observed when calculated per unit of fat-free dry matter.(ABSTRACT TRUNCATED AT 250 WORDS)

Poult Sci, 1985 May, 64(5), 963 - 8
Activation of hepatic microsomal mixed function oxidase system in broiler chicks by diet changes; Brenes A et al.; Two experiments were conducted to investigate the effect of diet composition on activity of the microsomal mixed function oxidase (MFO) system in broiler chicks . One-day-old chicks were fed for 10 days either a corn-soy (CS) diet or diets supplemented with a combination of fish meal, alfalfa meal, and torula yeast (FAY) or individually with fish meal (FM), alfalfa meal, distillers dried grains with solubles (DDGS), or torula yeast (TY) . Activities of hepatic microsomal aniline hydroxylase and aminopyrine N-demethylase were increased when chicks were fed FAY, FM, TY, or DDGS compared with those fed CS . In another experiment chicks were fed the CS and FAY diets to 4 weeks of age . Hepatic microsomal cytochrome P-450, cytochrome b5, and activities of aniline hydroxylase and aminopyrine N-demethylase were significantly increased in birds fed the FAY diet . Activities of NADPH and NADH-cytochrome C reductase were not affected . These results show that the hepatic microsomal MFO system is activated by changes in diet composition and suggest that this activation may be responsible for reducing estradiol and liver lipid levels when similar diets are fed to laying hens.

J Cell Physiol, 1985 May, 123(2), 288 - 96
Enhancement of colony-stimulating-factor--dependent clonal growth of murine macrophage progenitors and their phagocytic activity by retinoic acid; Goldman R; The effect of retinoic acid (RA) on the colony-stimulating-factor-dependent clonal growth of myeloid progenitors was assessed in semisolid agar cultures of mouse bone marrow cells using L-cell-conditioned medium that gave rise to macrophage colonies, granulocyte colonies, and mixed macrophage-granulocyte colonies and clusters . RA was found to enhance the overall formation of myeloid colonies (about 50%) and clusters in 7-day cultures . The increase was due to an enhanced formation of macrophage colonies (70-250%) and clusters which reached a maximal value at about 3 microM RA . In 4-day cultures, the effect of RA on macrophage colony formation was biphasic with a maximal enhancement at 10 nM . RA suppressed granulocyte-colony formation in 4-day cultures . RA increased the phagocytic activity of bone-marrow-derived macrophages at all stages of differentiation and/or maturation in culture . The Fc-receptor-mediated erythrophagocytosis as well as the phagocytosis of heat-killed yeast cells (HK-yeast) and starch particles increased by RA treatment in a dose-dependent manner, reaching an increase of 100-200% of the activity expressed in the absence of RA . Peritoneal exudate macrophages likewise exhibited an increased phagocytic response to a variety of particles, at both physiological and pharmacological concentrations of RA . Expression of an RA-mediated increase in phagocytic activity required a prolonged incubation with RA (greater than 19 hr) . The data suggest that RA may be of physiological relevance in the regulation of proliferation and function of hemopoietic cells . Therapeutic doses of RA may potentiate macrophage proliferation and function, elements that are crucial at all phases of the various defense mechanisms that the organism possesses.

Gastroenterology, 1985 May, 88(5 Pt 1), 1271 - 3
Blastomycosis of the esophagus presenting with gastrointestinal bleeding; McKenzie R et al.; A patient with abdominal discomfort and hematemesis was found to have lower esophageal inflammation on endoscopy . A biopsy specimen from this area showed yeast forms of Blastomyces dermatitidis with a polymorphonuclear infiltrate and granuloma formation . Extensive evaluation showed no other bleeding site . Sputum samples obtained earlier also showed B . dermatitidis . Treatment with amphotericin B led to resolution of bleeding and eradication of the fungus on repeat biopsy and sputum samples . An esophageal stricture subsequently developed and required dilation . Seven previously reported cases of esophageal blastomycosis are reviewed.

Proc Natl Acad Sci U S A, 1985 May, 82(10), 3192 - 6
Enzyme-catalyzed processes in organic solvents; Zaks A et al.; Three different lipases (porcine pancreatic, yeast, and mold) can vigorously act as catalysts in a number of nearly anhydrous organic solvents . Various transesterification reactions catalyzed by porcine pancreatic lipase in hexane obey Michaelis-Menten kinetics . The dependence of the catalytic activity of the enzyme in organic media on the pH of the aqueous solution from which it was recovered is bell-shaped, with the maximum coinciding with the pH optimum of the enzymatic activity in water . The catalytic power exhibited by the lipases in organic solvents is comparable to that displayed in water . In addition to transesterification, lipases can catalyze several other processes in organic media including esterification, aminolysis, acyl exchange, thiotransesterification, and oximolysis; some of these reactions proceed to an appreciable extent only in nonaqueous solvents.

J Am Acad Dermatol, 1985 May, 12(5 Pt 1), 852 - 6
Double-blind treatment of seborrheic dermatitis with 2% ketoconazole cream; Skinner RB Jr et al.; Thirty-seven patients with seborrheic dermatitis were treated topically with a 2% ketoconazole cream or its vehicle control in a double-blind study . The subjects were studied for numbers of Malassezia ovalis (Pityrosporum ovale) cells in their scalp scale; changes in numbers of yeast cells and morphology of M . ovalis were tabulated along with clinical assessment of improvement . The 2% ketoconazole cream, but not the placebo cream, reduced the numbers of viable yeast cells on the scalp . The clinical effect of 2% ketoconazole cream was good (75%-95% improvement) or better in eighteen of twenty subjects; the placebo cream produced good results in only three of seventeen subjects treated . Results of this study are consistent with the view that M . ovalis plays a central role in the pathogenesis of seborrheic dermatitis.

J Immunol, 1985 May, 134(5), 3346 - 55
Biochemical analysis and subcellular localization of a neutrophil-specific antigen, PMN-7, involved in the respiratory burst; Melnick DA et al.; The adherence of serum-opsonized yeast to neutrophils results in phagocytosis of these particulate stimuli and activation of the respiratory burst . Both events are mediated or modulated in part by the surface receptors for IgG and complement . The link between the binding of complex particulate stimuli to the cell surface, and the triggering of these neutrophil functions, is not completely understood . We have previously described an anti-human neutrophil, murine monoclonal antibody PMN7C3, which specifically inhibits the respiratory burst of neutrophils stimulated with serum-opsonized yeast . In the present study, we show that the antigen recognized by PMN7C3 (PMN7 antigen) is present on a number of neutrophil proteins, including the recently described group of related leukocyte membrane glycoproteins CR3, LFA-1, and p150,95 . The PMN-7 antigen differs from other antigens associated with the C3bi receptor complex (MAC 1, MO 1, OKM1, OKM10) in that it is present only on neutrophils among peripheral blood cells . Furthermore, the binding of PMN7C3 to the neutrophil surface inhibits the activation of the respiratory burst by serum opsonized zymosan without affecting phagocytosis of these particulate stimuli . The cross-linking of cell surface PMN7 antigen by multivalent antibody is associated with the capping and internalization of antigen-antibody complexes, and appears to be necessary for the expression of maximum inhibition of opsonized zymosan-triggered respiratory burst activity . PMN7C3 also binds to a group of granule-associated proteins biochemically distinct from CR3, LFA-1, and p150,95 . These granule-associated proteins containing PMN7 antigen can be mobilized to the cell surface with secretion . PMN7 antigen-bearing proteins may play a role in modulating the activation of the respiratory burst associated with phagocytosis of serum-opsonize zymosan.

Zh Mikrobiol Epidemiol Immunobiol, 1985 May, (5), 36 - 9
{Cultivation characteristics of Legionella pneumophila in a liquid medium}; Kostiuchenko VI et al.; The comparative analysis of liquid media for the cultivation of Legionella pneumophila, sterilized by filtration and autoclaving, has shown that, in contrast to media prepared on the basis of yeast extract, the capacity of media based on proteose peptone for supporting the growth of Legionella is not influenced by the method of their sterilization . The possibility of cultivating this organism in liquid media without ferric pyrophosphate has been demonstrated.

EMBO J, 1985 May, 4(5), 1093 - 102
A low copy number, copia-like transposon in maize; Johns MA et al.; Bs1, a transposable element that moved into the maize Adh1 gene following barley stripe mosaic virus infection, is shown to be present in 1-5 copies in all maize and teosinte lines tested . Bs1 sequences do not hybridize with the genome of barley stripe mosaic virus . The insertion of Bs1 is bounded by 304-bp perfect direct repeats, similar in structure to Ty1 in yeast, copia and related elements in Drosophila, and vertebrate pro-retroviruses, but different from all other known plant transposons . No free copies of the terminal sequences or large internal deletions of Bs elements could be detected . Bs1 is apparently not related to several transposons which moved into the Shrunken gene in lines made genetically unstable by barley stripe mosaic virus infection, suggesting that this virus may cause genome shock, resulting in a generalized liberation of transposons in response to environmental stress.

Proc Natl Acad Sci U S A, 1985 May, 82(10), 3340 - 4
Genes for respiratory chain proteins and ribosomal RNAs are present on a 16-kilobase-pair DNA species from Chlamydomonas reinhardtii mitochondria; Boer PH et al.; We have used heterologous hybridization and DNA sequence analysis to determine whether the 16-kilobase-pair (kbp) DNA from Chlamydomonas reinhardtii mitochondria is the functional equivalent of mtDNA in other eukaryotes . Restriction fragments corresponding to a continuous internal stretch spanning 75% of the 16-kbp DNA have been cloned and mapped, and regions hybridizing with probes specific for the cytochrome oxidase subunit I {CytOx I (acronym COI)} and apocytochrome b (Cyt b) genes of yeast and the mitochondrial 26S and 18S rRNA genes of wheat have been identified . Sequence analysis has verified the presence of CytOx I and the large and small subunit rRNA genes in the C . reinhardtii 16-kbp DNA . In the region of the 16-kbp DNA corresponding to exon 4 in the yeast CytOx I gene, the derived amino acid sequence is 61% and 63% identical with the CytOx I amino acid sequences of yeast and human mitochondria, respectively . Notably, tryptophan is specified by TGG rather than by TGA in this section of the C . reinhardtii CytOx I gene . A probe from the CytOx I region of the 16-kbp DNA hybridizes only with this 16-kbp DNA in Southern blots of total cellular DNA from C . reinhardtii but with a larger DNA species in the total cellular DNA of C . moewusii and C . eugametos--two species that lack a 16-kbp DNA . These observations provide evidence that C . reinhardtii 16-kbp DNA comprises at least part of the mitochondrial genome of this organism and that a homologous DNA exists in other species of Chlamydomonas.

Nucleic Acids Res, 1985 Apr 11, 13(7), 2485 - 502
Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene; Tso JY et al.; Full length cDNAs encoding the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from rat and man have been isolated and sequenced . Many GAPDH gene-related sequences have been found in both genomes based on genomic blot hybridization analysis . Only one functional gene product is known . Results from genomic library screenings suggest that there are 300-400 copies of these sequences in the rat genome and approximately 100 in the human genome . Some of these related sequences have been shown to be processed pseudogenes . We have isolated several rat cDNA clones corresponding to these pseudogenes indicating that some pseudogenes are transcribed . Rat and human cDNAs are 89% homologous in the coding region, and 76% homologous in the first 100 base pairs of the 3'-noncoding region . Comparison of these two cDNA sequences with those of the chicken, Drosophila and yeast genes allows the analysis of the evolution of the GAPDH genes in detail.

J Biol Chem, 1985 Apr 10, 260(7), 4384 - 9
Isolation and properties of recombinant DNA produced variants of human alpha 1-proteinase inhibitor; Travis J et al.; Using the glyceraldehyde-3-phosphate dehydrogenase promoter, nonglycosylated human alpha 1-proteinase inhibitor, representing 10% of the soluble cell protein, has been synthesized in yeast . Two forms of this protein were isolated with one being analogous to the human plasma protein and the other having the amino acid valine replacing methionine at position 358 (the P1 position) . Both proteins were more sensitive to heat inactivation than the plasma form, and both had shorter half-lives in rabbits . These differences were presumably due to the absence of carbohydrate . Each protein could bind neutrophil elastase at a rate only slightly slower than that of human plasma alpha 1-proteinase inhibitor . However, the valine variant was stable to oxidation, while the P1 methionine-containing protein was readily inactivated . The specificity of alpha 1-proteinase inhibitor (methionine) was identical to that of the plasma form; however, the valine form could only effectively bind to neutrophil or pancreatic elastase, "trypsin-like" serine proteinases not being inactivated at all . These data indicate the potential importance of mutant forms of proteinase inhibitors, produced by recombinant DNA technology, as therapeutic agents for the inactivation of excess proteinases of a specific type in tissues.

