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The neuropathological findings in a Japanese male with nephrosialidosis are reported. Clinically, coarse face, psychomotor retardation, macular cherry-red spot and proteinuria were noted at 1 year and 7 months. He was diagnosed to have nephrosialidosis on the basis of a deficiency of alpha-neuraminidase activity in both lymphocytes and cultured skin fibroblasts, and of severe glomerular and tubular involvement on renal biopsy. He died of multiple organ failure at 8 years and 6 months. There were numerous vacuoles and storage materials in visceral organs, particularly in the glomerular and tubular epithelial cells of the kidney and Kupffer cells as well as hepatocytes in the liver. Neuropathological examination revealed severe neuronal storage in the selected part of the central nervous system: lower motor neurons of the brain stem and spinal anterior horn cells, as well as neurons in the basal nucleus of Meynert. In the peripheral nervous system, sympathetic ganglia were severely affected. There was little or no neuronal storage in the basal ganglia, cerebral cortex or cerebellum, and demyelination was not found. Electron microscopic examination showed fine wavy multilamellar structures in the spinal anterior horn cells or Zebra body-like structures in the neurons of the Meynert's basal nucleus. Lectin histochemistry was positive for wheat germ agglutinin, Ricinus communis agglutinin-1 and peanut agglutinin within distended neurons. We conclude that the neuropathological feature in nephrosialidosis is not specific except for the selectiveness of the anatomical sites of involvement. It shares some aspects found in other types of sialidosis or galactosialidosis.

We investigated the presence of retroviral protein in the pancreatic islets of non-obese diabetic mice to prove that the virus-like particle observed specifically in the pancreatic Beta cell of these mice was retrovirus. Western blot analysis probed with anti-retrovirus antibody demonstrated the existence of retroviral gag (group specific antigen) protein p30 in the islets of female non-obese diabetic mice. Islets of non-obese diabetic mice which were treated with cyclophosphamide, known to accelerate the development of insulitis and diabetes mellitus, have shown both a significantly increased number of retrovirus-like particles (type C) and enhanced expression of gag protein p30, compared to those of mice not treated with cyclophosphamide. These results confirmed the presence of type C retrovirus in non-obese diabetic mouse Beta cells and suggest a role for retrovirus in the development of insulitis and diabetes in these mice.

Peptide YY (PYY) is thought to possess paracrine and endocrine functions. The highest concentrations of this peptide are in the colonic mucosa. The effect of PYY on electrolyte and water transport in the rat colon was studied in vivo. Under urethane anesthesia, rat colonic loops were perfused at a constant rate with physiological buffer solution containing phenol red as a nonabsorbable volume marker, and net movements of water, sodium, chloride and potassium in the perfused colon were determined every 10 min. Intravenous administration of PYY produced a dose-dependent increase in the net absorption of sodium chloride and water, as well as a decrease in the net secretion of potassium. PYY inhibited the reduction in net absorption of sodium chloride and water evoked by vasoactive intestinal peptide (VIP), but did not affect the VIP-evoked increase in net potassium secretion. These findings suggest that PYY acts as an enhancer of sodium chloride and water absorption and as an antagonist to VIP-induced secretion in the colon.

The effect of methoxamine, a specific alpha 1-adrenergic agonist, on the release of T3, T4 and cAMP from perifused mouse thyroid was studied to clarify the role of the alpha 1-adrenergic receptor in the regulation of thyroid hormone secretion. TSH-stimulated T3 and T4 release was inhibited significantly by methoxamine. With regard to cAMP release, methoxamine inhibited TSH-stimulated cAMP release in the presence of 4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone but did not inhibit TSH-stimulated cAMP release in the presence of 3-isobutyl-1-methylxanthine. Methoxamine did significantly suppress TSH-stimulated release of T3 and T4 in the presence of each phosphodiesterase inhibitor. Depletion of Ca2+ in the perifusion buffer abolished completely the inhibitory effect of methoxamine on TSH-stimulated T3 and T4 release. The present study suggests that activation of the alpha 1-adrenergic receptor inhibits TSH-stimulated T3 and T4 secretion through a Ca(2+)-dependent mechanism in the mouse thyroid gland.

