Use of Congo red as a microscopic fluorescence indicator of hyphal growth

Congo red was found to be feasible as a microscopic fluorescence indicator of hyphal growth at the single-hypha level. When 1 m Congo red was applied to mold of Aspergillus niger, the dye was found to a specific cell-wall component, chitin, without causing any inhibitory effect on hyphal growth. The bound Congo red emitted fluorescence at 614 nm. This binding reaction, however, proceeded more slowly than the growing speed of hypha. Consequently the fluorescence intensity was low at the apex where the surface area of the hypha was expanding rapidly. In contrast, as an apex where the growth was retarded, the fluorescence intensity became remarkably high. Therefore growing hyphae could be distinguished from non-growing hyphae by using Congo red.     

A missense mutation (G197→A) in theα-l-fucosidase gene of fucosidosis patients leads to loss ofα-l-fucosidase

Fucosidosis is an autosomal recessive lysosomal storage disease resulting from the absence of -l-fucosidase activity. Two natural missense mutations (G¹⁹⁷A) and (A⁸⁶⁰G) within the -l-fucosidase gene have been reported to be homozygous in four patients with fucosidosis. Expression of wild-type and mutated -l-fucosidase cDNAs in COS-1 cells revealed complete deficiency of -l-fucosidase for the G¹⁹⁷A transition and a normal level of enzyme for A⁸⁶⁰G. We therefore conclude that the change of G¹⁹⁷A is responsible for fucosidosis in the patients while A⁸⁶⁰G is a normal polymorphic variant of -l-fucosidase.     

Immunohistochemical localization of nerve growth factor and epidermal growth factor in guinea pig prostate gland

Prostate glands of adult guinea pigs were stained for nerve growth factor (NGF) and epidermal growth factor (EGF) by immunohistochemical methods. Both NGF and EGF were localized diffusely in the cytoplasm of the glandular epithelial cells, and also in their secretory products. These findings suggest that NGF and EGF are synthesized, stored, and secreted by the glandular epithelial cells of the prostate.     

High-performance liquid chromatographic determination of hippuric acid in human urine

A method is described for the determination of urinary hippuric acid by high-performance liquid chromatography. The method used ethyl acetate extraction for partial clean-up of the urine. The separation was carried out on a reversed-phase column using 20% methanol in 0.01 M aqueous potassium phosphate containing 0.5% acetic acid as a mobile phase. The column effluent was monitored with a UV detector at 254 nm. Hippuric acid was separated from other normal urine constituents in less than 10 min. Metabolites of xylene and styrene did not interfere with the assay. Analytical recoveries from urine were excellent and peak height and concentration were linearly related.     

Effect of sulpiride on serum growth hormone and prolactin concentrations following L-DOPA administration in man.

The effect of chronic administration of sulpiride on serum human growth hormone (hGH), prolactin and thyroid stimulating hormone (TSH) was examined in 6 normal subjects. Sulpiride was given orally at a dose of 300 mg (t.i.d.) for 30 days. Sulpiride raised serum prolactin levels in all subjects examined. In addition, sulpiride suppressed hGH release induced by L-dopa, although the basal hGH level was not changed. Sulpiride treatment appeared to antagonize partially the inhibitory effect of L-dopa on prolactin release. Following thyrotropin-releasing hormone (TRH) injection, the percent increment in prolactin levels from the baseline in sulpiride-treated subjects was less than in controls without sulpiride. In contrast, both the basal and TRH-stimulated TSH levels were not influenced by sulpiride. These observations suggest that sulpiride suppresses L-dopa-induced hGH release and stimulates prolactin release, presumably by acting against the dopaminergic mechanism either on the hypothalamus or on the pituitary. The decreased prolactin response to TRH after sulpiride treatment may indicate a diminished reserve capacity in pituitary prolactin release.     

A nuclear extract, prepared from mass-isolated germinal vesicles, retains a factor able to sustain a cytoplasmic cycle of starfish oocytes.

The germinal vesicle (GV) of starfish oocytes contains a factor which is required to drive the cytoplasmic cycle of the meiotic division. Biochemical investigation of this factor has been difficult due to the small quantities of obtainable GV materials. To overcome this, we have developed a mass-isolation procedure for the GVs of starfish oocytes, which depends on the softening of the cortex of the oocytes by cytochalasins to enable the GVs to pass through the cortex by centrifugation. From the isolated GVs, we have prepared a soluble fraction which retains the activity to induce the cytoplasmic cycle in the meiotic division of oocytes. The factor was sensitive to both heat and papain, suggesting that it is a protein.     