Biochemistry, 1985 Apr 9, 24(8), 1904 - 9
Fusion of phospholipid vesicles induced by muscle glyceraldehyde-3-phosphate dehydrogenase in the absence of calcium; Morero RD et al.; Ca2+-induced fusion of phospholipid vesicles (phosphatidylcholine/phosphatidic acid, 9:1 mol/mol) prepared by ethanolic injection was followed by five different procedures: resonance energy transfer, light scattering, electron microscopy, intermixing of aqueous content, and gel filtration through Sepharose 4-B . The five methods gave concordant results, showing that vesicles containing only 10% phosphatidic acid can be induced to fuse by millimolar concentrations of Ca2+ . When the fusing capability of several soluble proteins was assayed, it was found that concanavalin A, bovine serum albumin, ribonuclease, and protease were inactive . On the other hand, lysozyme, L-lactic dehydrogenase, and muscle and yeast glyceraldehyde-3-phosphate dehydrogenase were capable of inducing vesicle fusion . Glyceraldehyde-3-phosphate dehydrogenase from rabbit muscle, the most extensively studied protein, proved to be very effective: 0.1 microM was enough to induce complete intermixing of bilayer phospholipid vesicles . Under conditions used in this work, fusion was accompanied by leakage of internal contents . The fusing capability of glyceraldehyde-3-phosphate dehydrogenase was not affected by 5 mM ethylenediaminetetraacetic acid . The Ca2+ concentration in the medium, as determined by atomic absorption spectroscopy, was 5 ppm . Heat-denatured enzyme was incapable of inducing fusion . We conclude that glyceraldehyde-3-phosphate dehydrogenase is a soluble protein inherently endowed with the capability of fusing phospholipid vesicles.

J Clin Microbiol, 1985 Apr, 21(4), 641 - 2
A new biotype of Legionella dumoffii; Edelstein PH et al.; A new biotype of L . dumoffii was isolated from lung and sputum samples of an immunosuppressed patient with pneumonia . This strain differs from other described strains of L . dumoffii in that it fails to produce browning of tyrosine-containing buffered yeast extract medium.

Biochem Pharmacol, 1985 Apr 1, 34(7), 977 - 83
Enhanced production of ethylene from methional by iron chelates and heme containing proteins in the system consisting of quinone compounds and NADPH-cytochrome P-450 reductase; Komiyama T et al.; The addition of iron chelates or heme containing proteins to the systems consisting of NADPH-cytochrome P-450 reductase and quinone compounds, such as vitamin K3 (menadione), adriamycin, tetrahydropyranyladriamycin, daunomycin, aclacinomycin A, carbazilquinone, and mitomycin C, showed the enhanced production of ethylene from methional . In the vitamin K3 system, the effective iron chlates were Fe(II)-EDTA, Fe(II)-ADP, Fe(II)-bleomycin A2, and hemin, and the effective iron containing proteins were methemoglobin, myoglobin, ferredoxin, and partially purified cytochromes P-450, P-420, and b5, and the reversed effects were observed by horse radish peroxidase and sulfite reductase from yeast . In the system consisting of aclacinomycin A and methemoglobin, the ethylene production was potently inhibited by radical scavengers, such as Tiron, Tris, thiourea, and KI, and weakly inhibited by some other scavengers . In the system containing vitamin K3 and methemoglobin, the ethylene production was potently inhibited by catalase, but partially by superoxide dismutase, KCN, and NaN3 . In this system, the absorption spectrum of methemoglobin was immediately changed to oxyform and quenched with time, and catalase protected the decrement of the spectrum . The addition of hydrogen peroxide or cumene hydroperoxide to methemoglobin also produced ethylene from methional.

J Cell Sci, 1985 Apr, 75, 131 - 47
Immunofluorescence microscopy of microtubules in intact cell lineages of the moss, Physcomitrella patens . I . Normal and CIPC-treated tip cells; Doonan JH et al.; Monoclonal antibodies to yeast tubulin have been used to visualize the distribution of microtubules in the intact filamentous protonemata of the moss Physcomitrella patens . Protonemata were prepared for immunofluorescence by fixation in formaldehyde and cells were made permeable with Driselase . Extensive cell files were preserved by 'blotting' the moss onto glutaraldehyde-derivatized coverslips . Problems due to fluorescence from chloroplasts were obviated by extraction with dimethyl sulphoxide and the non-ionic detergent, Nonidet NP40 . These improvements allowed us to determine that microtubules were present throughout the cell cycle in the apical dome of caulonemal tip cells, that was a pronounced association of microtubules with the nucleus, that 'astral' microtubules were associated with the mitotic spindle and during anaphase may be involved in reorientation of the spindle before an oblique cytokinesis in caulonemata and that the cytokinetic phragmoplast appeared identical to the structure described for higher plants . Microtubules appeared to converge at the very tip of apical caulonemal cells and this was studied further by treating cells with CIPC--a drug that is known to produce multiple microtubule-organizing centres--and which here produces multiple foci for microtubules at the tip . These observations emphasize the involvement of microtubules in tip growth, alignment of the cell plate and nuclear migration--processes that are fundamental to the morphogenesis of filamentous organisms.

Photodermatol, 1985 Apr, 2(2), 80 - 5
The Candida phototoxicity test . The sensitivity of different strains and species of Candida, standardization attempts and analysis of the dose-response curves for 5- and 8-methoxypsoralen; Knudsen EA; The purpose of this study was to answer the question as to whether PUVA-sensitivity of different Candida strains and species varies, to investigate the possibilities for standardization of the test and to produce calibration curves suitable for an approximate quantification of psoralens in plants . In the test, paper discs with different concentrations of 5- or 8-MOP in ethanol 60% were placed on yeast-seeded Sabouraud-agar dishes that were irradiated with UVA . The results were based on the diameter of growth inhibition zones . Small but significant differences in sensitivity were found between the 12 Candida strains tested, somewhat greater between the 15 species . Standardization of the method implies the use of the same test organism, depth of agar, time of pre-diffusion, medium, solvent and method for application of psoralen-extract and UVA dose . Dose-response curves for 5-MOP and 8-MOP obtained under standardized conditions demonstrated a linear relationship between the diameter of growth inhibition zones and the logarithm of the psoralen concentration in the range from 1 to 400 micrograms/ml . They were suitable for estimating the content of ethanolsoluble phototoxic substances in terms of 5- or 8-MOP equivalences.

Cell Biol Int Rep, 1985 Apr, 9(4), 307 - 14
A prominent microtubule cytoskeleton in Acanthamoeba; Preston TM; A method for preparing by detergent extraction the cytoskeletons of substrate-attached, motile Acanthamoeba castellanii is described . A monoclonal antibody to yeast alpha tubulin has been used to demonstrate the presence of abundant microtubules in the cytoskeleton of this amoeba by fluorescence and whole-mount electron microscopy . Individual microtubules, often more than 10 micron long, interweave to form a well-developed 3-D network pervading the cytoplasm and embracing the nucleus . In some cases immunofluorescent staining reveals distinct nodes in the perinuclear region of this microtubular network.

Arch Pathol Lab Med, 1985 Apr, 109(4), 330 - 2
Hyphal forms of Histoplasma capsulatum . A common manifestation of intravascular infections; Hutton JP et al.; The usual tissue form of Histoplasma capsulatum is a small intracellular yeast, though extracellular organisms of unusually large size have also been described . The production of hyphae in tissue has been regarded as rare and we believe not generally appreciated . We report a case of probable Histoplasma endocarditis from which an embolus demonstrated abundant mycelial forms on histologic examination . Of previously reported cases of H capsulatum endocarditis with detailed pathologic description, nine of 27 cases are found to demonstrate hyphal forms . Hyphal structures should be considered a common morphologic variant of H capsulatum in intravascular lesions.

J Biol Chem, 1985 Mar 25, 260(6), 3658 - 65
The carbohydrate structure of porcine uteroferrin and the role of the high mannose chains in promoting uptake by the reticuloendothelial cells of the fetal liver; Saunders PT et al.; Uteroferrin, the iron-containing, progesterone-induced phosphatase of the porcine uterus, is a glycoprotein carrying a single oligosaccharide chain . Most of the uteroferrin isolated from either uterine secretions or allantoic fluid has endoglycosidase H-sensitive carbohydrate chains with either five or six mannose residues . As determined by 1H-NMR spectroscopy, the Man6 oligosaccharide has the following structure . (Formula: see text) The Man5 species lacks the terminal alpha 1,2-linked residue . Uteroferrin is transported across the pig placenta and has been proposed to be involved in iron transfer to the fetus (see Buhi, W . C., Ducsay, C . A., Bazer, F . W., and Roberts, R . M . (1982) J . Biol . Chem . 257, 1712-1721) . Injection of 125I-labeled uteroferrin into the umbilical vein of midpregnant fetuses resulted in incorporation of label into the liver, the major site of fetal erythropoiesis . Light and electron microscope autoradiography revealed that the primary sites of uteroferrin uptake were the reticuloendothelial cells lining the liver sinusoids . Reticuloendothelial cells isolated from either fetal pig or adult rat livers were shown to accumulate uteroferrin when cultured in vitro . Uptake was inhibited by yeast mannan and by glycopeptides isolated from either ovalbumin or uteroferrin . Rat cells did not accumulate uteroferrin whose high mannose chains had been removed using endoglycosidase H . Moreover, the K uptake values (3 X 10(-7) M), specific competition by D-mannose and L-fucose bovine serum albumin, and inhibition by EDTA are consistent with an uptake mechanism involving a receptor for high-mannose oligosaccharides on the liver sinusoidal cells . It is suggested that one function of this receptor in the fetal pig is to remove maternally derived uterine glycoproteins from the fetal circulation . In the case of uteroferrin this process provides iron to the fetal liver.

Eur J Biochem, 1985 Mar 15, 147(3), 489 - 93
Biosynthesis and secretion of tremerogen A-10, a polyisoprenyl peptide mating pheromone of Tremella mesenterica; Miyakawa T et al.; The biosynthetic pathway of tremerogen A-10, a polyisoprenyl peptide mating pheromone produced by mating type AB cells of the heterobasidiomycetous yeast Tremella mesenterica, was investigated by immunological techniques with antibody specific to the peptide moiety of the pheromone . Using the biological assay and the radioimmunoassay of the pheromone and its related substances, it was suggested that the peptide is synthesized near the end of logarithmic phase of growth with a temporary accumulation of precursors in the cell . The precursors initially appeared in membrane-bound form and were subsequently converted to soluble forms prior to the secretion . The pheromone acquired its biological activity during the secretion . In the presence of tunicamycin or compactin, pheromone production was blocked with accumulation of membrane-bound precursors . Monensin, however, blocked pheromone production with accumulation of soluble precursors . The molecular species which accumulated in the presence of the antibiotics were analyzed by immunoprecipitation followed by sodium dodecyl sulfate/urea/polyacrylamide gel electrophoresis . In the absence of the inhibitors, membrane-bound precursors with molecular masses of 28 kDa, 12 kDa, 7.8 kDa and 2.8 kDa were found . The precursors which accumulated in the presence of tunicamycin and compactin were the 12-kDa and 28-kDa species, respectively . The results suggested that membrane-bound very high-molecular-mass precursors were initially formed and their extensive modifications, including glycosylation, farnesylation and proteolytic digestion, occur in the membrane . Based on these data, a biosynthetic and secretory pathway was postulated.

Biochem J, 1985 Mar 15, 226(3), 653 - 9
Purification of nucleotide-requiring enzymes by immunoaffinity chromatography; Stapleton SR et al.; Monospecific (affinity-purified) anti-(yeast glucose-6-phosphate dehydrogenase) IgG inhibits three different NADPH-requiring enzymes, chicken liver dihydrofolate reductase, pigeon liver fatty acid synthetase and chicken liver malic enzyme . The inhibition of all three enzymes was approx . 50% in a 2h incubation with 100 micrograms of IgG . Similarly, with several different NADH-requiring enzymes, an immunocrossreactivity was observed . Monospecific anti-(rabbit muscle glyceraldehyde-3-phosphate dehydrogenase) IgG inhibited yeast alcohol dehydrogenase and pig heart malate dehydrogenase by 39% and 55% respectively . The cross-reactivity observed was tested by affinity chromatography . Immunoaffinity columns made with each monospecific IgG were able to bind each of the enzymes it immunotitrated . Enzymes were eluted with a nondenaturing solvent with little loss of activity . The immunoaffinity column with monospecific anti-(glucose-6-phosphate dehydrogenase) IgG as the bound ligand was also used to purify partially (over 150-fold) both isocitrate dehydrogenase and dihydrofolate reductase from crude rat liver homogenate.

Experientia, 1985 Mar 15, 41(3), 385 - 7
IgG purification to measure the level of an iodinated thyroglobulin peptide, the 3,5,3',5' tetraiodo-l-tyrosyl-l-tyrosine in human serum; Bernard D et al.; Antibodies reacting with 3,5,3',5' tetraiodo-l-tyrosyl-l-tyrosine (I2Tyr-I2Tyr) were elicited in rabbits by immunization with an oxidized yeast conjugate coupled with I2Tyr-I2Tyr . Ion-exchange chromatography was used to purify immunoglobulins, in order to improve the specificity in measurement of I2Tyr-I2Tyr level in patient serum . IgG binding capacity versus I2Tyr-I2Tyr was considerably increased after immunoglobulin purification.

