Role of mitogen-activated protein kinase pathway in reactive oxygen species-mediated endothelin-1-induced β-myosin heavy chain gene expression and cardiomyocyte hypertrophy

Endothelin-1 (ET-1) has been found to increase cardiac -myosin heavy chain (-MyHC) gene expression and induce hypertrophy in cardiomyocytes. ET-1 has been demonstrated to increase intracellular reactive oxygen species (ROS) in cardiomyocytes. The exact molecular mechanism by which ROS regulate ET-1-induced -MyHC gene expression and hypertrophy in cardiomyocytes, however, has not yet been fully described. We aim to elucidate the molecular regulatory mechanism of ROS on ET-1-induced -MyHC gene expression and hypertrophic signaling in neonatal rat cardiomyocytes. Following stimulation with ET-1, cultured neonatal rat cardiomyocytes were examined for ³H-leucine incorporation and -MyHC promoter activities. The effects of antioxidant pretreatment on ET-1-induced cardiac hypertrophy and mitogen-activated protein kinase (MAPKs) phosphorylation were studied to elucidate the redox-sensitive pathway in cardiomyocyte hypertrophy and -MyHC gene expression. ET-1 increased ³H-leucine incorporation and -MyHC promoter activities, which were blocked by the specific ET_A receptor antagonist BQ-485. Antioxidants significantly reduced ET-1-induced ³H-leucine incorporation, -MyHC gene promoter activities and MAPK (extracellular signal-regulated kinase, p38, and c-Jun NH₂ -terminal kinase) phosphorylation. Both PD98059 and SB203580 inhibited ET-1-increased ³H-leucine incorporation and -MyHC promoter activities. Co-transfection of the dominant negative mutant of Ras, Raf, and MEK1 decreased the ET-1-induced -MyHC promoter activities, suggesting that the Ras-Raf-MAPK pathway is required for ET-1 action. Truncation analysis of the -MyHC gene promoter showed that the activator protein-2 (AP-2)/specificity protein-1 (SP-1) binding site(s) were(was) important cis-element(s) in ET-1-induced -MyHC gene expression. Moreover, ET-1-induced AP-2 and SP-1 binding activities were also inhibited by antioxidant. These data demonstrate the involvement of ROS in ET-1-induced hypertrophic responses and -MyHC expression. ROS mediate ET-1-induced activation of MAPK pathways, which culminates in hypertrophic responses and -MyHC expression. Tzu-Hurng Cheng, Neng-Lang Shih: These authors have equally contributed to this work     

Diagnosing lactic acid fermentation based on specific rates of growth,substrate consumption and product formation

The on-line calculated specific rates of growth, substrate consumption and product formation were used to diagnose microbial activities during a lactic acid fermentation. The specific rates were calculated from on-line measured cell mass, and substrate and product concentrations. The specific rates were more sensitive indicators of slight changes in fermentation conditions than such monitored data as cell mass or product concentrations.List of Symbols 1/h specific rate of cell growth - 1/h specific rate of substrate consumption - 1/h specific rate of product formation - ^* dimensionless specific rate of cell growth - ^* dimensionless specific rate of substrate consumption - ^* dimensionless specific rate of product formation - _max 1/h maximum specific rate of cell growth - _max 1/h maximum specific rate of substrate consumption - _max 1/h maximum specific rate of product formation - X g/l cell mass concentration - S g/l substrate concentration - S ^* dimensionless substrate concentration - S ₀ g/l initial substrate concentration - P g/l product concentration     

State and time delay estimation of continuous microorganisms cultivation

A continuous fermentation model taking into account the culture memory is used for a state estimation design. The influence of the culture memory on the process dynamics is accounted for by a time delay parameter. The proposed procedure of on-line state estimation in the case when the delay has a constant value is based on the extended Kalman observer. The case when the delay parameter is evaluated on-line is also considered. An adaptive state and parameter algorithm on the base of the extended Kalman filter is proposed. The theoretical results are applied to continuous culture for growth of a strain of Saccharomyces cerevisiae.List of Symbols X, S mg/l Biomass concentration and substrate concentration respectively - S ⁰ mg/l Feed substrate concentration - Z mg/l Past substrate concentration - µ h^–1 Specific growth rate taking into account culture memory - h^–1 Specific consumption rate - h Time delay parameter denoting culture memory - D h^–1 Dilution rate - State variables vector - W _ij Gain coefficient for on-line state and parameter estimation - F Substrate feed rate vector - () Gain coefficient matrix - R Square symmetric Riccati matrix - K Matrix of coefficients - K(t) Delay kernel taking account of culture memory - Denote an estimation value The partial support by Bulgarian National Science Research Foundation under Grant SRTS 428/94 Modeling and Control of Fermentation Processes Taking the Memory Effect into Account is gratefully acknowledged.     