Functional and structural characteristics of interleukin 2 (IL-2) receptors on B-cell chronic lymphocytic leukemia (B-CLL) cells were analyzed by a proliferation assay, IL-2 binding assay and cross-linking study. In the 3H-thymidine incorporation assay, purified B-CLL cells from four out of sixteen cases, in which the percentage of Tac antigen (Tac Ag) positive cells in peripheral blood lymphocytes ranged from 0 to 48.8%, responded to IL-2 (100 U/ml) after both 3- and 6-day incubation. No relationship was found between the responsiveness to IL-2 and the percentage of Tac Ag positive cells. In the radiolabeled IL-2 binding assay, however, B-CLL cells from all seven cases examined, including three cases with mitogenic response to IL-2 and four cases without mitogenic response, were shown to have both high- and low-affinity receptors. The number of high- and low-affinity receptors per cell ranged from 29-186 and from 420 to 1,800, respectively. Furthermore, with the affinity cross-linking method p55 (Tac Ag) and p70/75 were found even in cases without mitogenic response in their B-CLL cells. In conclusion, the B-CLL cells so far examined possessed high-affinity IL-2 receptors consisting of p55 and p70/75; nevertheless, this was not sufficient to respond to the mitogenic signal of IL-2.

A sensitive and specific radioimmunoassay system for rat plasma cholecystokinin (CCK) was employed to study the effect of trypsin inhibitor on plasma CCK levels. Feeding the trypsin inhibitor, FOY-305 200 mg/kg, to rats for 10 days stimulated the pancreatic growth. However, there was no significant difference in fasting plasma concentrations and duodenal contents of CCK and secretin. On the other hand, acute ingestion of same dosage of FOY-305 caused a marked (20-fold) and sustained elevation of plasma CCK levels. The principal form of circulating CCK was CCK-22 like on Sephadex G-50, in contrast to that of the cerebrum and duodenum where CCK-8 like form, or CCK-8 and CCK-22 like forms were main components, respectively. A significant rise in plasma levels of immunoreactive secretin was also found. These results suggest that the pancreatic growth observed by feeding the trypsin inhibitor to rats is caused by an excessive amount of CCK in conjunction with secretin.

Using our electron microscopic method for polysome analysis and an immunofluorescent technique we studied Ig production and secretion by tumor cells in seven BJP myeloma patients and seven "nonsecretory" myeloma patients. In BJP myeloma Ig production and secretion is of three types: Type 1, only L-chains are synthesized and secreted; Type 2, the myeloma cells show fluorescence for H-chains, but upon polysome analysis there is no peak at polysomes corresponding H-chain production; Type 3, the myeloma cells show fluorescence for H-chains, and polysome analysis shows two peaks corresponding to L- and H-chain production. Polysome analysis of "nonsecretory myelomas show the presence of only very few membrane-bound polysomes and their distribution curves are entirely different from those of "ordinary" myeloma. Furthermore, the distribution patterns vary among seven cases. Results obtained by polysome analysis and immunofluorescent technique suggest that the "nonsecretory" myeloma could be divided into several subtypes.

Congenital dyserythropoietic anemia type I was diagnosed in a Japanese patient. Electron microscopic examination of thin sections revealed the features of erythroblasts peculiar to CDA type I; namely "spongy appearance" of nucleus and "internuclear chromatin bridge" etc. However, the widening of nuclear pores, which has been reported as another peculiar feature of CDA type I erythroblasts, were not confirmed. Aberrations of erythroblast nuclear membrane which have not been hitherto noticed were as follows: The nuclear envelopes of erythroblasts frequently lacked heterochromatin application in wide areas where they often showed a marked invagination or evagination. This is associated with the invasion of cytoplasmic organelles into the nuclear territory and occasionally with the extrusion of the nucleolus into the cytoplasm. There were no nuclear pores on the invaginated or the evaginated portions of the nuclear envelope where the perinuclear cistern was compressed to lose the cisternal space, or widened to contain some membrane debris. The findings with thin sectionings were confirmed with freeze-fracturing. Both findings suggest that an aberration may reside in the erythroblast nuclear membrane itself, but not in nuclear pores.

