Different effects of concanavalin A and its succinylated derivative on superoxide release in peritoneal macrophages.

The effects of tetravalent conconavalin A and its succinylated derivative on the intracellular production of superoxide anion (O-2) and its release into cell exterior of peritoneal macrophages were observed. Both tetravalent concanavalin A and its succinylated derivative induced marked enhancement of intracellular reduction of nitroblue tetrazolium, which could be inhibited by alpha-methyl-D-glucoside. The extent of activation of nitroblue tetrazolium reduction induced by both types of the lectin paralleled the activation ratio of oxygen consumption. There was little difference in the extent of intracellular O-2 production induced by two types of the lectin. Nitroblue tetrazolium reduction was not affected significantly by pretreatment with colchicine, rotenone or malonate, inhibitors of the cytoskeletal system and of the electron transport system. In contrast, tetravalent concanavalin A induced a higher rate of superoxide release than did succinylated divalent concanavalin A, which lacks the cross-linking activity of surface glycoproteins. These results indicate that superoxide production following oxygen consumption and superoxide release into cell exterior are controlled independently by a separate membrane mechanism and that superoxide production system is not essentially dependent on the involvement of the cytoskeletal system.

Effect of concanavalin A and its succinylated derivative on the oxidative metabolism of guinea pig peritoneal macrophages.

The effect of concanavalin A and its succinylated derivative on the metabolic regulation of guinea pig peritoneal macrophages was observed. The binding of tetravalent concanavalin A to the surface glycoproteins of macrophages caused a marked increase in the rate of oxygen consumption due to the activation of the hexose monophosphate shunt. Divalent succinylated concanavalin A, also induced a similar change in the rate of oxygen metabolism. The metabolic change induced by these two types of lectin was reversibly inhibited by alpha-methyl-D-glucoside, a haptenic inhibitor of these lectins, and was temperature dependent (observed at above 15 degrees C). It is suggested that the binding of these lectins to the surface glycoproteins, and not their cross-linking into caps, is required for the activation of oxygen metabolism of macrophages, and that highly fluid state of the plasma membrane seems to be an essential requirement for the transduction of glycoprotein perturbation on the macrophage surface into cellular interior via transmembrane control mechanism.

Action of neuropeptide Y on nociceptive transmission in substantia gelatinosa of the adult rat spinal dorsal horn.

Effects of neuropeptide Y (NPY) on substantia gelatinosa neurons were investigated in adult rat spinal cord slices using blind whole-cell patch-clamp technique. Bath application of NPY (1 microM) induced a membrane hyperpolarization, resulting in a suppression of the dorsal root stimulation-induced action potentials in 24% of the substantia gelatinosa neurons tested. In voltage clamp mode, NPY produced an outward current dose-dependently in about one third of substantia gelatinosa neurons at the holding potential of -60 mV, which was not affected by tetrodotoxin (1 microM). The NPY-induced current was suppressed by perfusion with a Ba2+-containing external solution and a Cs2SO4 or tetraethylammonium-containing pipette solution. In addition, The NPY-induced outward currents reversed its polarity near the equilibrium potential of K+ ions (-93 mV). The response to NPY recorded with guanosine-5'-O-(2-thiodiphosphate)-beta-S (GDP-beta-S) containing pipette solution was abolished 30 min after patch formation, suggesting that the response was mediated by the G-protein-coupled receptors. Application of an NPY-Y1 selective agonist, [Leu(31), Pro(-34)]-NPY (1 microM), for 30 s also induced an outward current with a similar time course and amplitude to that induced by NPY. On the other hand, the NPY response was blocked by a simultaneous application of NPY-Y1 selective antagonist, BIBP 3226 (1 microM). No significant changes were found in amplitude and frequency of miniature excitatory postsynaptic currents and dorsal root evoked excitatory postsynaptic currents by NPY. In addition, NPY did not affect both of the miniature inhibitory postsynaptic currents and evoked inhibitory postsynaptic currents, mediated by either the GABA or glycine receptor. These findings, taken together, suggest that NPY produces an outward current in substantia gelatinosa neurons through G-protein coupled, and NPY-Y1 receptor-mediated activation of K+ channels without affecting presynaptic components. The inhibition of the synaptic transmission from the primary fibers to the substantia gelatinosa neurons is considered to contribute to the antinociceptive effects of NPY.