Effect of pH on the activity of nervous cholinesterases of the rat towards different biochemical and histochemical substrates and inhibitors

Summary Cholinesterase activities of homogenates of rat brain and superior cervical ganglion were determined by automatic titration using several biochemical and histochemical substrates. High hydrolysis rates were observed when acetylcholine, acetylthiocholine, propionylcholine or propionylthiocholine was used as substrate; -naphthyl acetate and acetyl--methylcholine were hydrolyzed at a moderate rate, and activities were low towards butyrylcholine, butyrylthiocholine and benzoylcholine. With most substrates, the enzyme activity increased from pH 5 to pH 10 and decreased at pH 11. Acetylcholine and acetyl--methylcholine showed an activity maximum at pH 7 or 8. Inhibition by the selective inhibitor of specific cholinesterase 284 C 51 was not markedly affected by pH. On the other hand, the inhibiting power of the selective inhibitor of non-specific cholinesterase iso-OMPA markedly decreased when the pH was lowered. The inhibitor data at different pHs and with different concentrations of eserine, 284 C 51 or iso-OMPA at pH 6 indicated that acetylcholine, propionylcholine and propionylthiocholine are readily hydrolyzed by both specific and non-specific cholinesterase, while acetyl--methylcholine is mainly split by specific cholinesterase and butyrylcholine mainly by non-specific cholinesterase. The significance of propionylcholine and propionylthiocholine as substrates of specific cholinesterase is emphasized.     

The nervous system of Microstomum lineare (Turbellaria,Macrostomida)

Summary The ultrastructure of release sites of neurochemical messenger substances in the microturbellarian Microstomum lineare was examined. Aminergic neurites form conventional synapses and synapse-like structures (SLS). Variants of true synapses include: single synapses with symmetric pre- and postsynaptic densities, shared synapses, i.e., contacts between 1 pre- and 2 postsynaptic fibres, en passant synapses between parallel axonal membranes, and synapses without thickenings having only clustered vesicles in the presynaptic terminal. SLS on a nerve cell soma or facing an intercellular stromal channel near muscles are described. Peptidergic neurites containing large granular vesicles (LGV) form synaptoids and signs of putative neurosecretory release. Synaptoids between neurites and between neurite and muscle have lucent vacuoles (about 100nm) and dense material at the contact site. In en passage synaptoids dense-core vesicles are embedded in electron-dense material at the contact site. Putative signs of release of neurosecretory material other than typical exocytosis have been observed.     

Genetic Distance in Housekeeping Genes Between <Emphasis Type="Italic">Plasmodium falciparum</Emphasis> and <Emphasis Type="Italic">Plasmodium reichenowi</Emphasis> and Within <Emphasis Type="Italic">P. falciparum</Emphasis>

The time to the most recent common ancestor of the extant populations of Plasmodium falciparum is controversial. The controversy primarily stems from the limited availability of sequences from Plasmodium reichenowi, a chimpanzee malaria parasite closely related to P. falciparum. Since the rate of nucleotide substitution differs in different loci and DNA regions, the estimation of genetic distance between P. falciparum and P. reichenowi should be performed using orthologous sequences that are evolving neutrally. Here, we obtained full-length sequences of two housekeeping genes, sarcoplasmic and endoplasmic reticulum Ca²⁺-ATPase (serca) and lactate dehydrogenase (ldh), from 11 isolates of P. falciparum and 1 isolate of P. reichenowi and estimate the interspecific genetic distance (divergence) between the two species and intraspecific genetic distance (polymorphism) within P. falciparum. Interspecific distance and intraspecific distance at synonymous sites of interspecies-conserved regions of serca and ldh were 0.0672±0.0088 and 0.0011±0.0007, respectively, using the Nei and Gojobori method. Based on the ratio of interspecific distance to intraspecific distance, the time to the most recent common ancestor of P. falciparum was estimated to be (8.30±5.40) × 10⁴ and (11.62±7.56) × 10⁴ years ago, assuming the divergence time of the two parasite species to be 5 and 7 million years ago, respectively.This article contains an online supplementary table.Reviewing Editor: Dr. Martin Kreitman     

Properties of β-glucosidase purified from Aspergillus niger mutants USDB 0827 and USDB 0828

