Increased expression of hippocampal cholinergic neurostimulating peptide-related components and their messenger RNAs in the hippocampus of aged senescence-accelerated mice.

Hippocampal cholinergic neurostimulating peptide stimulates cholinergic phenotype development by inducing choline acetyltransferase in the rat medial septal nucleus in vitro. Adult senescence-accelerated-prone mice/8, a substrain of the senescence-accelerated-prone mouse, show a remarkable age-accelerated deterioration in learning and memory. We cloned mouse hippocampal cholinergic neurostimulating peptide precursor protein complementary DNA. The deduced amino acid sequence showed that the neurostimulating peptide itself is the same as that found in the rat. In situ hybridization revealed that the highest expression of the precursor protein messenger RNA was in hippocampal pyramidal neurons. Compared with a strain of senescence-accelerated-resistant mouse (control mouse), adult senescence-accelerated-prone mice/8 showed increased expression of both the precursor messenger RNA and the neurostimulating peptide-related immunodeposits in the hippocampal CA1 field. The deposits were intensely and diffusely precipitated in neuropils throughout the strata oriens and radiatum in senescence-accelerated-prone mice/8, but not in control mice. The neurostimulating peptide content in the hippocampus was higher in senescence-accelerated-prone mice/8 than in control mice, while its precursor protein itself was not different between the two strains. Furthermore, our previous and present data show that the medial septal and hippocampal choline acetyltransferase activity was significantly lower in senescence-accelerated-prone mice/8 than in control mice. The data suggest that, in hippocampal neurons in adult senescence-accelerated-prone mice/8, the production of hippocampal cholinergic neurostimulating peptide precursor protein in neuronal somata, which is associated with an increased expression of its messenger RNA in the CA1 field, occurs as a consequence of low activity in their presynaptic cholinergic neurons. This is followed by accelerated processing to generate bioactive peptide and transport to its functional fields. However, certain mechanisms reduce the release of the peptide and lead to its accumulation in the neuropil. These disturbances of the septohippocampal cholinergic system might be the biochemical mechanism underlying the characteristic deterioration of senescence-accelerated-prone mice/8.

Demonstration of deacetylated hippocampal cholinergic neurostimulating peptide and its precursor protein in rat tissues.

This report concerns the demonstration of hippocampal cholinergic neurostimulating peptide (HCNP), its deacetylated analogue (free HCNP) and HCNP precursor protein in rat tissues. To avoid possible enzymatic degradation during sample manipulation, tissue extracts were prepared under acidic conditions using trifluoroacetic acid. The tissue contents of free HCNP and of precursor protein were determined by radioimmuno-assay (RIA) using two antibodies with different specificities, and by a combination of HPLC and RIA. Free HCNP was detected in neuronal and renal tissues, but not in liver. All tissues examined had measurable amounts of HCNP precursor protein. The concentrations of free HCNP and precursor in neuronal tissues were inversely related to the age. These results suggest that the deacetylated analogue of HCNP and its precursor protein may have significant physiological functions, especially in the central nervous system of young animals.

NMDA receptor activation enhances the release of a cholinergic differentiation peptide (HCNP) from hippocampal neurons in vitro.

Hippocampal cholinergic neurostimulating peptide (HCNP) is a novel undecapeptide purified from the hippocampus of young rats. The peptide stimulates cholinergic phenotype development in the rat medial septal nucleus in vitro. Here, we have focused on the mechanism of release of the peptide from the hippocampus, by applying tissue culture techniques. Quantitation of HCNP in the culture supernatant after chemical stimulation was carried out by RIA, and by a combination of HPLC and RIA. We found that the N-methyl-D-aspartate (NMDA) receptor specifically mediates release of the deacetylated form of HCNP from the culture. Our results suggest that during the early development of hippocampal neurons, the peptide is released by NMDA receptor activation, and that it may be involved in mediating the effect of activity-dependent cues on developing septal cholinergic neurons.

Demonstration of the biological activity of peptide fragments related to human and rat hippocampal cholinergic neurostimulating peptide (HCNP).

Human and rat hippocampal cholinergic neurostimulating peptides (HCNPs) are 54.5% homologous; both stimulate acetylcholine synthesis in rat medial septal nuclei cultures. This in vitro system was used to test the bioactivity of short peptides containing human or rat HCNP sequences. Peptides with sequences corresponding to the N-termini and middle regions of both, and to the shared three C-terminal residues were not active. Tetrapeptides and hexapeptides whose C-terminus is this common sequence enhanced acetylcholine production, indicating that the minimum consensus sequence for HCNP activity is X-Gly-Pro-Leu.

High levels of hippocampal cholinergic neurostimulating peptide (HCNP) in the CSF of some patients with Alzheimer's disease.