Acute renal failure in primary macroglobulinemia with small-molecule IgM.

Primary macroglobulinemia with acute renal failure developed in a 46-year-old man after administration of contrast medium (containing diatrizoate sodium and meglumine [Urografin]) for intravenous pyelography. On admission, the monoclonal protein peak could not be detected in serum by cellulose acetate electrophoresis, but the serial electrophoresis revealed a gradual increase in monoclonal protein peak, which later became two peaks. The serum of the patient contained the monoclonal small-molecule IgM kappa-chain protein with 13 to 14 Svedberg units in sedimentation constants, which is antigenetically the same as native IgM heavy chain. Although acute renal failure is extremely rare in primary macroglobulinemia, the small-molecule IgM seems to be one factor that induces acute renal failure.

Acceleration of lipid peroxidation in human paraquat poisoning.

A 20-year-old man ingested approximately 15 to 20 mL of a 24% solution of paraquat. Acute renal and liver failure developed. The patient began undergoing hemodialysis and receiving vitamin E, but he died of pulmonary fibrosis the 27th day after the ingestion. During a two-week period of observation, evidence of lipid peroxidation in his serum was noted by determining malondialdehyde levels. Guinea pigs exposed to paraquat showed similar patterns of malondialdehyde presence in plasma. These results indicate that increased serum or plasma levels of lipid peroxide occurring in paraquat poisoning might reflect production of highly reactive oxygen free radicals and that, possibly, levels of lipid peroxides could serve as indicators of the efficacy of therapy directed toward scavenging free radicals.

Further studies of lipid peroxidation in human paraquat poisoning.

In patients with subacute toxic reactions from paraquat poisoning (death within 11 to 41 days), the extent of lipid peroxidation, expressed as serum malondialdehyde level, was 2.7-fold higher (12.33 +/- 4.42 nmole/mL) before pulmonary fibrosis than that in normal controls (4.55 +/- 1.23 nmole/mL). The extent of lipid peroxidation in patients with acute toxic reactions (death within one to three days) was not elevated; these patients died of pulmonary edema and hemorrhage (acute respiratory distress), liver failure, renal failure, and adrenal necrosis. Remarkable high levels of paraquat (greater than 5 mg/L) were found in the urine, serum, and tissues of patients with acute toxic reactions; a small amount of paraquat was found in the serum or urine of patients with subacute toxic reactions five to 11 days after ingestion. Patients who survived had no elevation in lipid peroxidation. Administration of vitamin E (100 to 4,000 mg/day from the first hospital day) had no effect on survival.

Prolonged effects of polyriboinosinic:polyribocytidylic acid on spontaneous running wheel activity and brain interferon-alpha mRNA in rats: a model for immunologically induced fatigue.

Following 2 weeks acclimation to the running wheel in the home cages, an i.p. injection of a synthetic double-stranded RNA, polyriboinosinic:polyribocytidylic acid (poly I:C, 3 mg/kg), was performed to produce the immunologically induced fatigue in rats. The daily amounts of spontaneous running wheel activity decreased to about 40-60% of the preinjection level until day 9 with normal circadian rhythm, then gradually returned to the baseline level by day 14. Rats given a heat exposure (36 degrees C for 1 h) for the consecutive 3 days showed an increase in activity except for the first day. In the open field test, the total moving distance and the number of rearing of the poly I:C-injected rats decreased on day 1, but they were not different from the saline-injected group on day 7, suggesting that the poly I:C-induced fatigue on day 7 was not due to the peripheral problems such as muscle/joint pain, but involved the CNS. Quantitative analysis of mRNA levels using a real-time capillary reverse transcriptase-polymerase chain reaction (RT-PCR) method revealed that interferon-alpha (IFN-alpha) mRNA contents in the cortex, hippocampus, hypothalamic medial preoptic, paraventricular, and ventromedial nuclei were higher in the poly I:C group than those in the saline and heat-exposed groups on day 7, although the amount of interleukin-1 beta mRNA showed no differences. Serum adrenocorticotropic hormone and catecholamine levels were not significantly different between groups. The present results indicate that the prolonged fatigue induced by poly I:C, which is evaluated by the spontaneous running wheel activity, can be used as an animal model for the immunologically induced fatigue associated with viral infection, and suggest that brain IFN-alpha may play a role in this model.