Fasting (24 h) normal dogs and depancreatized dogs were injected intravenously with highly purified porcine insulin (Actrapid) in the doses of 0.2 U/kg and 0.5 U/kg, respectively. Blood glucose decreased from 152 +/- 41 (SEM) mg/100 ml to 39 +/- 7 mg/100 ml in the depancreatized dog and from 95 +/- 3 mg/100 ml to 42 +/- 4 mg/100 ml in the normal animal. Using a specific antiserum for "pancreatic" glucagon, the circulating level of glucagon immunoreactivity did not rise from the basal value of 247 +/- 31 pg/ml in the depancreatized group whereas it rose significantly from 223 +/- 24 pg/ml to 321 +/- 41 pg/ml in the normal group. In contrast intravenous infusion of 7 g of arginine increased "pancreatic" glucagon immunoreactivity in both groups. Thus, extrapancreatic glucagon of the pancreatic type does not respond to hypoglycaemia but to arginine infusion.

We have noticed that functional disorders of the liver characterized by hepatomegaly and an increase in serum gamma-glutamyl transpeptidase develop in patients with prostatic cancer who are placed under longterm therapy with massive doses of estrogen after castration. We performed laparoscopy in six cases of prostatic cancer with hepatomegaly so that we could study the morphology of the liver. Our findings were as follows. In five, the histological features of the liver biopsies were very similar to those seen in alcoholic hepatitis. In spite of this fact, two of the five had no history of alcohol consumption. Furthermore, in one other case, liver damage resembling alcoholic hepatitis developed during abstinence. The findings in these three cases suggested that long-term, massive doses of synthetic estrogen may lead to liver injury similar to alcoholic hepatitis in nonalcoholics. The ultrastructural findings of the liver cells were also suggestive of the adverse effect of treatment. All cases were negative for hepatitis B surface antigen. Recent reports have demonstrated some nonalcoholics with histological features of the liver indicative of alcoholic hepatitis. This particular condition was termed "nonalcoholic steatohepatitis" by Ludwig et al. It is quite likely that synthetic estrogen is also responsible for "nonalcoholic steatohepatitis" when it is used in massive doses.

To examine a possible cause of axonal dystrophy in the nucleus gracilis, dorsal root ganglion (DRG) neurons of rats were investigated by means of electron-microscopic autoradiography and horseradish peroxidase (HRP) tracing method. Following injections of tritiated amino acids into the L6 and S1 DRG, labeling was observed on the initial and halfway developed dystrophic terminals in the ipsilateral gracile nucleus. However, no grains or few, if any, were found on the well developed huge dystrophic endings. Compared with the thoracic and upper lumbar DRG, a decrease in velocity and amount of retrograde HRP transport was demonstrated in the lower lumbar and sacrococcygeal DRG neurons, especially of large cell diameter, irrespective of age of rats. These findings led us to conclude that the axonal dystrophy reflects a state of an anterograde overtransport of the axoplasm caused by a diminished retrograde transport which is specific to lower lumbar and sacrococcygeal DRG large neurons.

We have studied the effect of ionophore A23187 on the release of T3 and cAMP from perifused rat thyroid glands and on the morphological changes of the follicle epithelial cells. Ionophore A23187 at a concentration of 5 microM significantly inhibited TSH-stimulated T3 release. Depletion of Ca2+ in the infusion buffer completely abolished the inhibitory effect of ionophore A23187 on TSH-stimulated T3 release. In the presence of TSH and 3-isobutyl-1-methylxanthine, ionophore A23187 at 5 microM did not significantly affect either the release of cAMP during a 3-h perifusion or the cAMP content in the thyroid tissues after 1-h perifusion. By electron microscopy, the follicle epithelial cells of the rat thyroid tissues showed marked responses to TSH after 3-h stimulation. By the addition of 5 microM ionophore A23187, the ultrastructural changes such as phagocytotic uptake of luminal colloid and the formation of intracellular colloid droplets were rarely observed in spite of the presence of TSH. The present study indicates that ionophore A23187 at a concentration of 5 microM inhibits both TSH-induced morphological changes of the follicle epithelial cells and TSH-stimulated T3 release without reducing cAMP level stimulated by TSH in rat thyroid glands.

A 50-year-old male with unresectable hepatocellular carcinoma (HCC) had a hypercalcemic crisis with a serum calcium concentration of 7.8 mEq/zeta, without any evidence for bone metastases or parathyroid lesions. The hypercalcemia was thought to be due to increased renal reabsorption of calcium and increased bone resorption, which was probably caused by humoral factors derived from the HCC, some being parathyroid hormone-like factors. Since conservative therapy for hypercalcemia was not sufficiently effective and was accompanied by progressive exacerbation of ascites and leg edema, transcatheter arterial chemo-embolization (TACE) was performed. On the following day, serum calcium concentration decreased from 6.3 mEq/zeta to the normal range, although serum alpha-fetoprotein levels decreased only slightly. Thereafter hypercalcemia did not develop for about 4 weeks. The results demonstrated that TACE can be effective for humoral hypercalcemia of HCC.