Biochem J, 1985 Mar 15, 226(3), 733 - 40
Comparison between 1-deoxynojirimycin and N-methyl-1-deoxynojirimycin as inhibitors of oligosaccharide processing in intestinal epithelial cells; Romero PA et al.; The alpha-glucosidase inhibitor N-methyl-1-deoxynojirimycin (MDJN) inhibits the synthesis of N-linked complex oligosaccharides in rat intestinal epithelial cells to the same extent as reported previously for 1-deoxynojirimycin (DJN) {Saunier, Kilker, Tkacz, Quaroni & Herscovics (1982) J . Biol . Chem . 257, 14155-14161} . Analysis of each of the endo-beta-N-acetylglucosaminidase H (endo H)-sensitive oligosaccharides separated by h.p.l.c . with yeast glucosidase I, which specifically removes the terminal glucose residue from oligosaccharides containing three glucose residues, and with jack-bean (Canavalia ensiformis) alpha-mannosidase, indicates that both inhibitors cause the accumulation of a mixture of glucosylated oligosaccharides containing one to three glucose residues and seven to nine, and even possibly six, mannose residues . About 70% of the endo H-sensitive oligosaccharides formed in the presence of MDJN contain three glucose residues, compared with only about 20% of the corresponding oligosaccharides of the DJN treated cells . It is concluded that both compounds inhibit the formation of N-linked complex oligosaccharides by interfering with the processing glucosidases . These compounds are valuable in the study of the role of oligosaccharides in glycoproteins.

Mycopathologia, 1985 Mar, 89(3), 139 - 45
In vitro phagocytosis of several Candida berkhout species by murine leukocytes; Fontenla de Petrino SE et al.; In vitro phagocytosis of thirteen Candida berkhout species by rat leukocytes was studied to assess a possible correlation between pathogenicity and phagocytosis Yeast-leukocyte suspensions were mixed up for 3 h and phagocytic index, germ-tube formation and leukocyte candidacidal activity were evaluated . Highest values for phagocytosis were reached in all cases at the end of the first hour . Leukocyte candidacidal activity was absent . Only C . albicans produced germ-tubes . The various phagocytosis indices were determined depending on the Candida species assayed . Under these conditions, the more pathogenic species presented the lower indices of phagocytosis . It is determined that the in vitro phagocytic index may bear a close relationship with the pathogenicity of the Candida berkhout.

Inflammation, 1985 Mar, 9(1), 81 - 90
Influence of fluid-phase chemoattractants on polymorphonuclear leukocyte chemotaxic responsiveness to a surface-bound attractant; Dahlgren C; Polymorphonuclear leukocytes (PMNLs) were allowed to migrate on slides with fixed yeast particles dotted about on the surface . Locomotion was quantified by counting the number of yeast particles in association with a PMNL . Addition of a complement source to yeast particles able to activate the complement system resulted in a chemotactic response even when fluid-phase attractants were removed prior to the measurement of PMNL chemotaxis, indicating that surface-bound attractants guided the PMNLs to the yeast particles . The presence of high concentrations of fluid-phase chemoattractants resulted in a reduced PMNL chemotactic response to the surface-bound gradient . From comparisons between the yeast-slide system and the locomotion-under-agarose assay, it could be concluded that PMNL chemotaxis in response to a surface-bound gradient is less influenced by factor-specific deactivation than the response to a fluid-phase attractant . The PMNL chemotactic response is reduced to both surface-bound and fluid-phase gradients as a result of a non-factor-specific deactivation.

Biochimie, 1985 Mar-Apr, 67(3-4), 417 - 22
Photochemotherapy using pyridopsoralens; Dubertret L et al.; Aiming to decrease the acute side effects and genotoxic hazards of PUVA, pyrido (3,4-C) psoralen (PP) and 7-methyl pyrido (3,4-C) psoralen (MPP) were synthesized and studied . Their UVA maximum absorption lies at 325 and 330 nm, respectively . Their photostability is comparable to that of 8-MOP . They complex to DNA in the dark, and, in the presence of UVA, produce only monoadditions to DNA, as shown by fluorescence and DNA denaturation-renaturation studies . In diploid eukaryotic yeast they are more effective than 8-MOP for the induction of lethal effects and mitochondrial damage . Their mutagenic activity per unit dose of UVA is in the same range as that of 8-MOP . However, per viable cell they are clearly less mutagenic than 8-MOP . This difference is also observed for recombinogenic activity . No oxygen effect is observed . In mammalian cells the following ranges of effectiveness are found: inhibition of DNA synthesis in human fibroblasts: MPP greater than PP greater than 8-MOP; mutagenic activity in V79 Chinese hamster cells: MPP greater than PP greater than 8-MOP; cell transforming ability in C3H embryonic mouse cells: MPP greater than 8-MOP greater than PP as a function of UVA dose, and: 8-MOP greater than MPP greater than PP as a function of survival; induction of sister chromatic exchanges (SCE) per unit dose: MPP greater than PP greater than 8-MOP in the linear part of the induction curve, and : 8-MOP greater than PP greater than MPP at the maximum level of SCE obtained.(ABSTRACT TRUNCATED AT 250 WORDS)

J Pharmacol Exp Ther, 1985 Mar, 232(3), 722 - 4
Kinetics of drug action in disease states . IX . Effect of experimental fever on phenobarbital concentrations at onset of loss of righting reflex in rats; Hisaoka M et al.; The purpose of this investigation was to determine the effect of fever on the concentration-pharmacologic activity relationship of phenobarbital (PB) . Fever was produced in adult female Lewis rats by either bacterial endotoxin or brewer's yeast . Endotoxin elevated body temperature by 0.9 +/- 0.6 degrees C in one study and by 1.0 +/- 0.4 degrees C in another . Brewer's yeast caused a more pronounced and protracted elevation of temperature averaging 1.8 +/- 0.3 degrees C at the time of the pharmacodynamic measurements . PB was administered by slow i.v . infusion until the rats lost their righting reflex . The concentrations of PB at that time in serum, cerebrospinal fluid and brain were appreciably lower in the hyperpyrexic than in control (saline-treated) animals, irrespective of the method used to produce fever . Thus, fever is associated with an increased sensitivity of the central nervous system to the depressant effect of PB . This observation may be of particular relevance to the use of PB for the treatment and prevention of febrile convulsions.

Cell Immunol, 1985 Mar, 91(1), 193 - 200
Modulation of human natural killer cell activity by recombinant human interleukin 2; Shaw AR et al.; Recombinant human IL-2, secreted by yeast harboring a plasmid containing a synthetic IL-2 gene, is biologically active in augmenting human natural killer (NK) cell activity . A dose-dependent linear stimulation of NK activity was obtained against the chronic myelogenous leukemia cell line K562 over the range 3 to 300 units/ml of IL-2 . Enhancement of NK activity was similarly demonstrable against the less NK-sensitive carcinoma cell lines LoVo and SKOSC . IL-2 could also be demonstrated to augment antibody-dependent cellular cytotoxicity (ADCC) against SKOSC targets . IL-2 responsiveness segregated with a non-E-rosetting fraction comprising 11% of postfractionation lymphocytes and containing 94% of the recoverable NK activity, suggesting that IL-2 might operate directly upon the NK cell rather than through an accessory cell . This is believed to be the first demonstration of NK stimulatory activity by the product of a totally synthetic human IL-2 gene . The availability, purity, and NK-enhancing properties of the recombinant IL-2 make it a potentially important agent for clinical trial.

Int J Cell Cloning, 1985 Mar, 3(2), 65 - 80
1,25-Dihydroxyvitamin D3 and the regulation of the differentiation and function of macrophages and granulocytes; Shezen E et al.; 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) exerts a differential inhibitory effect on the formation of granulocyte, granulocyte/macrophage, and macrophage colonies grown from mouse bone marrow precursor cells; 50% inhibition was attained at 1.1, 2.3, and 23 nM 1,25(OH)2D3, respectively . The inhibition of colony formation, as well as phagocyte proliferation in liquid cultures, requires the presence of 1,25(OH)2D3 in the early stages of culture (up to 72 h after culture initiation) . 1,25(OH)2D3 induces a dose- and time-dependent augmentation of the phagocytic capability of mononuclear phagocytes (up to 100%) towards both heat-killed yeast cells and IgG-coated sheep red blood cells . The augmentation of the phagocytic capability of the mononuclear phagocytes depends critically on when 1,25(OH)2D3 is added . It is effective when added up to 72 h after culture initiation, while at later stages (greater than or equal to 96 h) the cells are no longer induced to express enhanced phagocytic capability . We suggest that these phenomena may be relevant to hemopoietic processes.

Arch Pathol Lab Med, 1985 Mar, 109(3), 273 - 6
Filamentous Histoplasma capsulatum endocarditis involving mitral and aortic valve porcine bioprostheses; Svirbely JR et al.; We report a case of histoplasmosis capsulati endocarditis simultaneously infecting a mitral and aortic porcine bioprostheses . Histologically, the fungus demonstrated a diversity of morphologies, ranging from typical, intracellular, 2- to 5-micron yeast cells, to septate and branching hyphae, to bizarre hyphae with vesicular swellings . Cultures obtained from both valves were positive in four days . The combination of unexpected histologic appearance and rapid growth in culture may pose problems in diagnosis.

Eur J Biochem, 1985 Mar 1, 147(2), 241 - 7
Preparation and characterization of NADP derivatives alkylated at 2'-phosphate and 6-amino groups; Okuda K et al.; Reaction of NADP with 3-propiolactone at pH 6 gave new NADP derivatives carboxyethylated at the 2'-phosphate or 6-amino group, or both: 2'-O-(2-carboxyethyl)phosphono-NAD (I), N6-(2-carboxyethyl)-NADP (II), and 2'-O-(2-carboxyethyl)phosphono-N6-(2-carboxyethyl)-NAD (III) . Their structures were assigned on the basis of ultraviolet, 1H-NMR and 31P-NMR spectra, and also treatment with nucleotide pyrophosphatase or alkaline phosphatase . Carbodiimide-promoted reaction of derivative I with 1,2-diaminoethane gave 2'-O-{N-(2-aminoethyl)carbamoylethyl}phosphono-NAD (IV); derivative III gave 2'-O-{N-(2-aminoethyl)carbamoylethyl}phosphono-N6-{N-(2-aminoethyl ) carbamoylethyl}-NAD (IV) . The same reaction of derivative II, on the other hand, gave a mixture of N6-{N-(2-aminoethyl)carbamoylethyl}-NADP (Va) and its 3'-phosphate isomer (Vb) . The mixture was converted to Va via the 2',3'-cyclic derivative (Vc) . Their structures were assigned on the basis of ultraviolet and 1H-NMR spectra, and also treatment with alkaline phosphatase or 3'-nucleotidase . All the NADP derivatives obtained in this work could be reduced with yeast glucose-6-phosphate dehydrogenase.

J Leukoc Biol, 1985 Mar, 37(3), 349 - 58
Enhancement of hamster alveolar macrophage phagocytic activity by lymphokines; Widen RH et al.; Modulation of phagocytic activity of resident hamster pulmonary alveolar macrophages was accomplished by incubation of the cells in lymphokines prepared by stimulation of hamster splenocytes with concanavalin A or alloantigens in mixed lymphocyte cultures . Alveolar macrophages preincubated in either of these lymphokine preparations possessed significantly greater ability to ingest IgG or IgM plus complement-coated sheep erythrocytes, via their Fc or complement receptors, respectively, than macrophages exposed to control preparations . Ingestion of yeast particles also was enhanced with macrophages incubated in supernatants from cultures of stimulated splenocytes . Supernatant fluids from either mitogen- or alloantigen-stimulated splenocytes possessed migration inhibitory activity with characteristics similar to MIF from other animals; the phagocytosis-enhancing activity shared some of these characteristics.

Biochem J, 1985 Feb 15, 226(1), 331 - 4
Purification and properties of plant cytochrome b5; Bonnerot C et al.; Microsomal cytochrome b5 was 352-fold purified from potato tubers with a yield of 10.4% . To our knowledge, this is the first report relating the purification of higher-plant cytochrome b5 . Its Mr (16 700) and absorption spectrum are similar to those of animal and yeast cytochrome b5.

Nature, 1985 Feb 14-20, 313(6003), 590 - 2
Autonomous chaotic behaviour of the slime mould Dictyostelium discoideum predicted by a model for cyclic AMP signalling; Martiel JL et al.; How sustained oscillations lose their periodicity and thus give rise to chaos was first analysed in mathematical models, then observed in chemical systems such as the Belousov-Zhabotinsky reaction where chaos is autonomous because it originates from endogenous kinetic mechanisms . In contrast, chaos can also be obtained by periodically forcing an oscillatory system, as shown, for example, in cardiac cells and yeast glycolysis . Biochemical evidence for autonomous chaos has been obtained both in vitro for the peroxidase reaction and in enzymatic models not based directly on experimental systems . We report here the occurrence of autonomous chaos in a realistic model for the cyclic AMP signalling system of the slime mould Dictyostelium discoideum, based on receptor modification . This model is also capable of bursting, a phenomenon characteristic of some pacemaker neurones such as R15 in Aplysia . Whereas bursting has not been observed in D . discoideum, our model suggests that 'aperiodic signalling' in the mutant Fr17 provides the first example of autonomous chaos occurring spontaneously at the cellular level.

