Neuropathological analysis in spinal muscular atrophy type II.

We performed a neuropathological analysis, including in situ nick end labeling (ISEL) and immunohistochemistry, of two cases of clinicogenetically confirmed infantile spinal muscular atrophy (SMA) type II. Both cases showed severe reduction of the motor neurons and gliosis in the spinal cord and brain stem, although the occurrences of central chromatolysis and ballooned neurons were not frequent. Clark's and lateral thalamic nuclei, which are usually altered in SMA type I, were spared, whereas Betz cells in the precentral gyrus and large myelinated fibers in the lateral funiculus were reduced in number. Regarding apoptosis, only the younger case demonstrated a few ISEL-positive nuclei in the dorsal horn, with reduced Bcl-x expression level in the Purkinje cells. Unlike SMA type I, the expression of neurofilaments was not disturbed and the reduction in synaptophysin expression level in the anterior horn was mild. An oxidative stress-related product was deposited in atrophic motor neurons in the spinal cord, and neurons with nuclei immunoreactive for 8-hydroxy-2'-deoxyguanosine were found in the lateral thalamus. In contrast, the expression of glial glutamate transporters was not altered. These data suggest that oxidative stress and, to a lesser extent, apoptotic cell death, but not disturbed neurofilament metabolism or excitotoxicity, may be involved in neurodegeneration in SMA type II.

Effects of repeated stress on expression of interleukin-6 (IL-6) and IL-6 receptor mRNAs in rat hypothalamus and midbrain.

We examined the effects of single and repeated stress on the expression of interleukin-6 (IL-6) and IL-6 receptor (IL-6R) mRNAs in the rat midbrain and hypothalamus using reverse transcriptase-polymerase chain reaction (RT-PCR). Following a single episode of restraint stress for 4 hours (1R) or 4 hours per day on two (2R) or three (3R) consecutive days, the hypothalamus and midbrain were removed immediately and the levels of IL-6 and IL-6R mRNAs in both regions were determined. Regional differences in stress-related changes in mRNA levels were noted. The expression of IL-6 mRNA in the hypothalamus did not change in 1R group but decreased in 2R and 3R groups. The expression of IL-6R mRNA in the same region significantly diminished in all groups. In the midbrain, the expression of IL-6 mRNA increased in 1R group and decreased in 2R and 3R, while the expression of IL-6R mRNA significantly diminished in 1R and 3R groups but was not different from control in 2R group. Our findings indicate that repeated stress in rats produce changes in IL-6 and IL-6R mRNAs in the midbrain and hypothalamus that are different than those of a single stress episode.

Cbfa1 is a positive regulatory factor in chondrocyte maturation.

Cbfa1 is a transcription factor that belongs to the runt domain gene family. Cbfa1-deficient mice showed a complete lack of bone formation due to the maturational arrest of osteoblasts, demonstrating that Cbfa1 is an essential factor for osteoblast differentiation. Further, chondrocyte maturation was severely disturbed in Cbfa1-deficient mice. In this study, we examined the possibility that Cbfa1 is also involved in the regulation of chondrocyte differentiation. mRNAs for both Cbfa1 isotypes, type I Cbfa1 (Pebp2alphaA/Cbfa1) and type II Cbfa1 (Osf2/Cbfa1 or til-1), which are different in N-terminal domain, were expressed in terminal hypertrophic chondrocytes as well as osteoblasts. In addition, mRNA for type I Cbfa1 was expressed in other hypertrophic chondrocytes and prehypertrophic chondropcytes. In a chondrogenic cell line, ATDC5, the expression of type I Cbfa1 was elevated prior to differentiation to the hypertrophic phenotype, which is characterized by type X collagen expression. Treatment with antisense oligonucleotides for type I Cbfa1 severely reduced type X collagen expression in ATDC5 cells. Retrovirally forced expression of either type I or type II Cbfa1 in chick immature chondrocytes induced type X collagen and MMP13 expression, alkaline phosphatase activity, and extensive cartilage-matrix mineralization. These results indicate that Cbfa1 is an important regulatory factor in chondrocyte maturation.

Effects of restraint stress on alpha(1) adrenoceptor mRNA expression in the hypothalamus and midbrain of the rat.