Long-term results of hepaticojejunostomy for hepatolithiasis.

The results of a hepaticojejunostomy as a biliary-enteric bypass for benign disease are usually excellent. On the other hand, hepatolithiasis features a high rate of residual and recurrent stones with cholangitis after surgery. This study aims to evaluate the long-term results of a hepaticojejunostomy for hepatolithiasis regarding both the degree of the occurrence of postoperative cholangitis and the outcome. The clinical records of 159 patients with hepatolithiasis who underwent surgical treatment over a 23-year period were also retrospectively reviewed. Ninety-four of 159 patients underwent a hepatecetomy and 65 patients were subjected to liver-preserving surgery by means of intra- and postoperative endoscopic lithotripsy. In addition 72 patients underwent a hepaticojejunostomy. The rate of residual or recurrent stones was 31.4 per cent after complete stone removal. Twenty-two (30.6%) of the 72 patients developed some kind of cholangitis. This rate was significantly higher than that (three of 87 patients) of the non-biliary-enteric anastomosis group regarding the occurrence of biliary complications. We conclude that the use of a hepaticojejunostomy for patients with possible residual stones or intrahepatic bile duct lesions remains controversial.

[Selection of treatment in primary hepatolithiasis (PHL)].

Two hundred and three patients with PHL were encountered for the last 10 years. In order to select treatment of PHL, we classified PHL on the basis of atrophy of hepatic parenchyma (AHP), existence of extrahepatic gallstone and dilatation of extrahepatic bile duct (DEBD). For the case with AHP hepatic resection must be performed, because there is often chronic proliferating cholangitis and improvement in hepatic function can not be expected in view of AHP. For the case with extrahepatic gallstone without AHP hepatic resection. For the case with DEBD and without AHP extrahepatic gallstone must be removed. For the patient with extrahepatic gallstone and DEBD without AHP, indication of the removal of stones, depends on the existence of symptoms. Twenty-two cases treated by biliary drainage procedure often had some complications in postoperative course, that is 5 patients had cholangitis and 7 patients liver abscess. It was suggested, therefore, that biliary drainage procedure was contraindicated in PHL.

On the calculation of a "reference change" for comparing two consecutive measurements.

We describe a statistical method for calculating a "reference change," defined as that difference between two consecutive test results in an individual that is statistically significant in a given proportion of all similar persons. By allowing for variation in within-person variances, this procedure computes a reference change that is more specific (i.e., less prone to false positives) than that obtained directly from the distribution of observed differences between measurements. Moreover, the method may easily be extended to a test for trend in three successive measurements. The method has been applied to semi-annual measurements of serum calcium and alkaline phosphatase in 698 men and women enrolled in a large health-maintenance program. We believe that these ideas may also be usefully applied to successive laboratory tests in carefully defined patient populations--but this introduces special problems, which are discussed briefly.

Clinical data evaluation and the PL health control information system.

In a health control service environment, that is, a periodic, membership AMHTS type of comprehensive health check-up system, where clinical data evaluation especially an evaluation in terms of subject-specific normal ranges, is most important, the medical information system is required to handle: (1) Various network types files; (2) real-time immediacy; (3) an asserted reliability to meet personal health control purposes. As in other computer applications already successfully used, an indexed direct assess method (IDAM) developed is our solution. It allows us to provide multiple indices for the file network, instead of inverted files, a unique index-to-record relationship, preventing any unrecoverable chaining destruction and, thereby, provides any network type access a stable access time. Furthermore, for research purposes, a data integrity for on-line access and batch access was attained as well as a retrieval language system with a multiple key retrieval function.

The role of polymorphonuclear leukocytes in delayed hypersensitivity skin reactions: suppressive effects of anti-polymorphonuclear leukocyte serum.