Hippocampal cholinergic neurostimulating peptide (HCNP), originally purified from the hippocampus of young rats, enhances the cholinergic development of rat medial septal nuclei in vitro. This report concerns the determination of the HCNP content of the cerebrospinal fluid (CSF) of 173 clinically, and of 22 clinico-pathologically defined patients. A radioimmunoassay was used throughout. The HCNP level was relatively uniform among the clinically defined patients; for almost all non-Alzheimer's patients, the level fell within the range delimited by +/- 2 SD of the mean for all patients taken together, and none of them had a level above this range. By contrast, the early-onset Alzheimer's disease patients could be divided on the basis of their HCNP level into two groups, one with high levels (markedly above the mean +/- 2SD range), and the other with levels similar to those of the other patients. The analysis of the CSF samples obtained postmortem revealed that Group I Alzheimer-type dementia (ATD) patients with clinico-pathologically established diagnoses had a strikingly higher level of HCNP than patients with either Group II ATD or cerebral vascular disease. These results suggest that HCNP is involved in certain pathophysiological alterations associated with dementia, and that its determination may be useful in patient evaluation. Copyright 1998 Lippincott Williams & Wilkins

[Effects of buprenorphine on gastrointestinal interdigestive myo-electric complex in dogs].

The effect of buprenorphine on the interdigestive myoelectric complex (IMC) was studied by gastrointestinal electromyogram in 3 mature mongrel dogs. The electromyograms were used to study the IMC cycle and total elapsed time (TET). Buprenorphine had very strong hyperkinetic action on the gastrointestinal tract at doses of 1 and 2 micrograms/kg which revealed phasic contraction, but since this action disappeared when atropine was administered. The phasic contraction originated in the upper jejunum just under the ligament of Treitz and then showed a migration pattern of J-->D and J-->I in 11 out of 23 cases (47.8%), D-->J-->I in nine cases (39.1%), J-->D-->J-->I in one case and no response in two cases. TET (min) showed a mean value of 90.3 +/- 11.2 (n = 23), which was not significantly different from the controls (106.1 +/- 14.3) (n = 24), but tended to be longer (0.1 > p > 0.05). After administration of buprenorphine at a dose of 20 micrograms/kg, phasic contraction migrated to ileum, which was disappeared an interval of 6-7 hours until next IMC. At low doses of buprenorphine were administered after a meal, the postprandial pattern was restored and converted to fasted pattern only one IMC cycle. Buprenorphine is partial agonist for the mu receptor. Ahmad et al. (1989) reported that alpha 2 adrenergic receptor and opiate receptors (mainly mu) were presented in submucosal plexus. In concludes that alpha 2 adrenergic activity and other humoral factors seems to take an important role for the initiation of phasic contraction and to control maintenance of IMC cycle.

Mixed connective tissue disease associated with acute polyradiculoneuropathy.

A rare case of mixed connective tissue disease (MCTD) with acute polyradiculoneuropathy is reported. A 23-year-old woman presented with high body temperature, arthralgia and a headache, and developed gait disturbance two weeks later. She had many clinical features common to patients with MCTD. Her neurological manifestations were diagnosed as acute polyradiculoneuropathy based on the clinical picture, combined with supportive ancillary data, including cerebrospinal fluid (CSF) analysis, electrophysiological evaluation, sural nerve biopsy, peroneus brevis muscle biopsy, and magnetic resonance imaging (MRI). Her neurologic deficits, as well as associated laboratory findings, were improved by corticosteroid therapy.

[Experimental research on the interdigestive migrating electric complex after transection and anastomosis of the small intestine].

To study the motility functions after transection and anastomosis of the intestinal tract, the interdigestive action potential was induced experimentally and observed in the small intestines of conscious fasted dogs. When the intestinal tract was transected, the frequency gradient of the basic electric rhythm was disturbed even in cases of subsequent anastomosis. Differences arose in the frequency as upper and lower part of anastomosis, and it did not return to normal. The spike potential which appeared with the building up of the basic electric rhythm was propagated to the distal intestine in a normal course after transection of the intestinal tract according to observations centered on propagation of the IMEC. The propagation velocity of the IMEC gradually drops from the proximal jejunum toward the terminal ileum in the nonmanipulated intestine, while in the intestine undergoing transection and anastomosis, the velocity of propagation was reduced by half in the area of anastomosis, and tended to be even lower in cases of a double section and two anastomoses of the distal intestine. However, the total elapsed time in the small intestine was not extended by much. A new electric complex appeared from the stump of the distal intestine when the intestine was subjected to transection, and a stimulation which is involved in the appearance of the spike potential was formed.

[Gastric carcinoma with prominent reactive lymphoid hyperplasia; report of two cases with an immunohistological study].

An immunohistological study using a Leu 7 antibody, which is a marker for human NK and K cells, was carried out in two cases of gastric carcinomas with prominent reactive hyperplasia. The patients were a 72-year-old man and a 55-year-old man who had shown good postoperative courses in spite of the advanced stage of their tumors. There were many scattered Leu 7 positive cells in the lymphoid hyperplasia, predominantly in the germinal centers and the peri-follicles. The number of Leu 7 positive cells were even more pronounced in a patient whose condition was more advanced. These findings suggest that lymphoid hyperplasia, particularly with regard to the NK and K cell response, plays a significant role in the clinical benignity of the tumors.

