Lack of effects of thyroid hormones on human erythrocyte acetylcholine esterase

Effects of thyroid hormones and their metabolites such as L-T1, L-T2, L-T3 and L-T4 on human erythrocyte acetylcholine esterase were studied. The activity of the enzyme of intact erythrocytes was not affected by these hormones, though studied under various conditions. The physiological significance of the binding of these hormones to erythrocyte membranes remains unclear. Our results indicate that the acetylcholine esterase is not a suitable enzyme for cytochemical bioassay for thyroid hormones.     

Lytic β-1,3 Glucanase from Arthrobacter: Pattern of Action

The action pattern of lytic β-1,3 glucanase (glucanase I) from Arthrobacter which liberates predominantly laminaripentaose from various β-glucans has been studied. The enzyme was not active on short linear laminaridextrins, but was active on an enzymatically synthesized, linear β-1,3 oligoglucan preparation. Any intactness of the glucose residues of the chain ends of a substrate did not seem to be necessary for the action of the enzyme. The results of determination of laminaripentaose during a relatively early phase of the reaction suggested that about half of the reducing power liberated in the medium might be explained by the formation of the sugar. It seems that the formation of laminaripentaose relates to the initial attack of glucanase 1 on β-1,3 glucan chains.     

Energy Transducing Roles of Antiporter-like Subunits in Escherichia coli NDH-1 with Main Focus on Subunit NuoN (ND2)

The proton-translocating NADH-quinone oxidoreductase (complex I/NDH-1) contains a peripheral and a membrane domain. Three antiporter-like subunits in the membrane domain, NuoL, NuoM, and NuoN (ND5, ND4 and ND2, respectively), are structurally similar. We analyzed the role of NuoN in Escherichia coli NDH-1. The lysine residue at position 395 in NuoN (_NLys³⁹⁵) is conserved in NuoL (_LLys³⁹⁹) but is replaced by glutamic acid (_MGlu⁴⁰⁷) in NuoM. Our mutation study on _NLys³⁹⁵ suggests that this residue participates in the proton translocation. Furthermore, we found that _MGlu⁴⁰⁷ is also essential and most likely interacts with conserved _LArg¹⁷⁵. Glutamic acids, _NGlu¹³³, _MGlu¹⁴⁴, and _LGlu¹⁴⁴, are corresponding residues. Unlike mutants of _MGlu¹⁴⁴ and _LGlu¹⁴⁴, mutation of _NGlu¹³³ scarcely affected the energy-transducing activities. However, a double mutant of _NGlu¹³³ and nearby _KGlu⁷² showed significant inhibition of these activities. This suggests that _NGlu¹³³ bears a functional role similar to _LGlu¹⁴⁴ and _MGlu¹⁴⁴ but its mutation can be partially compensated by the nearby carboxyl residue. Conserved prolines located at loops of discontinuous transmembrane helices of NuoL, NuoM, and NuoN were shown to play a similar role in the energy-transducing activity. It seems likely that NuoL, NuoM, and NuoN pump protons by a similar mechanism. Our data also revealed that _NLys¹⁵⁸ is one of the key interaction points with helix HL in NuoL. A truncation study indicated that the C-terminal amphipathic segments of _NTM14 interacts with the _Mβ sheet located on the opposite side of helix HL. Taken together, the mechanism of H⁺ translocation in NDH-1 is discussed.     

Partial Sequences of Large Subunit Ribosomal DNA of a New Yeast Species,Trichosporon domesticum and Related Species

We determined the partial sequences of large subunit rDNA of a new yeast species, Trichosporon domesticum, which was isolated from the house of a summer-type hypersensitivity pneumonitis patient. Phylogenetically, T. domesticum was positioned in the taxonomic group containing T. montevideense and T. brassicae, which indicated an identical serotype. A phylogenetic relationship among all species of the genus Trichosporon which belong to the basidiomycetous yeast is clarified.     