Influence of livestock grazing on vegetation in a saline area in Northeast Thailand

The influence of grazing by water buffalo (Bubalas bufalis) and cattle (Bos taurus) was estimated for vegetation inside and outside cages in a saline area at Khon Kaen, Northeast Thailand. The home range of water buffalo and cattle shifted in response to the period of rice cultivation: during the rice-growing season these animals grazed on roadsides and abandoned places such as the study area; after rice harvest they grazed mainly rice stubble on the paddy. The vegetation in the study area was divided into three types: 1) dominated by the annual grassesChloris barbata andIschaemum rugosum; vegetational cover and plant height in the cage increase due to the increase of these grasses; 2) thorny shrub patch ofMaytenus mekongensis; other species in this patch almost died a year after experimental elimination of this shrub; this salt-tolerant shrub not only protected the co-existing species from grazing, but also suppressed salt accumulation; 3) almost pure stand ofPanicum repens; livestock preferred this perennial grass over others. Electrical conductivity (EC) of surface soil did not increase under the vegetation protected from grazing. Litter and other organic matter in the soil suppressed the upward movement of brine. Grazing led to an increase of bare ground where NaCl accumulated, and modified the heterogeneity of vegetation, which was reflected in the degree of salt accumulation.     

A possible clinical application of multicytokine-producing cytotoxic mononuclear cell (MCCM) therapy

When peripheral blood mononuclear cells (PBMC) were incubated with a streptococcal preparation, OK-432, for 24 h, PBMC acquired cytolytic activity against cultured and fresh human tumor cells. Such PBMC were called OK-432-activated mononuclear cells (OK-MC). OK-MC produce several kinds of cytokines such as interferon (IFN), IFN, and tumor growth inhibitory factor (TGIF) bothin vitro andin vivo. OK-MC-produced cytokines also inhibited the growth of cultured and fresh human tumor cells. The growth inhibition was examined by human tumor clonogenic assay using a double-layer agar technique. The results indicate that two pathways of anti-tumor activity are induced in OK-MC, i.e., cell-mediated and cytokine-mediated.     

Activation of the dioxin and glucocorticoid receptors to a DNA binding state under cell-free conditions

The activation in vitro of dioxin and glucocorticoid receptors from a non-DNA binding to a DNA binding state was characterized. Ligand-free dioxin and glucocorticoid receptors were partially co-purified from rat liver cytosol, and both receptors sedimented at 9 S following labeling with the respective ligand. The 9 S forms of the dioxin and glucocorticoid receptors have previously been shown to represent heteromeric complexes containing the Mr approximately equal to 90,000 heat shock protein. The 9 S ligand-free or ligand-bound glucocorticoid receptor was converted to the monomeric 4-5 S form upon exposure to 0.4 M NaCl even in the presence of the stabilizing agent molybdate. Under identical conditions, the 9 S ligand-free and ligand-bound dioxin receptor forms remained essentially intact. However, in the absence of molybdate, the dioxin receptor could be converted to a 4-5 S form upon exposure to high concentrations of salt. These results indicate that the glucocorticoid receptor readily dissociates from the 9 S to the 4-5 S form even in the absence of hormone, whereas both the ligand-free and ligand-occupied 9 S dioxin receptor forms represent more stable species. Gel mobility shift experiments revealed that the 4-5 S glucocorticoid receptor interacted with a glucocorticoid response element both in the absence and presence of ligand. On the other hand, occupation of the dioxin receptor by ligand greatly enhanced the ability of the receptor to be activated to a form that binds to its target enhancer element. Once dissociated, the monomeric form of the dioxin receptor was also able to interact with its DNA target sequences even in the absence of ligand. Thus, ligand binding efficiently facilitates subunit dissociation of the dioxin receptor but is not a prerequisite for DNA binding per se. Given the apparent stability of its non-DNA binding 9 S form, the dioxin receptor system might be a useful model for the investigation of the mechanism of activation of soluble receptor proteins.