J Biol Chem, 1985 Feb 10, 260(3), 1952 - 8
Characterization of an RNA polymerase activity from HeLa cell mitochondria, which initiates transcription at the heavy strand rRNA promoter and the light strand promoter in human mitochondrial DNA; Shuey DJ et al.; An RNA polymerase activity capable of initiating transcription at both the heavy strand rRNA promoter and the light strand promoter of human mitochondrial DNA has been partially purified from HeLa cell mitochondria and characterized in its requirements and products . The ratio of the two transcription initiating activities varied considerably from preparation to preparation . The human mtRNA polymerase partially purified by DEAE-cellulose and heparin-agarose chromatography exhibits a great sensitivity to ionic strength and to Mn2+, characteristics which clearly differentiate this enzyme from bacterial and eukaryotic nuclear RNA polymerases, and in contrast resemble the behavior of the yeast mtRNA polymerase . The human mtRNA polymerase exhibits a requirement for ATP which is 15- to 20-fold higher than that for the other NTPs, a low optimum template DNA concentration, and a marked susceptibility to inhibition by non-mitochondrial DNA.

J Protozool, 1985 Feb, 32(1), 12 - 9
Amebostomes of Naegleria fowleri; John DT et al.; The strain of ameba, culture incubation temperature, and phase of ameba growth affected the number of amebostomes present on amebae of Naegleria fowleri . Serial passage of N . fowleri through mice decreased the average number of amebostomes . Amebostomes were shown to be functional by their ability to engulf yeast cells.

Br J Exp Pathol, 1985 Feb, 66(1), 1 - 5
The effect of adjuvant arthritis on the subsequent local inflammatory response in rats; Mangan FR et al.; An assessment was made of the cellular inflammatory response to the subcutaneous implantation of sterile nitrocellulose discs and polyvinyl sponges in both normal rats and rats with adjuvant-induced arthritis . It was found that from 3 days after adjuvant injection the treated animals exhibited a reduced accumulation of inflammatory cells onto the nitrocellulose discs and that this impairment in response was not apparent until the discs had been retained for longer than 24 h . When discs were implanted after non-arthritogenic doses of adjuvant constituents or injection of brewer's yeast no effect was seen on the subsequent response . When polyvinyl sponges were used in adjuvant-treated animals a similar initial reduction in cellular accumulation was observed, which was later followed by increased cell numbers associated with enhanced granuloma formation . Differential cell counts revealed that both neutrophil and mononuclear cell types were affected . Some of the possible mechanisms involved in these observations are discussed.

EMBO J, 1985 Feb, 4(2), 475 - 9
Transformation of Aspergillus niger by the amdS gene of Aspergillus nidulans; Kelly JM et al.; Aspergillus niger grows poorly on acetamide as a nitrogen or carbon source and lacks sequences detectably homologous to the amdS gene encoding the acetamidase of Aspergillus nidulans . We have taken advantage of these observations to develop a transformation system for A . niger using the amdS gene as a dominant heterologous marker for selecting transformants on the basis of acetamide utilization . Transformants varied in their ability to grow on amide media and the number of integrated copies of the amdS plasmid ranged from 1 or 2 to greater than 100 . Southern analysis of transformants revealed that the multiple copies were integrated into the chromosome in tandem arrays . This result indicates that transformation of A . niger is more similar to mammalian cells than to yeast . Analysis of enzyme activity levels and RNA levels showed that most of the copies of amdS were expressed . Mitotic stabilities of transformants were found to be high . A transformant containing greater than 100 copies of the amdS gene was impaired in omega-amino acid utilization, a result that has also been found in A . nidulans . Since, in A . nidulans, omega-amino acids induce acetamidase via a characterizied regulatory gene (amdR/intA) this observation implies that titration of an analogous A . niger regulatory gene product by multiple amdS copies has occurred . Additional evidence suggested that the amdS gene is regulated in A . niger . It has also been shown that an unselected plasmid can be co-transformed with the amdS plasmid into A . niger.

J Prosthet Dent, 1985 Feb, 53(2), 214 - 6
Ability of enzymes to remove Candida; Tamamoto M et al.; Infection by C . albicans is a significant cause of denture stomatitis . Therefore, the results of this study, which demonstrated that yeast lytic enzymes and proteolytic enzymes removed C . albicans from acrylic resin surfaces, suggest that these compounds are potentially useful denture cleansers.

Z Hautkr, 1985 Feb 1, 60(3), 261 - 4, 269
{Trichomoniasis of the urogenital tract in patients hospitalized in 1976-1983}; Szarmach H et al.; We report on investigations carried out on T . vaginalis and mixed infection (T . vaginalis and yeast-like fungi) . Special attention was paid to metronidazol resistance on T . vaginalis strains isolated from patients who had been repeatedly treated with this medicament . Therefore we point to the necessity of defining the patients' susceptibility to this remedy.

Arch Ophthalmol, 1985 Feb, 103(2), 250 - 6
Antifungal synergism . A proposed dosage for corneal storage medium; Kowalski RP et al.; Fungal infections from eye bank-preserved corneas have led us to search for antifungal agents that will eliminate yeast and mold in McCarey-Kaufman (MK) medium and concurrently be nontoxic . Amphotericin B, natamycin, nystatin, and clotrimazole were tested in synergistic combinations in vitro against nine yeast and six mold specimens . For average yeast and mold concentrations of 3.4 X 10(4) and 1.36 X 10(4) colony-forming units (CFUs)/mL at 5 degrees C, and 3.36 X 10(4) and 2.3 X 10(3) CFUs/mL at 37 degrees C, respectively, a synergistic combination of all four drugs at one twelfth the minimal fungicidal concentrations proved fungicidal . This synergistic combination did not alter donor human corneal morphology under specular microscopy, nor did it inhibit rabbit corneal endothelial cell division preserved and propagated in antifungal supplemented MK medium . The synergistic drug mixture did prove to be fungicidal when the endothelial cells were challenged with fungal inoculum.

J Med Microbiol, 1985 Feb, 19(1), 109 - 14
Elastase activity of Coccidioides immitis; Nziramasanga P et al.; Twenty-two strains of Coccidioides immitis were tested for the ability to hydrolyse elastin . Screening assays with Czapek's or Tryptic Soy Agar supplemented with 0.5% elastin demonstrated that 21 strains (95%) were elastolytic . In broth cultures, elastase activity was induced by incorporation of insoluble elastin into the medium and induction was suppressed by supplementation with yeast extract . C . immitis appears to be unique amongst dimorphic fungal pathogens in its digestion of elastin.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1985 Feb, 180(2-3), 134 - 45
{Viral hepatitis and immunoprophylaxis}; Deinhardt F et al.; Today we distinguish 4 forms of viral hepatitis: hepatitis A, hepatitis B, hepatitis non-A, non-B and hepatitis occurring in the course of other viral diseases . The viruses of hepatitis A and hepatitis B have been identified but the agent(s) of hepatitis non-A, non-B remain unknown . Inoculation of normal pooled human immunoglobulin provides passive immunity for 2-3 months against hepatitis A but not against hepatitis B or hepatitis non-A, non-B . For passive protection against hepatitis B a special immunoglobulin with a high anti-HBs titer must be used whereas the protection against hepatitis non-A, non-B with immunoglobulin is uncertain . Live attenuated and noninfectious polypeptide vaccines for active immunisation against hepatitis A are currently developed and first clinical trials have begun with the live attenuated vaccine . A vaccine consisting of noninfectious highly purified HBsAg derived from the plasma of HBV carriers is in general use since two years and has proved safe and highly effective and vaccines are now developed from HBsAg obtained through molecular cloning of the HBsAg genome in plasmids and expression of the genome with HBsAg production in yeast cells . First clinical studies with this vaccine are encouraging and these as well as purely synthetic vaccines will in time replace the currently used vaccines . No vaccines could be developed so far against hepatitis non-A, non-B because the agent(s) of this disease are unknown.

J Bacteriol, 1985 Feb, 161(2), 615 - 9
Uptake of phenol by Trichosporon cutaneum; Mortberg M et al.; The soil yeast Trichosporon cutaneum, which is distinguished by having a strictly oxidative metabolism, can be induced to utilize phenol as a sole carbon source . The present paper shows that such phenol-induced cells contain a specific, energy-dependent uptake system for phenol . Phenol uptake is not directly linked to its o-hydroxylation inside the cell, the first step of phenol metabolism . The Km for uptake is 235 +/- 30 microM, that for hydroxylation only 4.5 +/- 0.5 microM . Further, the phenol analog 2,6-dimethylphenol, which can not be hydroxylated, competes with phenol for the uptake system . The pH dependence of uptake indicates that phenolate is an essential form during the uptake process . The energy requirement for phenol uptake is indicated by effects of various inhibitors of energy generation, including proton-conducting uncouplers . Direct monitoring of proton movements in a pH-stat during phenol uptake indicates a phenol-proton symport . One proton is cotransported with every phenol molecule . Phenol competes with the uptake of sucrose and glycerol by cells grown on these substrates . Under such conditions the uptake of phenol seems to proceed through a different system, with lower affinity for phenol than in phenol-grown cells.

J Biol Chem, 1985 Jan 25, 260(2), 880 - 6
Characterization of Leishmania donovani acid phosphatases; Remaley AT et al.; A crude membrane fraction from promastigotes of Leishmania donovani grown in a liquid culture medium containing 20% fetal calf serum was prepared by freeze-thawing, centrifugation (200,000 X g, 30 min), and extraction with 2% (w/v) sodium cholate . After removal of the bile salt by chromatography on a Sephadex G-75 column, the solubilized membrane protein fraction, rich in acid phosphatase activity, was chromatographed on columns containing concanavalin A-Sepharose, QAE-Sephadex, and Sephadex G-150 and G-100 . Three distinct acid phosphatases were resolved: the major phosphatase activity (70% of the total) was L-(+)-tartrate-resistant (designated ACP-P1) and corresponds to the acid phosphatase localized to the outer surface of the parasite's plasma membrane; the other two phosphatases (ACP-P2 and ACP-P3) account for the remaining 30% of the particulate acid phosphatase activity, and both of these enzymes are L-(+)-tartrate-sensitive . Using a combination of sucrose density gradient centrifugation, gel filtration chromatography, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, it was determined that ACP-P1 is a 128,000-dalton protein composed of two subunits of 65,000-68,000 daltons . ACP-P1 has an isoelectric point of 4.1, a pH optimum of 5.5, hydrolyzes fructose 1,6-diphosphate, but no other sugar phosphates and dephosphorylates phosphotyrosine, yeast mannan, and the phosphorylated form of rat liver pyruvate kinase . ACP-P2 (pI, 5.4) and ACP-P3 (pI, 7.1) with molecular masses of 132,000 and 108,000 daltons, respectively, are both tartrate-sensitive and are distinguished from each other on the basis of their sensitivity to inhibition by polyanionic molybdenum complexes . These two phosphatases also have their pH optima in the pH 5.0-6.0 range, but have a considerably broader substrate specificity than ACP-P1.

J Mol Biol, 1985 Jan 20, 181(2), 199 - 209
In phage lambda, cos is a recombinator in the red pathway; Stahl FW et al.; Among lambda particles carrying chromosomes that have failed to replicate during a lytic cycle cross there is a high frequency of Red-mediated recombination near the right-hand end . Earlier work has shown that this recombination is dependent on cos (cohesive end site), the packaging origin of lambda . In contrast to the prediction of the break-copy model proposed earlier, we find a high recombination rate near cos even when only one of the two participating parents has a functional cos at that locus . The exchange is accompanied by loss of the stimulating cos in the recombination product, irrespective of the marker configurations: a+b+cos- rather than a+b+cos+ is produced in the cross a+b-cos- x a-b+cos+ as well as in the cross a+b-cos+ x a-b+cos- . Further analyses of these and earlier data allow the formulation of a detailed model for cos-stimulated, Red-mediated genetic exchange . In this model, cos stimulates exchange by virtue of being a double-strand cut site . The model has several features like that proposed for yeast . This role of cos in the Red pathway contrasts with the role of cos in the RecBC pathway, in which cos serves as an entry site for a recombinase that stimulates exchanges far from cos.