[Early combined carcinoma and malignant lymphoma in the stomach associated with tonsillar malignant lymphoma--report of a case].

This report deals with a histologic demonstration of a combined tumor of early signet ring cell carcinoma and lymphoma of the stomach in a 52-year-old Japanese man. The Gastric carcinoma was found by a systemic work up for a tonsillar lymphoma. The relationship between the malignant lymphomas of the stomach and of the tonsil is not clear in this case. The carcinoma and lymphoma of the stomach were both small, and the depth of invasion was localized to the mucosa and submucosa, respectively. The lymphoma was observed just beneath the cancer and was a diffuse, large cell type, and of the same histologic typing as the tonsillar lymphoma. Based on the broader size of the carcinoma, it is considered that the carcinoma probably induced the lymphoma in the stomach. This of an early combined tumor of carcinoma and lymphoma of the stomach is, to our knowledge, the first to be reported in the literature.

Distribution of hippocampal cholinergic neurostimulating peptide (HCNP)-like immunoreactivity in organs and tissues of young Wistar rats.

This report concerns the distribution of the hippocampal cholinergic neurostimulating peptide (HCNP) in tissues and organs of 11-day-old Wistar rats. HCNP, originally isolated and purified from the hippocampus of young rats, is an undecapeptide (acetyl-Ala-Ala-Asp-Ile-Ser-Gln-Trp-Ala-Gly-Pro-Leu). HCNP distribution was investigated by using immunohistochemical techniques, employing an affinity-purified rabbit antibody that specifically recognizes HCNP and its 21-kDa precursor protein. Positively stained cells were detected in a variety of tissues and organs, including salivary gland, small intestine, colon, pancreas, bronchiole, adrenal gland, testis, as well as several others. The nerve fibres around blood vessels of almost all organs expressed HCNP. Our results suggest that HCNP or its precursor, or both, may have a specific function not only in the central nervous system, but also in the peripheral nervous system, and possibly in certain specialized duct and gland cells as well.

[Experimental investigations of the propagation mechanism and the nerve regulatory mechanism of the interdigestive migrating electric complex in intestinal movement].

Segments and Thirty-Vella type loops of the small intestine of dogs were prepared, and the excitation propagation pattern of the interdigestive migrating electric complex (IMEC) was observed by electromyography. The nerve regulatory mechanism in intestinal movement was investigated. The results indicated that first, the frequency of the basic electric rhythm (BER) is controlled so that a downward gradient is formed from the upper to the lower intestine, but this gradient continues to be controlled by a two-dimensional control mechanism involving parietal factors and extrinsic nerves due to transection of the intestines and damage to the extrinsic nerves in the dominant region. However, propagation of the IMEC in the small intestine appeared to be regulated by a combination of the two-dimensional control mechanism involving extrinsic and parietal nerves corresponding to the BER control mechanism. It was clear that the propagation mechanism of IMEC is related to the BER frequency, and propagation of the IMEC is downward in accordance with the frequency gradient of BER.

[An undifferentiated carcinoma of the stomach--a report of 2 cases].

Two cases of an undifferentiated carcinoma of the stomach have been investigated by conventional histologic, immunohistochemical, and ultrastructural methods. The patients were a 73-year-old man and 65-year-old man. A postoperative histological examination of the resected stomach of each patient disclosed large areas of an undifferentiated carcinoma which were relatively well demarcated from areas of the adenocarcinoma. In the undifferentiated carcinomatous areas, the reticulin fiber stain was epithelial in pattern, and mucin staining proved negative. Similarly both Grimelius and Fontana-Masson staining also negative. In like manner immunostains using cytokeratin , vimentin , IgG, IgA, s-100 protein, and NSE were all negative, and an electron microscopic study showed no neurosecretory granules or mucin secretory granules. Based on these findings, the diagnosis of either a malignant lymphoma or small cell anaplastic carcinoma could be excluded, and thus an undifferentiated carcinoma was the determination.

An unusual course of progressive multifocal leukoencephalopathy in a patient with idiopathic CD4+ T lymphocytopenia.

A case is reported of idiopathic CD4+T lymphocytopenia with progressive multifocal leukoencephalopathy and cervical lymph node tuberculosis. A 57 year old Japanese man presented with cervical lymphadenopathy and progressive neurological deficits, and six months later he developed akinetic mutism. He had a persistent severely depressed number of circulating CD4+T lymphocytes in the absence of human immunodeficiency virus infection. T1 weighted MRI showed a diffuse decreased signal intensity limited to the white matter without mass effect. A brain biopsy specimen had a morphology similar to that of progressive multifocal leukoencephalopathy. Polyomavirus antigen was detected in the brain lesion, and viral DNA was identified in nucleated blood cells and urine. Unusually this serious medical condition has lasted for more than three years without remission. To our knowledge this is the first patient with CD4+T lymphocytopenia with progressive multifocal leukoencephalopathy, suggesting that similar opportunistic infections should be considered even in previously normal people.