We examined the effects of restraint stress on alpha(1) adrenoceptor mRNA expression in the rat brain using reverse transcriptase-polymerase chain reaction (RT-PCR). After rats had been restrained for 10, 30, 60, 120 or 240 min, the hypothalamus and midbrain were removed immediately and alpha(1) adrenoceptor mRNA levels in these regions were determined by RT-PCR. Blood samples were also collected for simultaneous measurement of serum adrenocorticotropic hormone (ACTH) and corticosterone. Restraint stress resulted in a variety of changes in the hypothalamus and midbrain. In the hypothalamus, 30 and 60 min of stress resulted in a significant fall in the level of alpha(1) adrenoceptor mRNA relative to the control. This was associated with a rise in serum ACTH and corticosterone. In the midbrain, significant elevation of alpha(1) adrenoceptor mRNA was noted after 60, 120 and 240 min of restraint stress. Our findings indicated that the influence of restraint stress on alpha(1) adrenoceptor mRNA level in the hypothalamus is different to that of the midbrain region in rats.

Effects of single and repeated stresses on the expression of mRNA for alpha1-adrenoceptors in the rat hypothalamus and midbrain.

We examined the effects of single or repeated stress on the expression of mRNA for alpha1-adrenoceptors in the rat hypothalamus and midbrain using the reverse transcriptase-polymerase chain reaction (RT-PCR). Single stress significantly increased the mRNA level for alpha1-adrenoceptors in the midbrain, but had no effect on mRNA levels in the hypothalamus. Repeated stress significantly decreased mRNA levels for alpha1-adrenoceptors in both regions.

Effects of a long-term inhalation of fragrances on the stress-induced immunosuppression in mice.

The aim of this study was to determine the effects of the long-term application of various fragrances on the suppression of immune response induced by high-pressure stress in mice. The immune response was analyzed based on plaque-forming cell (PFC) count, using mice sensitized with sheep red blood cells. The decreased PFC involving thymic involution induced by high-pressure stress in mice was restored by exposing the stressed mice to tuberose, lemon, oakmoss and labdanum for 24 h following exposure to stress. The decreased PFC and thymic involution from stress were restored by exposure to lemon and oakmoss, but not to tuberose and labdanum when the mice were exposed to those fragrances continuously for 3 weeks before the stress was given, followed by exposure to the same fragrances for 24 h after the stress. The decreased PFC and thymic involution from stress were restored by exposure to lemon and labdanum for 24 h after the stress, but not to tuberose over 3 weeks before the stress was given. These data suggest that the neuroimmunomodulatory effects of fragrances may be affected by tolerance depending on the kinds of fragrances in the case of a long-term application.

The expressions of mRNAs for interleukin-6 (IL-6) and the IL-6 receptor (IL-6R) in the rat hypothalamus and midbrain during restraint stress.

Over the past few years, it has been reported that physical and psychological stress elevate plasma interleukin-6 (IL-6), and that neural cells can produce IL-6 and have receptors for IL-6 (IL-6R). However, it is unknown whether IL-6 plays a role in regulating the functions of neural cells in response to stress. We demonstrated recently, using the reverse transcriptase-polymerase chain reaction (RT-PCR), that the levels of mRNAs for IL-6 and IL-6R in the rat brain are changed by restraint stress for four hours. In the present study, we investigated the expression of mRNAs for IL-6 and the IL-6R in the rat hypothalamus and midbrain during restraint stress. After rats had been restrained for 10, 30, 60, 120 or 240 min, the hypothalamus and midbrain were removed immediately and levels of IL-6 mRNA and of IL-6R mRNA in these regions were determined by RT-PCR. The expression of mRNAs for IL-6 and IL-6R in both regions was reduced after short-term (30-60 min) restraint stress and tended to return toward the control level after 120 min restraint stress. After long-term (240 min) restraint stress, the level of IL-6 mRNA was significantly increased in the midbrain, while the level of IL-6R mRNA was significantly reduced in both regions. These findings suggest that the need for IL-6 might decline after short-term restraint stress and, moreover, that the synthesis and secretion of IL-6 might be enhanced and IL-6 might be needed as a neurotrophic factor in the midbrain after long-term stress.

The influence of restraint stress on the expression of mRNAs for IL-6 and the IL-6 receptor in the hypothalamus and midbrain of the rat.

Using the reverse transcriptase-polymerase chain reaction (RT-PCR), we investigated the influence of restraint stress on the expression of the mRNA for interleukin-6 (IL-6) and the mRNA for the IL-6 receptor (IL-6R) in the rat brain. After rats had been restrained for 4 hours, the hypothalamus and midbrain were removed at fixed intervals up to 24 hours, and levels of IL-6 mRNA and of IL-6R mRNA in these regions were determined by RT-PCR. Restraint stress significantly enhanced the expression of IL-6 mRNA and reduced that of IL-6R mRNA in the midbrain, whereas the stress caused the reduced expression of IL-6R mRNA without any change in the level of IL-6 mRNA in the hypothalamus. After the stress, the expression of mRNAs for IL-6 and IL-6R continued to diminish in both regions. These findings indicate that the levels of mRNAs for both of IL-6 and IL-6R in the rat brain can be influenced by restraint stress.