Nature, 1985 Jan 10-18, 313(5998), 147 - 9
A hypersensitive site in hsp70 chromatin requires adjacent not internal DNA sequence; Costlow NA et al.; Nuclease-hypersensitive sites in chromatin exist at the 5' side of many eukaryotic genes . To gain some understanding of the molecular basis of these hypersensitive sites, we have now examined the pair of sites upstream of the Drosophila hsp70 gene in a series of plasmids that contain deletions in the hypersensitive region and have been transformed into yeast cells . Hypersensitive sites 5' to a Drosophila hsp70 gene are preserved when this gene is introduced into yeast by transformation . We find that a yeast strain containing a plasmid in which the deletion extends through the first hypersensitive site still displays the normal pair of hypersensitive sites, so DNA sequences over which the first hypersensitive site is centred are not required for hypersensitivity at this position and the site can form over a foreign DNA sequence juxtaposed against this deletion end point . Deletions progressing further into the region bracketed by the pair of 5' hypersensitive sites eliminate the first hypersensitive site and alter the downstream site . We propose that the hypersensitive sites are generated through the binding of a protein that renders flanking sequences more accessible to nucleases, perhaps by preventing normal chromatin packaging.

Biosystems, 1985, 18(3-4), 241 - 7
An evaluation of tubulin as a molecular clock; Little M; The available sequence data for tubulin indicates that it cannot be used as a molecular clock . Apparent alpha-tubulin mutation rates, for example, vary from 0.16 to 3.8 PAMs per 100 million years depending on which two alpha-tubulins are compared . All animal alpha-tubulin mutation rates seem to be quite low, whereas those of non-animals are relatively high . A similar division is not present amongst the beta-tubulins; their apparent mutation rates, however, vary just as much . For any given tubulin, the largest number of amino acid sequence differences are obtained when comparing it to the tubulins of yeasts . Sequence comparisons with the tubulins of unicellular algae and chelates show far fewer differences . Cytochrome c data, however, show that the ciliates diverged from animals well before the yeasts . This means, therefore, that the average tubulin mutation rates in yeasts and ciliates since the time they shared a common ancestor must be quite different . The high mutation rate of yeast tubulins may possibly reflect the absence of cilia . Structural constraints imposed on tubulin by the large number of interactions with other components of the complex ciliary axoneme probably have a significant effect on its rate of mutation.

Nutr Cancer, 1985, 7(1-2), 25 - 36
Effects of selenium supplementation on hepatocarcinogenesis in rats; Aquino TM et al.; Four groups of weanling male Wistar rats (Groups A-D) received diethylnitrosamine (DEN, 40 ppm) in their drinking water for four weeks; after a recovery period of two weeks, they received (for the rest of the experiment) phenobarbital (PB, 500 ppm) added to a Torula yeast-based diet containing 0.17 ppm of selenium . Dietary selenium (2 ppm), as sodium selenite, was given to Group B one week before and during DEN treatment, to Group C one week before and during PB treatment, and to Group D during the entire experiment . Groups A and E received the unsupplemented diet, whereas Group E was not treated with DEN or PB . Pair-feeding conditions were used to minimize possible influences of differences in food intake and growth . Rats were killed at the 19th and 24th weeks after the experiment began . No significant differences were found in food and fluid intakes or in growth rates among the groups . Livers in Group E were histologically normal, whereas preneoplastic and neoplastic lesions were found in all other groups . In rats killed at the 19th and 24th weeks, the numerical and the volumetric densities of preneoplastic lesions did not differ significantly between all the groups . Similarly, the incidence of hepatocellular carcinomas only detected at 24 weeks was not significantly different between the groups . These results indicated that in this particular model of hepatocarcinogenesis, the dietary supplementation of 2 ppm of selenium did not modify the development of preneoplasia and carcinomas.

Drug Nutr Interact, 1985, 3(4), 229 - 38
Differential effects of dietary selenium on hepatic and renal glutathione metabolism in the rat; Davies MH et al.; Studies were undertaken to determine whether or not dietary selenium (Se), as sodium selenite, at suboptimal (unsupplemented torula yeast diets, 0.02 ppm Se) or supplemented up to dietary excess, but nontoxic, levels (5.0 ppm Se) could selectively modify hepatic and renal reduced glutathione (GSH) levels and the enzymes involved in GSH metabolism . Male rats were provided torula yeast semipurified diets containing approximately 0.02 ppm Se in the basal diet and supplements of 0.1, 2.0, or 5.0 ppm Se for 3 or 6 weeks . Hepatic GSH increased in a nonlinear manner, with increasing dietary Se at both time points . Renal GSH was not similarly influenced . Neither hepatic nor renal gamma-glutamylcysteine synthetase or gamma-glutamyl transpeptidase activities are altered by supplements of Se . This suggests that synthetic and degradation enzyme activities are not influenced by Se . The capacity for the maintenance of GSH in the reduced state by glutathione reductase activity increased with increasing levels of dietary Se in the liver but not in the kidney . In both tissues greater Se supplements yielded greater tissue burdens of Se . These results suggest that GSH metabolism in hepatic and renal systems is differentially mediated, and the basis for these differences could be influenced by the relative levels in glutathione metabolizing enzymes.

Gene, 1985, 35(3), 237 - 48
Tubulin genes of the African trypanosome Trypanosoma brucei rhodesiense:nucleotide sequence of a 3.7-kb fragment containing genes for alpha and beta tubulins; Kimmel BE et al.; Most tubulin genes of the African trypanosome Trypanosoma rhodesiense are contained in 3.7-kb tandemly repeating units . One member of the 3.7-kb repeat family has been isolated from a T . rhodesiense genomic library, cloned, and sequenced . The 3646-bp fragment contains a complete alpha-tubulin gene and portions of two beta-tubulin genes . No introns are present . The genes are separated by 634- and 333-bp intergenic regions, which lack typical eukaryotic promoter and poly(A) signal sequences . However, both intergenic regions exhibit some structural similarity with sequences proposed to be involved in transcription termination and poly(A) addition in yeast . The 634-bp intergenic region shows homology to the "mini-exon" sequence associated with variable surface glycoprotein (VSG) and other trypanosome mRNAs . A comparable sequence is not found in the 333-bp intergenic region . T . rhodesiense alpha and beta-tubulins exhibit about 84-85% amino acid (aa) sequence homology with tubulins of mammals; the genes show about 74-75% nucleotide sequence homology . The alpha-tubulin contains 451 aa and the beta tubulin 442 aa; both have tyrosine as the C-terminal aa.

Alcohol, 1985 Jan-Feb, 2(1), 117 - 21
Catalase mediated conversion of cyanamide to an inhibitor of aldehyde dehydrogenase; DeMaster EG et al.; A minor pathway for cyanamide metabolism catalyzed by catalase is responsible for the conversion of cyanamide to an inhibitor of aldehyde dehydrogenase . Catalase itself is also inhibited by cyanamide . Both the activation of cyanamide by catalase and the inhibition of catalase by cyanamide were blocked in vivo by ethanol pretreatment, suggesting that these two processes are closely linked . Like other catalase oxidation reactions, the catalase mediated activation of cyanamide was inhibited by 3-amino-1,2,4-triazole in vivo and sodium azide in vitro . The relative formation of the active cyanamide metabolite was assessed in vitro by following the loss of yeast aldehyde dehydrogenase activity with time . Inhibition of the yeast enzyme by activated cyanamide was dependent on NAD+ or NADP+, a requirement not fulfilled by NADH or NADPH . Although H2O2 inhibited yeast aldehyde dehydrogenase in vitro and cyanamide inhibited hepatic catalase in vivo, the possible in hepatic H2O2 concentration following cyanamide administration does not account for the effects of cyanamide on ethanol metabolism . While the cyanamide activating enzyme has been identified as catalase, the reaction products of this reaction and, in particular, the structure of the active metabolite involved in the inhibition of aldehyde dehydrogenase remain unknown.

Pathology, 1985 Jan, 17(1), 20 - 23
Trichosporon beigelii: survey of isolates from clinical material; Pritchard RC et al.; Two hundred and eight isolates of Trichosporon beigelii were identified over the period January 1973 to July 1983 . 45.7% of these were from skin, 25.0% from nail, 22.6% from tissues and fluids, 3.4% from hair and 3.4% from sputum . Tr . beigelii was isolated in association with a recognized pathogen in 23 cases, 9 with a yeast, 14 with a dermatophyte . In 38 cases, Tr . beigelii was the only organism isolated when direct microscopic examination of clinical material showed the presence of hyphae and/or yeast cells . Although Tr . beigelii could only be assigned a definite pathogenic role in 6 cases of genital white piedra, and in one case of peritonitis associated with chronic ambulatory peritoneal dialysis, we believe that this organism was pathogenic in many cases of skin infection . In most of the cases where it was isolated from tissue or fluids at Royal North Shore Hospital, Tr . beigelii was not considered to be significantly contributing to the disease process.

Drug Nutr Interact, 1985, 3(3), 129 - 40
Selenium deficiency and detoxication functions in the rat: effect of chronic dietary cadmium; Olsson U; Male rats from moderately selenium-deficient dams were fed a Torula yeast-based, selenium-deficient diet for 7 weeks, with or without added supplements of sodium selenite (0.2 ppm selenium) and cadmium chloride (50 ppm cadmium) in the drinking water . Cadmium caused about 10% body-weight loss in selenium-deficient, as well as in supplemented rats . Glutathione peroxidase activity in liver 105,000 g supernatant and in erythrocyte hemolysate from selenium-deficient rats was about 1% and 3%, respectively, of that in supplemented rats . A cadmium-induced decrease of glutathione peroxidase activity was found in erythrocyte and liver preparations from selenium-supplemented rats, while cadmium caused an increase of the liver activity in selenium deficiency . Selenium deficiency per se caused a significant decrease of cytochrome P-450 content, while cadmium treatment did not modify further the content of this enzyme . NADPH-cytochrome c reductase was not changed by selenium regimen or cadmium treatment, while cytochrome b5 was increased on cadmium treatment of the supplemented rat . The microsomal metabolism of N,N-dimethylaniline showed a decrease of the cytochrome P-450-dependent C-oxygenation in selenium-deficient groups . Cadmium treatment had no further significant effect . The flavin-containing monooxygenase, which performs N-oxygenation of N,N-dimethylaniline, was decreased significantly by cadmium treatment in selenium deficiency . Selenium deficiency seems thus to be connected with higher susceptibility to cadmium-induced impairments of liver detoxication functions, although progressive accumulation of cadmium in the liver appears to produce only modest effects.

Clin Lab Haematol, 1985, 7(1), 43 - 53
The morphology and viability of blood components processed by high-gradient magnetic separation; Paul F et al.; The effect of high-gradient magnetic separation on blood components has been investigated . For erythrocytes there is no significant change in the morphology, size distribution and in the in-vivo survival times at the confidence level P = 0.05 . For neutrophils stimulated nitro-blue tetrazolium reduction is maintained after HGMS . No change in chemotaxis and yeast cell phagocytosis was observed . Platelet distribution curves show no detectable changes before and after filtration . It is concluded that blood separated by HGMS is suitable for diagnostic analysis and may be considered for return to the patient.

Anal Biochem, 1985 Jan, 144(1), 258 - 66
Single-run high-performance liquid chromatography of nucleotides, nucleosides, and major purine bases and its application to different tissue extracts; Wynants J et al.; A high-performance liquid chromatography (HPLC) method is described for the separation and quantitation of nucleotides, nucleosides, purine bases, and related compounds in one single run . The separation of a standard mixture of at least 24 components is achieved within 35 min on glass columns (30 cm, 3-mm i.d.) with C-18 reversed-phase particles of 5 micron, and ammonium dihydrogen phosphate (0.15 M, pH 6.00) and a slow linear gradient of methanol/acetonitrile (to 15%) as eluting solvent . The method has been applied to microsamples of different cells and tissues . Samples (2.5 mg dry wt) were cooled in liquid nitrogen, lyophilized, and extracted with 0.6 N perchloric acid . After neutralization with potassium bicarbonate, the extract (20 microliter) was directly injected into the column . To illustrate the wide applicability of the method, representative chromatograms are shown of extracts of biopsies from heart tissue, skeletal muscle, and brain and liver and from hepatocytes, erythrocytes, and yeast cells, under different conditions, known to induce changes in purine metabolism.

Mol Biol (Mosk), 1985 Jan-Feb, 19(1), 144 - 61
{Primary organization of nucleosome core particles in active and repressed nuclei}; Bavykin SG et al.; A refined high-resolution map for the linear arrangement of histones along DNA in the nucleosomal core particles has been determined by DNA-protein crosslinking . Histones are aligned on one strand of the 145 bp core DNA in the following order: (5') H2B25,35--H455,65--H375,85,, 95/H488--H2B105, 115--H2A118--H3135, 145/H2A145 (3') (the subscripts indicate the approximate distance in nucleotides of the main histone binding sites from the 5'-end of the core DNA) . This suggests a symmetrical and rather autonomous arrangement of the histone tetramer (H3, H4)2 and two dimers (H2A, H2B) on the double-stranded core DNA: H2A/H3--(H2A, H2B)--(H3, H4)2--(H2B, H2A)--H3/H2A . The arrangement of histones on DNA was found to be very similar for the cores isolated from the repressed nuclei of sea urchin sperm and chicken erythrocytes and from the active in transcription and replication Drosophila embryo and yeast nuclei . This indicates that the core nucleosome structure is highly conserved through evolution and that the overall inactivation of chromatin does not affect the primary organization of the cores . A new binding site H2B58 was found for a sea urchin spermal variant of H2B which contains an additional basic segment within the N-terminal part of the molecule . The core isolation procedure was shown to introduce changes into the core structure which are reflected in the appearance of a new binding site H2A75.

