Immunoelectrophoretic Analysis of Wheat Flour Albumins

Albumin preparations from four kinds of wheat flour (Durum, Manitoba No. 2, Western White and Norin No. 26) were analyzed by Immunoelectrophoresis. Seven to eleven components were detected for each preparation. They were classified and designated on the basis of the electrophoretic mobility (R) and the character of precipitin lines formed by antigen-antibody reactions.     

Occurrence of methionine-enkephalin-like immunoreactive cells in the feline vagus nerve

The occurrence of peptidergic cellular elements in the feline vagus nerve was examined by an immunohistochemical method. Some methionine-enkephalin-like immunoreactive cells were present around a few capillaries within the vagus nerve, whereas substance-P- and vasoactive-intestinal-polypeptide-like immunoreactive cells were not observed. The methionine-enkephalin-like immunoreactive cells had an oval shape and their diameter was 7-10 micron. These cells appeared to be in contact with the pericyte of the capillaries. Though the physiological function of these enkephalin-positive cells is unclear, our findings suggest that they can act as endocrine cells and release methionine-enkephalin into the capillaries.     

Generation of Active Oxygen Species in Blood Platelets—Spin Trapping Analysis

Generation of active oxygen species by bovine blood platelets was examined by the electron spin resonance (ESR) spin trapping technique with 5,5-dimethyl-l-pyroline-l-oxide (DMPO). The hydroxyl spin-trapped adduct 5,5-dimethyl-2-hydroxy-l-pyrolidinyloxy (DMPO-OH) was formed in the presence of platelets, indicating the generation of hydroxyl radicals (· OH) by the platelets. Generation of · OH was observed even with platelets in the resting state, but was markedly enhanced when the platelets were activated with stimulants. Stronger stimulants such as the calcium ionophore ionomycin, induced greater radical gener-ation than the weaker stimulant ADP. When the platelets were stimulated by thrombin, generation of · OH was greatest after l.5 min, and depended on the dose of the stimulant. It was inhibited by inhibitors of platelet activation such as forskolin and phenolic antioxidants.     

Pharmacokinetic Analysis of Free Radicals by in vivo BCM (Blood Circulation Monitoring)-ESR Method

In pharmacokinetic studies, a variety of analytical method including radioisotopic detection and HPLC (high performance liquid chromatography) has been used. In the present investigation, we developed in vivo BCM (Blood Circulation Monitoring)-ESR method, which is a new technique with a conventional X-band ESR spectrometer for observing stable free radicals in the circulating blood of living rats under anaesthesia. Both 5-(PROXYL derivatives) and 6-(TEMPO derivatives) membered nitroxide spin probes with various types of substituent functional group were used. After physicochemical properties of the spin probes such as hyperfine coupling constant (A-value), g-value and partition coefficient as well as chemical stability of the compounds in the fresh blood were obtained, the in vivo BCM-ESR method was performed in normal rats. Several pharmacokinetic parameters such as half-life of the probes, distribution volume, total body clearance and mean residence time were obtained and discussed in terms of their chemical structures. In addition, clearance of a spin probe was related to the urine concentration. The BCM-ESR method was found to be very useful to observe free radicals at the real time. By time-dependent ESR signal decay of spin probes, pharmacokinetic parameters were obtained.     

Tissue changes in cryptococcosis: histologie alteration from gelatinous to suppurative granulomatous tissue response with asteroid body

The histologic variety and transformation in cutaneous cryptococcosis with acute lymphocytic leukemia before antifungal treatment and after the start of treatment were studied by the light and electron microscopic examinations. The initial cutaneous lesions before treatment revealed gelatinous tissue reactions, and Cryptococcus neoformans (serotype A) were isolated from the skin biopsy specimen and blood. However, later recurrent cutaneous lesions receiving antifungal treatment revealed suppurative granulomatous tissue reactions, and fungal cultures of the skin biopsy specimen changed to negative even though numerous yeasts stained with PAS were observed in skin lesions. Moreover, in the later lesion a few giant cells contained asteroid bodies without central spores. Ultrastructure of the later cutaneous lesions is presented.     

Influence of colchicine on the addition of a sugar to the enamel protein in secretory ameloblasts of cultured germs of rat molar tooth by 3H-galactose radioautography

Summary The influence of colchicine on the addition of ³H-galactose to the enamel protein in secretory amelloblasts of cultured germs of rat molar tooth was investigated by light- and electron-microscopic radioautography. In tooth germs cultured without colchicine, the reaction products of ³H-galactose were observed over Golgi cisternae at early chase times and then localized over the enamel with time. In tooth germs cultured with colchicine, the silver grains were seen over the Golgi cisternae, condensing granules and accumulated secretory granules. Some grains also appeared with time over the pale granular material precipitated in the intercellular space with colchicine treatment. In quantitative analysis with light microscopic radioautography, values of silver grain counts over the unit area (100 m²) on ameloblasts and enamel of colchicine-treated tooth germs were significantly lower at both 0 min and 30 min chase after 30 min pulse than those of control tooth germs, respectively. This finding indicates that colchicine diminished the incorporation of ³H-galactose into the secretory ameloblast of cultured tooth germs. It is suggested that colchicine decreases the activity of the Golgi apparatus with regared to the addition of sugar to the synthesizing glycoprotein in the secretory ameloblast.     

Axonal microtubules necessary for generation of sodium current in squid giant axons: I. pharmacological study on sodium current and restoration of sodium current by microtubule proteins and 260K protein

Summary Effects of the reagents suppressing or supporting axoplasmic microtubule assembly were studied on the Na ionic current of squid giant axons by perfusing the axon internally with the solution containing the reagent. Among the reagents suppressing the assembly, colchicine, vinblastine, podophyllotoxin, sulfhydryl reagents such as DTNB and NEM, and chaotropic anions such as iodide and bromide, were examined. These reagents reduced maximum Na conductance and shifted the voltage dependence of steady-state Na activation in a depolarizing direction along the voltage axis. They also made the voltage dependence less steep, but did not affect sodium inactivation appreciably. Effects on Na ionic current of reagents which support microtubule assembly (Taxol, DMSO, D₂O and temperature) were opposite the effects of those agents suppressing assembly. At the same time, we demonstrated that after Na currents were partially reduced, they could be restored by internally perfusing the axon with a solution containing microtubule proteins, 260K proteins and cAMP under conditions favorable for microtubule assembly. For full restoration, it was found that the following conditions were necessary: (1) The microenvironment within the axon is suitable for microtubule assembly. (2) Tubulins incorporated into microtubules are fully tyrosinated at their C-termini. (3) A peripheral protein having a molecular weight of 260,000 daltons (260K protein) is indispensable. These results suggest that axoplasmic microtubules and 260K proteins in the structure underlying the axolemma play a role in generating Na currents in squid giant axons.