Electrophoresis of mutant proteins in inherited diseases

The electrophoretic approaches for detection of mutant proteins in inherited diseases are briefly reviewed and discussed. Mutation of a protein, known to be associated with a specific inherited disease, is detected by immunoblotting, immunoprecipitation or enzyme staining, combined with various electrophoretic techniques. Some instrumental and technological devices for two-dimensional electrophoresis have been reported for the screening of mutant proteins in diseases of currently unknown etiology.     

Purification and properties of extracellularβ-fructofuranosidases fromAureobasidium sp. ATCC 20524

Summary Two extracellular -fructofuranosidases (E-1 andE-2) fromAureobasidium sp. ATCC 20524, producing 1-kestose (1^F--fructofuranosyl-sucrose) from sucrose, were purified to homogeneity. Molecular weights of the enzymes were estimated to be about 304000 (E-1) and 315000 (E-2) Da by gel filtration. The enzymes contained 33% (w/w) (E-1) and 27% (w/w) (E-2) carbohydrate. TheK _m values for sucrose ofE-1 andE-2 andE-2 were 0.34 and 0.28 M, respectively. were 0.34 and 0.28 M, respectively. The enzymatic profiles of these enzymes were almost identical to intracellular enzymesP-1 andP-2 except for the differences in carbohydrate content andK _m values ofE-2 andP-2.     

Application of enzyme-linked immunosorbent assay for mass examination of strongyloidiasis in okinawa, Japan

The enzyme-linked immunosorbent assay was tested to evaluate whether it could be applicable in screening for mass examination of strongyloidiasis. A total of 2906 inhabitants in three areas (858 in Gushikawa Village, 849 in Nakazato Village and 1199 in Sashiki Town) were screened by the enzymatic assay and approximately 11–30% (11.8% in Gushikawa, 17.0% in Nakazato and 27.7% in Sashiki) were considered to be antibody positive. In the parasitological follow-up examinations of those who were antibody positive, actual infection was found in more than half (51%) the subjects. The overall infection rates estimated from the results reached 5.8% in Gushikawa, 9.1% in Nakazato and 14.0% in Sashiki (mean = 10.4%). The infection rates were significantly higher than those in previous surveys conducted in the same areas. The ELISA technique was found to be useful for strongyloidiasis screening and for seroepidemiological purposes in Okinawa.     

Pulse sequences generated by a degenerate analog neuron model

The response characteristics of an electronic neuron model proposed by the authors are investigated. Periodic stimulating pulse sequences with a fixed frequency are applied to the analog neuron model and the response pulse sequences are studied. In the degenerate case, the state transition of the neuron model during one period of the stimulating pulse sequence is described by a first order piecewise linear difference equation with a jump. It is shown that the periodic response pulse sequences of the neuron model belong to a special class of pulse sequences generated by a simple algorithm, and that the relation between the pulse width (or amplitude) of the stimulating pulse and the firing rate of the neuron model takes the form of an extended Cantor function.     

Pathological studies on encephalitis in mice experimentally inoculated with bovine coronavirus

One- to 21-day-old mice were examined pathologically after inoculated intracerebrally or subcutaneously with the Kakegawa strain of bovine coronavirus. In 1- to 4-day-old mice inoculated intracerebrally, the brain contained a small number of neutrophils and lymphocytes having infiltrated diffusely and perivascularly and some degenerative neurons. In mice inoculated intracerebrally at 7 to 21 days of age, severe necrosis of pyramidal cells was shown in Ammon's horn. Perivascular infiltrations of neutrophils and lymphocytes were moderate to severe. Some neurons were degenerative in the cerebral cortex, thalamus and midbrain. Degeneration of some neurons and mild infiltration of neutrophils and lymphocytes were observed in the brain of mice inoculated subcutaneously at 1 to 7 days of age. Perivascular infiltration of neutrophils and lymphocytes was prominent in the cerebral cortex of mice inoculated subcutaneously at 14 days of age. Cellular infiltration was also seen in the thalamus, Ammon's horn, midbrain, cerebellum and medulla oblongata. All the mice, except one, inoculated subcutaneously at 21 days of age were free from neural changes. Electron-microscopically, virus particles were observed in and outside of the degenerative neurons. They had a core 70 nm in diameter and an envelope with spikes.     