J Am Acad Dermatol, 1985 Jan, 12(1 Pt 1), 56 - 61
Pityrosporum folliculitis: a common disease of the young and middle-aged; Back O et al.; Fifty-one patients, thirty-nine women and twelve men, with Pityrosporum folliculitis are described . This investigation clearly demonstrates that Pityrosporum folliculitis is a real entity . The diagnosis is based primarily on the clinical picture, direct microscopy, histopathology, and the effect of antimycotic treatment . The typical patient is a woman of 30 years with itching follicular papules and pustules localized to the upper trunk or upper arms . Direct microscopy reveals round yeast cells and sometimes even hyphae . In biopsy specimens, abundant round budding yeast cells and occasionally hyphae are seen in a dilated follicle . Yeast growth is obtained only on lipid-enriched media . Twenty-five patients were treated with selenium sulfide shampoo, twelve with 50% propylene glycol in water, and ten with topical econazole cream with good results . Cure or marked improvement was seen after 3 to 4 weeks, but symptoms and lesions recurred if treatment was not continued intermittently . Predisposing factors such as occlusion and greasy skin are probably important, and future studies should focus on fungal hypersensitivity, quantitative variations in the number of Pityrosporum orbiculare, lipid composition of the skin, and extended epidemiologic data.

J Infect Dis, 1985 Jan, 151(1), 57 - 64
Intracellular growth and phagocytosis of Blastomyces dermatitidis by monocyte-derived macrophages from previously infected and normal subjects; Bradsher RW et al.; Blastomyces dermatitidis evokes responses of human cellular immunity typical of other intracellular fungal pathogens . Differences in growth rates of intracellular Blastomyces yeast and the differences in amounts of yeast phagocytized by macrophages were determined for macrophages derived from peripheral blood monocytes from 11 persons with treated blastomycosis and 11 normal, healthy persons . Cellular immunity was examined by lymphocyte uptake of {3H}thymidine in response to a specific antigen of Blastomyces yeast . Yeast were more readily phagocytized by macrophages from the previously treated donors when compared with those from the normal donors; the yeast were confirmed to be intracellular by transmission electron microscopy . Likewise, a decrease in growth rates of yeast was demonstrable in cultures of macrophages from previously treated donors as compared with normal donors . This greater efficiency of phagocytosis and growth inhibition of B . dermatitidis reflects another mechanism of human cellular immunity to this fungal infection.

Dev Biol Stand, 1985, 62, 119 - 27
The use of tissue culture attenuated live vaccine for Theileria hirci; Hooshmand-Rad P; Malignant sheep theileriosis which is a tick-borne disease is caused by the protozoan parasite Theileria hirci . The pathogenic form of the parasite, schizonts, inhabits the lymphoid cells of the mammalian host, and if an immunity is engendered against them animals can resist the infection . Theileria hirci infected lymphoid cells were cultivated in vitro, using Eagle's MEM supplemented with 10% ovine serum, yeast extract and lactalbumine hydrolysate (YL), respectively 250 and 1250 mg/L . Schizonts of the parasite which propagated synchroneously with the lymphoid host cells, and had been attenuated in the course of time, were used as an immunizing live vaccine at a passage level (100) that did not produce the erythrocytic forms of the parasite and was harmless to the recipient animals . The vaccinal dose contained 5 X 10(5) infected cells . The vaccine was preserved at -70 degrees C and dispatched to the field under the 4 degrees C condition with a shelf-life of 5 days . Experimental and mass field vaccination have been carried out . The first and second challenge tests, performed on experimentally vaccinated animals, were carried out respectively two and six months post-vaccination . Ticks infected with a heterologous strain of T . hirci were used in the challenge tests . All controls died of acute theileriosis whereas the vaccinated animals resisted the challenge tests . Further mass vaccination is planned.

Prikl Biokhim Mikrobiol, 1985 Jan-Feb, 21(1), 3 - 17
{Isolation of protein hydrolysates with given properties (a review)}; Nekliudov AD et al.; Methods for obtaining protein hydrolysates with certain given properties are considered . It is shown that, besides food proteins, various yeast proteins can be used . Advantages and disadvantages of different types of hydrolysis, hydrolysis conditions, methods of hydrolysate purification and prevention of labile amino acid destruction are described.

Exp Hematol, 1985 Jan, 13(1), 44 - 50
Development of specific surface receptors recognizing mannose-terminal glycoconjugates in cultured monocytes: a possible early marker for differentiation of monocyte into macrophage; Kataoka M et al.; Surface receptors for mannose-terminal glycoconjugates have been reported in various macrophage populations and are thought to be involved in specific binding and internalization of mannose-rich substances . They thereby may serve a function in such phenomena as phagocytosis of yeast and tumor cell recognition . Little is known of mannosyl receptors in blood monocytes . We synthesized a probe by covalently linking D-mannose to bovine serum albumin (BSA) . Using this probe in fluoresceinated or latex minibead-derivatized forms, we searched the surface of human monocytes for the presence of mannosyl receptors . 125I-labeled probe was further used to quantify the number of receptors and the kinetics of the binding . Freshly isolated monocytes did not bind the probe, indicating the absence of mannosyl receptors . When placed in a culture system that preserves functional and morphological homogeneity of the cells, surface receptors for D-mannosyl glycoproteins developed within four days, reached a peak after one week, and then remained fairly stable . Binding parameters (Kd, Bmax, and receptor number) also remained stable and were not dissimilar to those reported for macrophages, although the Kd was consistently larger in cultured monocytes . When studied at 37 degrees C, the ligand-receptor complex was internalized through a system of coated pits and vesicles . The development of these receptors before evidence of morphological or functional differentiation suggests that these receptors may constitute an early marker for differentiation of monocytes into macrophages.

Scand J Immunol, 1985 Jan, 21(1), 43 - 7
The role of negative charge in the complement-dependent dissociation of IgG-mediated aggregation; Hed J et al.; The IgG-mediated aggregation of yeast particles was used as a model to study the dissociating effect of human serum complement on immune aggregates . A 92% decrease in non-aggregated particles was obtained by coating particles with IgG . After incubation of the aggregates in normal human serum (NHS) at 37 degrees C the number of free particles increased almost 10 times . The effect could be obtained with ethyleneglycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid-containing serum but not ethylenediaminetetraacetic acid-containing serum, indicating the importance of the alternate pathway of complement activation . Treatment with NHS did not release anti-yeast IgG from the particles, indicating that disruption of antigen-antibody bonds did not explain the phenomenon . The negative charge of the different particles was studied by measuring the interaction to diethylaminoethyl-Sephacel beads . Compared with non-coated and IgG-coated particles, the NHS-treated particles were not released until the ionic strength increased to 200 mM NaCl or the pH decreased to 4.4, indicating a strong negative charge of NHS-treated particles . We propose that complement increases the negative charge of immune complexes, thereby inducing repulsive forces that counteract Fc-Fc interactions and increase the solubility of the complexes.

Curr Top Med Mycol, 1985, 1, 313 - 51
Biochemical targets for antifungal azole derivatives: hypothesis on the mode of action; Vanden Bossche H; The selective interaction of low concentrations of azole derivatives and other nitrogen heterocycles with cytochrome P-450 may be at the origin of the inhibition of ergosterol biosynthesis . From the depletion of ergosterol and the concomitant accumulation of 14 alpha-methylsterols, alterations in membrane functions, the synthesis and activity of membrane-bound enzymes, mitochondrial activities, and an uncoordinated activation of chitin synthase may result . Since chitin synthesis is more important in the hyphal form than in the budding form of C . albicans, the uncoordinated activation of chitin synthesis may be more trouble for the hyphal growth than for yeast budding . The assumption is made that from this difference the greater sensitivity of hyphal growth to azole antifungal agents may originate . It is also assumed that the higher degree of lipid unsaturation may be related to an inhibition of ergosterol biosynthesis . The inhibition of fatty acid desaturation and elongation induced by higher doses of miconazole and ketoconazole and the longer contact times might be related to interference with membrane fluidity, or it might due to chelation of the iron used in the oxidation reduction sequence during desaturation . The decreased availability of ergosterol and the accumulation of 14 alpha-methylsterols also may provide the environment needed to inactivate membrane-bound enzymes; e.g., cytochrome c peroxidase . However, it is still too speculative to correlate effects on membrane components with miconazole-induced changes in properties of all oxidases; e.g., the NADH-dependent, cyanide-insensitive oxidase . The accumulation of toxic concentrations of hydrogen peroxide, resulting from an increased NADH-oxidase activity and disappearance of the peroxidase and catalase activity, may contribute to the degeneration of subcellular structures . The complete disappearance of catalase observed at concentrations of miconazole greater than or equal to 10(-5) M may originate from direct effects on the cell . At these high concentrations reached only by topical application, direct membrane damage resulting from interaction of miconazole with lipids was observed . These direct interactions result in an inhibition of membrane-bound enzyme and mitochondrial activities and in leakage of intracellular components . The direct interactions were much less pronounced in cells treated with ketoconazole . This correlates with the smaller area occupied in the membrane per ketoconazole molecule (30 A2), compared with that occupied in the membrane per miconazole molecule (90 A2).(ABSTRACT TRUNCATED AT 400 WORDS)

Curr Genet, 1985, 9(4), 293 - 8
Molecular cloning of the 3-phosphoglycerate kinase (PGK) gene from Aspergillus nidulans; Clements JM et al.; The Aspergillus nidulans 3-phosphoglycerate kinase gene (PGK) has been isolated from a phage lambda genomic library, using the equivalent yeast gene as a hybridization probe . The location of the PGK gene within the cloned DNA has been physically mapped . The DNA sequence of a small region of the putative PGK has been determined and found to code for amino acids corresponding to the N-terminal end of the PGK protein . In contrast to the yeast PGK gene the Aspergillus gene contains a 57 base pair intron occurring between the coding sequences for amino acid 22 and 23 . A DNA fragment encompassing the PGK gene was shown to hybridize a 1,700 base poly(A) mRNA, sufficient to encode the PGK polypeptide.

Biosystems, 1985, 18(3-4), 269 - 78
5 S and 5.8 S ribosomal RNA sequences and protist phylogenetics; Walker WF; More than 100 5 S 5.8 S rRNA sequences from protists, including fungi, are known . Through a combination of quantitative treeing and special consideration of "signature' nucleotide combinations, the most significant phylogenetic implications of these data are emphasized . Also, limitations of the data for phylogenetic inferences are discussed and other significant data are brought to bear on the inferences obtained . 5 S sequences from red algae are seen as the most isolated among eukaryotics . A 5 S sequence lineage consisting of oomycetes, euglenoids, most protozoa, most slime molds and perhaps dinoflagellates and mesozoa is defined . Such a lineage is not evident from 5.8 S rRNA or cytochrome c sequence data . 5 S sequences from Ascomycota and Basidiomycota are consistent with the proposal that each is derived from a mycelial form with a haploid yeast phase and simple septal pores, probably most resembling present Taphrinales . 5 S sequences from Chytridiomycota and Zygomycota are not clearly distinct from each other and suggest that a major lineage radiation occurred in the early history of each . Qualitative biochemical data clearly supports a dichotomy between an Ascomycota-Basidiomycota lineage and a Zygomycota-Chytridiomycota lineage.

Scand J Rheumatol, 1985, 14(4), 364 - 8
Selenium treatment in rheumatoid arthritis; Tarp U et al.; A low selenium level has been reported in rheumatoid arthritis and juvenile chronic arthritis . Selenium is an essential part of the enzyme glutathione peroxidase, which catabolizes peroxides, compounds which are suggested to be of pathogenetic importance in rheumatic diseases . To assess a possible antirheumatic effect of selenium, 40 patients with active RA were included in a 6-month double-blind clinical study of selenium versus placebo . The patients in the selenium group were given daily supplements of 256 micrograms selenium in selenium-enriched yeast . Although concentrations of selenium in serum and erythrocytes increased considerably, no significant antirheumatic effect of selenium could be demonstrated.

Cytobios, 1985, 43(174S), 335 - 47
M-phase promoting factors from eggs of Xenopus laevis; Gerhart J et al.; When an M-phase promoting factor (MPF) is injected into Xenopus oocytes, which are naturally arrested at the G2/prophase boundary, it induces rapid entry of the cells into M-phase . MPF is present in late G2 and in M-phase of a variety of cell types, such as Xenopus eggs (naturally arrested in M), cleaving embryos, yeast, HeLa, and CHO cultures . MPF has been purified approximately 50-fold from eggs . It is stabilized by gamma-thio-ATP and by phosphoprotein phosphatase inhibitors . It runs as a protein of approximately 100 kd size on gel filtration . Oocytes contain a precursor of MPF, which is activated by post-translational means when a small amount of purified MPF is injected into the cell . Thus, MPF appears to be an auto-activating cytoplasmic trigger of M-phase . At anaphase of the cell cycle, MPF is inactivated due to the appearance of an 'anti-MPF' activity . Monoclonal antibodies have been prepared to partially purified MPF stabilized by gamma-thio-ATP, and several preparations which inactivate MPF were obtained . The antibodies are directed against thio-phosphate groups carried by a set of proteins including MPF . This indicates that MPF is present in our active preparations as a thio-phosphoprotein . These and other data suggest that MPF is normally activated in the cell cycle by a phosphorylation reaction.

