Three new species ofBullera isolated from leaves in the Ogasawara Islands

Thirteen strains of ballistoconidium-forming yeasts were isolated from leaves collected in the Ogasawara Islands, Japan. They represent three different species in the genusBullera on the basis of morphological, physiological, and biochemical characteristics, analyses of the sequences of internal transcribed spacer regions and small subunit ribosomal DNA, and a nuclear DNA-DNA hybridization study. Three new species,Bullera boninensis (five strains),B. waltii (seven strains), andB. schimicola (one strain), are proposed for these 13 strains.     

Identification of Ca2+-Activated Neutral Protease in the Peripheral Nerve and Its Effects on Neurofilament Degeneration

Rat sciatic nerve segments were incubated in five different media. Disappearance of neurofilament (NF) triplet proteins (200K, 160K, and 68K MW) occurred in medium containing Ca²⁺ and was inhibited by the addition of E-64-c or leupeptin. Therefore, the presence in the peripheral nerve of an enzyme whose properties are similar to those of Ca²⁺-activated neutral protease (CANP) is suggested. The extraction of crude CANP from rat sciatic nerve was performed. CANP activity was completely recovered (0.129 ± 0.008 U/g) in the precipitate salted out by the addition of 0 to 50% saturated ammonium sulfate to the soluble fraction of the peripheral nerve (crude CANP). Properties of the crude CANP were examined using NF as a substrate and were found to be similar to those of the CANP extracted from skeletal muscle. Identification of the crude CANP with the CANP extracted from rat skeletal muscle was performed using the immunoreplica method. Bands corresponding to 73K were detected in both CANPs.     

Oxytocin in the cerebrospinal fluid and plasma of pregnant and nonpregnant subjects

The oxytocin concentration in the cerebrospinal fluid (CSF) and plasma of pregnant women at term with and without labor pain were measured by radioimmunoassay and compared with those of non-pregnant women of matched age. The oxytocin concentrations in the CSF were 4.9 +/- 4.1 microU/ml (mean +/- S.D.) in pregnant women with labor pain, 4.1 +/- 2.4 microU/ml in those without labor pain and 4.0 +/- 2.8 microU/ml in nonpregnant women, and the oxytocin concentrations in the plasma of these subjects were 45.2 +/- 19.6, 17.1 +/- 22.2 and 7.0 +/- 5.3 microU/ml, respectively. Thus the oxytocin level in the CSF did not change appreciably even when the level in the plasma was raised in the pregnant women with labor pain. These findings suggest that oxytocin does not penetrate the blood-brain barrier, and that oxytocin in the CFS has little or no central role in parturition in women.     

Histochemical study of the postnatal development of autonomic nerves in the mouse iris,using a whole-mount preparation method

Summary Postnatal development of autonomic nerves in the mouse iris was studied histochemically from one day to five months of age. For the demonstration of aminergic nerves the glyoxylic acid method was used, while for cholinergic nerves, Karnovsky and Roots' method was utilized on the whole-mount preparations of irises. The results obtained were as follows. In one-day-old mice, many aminergic nerves could already be seen, while cholinergic nerves were scarcely observable. Both types of nerve increased rapidly in the first 2 weeks. Both completed development between 3 and 4 weeks, although aminergic nerves were observed to develop earlier than the cholinergic.     

Heterogeneity of the minimum functional unit of hemocyanins from the spider (Argiope bruennichii), the scorpion (Heterometrus sp.), and the horseshoe crab (Tachypleus tridentatus).

The heterogeneity of arthropod hemocyanins was studied by polyacrylamide gel electrophoresis and immunochemical techniques. The spider (Argiope bruennichii), the scorpion (Heterometrus sp.), and the horseshoe crab (Tachypleus tridentatus) were found to have 4, 5, and 5 minimum functional units of hemocyanin, respectively, the apparent molecular weights of which were 79,000, 81000, and 80,000, respectively, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.     

Immunohistochemical detection of tyrosine hydroxylase and concentrations of monoamines in the substantia nigra and hypothalamus of hereditary microphthalmic rats.

We compared tyrosine hydroxylase immunoreactivity in the substantia nigra and hypothalamus of hereditary microphthalmic rats with that of normal rats. A considerable number of neuronal cell bodies expressing tyrosine hydroxylase were present in the substantia nigra of the microphthalmic mutant as well as normal rats. Neuronal cells positive for tyrosine hydroxylase in the hypothalamus were fewer than in the substantia nigra in both rats. The concentrations of monoamines (dopamine, noradrenaline, adrenaline, and serotonin) in the substantia nigra and hypothalamus in the microphthalmic mutant were approximately the same as those of normal rats, although the diurnal fluctuation of a few monoamines was observed in normal rats. These results suggest that the metabolic aspects of catecholamine in the substantia nigra and hypothalamus of the microphthalmic mutant rat do not markedly differ from those of normal rats.     

In Vitro Effects of Immunosuppressive Agents on Cytokine Production by HTLV-I-Infected T Cell Clones Derived from the Ocular Fluid of Patients with HTLV-I Uveitis

The present study was designed to investigate the in vitro effects of potential therapeutic agents on cytokine production by five HTVL-I-infected T cell clones (TCC) established from the ocular fluid of patients with HTLV-I uveitis. Each of the five HTLV-I-infected TCC was cultured at 1 × 10⁶ cells/ml with or without an immunosuppressive agent (hydrocortisone, FK506, rapamycin, indomethacin, or prostaglandin E₂) for 22 hr in humidified 5% CO₂ in air at 37 C. The production of various cytokines in the culture supernatant from each TCC was measured by ELISA. The HTLV-I-infected TCC produced high amounts of IL-1α, IL-3, IL-6, IL-8, TNF-α, IFN-γ, and GM-CSF, and low but significant levels of IL-2 and IL-10 without any stimuli. Hydrocortisone severely depressed the production by these TCC of all the cytokines except for IL-2, which was slightly increased. Prostaglandin E₂ depressed the production of IL-1α, while it up-regulated the production of IL-6, TNF-α, and IFN-γ. Rapamycin depressed the production of IL-6 and TNF-α, and FK506 depressed the production of TNF-α. Hydrocortisone also severely depressed the cytokine production by PHA-stimulated peripheral blood mononuclear cells obtained from healthy volunteers. Of the immunosuppressive agents tested, hydrocortisone exhibited the strongest suppression of cytokine production by HTLV-I-infected TCC. This result was in agreement with the in vivo effects of hydrocortisone in patients with HTLV-I uveitis. These TCC will be useful in investigating the effects of potential therapeutic agents for HTLV-I uveitis in vitro.     

Solubilization of active complement receptor type 1 (CR1) from human erythrocyte stroma with trypsin and its purification

Mild trypsinization of human erythrocyte stroma solubilized CR1 (complement receptor type 1, C3b/C4b receptor) without significant loss of decay-accelerating activity to C5 convertases on hemolytic intermediate cells (EAC 1-3b, P). The solubilized CR1 was purified using DEAE-Sephacel, C3-Sepharose, and anti-CR1-Sepharose column chromatographies. The purified material showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under non-reducing conditions, and its molecular weight was determined to be 175K, about 20K smaller than native CR1. Because the purified sample was separated into the several segments by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, the molecule is considered to be nicked and those segments are associated by disulfide bonds. These results mean that a large portion of the CR1 molecule is present outside of the plasma membrane of erythrocytes, and the intramembranous and cytoplasmic domains are not necessary for decay-accelerating activity.