Rabbit antisera (APS) against normal guinea pig peritoneal exudate polymorphonuclear leukocytes (PMN), when injected intraperitoneally three times within 24 h into guinea pigs, lead to a marked fall in the blood PMN count (below 100/mm3) 24-72 h after the injection and a mild depression in the number of circulating mononuclear leukocytes (MNC) at 36-72 h. APS treatment of guinea pigs led to a marked suppression of delayed hypersensitivity skin reactions in response to dinitrophenylated bovine gamma-globulin. The volume, induration, mean diameter of the reaction and leukocyte (both PMN and MNC) emigration into local sites were suppressed, as was, to a lesser extent, vascular permeability. Based on these observations it appears that circulating PMN are required for the full expression of delayed hypersensitivity reaction engendered by bovine gamma-globulin.

Reversed-phase high-performance liquid chromatographic investigation of mucosal nucleosides and bases and urinary modified nucleosides of gastrointestinal cancer patients.

Reversed-phase high-performance liquid chromatography (HPLC) was used to determine the levels of nucleosides, bases and their metabolites in perchloric acid extracts of gastrointestinal mucosa. By comparing the levels of these compounds in the normal portion with the neoplastic portion of mucosa resected from malignant cancer patients, it was found that there was significant elevation of the uracil level in the neoplastic mucosa of all eight patients with colorectal cancer (2.7-fold in normal mucosa), but only in the neoplastic mucosa of one out of four patients with gastric cancer. The levels of hypoxanthine and uridine in the colorectal cancer mucosa samples and the inosine in gastric cancer samples were also significantly higher than those in normal mucosa. The urinary modified nucleosides were prefractionated with a boronate affinity gel column, and their levels were determined by the same HPLC method. There was no significant difference in the concentrations of pseudouridine, 1-methylguanosine N2-methylguanosine and N2,N2-dimethylguanosine between urine samples taken before and after surgery from eight patients with malignant colorectal cancer. Contrary to other reports, no significant differences in modified nucleoside levels were observed between urine samples from patients with colorectal cancer and those from normal subjects.

Reversed-phase liquid chromatographic investigation of nucleosides and bases in mucosa and modified nucleosides in urines from patients with gastrointestinal cancer.

Reversed-phase high-performance liquid chromatography (HPLC) has been used to determine the level of nucleic acid metabolites in perchloric acid extracts of gastrointestinal mucosa. By comparing the levels of these compounds in the normal portion with the levels in the neoplastic portion of mucosa resected from patients with malignant cancer, it was found that uracil was significantly elevated in the neoplastic colorectal mucosa (adenocarcinoma) of eight patients with colorectal cancer (P less than 0.01, statistically significant with the paired t test). The mean level of uracil in neoplastic colorectal mucosa was 2.7-fold higher than that in normal mucosa. However, in neoplastic gastric mucosa, only one out of four patients with gastric cancer showed elevated uracil. In neoplastic mucosa, the levels of hypoxanthine and uridine for colorectal cancer, and inosine for gastric cancer, were also significantly higher than those in normal mucosa (P less than 0.05, with the paired t test). The urinary modified nucleosides were prefractionated with a boronate affinity gel column, and their levels determined by the same HPLC method. No significant differences in the concentrations of pseudouridine, 1-methylguanosine, N2-methylguanosine or N2,N2-dimethylguanosine were observed in pre- and post-operative urines from patients with colorectal cancer and normal urines.

A family of Gardner's syndrome with report of an autopsy case.

A family with Gardner's syndrome including one autopsy case is reported. A 22 year-old man had multiple polyposis of the colon and the small intestine, epidermal cysts, soft part tumor of the abdominal skin and osteomas in the calvarium and rib, and died of a carcinoma in the transverse colon and embolism of the brain. Histologically, all polyps were diagnosed as adenomatous polyp. The greater their diameter, the higher the tendency of atypism was observed. However, the polyps around the cancer were few, small, and with low atypism, and we could not obtain evidence showing direct transformation of adenomatous polyps into carcinoma. His two elder brothers had multiple polyposis of the colon, epidermal cysts and osteomas in the calvarium or mandibular bone, but are still alive.