Proc Natl Acad Sci U S A, 1985 Jan, 82(1), 14 - 8
13C NMR studies of carbon metabolism in the hyphal fungus Aspergillus nidulans; Dijkema C et al.; Natural-abundance high-resolution 13C NMR spectra (linewidth, 10 Hz) of the hyphal fungus Aspergillus nidulans have been obtained after growth on glycolytic or gluconeogenic carbon sources . Various polyols, some tricarboxylic acid-cycle intermediates and amino acids, and some phospholipids and fatty acyl compounds are present . The polyols found are mannitol, arabitol, erythritol, and glycerol . The nature of the carbon source has a pronounced effect on the pool sizes of the various polyols . All are present when the fungus is grown on sucrose or sucrose/acetate under strongly aerobic conditions . When grown on acetate, both arabitol and glycerol levels are low, whereas on glycerol erythritol is also hardly detectable . The effect of oxygen is most outspoken in glycolytically grown cultures . Limited oxygenation leads to low levels of arabitol and glycerol . Strong oxygenation changes the ratio of erythritol to mannitol, favoring the C4 polyol . An increase in oxygen supply leads to (i) stimulation of the fluxes through the pentose phosphate pathway and glycolysis, (ii) an overflow of reduced metabolites both at the pentose phosphate pathway level (erythritol and arabitol) and at the C3 level of the glycolytic pathway (glycerol), and (iii) the usual accumulation of mannitol . Upon starvation, glycerol, the other three polyols, and the tricarboxylic acid-cycle intermediates and their associated amino acids disappear in this order . As in yeast, gluconeogenic substrates lead to the synthesis of trehalose, which also occurs when mycelium is grown on acetate/sucrose under limiting aeration . A transient formation of trehalose has been observed upon incubation of starved mycelium, cultured on different substrates, with {13C}glucose.

Arzneimittelforschung, 1985, 35(4), 715 - 20
2-{3-(1,1-Dimethylethyl)-5-methoxyphenyloxazolo{4,5-b}pyridine, a new topical antiinflammatory and analgesic compound lacking systemic activity and gastric side effects; Bonney RJ et al.; The substituted oxazolopyridine 2-{3-(1,1-dimethylethyl)-5-methoxyphenyl}oxazolo{4,5-b}pyridine (OZP) inhibits phorbol myristate acetate-induced increases in vascular permeability and neutrophil accumulation in rat ears with ED50 of 253 and 200 micrograms, respectively . This compound is as potent as indomethacin to inhibit UV-induced erythema in guinea pig skin and is an effective analgesic when applied topically to the rat footpad in the yeast hyperalgesia model . OZP is a cyclooxygenase inhibitor with an IC50 of 0.06 mumol/l and inhibits prostaglandin E2, but not leukotriene C4 synthesis, by mouse peritoneal macrophages . This compound is inactive in the carrageenan paw edema assay at 90 mg/kg when administered orally or intraperitoneally, but is effective when injected into the paw . OZP is not a contact allergen and does not cause gastric irritation in rats at doses up to 180 mg/kg orally . OZP is rapidly metabolized by rat liver microsomes in a concentration and time dependent manner . Furthermore, when administered orally, OZP is cleared rapidly in rats with plasma levels being detected only at 5 and 30 min following a 2 mg/kg dose . There was no drug in the gastrointestinal tract of rats 3 h after an oral dose . Thus, this compound appears to be a new, potent and safe topical antiinflammatory and an analgesic agent lacking systemic effects.

Lymphokine Res, 1985 Spring, 4(2), 117 - 31
Biologic activities of recombinant human interleukin 2 on murine lymphocytes; Bleackley RC et al.; Recombinant human IL-2, produced by yeast cells, was tested in a number of in vitro responses with murine lymphocytes . The responses studied included proliferation of a cloned murine T lymphocyte line, generation of cytotoxic responses, recall of cytotoxic memory, and restoration of responses in spleen cells taken from cyclophosphamide-treated mice . In all cases, the recombinant human IL-2 had the same activity as purified murine IL-2 . The recombinant material represents a source of IL-2 free of other lymphokines . The responses described in this work can therefore be ascribed to the direct effects of human IL-2, of known sequence, on the cells of interest.

Arch Immunol Ther Exp (Warsz), 1985, 33(3), 423 - 8
Variations in sheep serum conglutinating complement activity and pathway involved in reactivity with sheep erythrocytes sensitized by rabbit antibody; Stankiewicz M et al.; Based on conglutination tests with the sheep E-rabbit A indicator system, three types of sheep sera were encountered . Type 1 sera failed to directly conglutinate or sensitize sheep E-rabbit A for conglutination by bovine conglutinin . Type 2 sera also failed to directly conglutinate sheep E-rabbit A but sensitized the indicator for conglutination by bovine conglutinin . Type 3 sera both directly conglutinated and sensitized sheep E-rabbit A for conglutination . Changes in serum type were induced in sheep by venepuncture (type 1 to type 2) or venepuncture and an intraperitoneal injection of yeast cells (type 2 to type 3) . Direct conglutinating activity of type 3 sera was inhibited by heating serum at 50 degrees C for 30 min and was not restored by alternative activation pathway factor B . Chelation of Ca2+ in type 2 and 3 sera blocked sensitization of sheep E-rabbit A for conglutination by bovine conglutinin, indicating that the classical activation pathway was involved.

Prog Clin Biol Res, 1985, 172B, 175 - 84
X-ray studies on the interaction of the anticancer agent cis-{Pt(NH3)2cl2} to tRNAphe . A mechanism for the formation of the intrastrand cross-link to adjacent guanines in DNA; Sundaralingam M et al.; The interaction of the potent anticancer agent cis-{Pt(NH3)2cl2} to phenylalanine tRNA from yeast has been investigated by x-ray crystallography . Two major binding sites were excavated from difference Fourier maps and both showed monodentate binding to guanine bases (G15 and G18) at N7 . The expected intrastrand cross-linked complex between adjacent guanines was not observed in tRNA . Although both bases G15 and G18 are in the dihydrouridine loop, they are engaged in tertiary base interactions viz . G15-C48 and G18-psi 56 . The high C1- ion concentration in the crystal made the reaction sluggish and was perhaps responsible for the formation of only the monodentate complex . This appears to be the initial complex that is formed during the interaction of the Pt drug with DNA, with a subsequent attack of the drug on the N7 of the 5'-side guanine base to generate the expected intrastrand cross-linked product involving adjacent guanines . The wedge-shaped Pt-GG complex that is formed perturbs not only the local geometry but also produces a kink or bend in the DNA duplex.

Gene, 1985, 35(3), 289 - 96
Molecular cloning and nucleotide sequences of cDNAs specific for rat liver ribosomal proteins S17 and L30; Nakanishi O et al.; cDNA clones coding for rat liver ribosomal proteins S17 and L30 have been isolated by positive hybridization-translation assay from a cDNA library prepared from 8-9S poly(A)+RNA from free polysomes of regenerating rat liver . The cDNA clone specific for S17 protein (pRS17-2) has a 466-bp insert with the poly(A) tail . The complete amino acid (aa) sequence of S17 protein was deduced from the nucleotide sequence of the cDNA . S17 protein consists of 134 aa residues with an Mr of 15 377 . The N-terminal aa sequence of S17 protein determined by automatic Edman degradation is consistent with the sequence data . The aa sequence of S17 shows strong homology (76.9%) to that of yeast ribosomal protein 51 {Teem and Rosbash, Proc . Natl . Acad . Sci . USA 80 (1983) 4403-4407} in the two-thirds N-terminal region . The cDNA clone specific for L30 protein (pRL30) has a 394-bp insert . The aa sequence of L30 protein was deduced from the nucleotide sequence of the cDNA . The protein consists of 114 aa residues with an Mr of 12 652 . When compared with the N-terminal aa sequence of rat liver L30 protein {Wool, Annu . Rev . Biochem . 48 (1979) 719-754}, pRL30 was found not to contain the initiation codon and 5'-noncoding region . The cDNA showed twelve silent changes in the coding region, one point mutation and one base deletion in the 3'-noncoding region, compared with mouse genomic DNA for L30 protein {Wiedemann and Perry, Mol . Cell Biol . 4 (1984) 2518-2528}.

Dev Biol Stand, 1985, 59, 113 - 20
Properties and relative immunogenicity of various preparations of recombinant DNA-derived hepatitis B surface antigen; Burnette WN et al.; The gene for hepatitis B surface antigen was cloned into vectors for expression in yeast and mammalian cells . These hosts assemble the surface antigen protein into spherical structures containing cell-derived lipids . Particles were isolated from these cell cultures and purified by standard biochemical and biophysical means . Protein micells were formed from such particles by removal of lipid with nonionic detergent . Both particle and protein micellar preparations were formulated with alum adjuvant and tested for their immunopotency in mice . All the materials so analysed proved to be highly immunogenic . Safety and regulatory aspects of these materials and other potential and current hepatitis B vaccines are discussed . It is concluded that the yeast-derived materials and certain mammalian cell production systems present the most suitable opportunities for new hepatitis B vaccines.

Biol Cell, 1985, 53(2), 149 - 54
Fluid phase and mannose receptor-mediated uptake of horseradish peroxidase in mouse bone marrow-derived macrophages . Biochemical and ultrastructural study; Lang T et al.; A detailed study of horseradish peroxidase (HRP) uptake by in vitro cultured bone marrow-derived macrophages was undertaken . Biochemical quantitations performed over a wide range of HRP concentrations, in the absence or presence of yeast mannan, showed that these macrophages pinocytose HRP by both fluid phase and mannose receptor-mediated uptake . The relative contribution of these two types of endocytosis varied with the concentration of enzyme in the extracellular medium . A morphological study at the light and electron microscope levels conducted in parallel confirmed the biochemical data.

Adv Pediatr, 1985, 32, 287 - 320
Viral hepatitis: implications to pediatric practice; Balistreri WF; A major frontier is the development of effective therapy for patients with chronic type B hepatitis, which currently remains refractory to all known antiviral agents and is associated with severe long-term consequences . Future research should focus on the relationship of HBV to hepatocellular carcinogenesis . This might answer the question as to when integration of viral DNA into the host cell occurs and whether this is an irreversible step in the progression toward hepatocellular carcinoma . The exciting work regarding vaccines against human hepatitis will continue in view of the worldwide impact of elimination of these viruses . Chemically synthesized vaccines may be safer, cheaper, and more effective, and also produce more persistent protective immunity . This technology requires determination of immunogenic protein molecules, identification and in vivo synthesis of the amino acid sequences important for cellular recognition, and the application of recombinant DNA technology to clone and propagate the critical antigenic determinants by incorporation into other hosts such as vaccinia . The potential of newer forms of hepatitis B vaccine has been demonstrated by the study of Scolnick et al., in which HBsAg, produced by a recombinant strain of yeast, was given to 37 healthy low-risk volunteers as a means of vaccination . The dose and schedule were the same as that used for the serum HBsAg-derived vaccine . The results were encouraging in that an 80%-100% anti-HBs positivity was documented at three months in these individuals . These experiences also suggest that a similar sequence of events, namely definition, elucidation of the molecular biology, and development of immunologic prevention of non-A, non-B viruses is at hand.

Z Erkr Atmungsorgane, 1985, 165(2), 149 - 62
{Modulation of alveolar macrophage activity by ambroxol, bromhexine and exogenous arachidonic acid}; Winsel K et al.; The increased generation of reactive oxygen metabolites (ROM) (O2-, H2O2, 1O2, X OH, OX-) by alveolar macrophages (AM) and neutrophils has been proposed as an important step in the pathogenesis of many acute and chronic lung disorders . With regard to a possible therapeutic application in this context the effect of bromhexine and ambroxol on the activity of AM of of guinea-pigs and of patients with lung diseases was investigated . The activity of AM, which were isolated by bronchoalveolar lavage, was determined by means of chemiluminescence (CL)-measuring . Ambroxol and bromhexine cause a depression of the spontaneous as well as of the induced CL {yeast cell walls, yeast glucan, exogenous arachidonic acid (AA)} of AM . AM from guinea-pigs and in some cases also from patients show primarily an increase of the CL-signal under the influence of bromhexine . These results suggest that ambroxol and bromhexine reduce the production of ROM by AM . The mechanism is possibly due to the activation of acyl-CoA-lysophosphatide acyltransferase . The increase of this enzyme activity lowers the intracellular concentration of the free AA, whereby the generation of ROM is diminished too . The investigations exhibit a new possibility of influence of respiratory burst and thus indirectly of AA-liberation from phospholipids . The probable reduction of AA-liberation by these drugs could also be of importance for other lung cells especially concerning the biosynthesis of eicosanoids (AA-metabolites), which play an important part as pathogenetic mediators in different lung diseases.