Kinetics of the Unrolling of the Second Leaves Detached from Rice Seedlings by Blue Light

Unrolling due to blue light (B) irradiation of the second leavesdetached from 8-day-old rice (Oryza saliva L.) seedlings wassimilar to that reported previously for nondetached leaves.The effect of B was counteracted by irradiation with red light(R). The counteracting effect of R was reversed by subsequentirradiation with far-red light (FR). When the detached leaf was irradiated with B passed througha 1-mm-wide slit 5, 8, 10, 12 or 15 mm from the leaf tip, irradiation10 mm from the leaf tip was the most effective. The effect of a 1 mm-wide-B irradiation 10 mm from the leaftip was counteracted by a 1 mm-wide-R irradiation at the sameposition, but not by irradiations at the other points. The counteractingeffect of R was reversed by a 1 mm-wide-FR irradiation at thesame position. This suggests that the excitation or the reactionof the B photoreceptor(s) is affected directly by the PFR formof phytochrome. The dose-response curve for the unrolling caused by B showeda simple Bunsen-Roscoe relation without two peaks, which differsfrom that for the phototropism in Avena caused by B. (Received August 21, 1980; Accepted December 20, 1980)     

Light and electron microscopical combined staining by immunohistochemical and enzyme histochemical methods using rat prolactin-secreting pituitary tumours

Summary Combined immunohistochemical staining (IHCS) and enzyme histochemical staining (EHCS) methods for light microscopy (LM) and electron microscopy (EM) are reported, using oestrogeninduced rat pituitary tumours. For LM, combined staining for alkaline phosphatase and acid phosphatase by EHCS, using the azo dye method, and for prolactin and ACTH by IHCS, using the enzyme-labelled antibody method, gave the best results on 1 m glycol methacrylate sections. For EM, combined staining by EHCS on 30 m tissue sections followed by IHCS for prolactin on ultrathin Epon sections (enzyme-labelled antibody method) provided acceptable results. By these combined staining methods, the neoplastic prolactin cells were shown to have close affinity to rich alkaline phosphatase-positive capillaries and to possess an alkaline phosphatase-positive cell membrane. Furthermore, they revealed acid phosphatase-positive lysosomal and secretory granules. These combined staining methods may be valuable in studies on the actual functional status of cells.     

Recessive and dominant genes controlling inducible sexual agglutinability in Saccharomyces cerevisiae

Summary We have characterized the two dominant genes, IND 1 and IND 2, responsible for inducible sexual agglutinability. The strains carrying these genes differ from the inducible strains carrying the recessive gene, saa 1 in the following points. The former strains produce agglutination substance at 22°, 28°, and 37° C only in response to sex pheromone of the opposite mating type, but the latter strains produce the substance constitutively without the pheromone at 22° C, only in response to the pheromone at 28° C, and do not produce the substance, even in the presence of the pheromone, at 37° C.We suggest that strains carrying one of the dominant, inducible genes are wild type and have a pheromone-controlled regulatory system of sexual agglutinability.     

Effect of modification on physicochemical and biological properties of haptoglobin. VI. Reaction with azlactone of p-nitrobenzoyl-valine.

The azlactone of p-nitrobenzoyl-valine (Nbz-Val) has been used for modification of xi-amino groups of lysine in haptoglobin type 1-1, in hemoglobin, and in the haptoglobin-hemoglobin complex. By the use of this reagent 95% of amino groups in haptoglobin and 90% in hemoglobin have been blocked without any changes in peroxidase activity of the formed complexes: Nbz-Val.haptoglobin with hemoglobin, Nbz-Val. hemoglobin with haptoglobin, and Nbz-Val.(haptoglonin-hemoglobin). After reduction and reoxidation, Nbz-Val.haptoglobin was found to retain 90% of peroxidase activity when complexed with hemoglobin. Beta chains separated either from haptoglobin or Nbz-Val.haptoglobin showed 15% of peroxidase activity in the complex with hemoglobin, alpha chains of the same origin were completely inactive. Whereas recombination of haptoglobin from alpha and beta chains resulted in 42% hemoglobin-binding capacity, renaturation of Nbz-Val.haptoglobin from separated subunits was found to proceed with almost 100% yield. In immunodiffusion with rabbit anti-haptoglobin or anti-Nbz-Val.haptoglobin sera, preparations of haptoglobin and Nbz-Val.haptoglobin after reduction and reoxidation or after recombination from separated subunits gave similar precipitation arcs showing the reaction of immunological identity.     

High-dose zolpidem abuse in a patient with insomnia comorbid with major depressive disorder

Sleep and Biological Rhythms - A 39-year-old married woman was referred to a psychiatry outpatient clinic for treatment of major depressive disorder (MDD). The dose of zolpidem she had been taking...