Summary Two extracellular -glucosidases (EC 3.2.1.21) were isolated from Aspergillus niger USDB 0827 and A. niger USDB 0828, and their physical and kinetic properties studied. Both enzymes were very similar in terms of molecular size (230000 Da), pH optimum (pH 4.6), temperature optimum (65° C), stability at high temperatures and substrate preferences. They were capable of hydrolysing -linked disaccharides, phenyl -d-glucoside, p-nitrophenyl -d-glucoside (PNPG), o-nitrophenyl -d-glucoside, salicin and methyl -d-glucoside but lacked activity towards -linked disaccharides, a range of p-nitrophenyl monoglycosides and p-nitrophenyl diglycosides. Both -glucosidases were better at hydrolysing cellobiose than cellotriose, cellotetraose or cellopentaose. For both enzymes, glucose showed competitive inhibition with PNPG as substrate but had no effect with cellobiose. However, the two -glucosidases differed in inhibition by glucono-1,5-lactone and affinity for cellobiose. -Glucosidase from A. niger USDB 0827 also gave lower specific activity, and was more susceptible to metal ions (Ag⁺, Fe²⁺ and Fe³⁺) inhibition than that of A. niger USDB 0828. Correspondence to: Y. K. Hoh     

Existence and uniqueness of a sharp travelling wave in degenerate non-linear diffusion Fisher-KPP equations

In this paper we use a dynamical systems approach to prove the existence of a unique critical value c ^* of the speed c for which the degenerate density-dependent diffusion equation u _ct = [D(u)u _x ] _x + g(u) has: 1. no travelling wave solutions for 0 < c < c ^*, 2. a travelling wave solution u(x, t) = (x - c ^* t) of sharp type satisfying (– ) = 1, () = 0 ^*; '(^*–) = – c ^*/D'(0), '(^*+) = 0 and 3. a continuum of travelling wave solutions of monotone decreasing front type for each c > c ^*. These fronts satisfy the boundary conditions (– ) = 1, '(– ) = (+ ) = '(+ ) = 0. We illustrate our analytical results with some numerical solutions.     

Effect of time delay in specific growth rate on the periodic operation of bioreactors with input multiplicities

The effect of time delay in specific growth rate () on the periodic operation of bioreactors with input multiplicities is theoretically analyzed for productivity improvement. A periodic rectangular pulse is applied either in feed substrate concentration (S_f) or in dilution rate (D). Periodic operation under feed substrate concentration cycling gives improvement in productivity at lower value of ¯S_f of the two steady-state multiplicities of S_f only when the time delay in is larger. Whereas the larger value of ¯S_f gives improvement in average productivity for all values of time delay. Dilution rate (D) cycling gives an improvement in average productivity particularly for larger time delay in . This improvement in average productivity is obtained only at smaller value of dilution rate out of the two steady-state input multiplicities of D.List of Symbols D 1/h dilution rate - F memory function - g dummy variable - K_i g/l substrate inhibition constant - K_m g/l substrate saturation constant - P g/l product concentration - P_m g/l product saturation constant - Q g/(hl) product cell produced per unit time - S g/l substrate concentration - S_f g/l feed substrate concentration - S_f,p g/l feed substrate concentration during fraction of a period - X g/l biomass concentration - Y_X/S g/g cell mass yield - w variable either S or Z - Z g/l weighted average of substrate concentration Greek Letters 1/h time delay parameter - ₁ , ₂ product yield parameters, g/g and 1/h - pulse width expressed as a fraction of a period - 1/h specific growth rate - _m 1/h maximum specific growth rate - h period of oscillation - – average value     

Development and parasitism in the genus Hemileia (uredinales)

Seventeen species ofHemileia were studied with emphasis on these characters: extent of colonization, specialization of parasitic mycelium, haustorial development, pathologic histology, soral morphology and sporulation capacity. An attempt was made to trace the varying degree of development and parasitism in this rust genus comprising of just over 40 reported species. It was noted that theGopalkrishnan's Subepidermal type more advanced than his Superstomal — B type.Fourth part of my thesis submitted to the University of Poona, India, accepted in 1968 for Ph.D. Not published.     