Int J Biochem, 1985, 17(1), 1 - 11
NAD+ kinase--a review; McGuinness ET et al.; NAD+ kinase catalyzes the only (known) biochemical reaction leading to the production of NADP+ from NAD+ . Most evidence indicates it is found in the cytoplasm, but reports of its presence in (other) cell bodies can not be discounted . Viewed as a protein, our knowledge of NADK composition and architecture is rudimentary . Though recognized as a large multimeric protein, no agreement is evident for the molecular weight (Mr = approximately 4-65 X 10(4} of the native protein . Is calmodulin an integral subunit of (some, all) NAD+ kinases (analogous to phosphorylase kinase in skeletal muscle)? Or is it an external modulator? Consensus is evident that a subunit of molecular weight 30-35 X 10(3) is a component of the mammalian and yeast kinase . In one case (rabbit liver) two types of subunits are reported to give rise to oligomers differing in molecular weight and catalytic activities . Viewed as an enzyme it is not known why such a complex aggregate is needed for what might otherwise appear to a routine phosphorylation reaction . Rapid equilibrium random (for pigeon liver and C . utilis preparations) and ping-pong (for A . vinelandii kinase) mechanisms have been proposed for the reaction, with multiple reactant binding sites indicated for the random cases . From the perspective of enzyme modulation, the demonstration that green plant and sea urchin egg kinases are targets for calmodulin regulation by intracellular Ca2+ links NADP+ production in these sources to the multi-level discriminatory control functions inherent to this Ca2+-protein complex . Significant questions arise from the results of various investigators considered in this review . These queries offer fertile ground for the selective design of key experiments directed to a better understanding of NAD+ kinase function and pyridine nucleotide biochemistry.

Curr Genet, 1985, 10(4), 321 - 8
The mitochondrial genome of fertile maize (Zea mays L.) contains two copies of the gene encoding the alpha-subunit of the F1-ATPase; Isaac PG et al.; In contrast to the situation in animals and fungi the alpha-subunit of the mitochondrial F1-ATPase is encoded by two identical mitochondrial genes (ATP A) in male fertile maize (Zea mays L.) . Cytoplasmic male sterile (T, C and S) maize mitochondrial genomes only contain a single copy of the gene . Sequence analysis reveals that the uninterrupted coding region of both copies of the gene is 1,524 bp long and encodes a polypeptide of 508 amino acids with a molecular weight of 55,117 . The predicted amino acid sequence shares over 60% homology with the nuclear encoded alpha-subunit from yeast and bovine ATPase and approx . 50% with the corresponding chloroplast and bacterial polypeptides.

Comp Biochem Physiol A, 1985, 82(1), 233 - 7
Plasma essential amino acids changes in sea-bass (Dicentrarchus labrax) after feeding diets deficient and supplemented in L-methionine; Thebault H; A fishmeal-based reference diet, a soybean-yeast-based methionine (MET)-deficient diet and two MET-supplemented diets were fed to groups of yearling sea-bass, reared in sea-water at 21 +/- 1 degrees C . Levels of most free essential amino acids (EAA) reached a maximum concentration in plasma between 4 and 8 hr after a single meal . Plasma MET level remained very low with the deficient diet, exhibiting by 4 hr a 10-fold reduction when compared to the reference diet, although MET content was divided only by two in the former . Daily MET intake was then maintained under minimum requirement in fish fed the deficient diet . Supplemented MET reached peak level in plasma substantially sooner than the rest of the EAA, indicating faster absorption and degradation when compared to MET incorporated in feed proteins . Evolution of methionine sulfoxide (MSO) levels in plasma after feeding MET-supplemented diets gives evidences that a degradation pathway of MET into MSO exists in sea-bass.

Int Arch Allergy Appl Immunol, 1985, 78(2), 182 - 9
Analytic study of the differential anticomplementary effects of dextran sulphate and heparin in the assay for the mouse alternative pathway; Klerx JP et al.; In a recent paper, a linkage between immunological adjuvant activity in mice and in vitro anticomplementary (alternative pathway assay) effects was described for different polyanions . This connection was found only if mouse serum was used as a complement (C) source . In order to investigate the possible role of C in adjuvant activity, the differential effects of polyanions on mouse C were studied in detail . For this study, substances with different activity were selected, namely dextran sulphate with strong C-regulatory and immunoadjuvant activities, and heparin, which was weakly anticomplementary and devoid of adjuvant effect . In general, studies of mouse C are complicated by the unavailability of isolation procedures for the C-components involved . This difficulty was circumvented by making use of C5-deficient serum and of the haemolytic activity of mouse membrane attack complexes formed in the fluid phase . With yeast cells as alternative pathway activators it was shown that the effect of heparin on this pathway was restricted to activation of the terminal route . In contrast, dextran sulphate also caused a functional decay of a yeast-bound alternative pathway C5-convertase and interfered with the haemolytic activity of fluid-phase membrane attack complexes as well . Further studies will be needed to decide whether these specific effects of dextran sulphate are related to the immunological adjuvant activity of the substance in mice.

J Biol Chem, 1984 Dec 10, 259(23), 14637 - 41
Swainsonine and castanospermine blockade of mannose glycoprotein uptake by macrophages . Apparent inhibition of receptor-mediated endocytosis by endogenous ligands; Chung KN et al.; Receptor-mediated uptake of mannose-terminated glycoproteins by macrophages is blocked by treating the cells with swainsonine, an inhibitor of alpha-mannosidase II, and by castanospermine, an inhibitor of the endoplasmic reticulum processing enzyme alpha-glucosidase . Both inhibitors are known to cause accumulation of unprocessed oligosaccharide chains terminating in mannose . Inhibition of ligand uptake by the drugs was time- and dose-dependent . Swainsonine produced a maximal effect after 2 h; castanospermine required 5-6 h . Following swainsonine treatment, complete recovery of mannose receptor activity required 24 h and was blocked by cycloheximide suggesting that new receptor synthesis was necessary . Tunicamycin, an inhibitor of oligosaccharide assembly, had no effect on uptake of mannosylated ligands, but tunicamycin pretreatment reduced the sensitivity to swainsonine . These effects of swainsonine and castanospermine appear to be specific, other macrophage pinocytosis receptors (e.g . mannose phosphate) or phagocytosis of yeast particles were unaffected . Moreover, swainsonine had no effect on the fibroblast mannose phosphate receptor . The ability of macrophages to process newly synthesized oligosaccharides was blocked following treatment with swainsonine . Normal processing was fully recovered 24 h after removal of the drug . Mannosidase II was partially inactivated by swainsonine treatment and only a portion was recovered after 24 h . Treatment of macrophages with swainsonine also resulted in an increase in net lysosomal enzyme secretion . Inhibition of mannose-specific receptor-mediated endocytosis in macrophages by swainsonine and castanospermine appears to be due to the formation of mannose-terminated membrane glycoproteins which engage the mannose receptor thereby preventing function . These results suggest a novel mechanism for regulation of receptor-mediated endocytosis.

Hum Nutr Appl Nutr, 1984 Dec, 38(6), 421 - 34
A holistic view of allergic disease; Morrow-Brown H; A personal approach to the diagnosis and treatment of patients with food allergy or intolerance is presented in this paper . Patients with these problems may prove difficult to treat because of intolerance to several different foods, and the situation may be further complicated by allergy to inhaled substances . Fasting and elimination and rotation diets may be used for diagnosis, together with open food challenges . Although 'blind' food challenge may be useful as a research procedure, it can be unhelpful in day-to-day treatment . Examples of the use of this approach in patients sensitive to cow's milk protein, egg, wheat, yeast and food additives are presented and discussed . The author concludes that there is enormous suffering from ingestant allergy which could be relieved by recognizing and avoiding the causative factors.

J Bacteriol, 1984 Dec, 160(3), 1171 - 4
Assimilation of chlorinated alkanes by hydrocarbon-utilizing fungi; Murphy GL et al.; The fatty acid compositions of two filamentous fungi (Cunninghamella elegans and Penicillium zonatum) and a yeast (Candida lipolytica) were determined after the organisms were grown on 1-chlorohexadecane or 1-chlorooctadecane . These organisms utilized the chlorinated alkanes as sole sources of carbon and energy . Analyses of the fatty acids present after growth on the chlorinated alkanes indicated that 60 to 70% of the total fatty acids in C . elegans were chlorinated . Approximately 50% of the fatty acids in C . lipolytica were also chlorinated . P . zonatum contained 20% 1-chlorohexadecanoic acid after growth on either substrate but did not incorporate C18 chlorinated fatty acids.

J Immunol, 1984 Dec, 133(6), 3415 - 23
Studies on the mechanism of natural killer cell-mediated cytotoxicity . V . Lack of NK specificity at the level of induction of natural killer cytotoxic factors in cultures of human, murine, or rat effector cells stimulated with mycoplasma-free cell lines; Wright SC et al.; We have proposed that lysis of target cells by NK cells is mediated by NK cytotoxic factors (NKCF) . According to our model, for a target cell to be NK-sensitive, it must be recognized by the NK cell, it must stimulate the release of NKCF, and it must be sensitive to lysis by these factors . This report examines whether the ability to stimulate release of NKCF is a characteristic restricted to NK-sensitive tumor cells or whether it is also a property of NK-resistant target cells . Many different types of cell lines were tested for their ability to stimulate release of NKCF in the human, rat, and murine systems . It was found that mycoplasma-free NK-sensitive cell lines, resistant cell lines, and Con A could stimulate the release of NKCF . Many different types of cell lines grown in suspension or in monolayers were found to be effective stimulators, including T or B lymphoid, myeloid, and those of histiocytic origin . Cells cultured in the absence of serum stimulated NKCF release, thus ruling out the possible involvement of serum components in stimulation . NKCF was also produced by xenogeneic combinations of effector and stimulator cells, demonstrating lack of species specificity in NKCF production . Factors stimulated by NK-resistant cell lines or by Con A exhibited the same NK target specificity as supernatants stimulated by NK-sensitive tumor cells . The finding that many different NK-resistant cell lines can stimulate the release of NKCF indicates that there is no apparent NK specificity at the level of induction of NKCF release from human, rat, or murine effector cells . Therefore, the NK specificity of a target cell is determined ultimately by its sensitivity to lysis by NKCF.

J Biomol Struct Dyn, 1984 Dec, 2(3), 525 - 30
Crosslinking of tRNAs by the carcinostatic agent dirhodium tetraacetate; Rubin JR et al.; A crystalline complex of yeast tRNA(phe) and dirhodium tetraacetate (DRTA) was prepared and its X-ray structure determined . The bifunctional DRTA forms an intermolecular cross-link between the N(1) position of adenine A36 in the anticodon triplet and possibly a ribose hydroxyl group of residue A76 at the 3' terminus of a symmetry related tRNA molecule . The rhodium complex apparently shows a preference for binding to the N(1) position of adenine in a single strand region of the tRNA molecule.

Can J Psychiatry, 1984 Dec, 29(8), 707 - 11
Diet and monoamine oxidase inhibitors: a re-examination; Sullivan EA et al.; Monoamine oxidase inhibitors (MAOIs) are attracting renewed attention as effective antidepressants for refractory depressions, particularly among the elderly . However, widespread fears concerning the interactions of MAOIs with tyramine-containing foods have led to the development of long and complicated diets . These diets have served as an obstacle to the ready use of MAOIs, yet very little systematic or critical review of the basis for food restriction has been undertaken . An international survey of MAOI diets was conducted and from the diets collected, foods were categorized according to frequency of restriction on the diet lists . On the basis of this survey and a critical review of the literature it was determined that only four foods clearly warrant absolute prohibition: aged cheese, pickled fish (herring), concentrated yeast extracts and broad bean pods . While there is insufficient evidence to prohibit alcohol completely (even chianti wine) true moderation must apply . It is suggested that a radically simplified diet should be investigated on a prospective basis.

Proc Natl Acad Sci U S A, 1984 Dec, 81(23), 7417 - 20
Intron splicing: a conserved internal signal in introns of animal pre-mRNAs; Keller EB et al.; Splicing of introns of yeast pre-mRNAs requires an internal conserved sequence T-A-C-T-A-A-C that is located 20-55 nucleotides from the 3' intron boundary . Sequences differing only in certain positions from this yeast signal have now been identified in the corresponding internal region of pre-mRNA introns of a variety of animal genes . A computer program that searches for homologues to a consensus structure and calculates the accuracy of match of each homologue is used to locate these sequences . We list here the signals found by this search in introns of sea urchin, mouse, rat, and human genes and give the consensus for each species . We also give the consensus found for Drosophila and chicken and duck signals . We then discuss the accumulating evidence that these internal signals are required for splicing in animals . It is also noted that a single-stranded region of small nuclear RNA U2 contains sequences complementary both to the proposed mammalian internal signal and to the neighboring CT-A-G at the 3' intron boundary . A role for U2 ribonucleoprotein in intron splicing is thus suggested.

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