In order to clarify morphological changes of the liver in the uremic state, 16 uremic patients treated with chronic hemodialysis were studied. Biopsy was performed in 14 cases under laparoscopic observation and in two on the occasion of renal transplantation. One uremic patients not being treated with dialysis was also studied for comparison. All biopsy specimens were examined by light and electron microscopy. The liver usually appeared mildly or moderately swollen under laparoscopic observations, which was considered at least partially due to the enlargement of the hepatocytes. All patients had hepatocytes with an Orcein-negative "ground glass" appearance, in which marked proliferation of smooth endoplasmic reticulum (SER) was found by electron microscopy. Since the patient not being on dialysis also had such hepatocytes, this finding may be characteristic of uremia. With electron microscopy, in addition to proliferation of SER, alteration of mitochondria and rough endoplasmic reticulum (RER) and an increase in cytoplasmic lipid droplets were observed. Hypertrophy of the Golgi apparatus containing electron-dense particles (VLDL) was often found in patients associated with hypertriglyceridemia. Amorphous electron-dense inclusions in microbodies were occasionally observed. Siderosis was observed in nine patients including three having parenchymal siderosis. With electron microscopy, various siderosomes were seen in the cytoplasm of hepatocytes in patients with parencymal siderosis. Conclusively, these histological and ultrastructral features of hepatocytes are rather associated with several metabolic abnormalities in uremia.

Clinical and electrophysiological studies were carried out on 39 patients with the Guillain-Barré syndrome to evaluate which elements were of prognostic value during the acute phase. Residual clinical signs such as motor weakness and absent patellar tendon reflexes were found in 16 (52%) of those patients who had had a preceding illness. Persistence of deficit was significantly correlated to age at onset, the degree of quadriparesis and loss of deep sensation in the acute phase. Of the 10 patients who showed a reduction in motor nerve conduction velocity (MCV) in the early stage, 8 (80%) revealed significantly residual clinical symptoms at follow-up. There was a tendency for the incidence of residual signs to be more common in the patients with slowing of mixed nerve conduction velocity, and prolonged latency of H-wave and the residual latency. Nerve conduction studies, especially measurement of MCV, were of value as a reliable prognostic indicator in this syndrome.

A case of "nonsecretory" myeloma is described. The patient had typical osteolytic lesions and marked infiltration of myeloma cells in the bone marrow, and plasma cell leukemia. A good partial remission was obtained with Melphalan, but the patient relapsed and died one year later. Immunofluorescent and immunoelectroscopic studies on the myeloma cells demonstrated the presence of cytoplasmic gamma-and kappa-chains at the initial stage and of only kappa-chains at a relapse. The electron microscopic method for polysome analysis indicated that both L-and H-chains were synthesized on membrane-bound polysomes initially, but the ability to produce H-chain was missing at the relapse.

We established a perifusion system using mouse thyroid glands. In this system, TSH increased the release of T3 and T4 significantly, and the response of thyroglobulin to TSH was delayed in comparison with that of T3 and T4.

Immunohistochemical localization of cytochrome P-450 in the colonic mucosa of 3-methylcholanthrene-pretreated and untreated rats was studied by indirect fluorescent antibody staining technique. A polyclonal antibody for cytochrome P-450MC purified from hepatic microsomes of 3-methylcholanthrene-pretreated rats was used for this experiment. A strong immunofluorescence was found to be localized in the cytoplasm of the surface epithelium of the mucosa in the colon of 3-methylcholanthrene-pretreated rats. A faint immunofluorescence was also observed in the epithelium of untreated rats. 7-Ethoxycoumarin O-deethylase activity of colonic microsomes was significantly enhanced by 3-methylcholanthrene-pretreatment in parallel with an increase in the intensity of immunostaining for cytochrome P-450MC in Western blotting analysis. This is the first report on the localization of cytochrome P-450 in the colonic mucosa.