Comparative studies on the properties of spinach chloroplasts prepared by media containing choline chloride or sucrose

Spinach chloroplasts were isolated and stored in a medium containing0.5 M choline chloride. The properties of these chloroplastswere compared with those of chloroplasts prepared in an ordinarysucrose medium. In marked contrast to sucrose-prepared chloroplasts, choline-preparedchloroplasts did not show "conventional uncoupling" (stimulationof ferricyanide reduction paralleling the inactivation of phosphorylation)after transient warming in the temperature range between 20?and 55?C. The thermal stability of the oxygen evolving system, the vulnerabilityof the lipophilic structure in the thylakoids to alcohol duringthe warming treatment, and the effects of amine on photophosphorylationwere similar in both chloroplast preparations. After the warming treatment, the degree of swelling in chloroplastswas larger and the intermediate electron transport systme (fromreduced 2,6-dichlorophenolindophenol to methyl viologen) wasmore stable against uncoupling, in choline-prepared chloroplaststhan in sucrose-prepared ones. The presence and absence of "conventional uncoupling" in twochloroplast preparations were ascribed to differences in thermalstability. In choline-prepared chloroplasts the uncoupling temperatureof the electron transport system was higher than the inactivationtemperature of the oxygen evolving system, but it was lowerin the sucrose-prepared ones. (Received February 13, 1974; )     

Lipophilic interaction between thylakoid membranes and aliphatic compounds

Spinach chloroplasts pre-incubated at various temperatures, changed their photosynthetic activities (measured at 15°C) as the incubation temperature was raised; these activities showed characteristic activitytemperature profiles as follows:

1. The profiles were shifted to lower temperatures if various aliphatic compounds were present in the incubation mixture.
2. In general, the extent of the shift was proportional to the product of the concentration and the partition coefficient of the compound, i.e. to the concentration of the compound partitioned in the lipophilic chloroplast phase.

The combined effects of heat and the aliphatic compounds tested indicated that the lipophilic groups in these compounds play a determinative role in the heat denaturation of thylakoids. The consequence of such structure alterations is inactivation of photosynthetic functions. The lipophilic properties of the spinach thylakoid membrane as compared with certain artificial and natural membranes are described.     

Knockdown of Carotenoid Cleavage Dioxygenase 4 (CCD4) via Virus-Induced Gene Silencing Confers Yellow Coloration in Peach Fruit: Evaluation of Gene Function Related to Fruit Traits

Plant Molecular Biology Reporter - Transgenic approach is an excellent way for the clarification of gene function, but it is generally difficult to create transgenic plants for most of the fruit...     

Water-soluble fractions from defatted sesame seeds protect human neuroblast cells against peroxyl radicals and hydrogen peroxide-induced oxidative stress

Oxidative stress is involved in the development of aging-related diseases, such as neurodegenerative diseases. Dietary antioxidants that can protect neuronal cells from oxidative damage play an important role in preventing such diseases. Previously, we reported that water-soluble fractions purified from defatted sesame seed flour exhibit good antioxidant activity in vitro. In the present study, we investigated the protective effects of white and gold sesame seed water-soluble fractions (WS-wsf and GS-wsf, respectively) against 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH) and hydrogen peroxide (H₂O₂) induced oxidative stress in human neuroblast SH-SY5Y cells. Pretreatment with WS-wsf and GS-wsf did not protect cells against AAPH-induced cytotoxicity, while simultaneous co-treatment with AAPH significantly improved cell viability and inhibited membrane lipid peroxidation. These results suggest that WS-wsf and GS-wsf protect cells from AAPH-induced extracellular oxidative damage via direct scavenging of peroxyl radicals. When oxidative stress was induced by H₂O₂, pretreatment WS-wsf and GS-wsf significantly enhanced cell viability. These results suggest that in addition to radical scavenging, WS-wsf and GS-wsf enhance cellular resistance to intracellular oxidative stress by activation of the Nrf-2/ARE pathway as confirmed by the increased Nrf2 protein level in the nucleus and increased heme oxygenase 1 (HO-1) mRNA expression. The roles of ferulic and vanillic acids as bioactive antioxidants in these fractions were also confirmed. In conclusion, our results indicated that WS-wsf and GS-wsf, which showed antioxidant activity in vitro, are also efficient antioxidants in a cell system protecting SH-SY5Y cells against both extracellular and intracellular oxidative stress.