Enzymic synthesis of the trisaccharide core region of the carbohydrate chain ofN-glycoprotein

Transmannosylation from mannotriose (Man1-4Man1-4Man) to the 4-position at the nonreducing end N-acetylglucosaminyl residue ofN,N-diacetylchitobiose was regioselectively induced through the use of -d-mannanase fromAspergillus niger. The enzyme formed the trisaccharide Man1-4GlcNAc1-4GlcNAc (3.7% of the enzyme-catalysed net decrease ofN,N-diacetylchitobiose) from mannotriose as a donor andN,N-diacetylchitobiose as an acceptor. Mannobiose (Man1-4Man) was also shown to be useful as a donor substrate for the desired trisaccharide synthesis.Abbreviations Man d-mannose - (M _n) (n=1–5) -linkedn-mer of mannose - GlcNAc₂ 2-acetamido-2-deoxy--d-glucopyranosyl-(1–4)-2-acetamido-2-deoxy-d-glucose     

Mode of uptake of sterol by Arthrobacter simplex

The mechanism of uptake of water-insoluble -sitosterol by a newly isolated strain of Arthrobacter simplex SS-7 was studied. The production of an extracellular sterol-pseudosolubilizing protein during growth of A. simplex on -sitosterol was demonstrated by isolating the factor from the cell-free supernatant and its subsequent purification by Sephadex G-150 column chromatography. The M _r of the purified sterol-pseudosolubilizing protein determined by SDS–PAGE was 19kDa. The rate of sterol pseudosolubilization (5.2×10^–3g l^–1h^–1) could not adequately account for the rate of sterol uptake (72×10^–3g l^–1h^–1) and the specific growth rate (56×10^–3 h^–1). However in the unfavourable growth condition, when the cells were treated with sodium azide at the level of 30–60% of MIC, the sterol pseudosolubilization accounted for nearly 74% of the total growth containing 96% free cells. Cellular adherence to substrate particles was found to play an active role in the normal growth of the strain on -sitosterol. Unlike sodium acetate-grown cells, whose surface activity was negligible (60mNm^–1), the sterol-grown cells had strong surface activity (40mNm^–1). The high lipid content and long chain fatty acids in the cell-wall of -sitosterol-grown cells probably contribute to the high sterol adherence activity of the cells.     

Role of the N-terminal region of the A chain in beta 1-bungarotoxin from the venom of Bungarus multicinctus (Taiwan-banded krait)

The N-terminal -amino groups of ₁-bungarotoxin (₁-Bgt) fromBungarus multicinctus venom were modified with trinitrobenzene sulfonic acid and the modified derivative was separated by high performance liquid chromatography. The trinitrophenylated (TNP) derivative contained two TNP groups at the -amino groups of A chain and B chain and showed a marked decrease in enzymatic activity. Methionine residues at positions 6 and 8 of the A chain were oxidized with chloramine T or cleaved with cyanogen bromide to remove the N-terminal octapeptide. Oxidation of methionine residues and removal of the N-terminal octapeptide caused a precipitous decrease in enzymatic activity, whereas antigenicity remained unchanged. The presence of dihexanoyllecithin influenced the interaction between ₁-Bgt and 8-antilinonaphthalene sulfonate (ANS) and revealed that ₁-Bgt consists of two types of ANS-binding sites, one at the substrate binding site of the A chain and the other might be at the B chain. The modified derivatives still retained their affinity for Ca²⁺ and ANS, indicating that the N-terminal region is not involved in Ca²⁺ and substrate binding. A fluorescence study revealed that the -amino group of the A chain was in the vicinity of substrate binding site and that the TNP -amino groups were in proximity to Trp-19 of the A chain. In addition, the study showed that the N-terminal region is important for stabilizing the architectural environment of Trp-19. The results, together with the proposal that Trp-19 of the A chain is involved in substrate binding, suggest that the N-terminal region of the A chain plays a crucial role in maintaining a functional active site for ₁-Bgt.     

Two rare GPT phenotypes in a Norwegian family

Summary Phenotypes of members of one Norwegian family have revealed a new allele in the GPT system: Gpt ⁷. Quantitative data of the phenotypes are given.

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Zusammenfassung Phenotypen bei Mitgliedern einer norwegischen Familie haben ein neues Allel im GPT-System gezeigt: Gpt ⁷. Es werden quantitative Daten dieser Phenotypen gegeben.

     

A fluorometric assay of ceramide glycanase with 4-methylumbelliferyl β-d-lactoside derivatives