Myogenic hyperuricemia is caused by increased production of uric acid secondary to enhanced release of inosine and hypoxanthine from exercising muscles into blood. This novel mechanism is evident in glycogenosis types VII (muscle phosphofructokinase deficiency), III (debranching enzyme deficiency), and V. In this study, we examined the effect of muscular exercise on the glycolysis of circulating erythrocytes in glycogenosis types VII and III. The concentration of erythrocyte 2,3-bisphosphoglycerate, which had decreased because of genetic partial deficiency of erythrocyte phosphofructokinase, was further decreased after prolonged bed rest in patients with type VII. Ergometric exercise rapidly increased fructose-1,6-P2, dihydroxyacetone-P plus glyceraldehyde-3-P, and 2,3-bisphosphoglycerate in circulating erythrocytes. Similar changes were observed after exercise in patients with type III. The exercise-induced metabolic alteration of erythrocytes was reproduced in vitro by incubating normal erythrocytes in the presence of inosine. We conclude that physical activity affects glycolysis in erythrocytes in glycogenosis types VII and III, and that myogenic factors including inosine are responsible for this change.

Degradation of purine nucleotides was evaluated in different types of rat skeletal muscle during ischemic and non-ischemic contraction. Extensor digitorum longus (EDL, fast) and soleus (slow) muscles were stimulated electrically via the sciatic nerve (5 Hz, 10 min). Under non-ischemic condition, the concentrations of IMP, inosine, adenosine, and hypoxanthine increased in EDL muscles but not in soleus muscles during stimulation. Under ischemic condition, these metabolites increased in both EDL and soleus muscles and the changes in concentrations of IMP and inosine were greater in ischemic EDL muscles. The increase in inosine had a strong positive correlation with that in IMP in ischemic EDL and soleus muscles, but the ratio, Δinosine/ΔIMP was smaller in EDL muscles. The increase in adenosine under ischemic condition was not significantly different between the two muscles. These findings suggest that ischemia enhances overall degradation of purine nucleotides in contracting fast and slow muscles, and that although the degradation of adenine nucleotides to IMP is greater in fast muscles than in slow muscles, the relative degradation rate of IMP to inosine with respect to the intramuscular IMP concentration is rather smaller in fast muscles.

We studied two brothers with hereditary xanthinuria (xanthine oxidase deficiency) and their family members. The two brothers had extremely low concentrations of urate but markedly high concentrations of xanthine and hypoxanthine in plasma and urine. Xanthine oxidase activities were virtually absent in the duodenal mucosa. In their parents (presumed obligate heterozygotes), the activities of xanthine oxidase were about half that of normal subjects. Although plasma xanthine and hypoxanthine concentrations of the parents were normal, urinary xanthine and hypoxanthine excretions were significantly higher than those of normal subjects (xanthine, father 17.1 mg/g creatinine and mother 27.4 vs. normal controls 5.7 to 11.0; hypoxanthine, father 14.0 and mother 27.3 vs. controls 4.0 to 8.4). Similar changes in the metabolite concentrations were seen in at least 6 other relatives, suggesting they were heterozygotes. This study indicates that the presumed obligate heterozygotes of xanthine oxidase deficiency retained about half normal enzyme activities causing the partial metabolic blockage in vivo at this enzyme step.

Excess purine degradation in exercising muscle due to impaired glycolysis causes hyperuricemia in muscle glycogenoses (myogenic hyperuricemia). However, hyperuricemia has been seen more frequently in Type VII than in Type V. This study was designed to clarify a metabolic basis for the different frequency of hyperuricemia between the diseases. 5 patients (2 males, 3 females) with Type V and 4 (3 males, 1 female) with Type VII participated in the study. 1) In every patient, semi ischemic forearm exercise caused no increase in lactate but exaggerated increases in ammonia, inosine and hypoxanthine in cubital venous blood. The ammonia increase was not different between Type V and Type VII, but it was greater in male than female patients. 2) Semiischemic forearm exercise performed after glucagon injection showed that lactate production was induced by exercise in Type V but not in Type VII. 3) Saline or glucose solution was infused in a patient with Type V during exercise on a bicycle ergometer. With saline infusion, ammonia and hypoxanthine in systemic circulation increased greatly after exercise. Conversely, with glucose infusion there were no increases in these metabolites. These findings indicated that blood glucose can be available as metabolic fuel through muscle glycolysis in Type V but not in Type VII. Thus, muscle purine degradation may be accelerated more in Type VII than in Type V, leading to a higher frequency of hyperuricemia.