Immunochemical study on PHI/PHM with use of synthetic peptides

We have synthesized PHI and PHM (human PHI) as well as their fragments, PHI (1-6), PHI (1-15), PHI (14-19), PHI (14-27), PHI (20-27), PHM (1-15) and PHM (13-27), by the solution or solid-phase method for peptide synthesis. Using the highly purified synthetic peptides as immunogens or haptenic immunogens, five kinds of PHI/PHM specific antisera were produced. The major antibody-recognition sites of the five antisera were located respectively in the PHI C-terminal (R8201), in the PHI N-terminal (R8403), in the PHM C-terminal (R8502), and in the PHM whole molecule (R8702 and R8703). Radioimmunoassays (RIAs) with antisera R8201, R8403 and R8502, respectively, showed a wide distribution of immunoreactive (IR) PHI/PHM in porcine and human gastrointestinal and brain tissues. The concentrations of IR-PHI in the porcine gastrointestinal tissues, however, differed between the R8201 and R8403 RIAs employed for measurement. By using these two different PHI RIAs, the IR-PHI in the porcine brain tissue extract was shown to be almost a single component coeluting with synthetic PHI in gel filtration. The IR-PHI in the extract of porcine lower intestine on the other hand, contained, besides a PHI-like component, unidentified component(s) eluting immediately after synthetic PHI in gel filtration; this crossreacted with the PHI C-terminal specific R8201 antiserum but not with the N-terminal specific R8403 antiserum, suggesting the presence of the C-terminal-related fragment(s) of PHI in the tissues.     

Effects of Preillumination on Dark Nitrogen Fixation and Respiration by Anabaena Cylindrica

In whole filaments of Anabaena cylindrica dark nitrogen-fixingactivity (measured as acetylene reduction) and respiration increasedwith the light intensity of a fixed period of preillumination,saturating at ca. 10,000 lux. With saturating light during preillumination,the amount and duration of dark nitrogen-fixing activity increasedwith length of preillumination, but respiration declined rapidlyin the dark. At dark respiration rates below 250 nmol O₂ uptake mg protein^–1?h^–1(State 1) no significant nitrogen-fixing activity is observed.From 250 to 550 nmol O₂ uptake?mg protein^–1?h^–1(State 2), nitrogen-fixing activity depends on O₂ uptake whileabove 550 nmol O₂ uptake?mg protein^–1?h^–1 (State3), nitrogen-fixing activity no longer increases with furtherincrease in O₂ uptake rate. (Received June 18, 1983; Accepted November 10, 1983)     

Effect of High-intensity Light Given Prior to Low-temperature Treatment on the Long-day Flowering of Pharbitis nil

Pharbitis nil, strain Violet which had been exposed to high-intensitylight (18,000 lux at 23?C) for 7 days followed by a low-temperaturetreatment (13–14?C) for 7 days initiated flower buds evenunder continuous light, but plants given these treatments inreverse order failed to bud. Three days of high-intensity lightat 23?C was most effective in promoting the flower-inducingeffect of the subsequent low-temperature period. Six days oflow temperature following the 3-day high-intensity light periodinduced near-maximum flowering response. DCMU (5?10^–6M) given during the high-intensity light period inhibited flowering,but when given during or after the low-temperature period itwas ineffective. DCMU at the same concentration given before,during or after an inductive 16-hr dark period at 26?C did notinhibit flowering. Sucrose, ATP, NADPH and some other reducingagents tested did not nullify the DCMU effect nor substitutefor the effect of high-intensity light. But, the high-intensitylight effect could be substituted, at least partly, by 5-chlorosalicylicacid, 3,4-dichlorobenzoic acid and some other benzoic acid derivatives,which are highly effective in inducing long-day flowering inthe short-day plant, Lemna paucicostata. (Received October 20, 1981; Accepted February 3, 1982)     

Electron paramagnetic resonance properties and oxidation-reduction potentials of the molybdenum,flavin, and iron-sulfur centers of chicken liver xanthine dehydrogenase

The oxidation-reduction potentials of the various prosthetic groups in the native and desulfo forms of chicken liver xanthine dehydrogenase, determined by potentiometric titration in 0.05 m potassium phosphate buffer, pH 7.8, are: Mo(VI)/Mo(V) (native), ?357 mV; Mo(VI)/Mo(V) (desulfo), ?397 mV; Mo(V)/Mo(IV) (native), ?337 mV; Mo(V)/Mo(IV) (desulfo), ?433 mV; FAD/FADH · ?345 mV; FADH · FADH₂, ? 377 mV; (Fe/S)I_ox/(Fe/S)I_red, ?280 mV; (Fe/S)II_ox/(Fe/S)II_red, ? 275 mV. Titration at pH 6.8 revealed that the Mo and FAD centers but not the Fe/S centers are in prototropic equilibrium. Spectroscopic studies on the native and deflavinated enzymes show that environment of the flavin in xanthine dehydrogenase differs from that in bovine milk xanthine oxidase.     

Presence of immunoreactive alpha-endorphin in rat pituitary gland.

Utilizing radioimmunoassay for α-endorphin, we attempted to demonstrate immunoreactive α-endorphin in acid extracts of pars distalis and combined pars intermedia and pars nervosa of the rat pituitary gland after chromatography on Sephadex G-25. β-Lipotropin, β-endorphin and γ-endorphin were not converted into α-endorphin during the extraction and gel chromatographic procedures. Concentrations of immunoreactive α-endorphin determined after gel chromatography of extracts from pars distalis and combined pars intermedia and pars nervosa were 1.1±0.6 and 130±17 ng/mg wet tissue (mean±SE), respectively. Serial dilution of these extracts gave parallel lines to the standard curve of synthetic α-endorphin, but not to that of γ-endorphin or δ-endorphin. These results suggest the existence of immunoreactive α-endorphin indistinguishable in molecular size from synthetic α-endorphin in the rat pituitary gland.     

Differential effects of oxygen on N2 fixation and C2H2 reduction by Anabaena cylindrica

Both nitrogen fixation and acetylene reduction by intact cellsof Anabaena cylindrica were inhibited by oxygen, but nitrogenfixation was invariably less sensitive than acetylene reduction.The C₂H₂/N₂ ratio ranged from 6 to 8 in the absence of oxygen,and it decreased with increase in partial pressure of oxygento 2 at a pO₂ of 0.3 atm. (Received June 5, 1979; )