Case reports of epulis treated by CO2 laser without anesthesia.

This study reports seven cases of epulis excised by the CO2 laser without anesthesia. The laser was used in a continuous wave mode at power setting of 2-4 W. All lesions were excised completely, and no patients felt pain during laser irradiation. The operation was successful without using surgical packs, and there was no postoperative discomfort. There has been no evidence of recurrence after operation.

An unclassified cerebral small cell tumor in a patient with human T-cell lymphotropic virus type 1-induced primary extranodal lymphoma.

We describe an autopsy case of a thalamic tumor in a patient with human T-cell lymphotropic virus type 1 (HTLV-1)-induced extranodal lymphoma of the skull. Neither brain tumors associated with adult T-cell leukemia nor HTLV-1-induced extranodal lymphoma of bone have previously been reported. The tumor, which resembled an oligodendroglioma or dysembryoplastic neuroepithelial tumor, was composed of medium-sized cells with dark, round-to-polygonal nuclei and a mucinous matrix that formed pericellular lakes. Tumor cells were immunoreactive with S-100 protein and neuron-specific enolase but not with glial fibrillary acid protein, synaptophysin, or vimentin. Tumor cells had prominent rough endoplasmic reticulum and free ribosomes with short processes, compatible with an oligodendroglial or small neuronal nature. Many atypical lymphocytes had infiltrated the leptomeningeal space, subependymal zone along the ventricular walls, fornices, corpus callosum, and right frontal lobe. Multinucleated or bizarre astrocytes and macrophages were found exclusively in the right frontal white matter. The coexistence of this unclassified peculiar brain tumor with bizarre astrocytosis suggests a cytopathic effect of HTLV-1 on human glioneuronal cells.

Effects of citrus fragrance on immune function and depressive states.

In our previous experiments on animals evidence was found that citrus fragrance can restore the stress-induced immunosuppression, suggesting that citrus fragrance may have an effect on restoring the homeostatic balance. Since a dysregulation of the neuroendocrine and immune function is thought to be associated with psychosomatic or psychiatric disorders an attempt was made to restore their mental health by stimulation of one of the sensory systems. Fragrance (citrus was our choice) which comforts through stimulation of the olfactory system was applied to depressive patients. It was given to 12 depressive subjects and the results indicated that the doses of antidepressants necessary for the treatment of depression could be markedly reduced. The treatment with citrus fragrance normalized neuroendocrine hormone levels and immune function and was rather more effective than antidepressants.

Epstein-Barr virus early antigen induction in nasopharyngeal hybrid cells by Chinese medicinal herbs.

The effects of Chinese medicinal herbal drugs (CMH; Daphne genkwa, Wikstroemia indica, Croton oil) were studied for Epstein-Barr virus (EBV) early antigen (EA) induction in established nasopharyngeal hybrid cells. Both ether and water extracts of CMH were found to have inducing activity. However water extracts of the same herbs were not as strong as that of other extracts. The EA positive-cell rate was from 18.2 to 42.2% in ether extracts and 1.0 to 3.8% in water extracts at 10 microgram/ml of the concentration. N-Butyrate alone showed a 40.2% positive rate and in the both treatment of water extracts, a combination effects was seen in induction of the EBV-EA. This in vitro system for the induction of EBV-EA was thought to be useful to determine what is the causal factors for activation of EBV in vivo.

Studies on a new antibiotic FR-900336 taxonomy, isolation and characterization.

A new lipophilic antibiotic, FR-900336 was isolated from a culture of Streptomyces sioyaensis subsp. tanegashimaensis . FR-900336 is light yellow and has a molecular formula C30H30NO13C1 . The characterization by IR, UV, 1H NMR and 13C NMR spectra makes a quinone structure very probable. FR-900336 is active against Gram-positive bacteria and fungi.

Studies on a new antibiotic FR-900109. 1. Taxonomy, isolation and characterization.

FR-900109 is a new antibiotic obtained from fermentation broth of a streptomyces which was identified as Streptomyces prunicolor. Its elementary analysis and mass spectroscopic measurement suggest that the molecular formula is C27H32O9. It has an ultraviolet absorption maximum at 254 nm. The antibiotic is active against Gram-positive bacteria. Acute toxicity in mice is very low.

Confirmation of new, high-mass saponins from Gleditsia japonica by field desorption mass spectrometry.

The molecular weights and complete sugar sequences have been determined in saponins carrying 3 to 8 free sugars. The results are in agreement with parallel chemical and instrumental analyses of the triterpenoid saponins derived from Gleditsia japonica. The capacity and limitations of field desorption mass spectrometry in structural elucidation of substituted oligoglycosides are described.