4-Methylumbelliferyl 6-O-benzyl--d-lactoside (6Bn-MU-Lac) and some related compounds were synthesizedvia different selective reactions including phase-transfer glycosylation. Their suitability as substrates for a fluorometric assay of ceramide glycanase (CGase) was evaluated. Among others, the 6Bn-MU-Lac, which is resistant to exogalactosidase, was found to be a suitable substrate for routine assay of the CGase activity. For American leech CGase, theK _m value is 0.232 mM at pH 5. Abbreviations: CGase, ceramide glycanase; Gal, galactose; Glc, Glucose; Lac, lactose; MU, 4-methylumbelliferone; MU-Lac, 4-methylumbelliferyl -d-lactoside; bBn-Lac, 6-O-benzyl-lactose; 6Bn-MU-Lac, 4-methylumbelliferyl 6-Obenzyl--d-lactoside; 46Bd-MU-Lac, 4-methylumbelliferyl 4,6-O-benzylidene--d-lactoside; MU-Cel, 4-methylumbellifery -d-cellobioside; 46Bd-MU-Cel, 4-methylumbelliferyl 4,6-O-benzylidene--d-cellobioside; TLC, thin layer chromatography;¹H-NMR, proton nuclear magnetic resonance; GSL, glycosphingolipids; CSA, 10-camphorsulfonic acid. See Scheme 1 for chemical structures.     

A Cytogenetic Study of the Leaf Beetle Genus Cyrtonus (Coleoptera, Chrysomelidae)

The chromosomes of ten species of Cyrtonus and the genome sizes of six are surveyed. Among the total of 15 chromosomally studied species, 11 have 2n=28 chromosomes and a 13+Xy_p male meioformula, three have 2n=40 and 19+Xy_p and one 2n=46 and 22+Xy_p. All but one species with 28 chromosomes show only metacentric or submetacentric chromosomes, whereas the species with 40 and 46 chromosomes display some telocentrics or subtelocentrics, that are probably derived from the former by centric fissions. However, since the number of major chromosome arms is strikingly higher in these latter species (NF=70 and 78) than in the 28-chromosome species (mostly NF=56), other chromosomal rearrangements such as pericentric inversions or heterochromatin accretions could also be involved. The genome sizes display a narrow range, from 1C=0.6–1.22pg, and they are not significantly correlated with the chromosome numbers. Some possible factors implied in the rough chromosomal evolution of Cyrtonus are discussed in relation to a few other genera of the subfamily Chrysomelinae.     

Requirements for in vitro propagation of seven nitrogen-fixing Alnus species

Studies on the in vitro propagation of Alnus crispa, A. glutinosa, A. incana, A. japonica, A. rubra, A. sinuata and A. viridis indicated interspecific as well as intraspecific variations in their requirements for in vitro culture. The WPM and Blaydes media supported, respectively, growth of A. glutinosa and A. crispa but not that of both species, while the MS medium induced equal or significantly better growth than WPM and Blaydes media for both species. The optimum type and concentration of sugar to be used in the multiplication medium varied with species. Only A. glutinosa showed good growth on sucrose while glucose was optimum for all other species but at different concentrations. All species rooted in 3 weeks on half-strength MS medium including 1 M IBA. All clones of A. glutinosa and A. rubra rooted 100%, whereas easy-to-root and difficult-to-root clones were observed in the other species. In the rooting medium, glucose promoted rooting of the difficult-to-root clones better than sucrose. Survival following transfer to an artificial substrate was 100% for all species. Nodulation tests using pure cultures of two Frankia strains showed 100% nodulation on all Alnus clones.     

A proposito del llamado micetoma maduro-micosico del pulmon

Conclusiones Se efectúa el estudio de 5 observaciones del llamado Micetoma maduromicósico de pulmón en sus aspectos, histopatológico, micológico y clínico.Todas ellas pertenecen a mujeres y configuraron el cuadro de cavidad bronquial empastada, anotando el predominio de su localización en el lóbulo superior izquierdo.Se señala la uniformidad de los caracteres morfológicos que presenta la masa miceliana llamada grano en todos los casos estudiados, en los cuales no fué posible individualizar la existencia de órganos de fructificación que permitiéran una clasificación, cuando más no fuera, genérica del hongo observado.Se critica la aplicación del término Micetoma para éstos casos con igual criterio que el clásico, que supone una enfermedad micótica primitiva, razón por la cual se prefiere hablar de cavidad con contenido micótico o maduromicótico.En el único caso que se logró cultivar al hongo parásito, el estudio micológico del mismo permitió aislar una especie del GéneroAspergillua con caracteres morfológicos sumamente atípicos.

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Summary The author presents the study of five observations of the so-called Maduromycosis-mycetoma of the lungs in their clinical, histopathologic and mycologic aspects.The pathologic features in all these cases have been found in bronchial cavities of women, situated in the upper lobe of the lung. A compact mycelial mass, the grain, filled up these cavities.In one of the five cases a fungus was cultivated which was classified as belonging to the GenusAspergillus Michelii, with abnormal and atypical features.Short criticism is presented about the concept Maduromycosis mycetoma of the lung, applied by authors designating this process.