Effect of Zena F-III, a liquid nutritive and tonic drug, on the neurochemical changes elicited by physical fatigue in mice.

The effects of a liquid nutritive and tonic drug (NTD) on the neurochemical changes elicited by physical fatigue in mice were investigated in terms of the calcium-dependent dopamine synthesizing function of the brain. In this study, Zena F-III (Taisho Pharmaceutical Co., Ltd., Japan), one of the most popular NTDs in Japan, containing 15 crude drug extracts together with taurine, caffeine, and vitamins, and formulated based on the precepts of traditional Chinese medicine, was used. Male mice were forced to walk for 0-6 h at a speed of 3 m/min using a programmed motor-driven wheel cage. The serum and brain calcium levels in the mice were significantly increased following forced walking. The increase in brain calcium level began later and was more gradual than that in the serum calcium level, and reached its maximum value following forced walking for 3 h. The neostriatal dopamine level was also significantly increased, and locomotor activity significantly decreased following forced walking for 3 h. Prior oral administration of F-III (10 ml/kg) attenuated the increases in the serum and brain calcium levels, the increase in the brain dopamine levels, and the decrease in locomotor activity induced by forced walking. Taking into consideration these findings with our previous reports, it is suggested that physical fatigue leads to an increase in dopamine synthesis in the brain through a calcium/calmodulin-dependent system, thereby inducing behavioral changes, and that F-III inhibits this pathway and may alleviate overwork-induced physical fatigue.

An autoradiographic study of calcium movement in the enamel organ of rat molar tooth germs.

The distribution and movement of calcium through the enamel organ and into the forming enamel was studied in 6-day-old rats, intravenously injected with 45Ca. To prevent dislocation of radiocalcium in the specimens, the tooth germs were rapidly frozen/freeze-substituted and processed for 45Ca autoradiography under dry conditions. At 30 s after the 45Ca injection, there was a decrease in labelling intensity progressing from the overlying connective tissue to the enamel organ and, in the secretory ameloblasts, from the proximal to distal cytoplasm. The most intense labelling was in the enamel matrix, where it was restricted to the superficial layer extending approx. 15 microns below the surface. At later times the density of silver grains over the connective tissue decreased considerably, whereas secretory ameloblasts showed an increasing intensity in the distal portions. Enamel had the heaviest labelling: the width of the labelled enamel increased gradually to only 40 microns from the surface 60 min after the injection. The use of wet emulsion over similarly prepared sections caused a severe dislocation of radiocalcium in the specimens. These findings confirm the rapid penetration of systemically administered calcium to newly formed enamel, probably due to isotopic exchange. A relatively slow diffusion through the enamel organ and into the surface layer of enamel suggests that net transport of calcium through the enamel organ is transcellular.

Proposed standard for human blood vitamin B1 value using HPLC. The Committee for Vitamin Laboratory Standards, Japan.

Standard reference ranges for all laboratory test values are mandatory. This study was designed to establish a reference range for blood vitamin B1 levels, since the normal range has not been determined in the Japanese population. We founded the Japan Committee for Vitamin Laboratory Standards, which was incorporated with the Vitamin Society of Japan and the Japanese Society of Nutrition and Food Science. We standardized whole blood vitamin B1 levels using three HPLC techniques (post-column reverse-phase HPLC, pre-column reverse-phase HPLC, and precolumn GP-HPLC). The reference range was obtained in 54 volunteers administered a 1,800 kcal diet with 2 mg of vitamin B1 (1.74 mg measured) daily to avoid marginal vitamin B1 deficiency in the population. The range for each assay was 26-47, 28-51, and 28-56 ng/ml, respectively. Our data suggest that 26-28 ng/ml is the lower limit of normal for whole blood vitamin B1, but further studies in a larger population are needed in order to obtain more definitive results.

[Benefits of totally implanted central venous access in patients with cancer].

A totally implanted system for improved central venous access has been investigated during 44 procedures in 38 patients (37 with cancer and one with cerebral infarction). Most of them lacked peripheral venous sites, and ten per cent of the patients had a prior chronic external central venous catheter. This system is implanted using local anesthesia and consists of a port connected to a central venous catheter threaded through the subclavian vein into the superior vena cava. The system was used mainly for intermittent administration. Injections and infusions of virtually all classes of antineoplastic drugs or antibiotics (81%), blood components (26%), hyperosmolar total parenteral nutrition (47%), and intravenous solutions (53%) were administered through the system. Twenty-one patients died with the system still functioning. Ten of the systems were eventually removed, either after completion of therapy (1) or because of a complication (9). In another 13 cases the devices were working at the completion of the study. Infections accounted for nine per cent of port complications. Four systems became occluded and two of them were replaced. Overall patient acceptance was excellent. This system can be assigned to an appropriate setting that facilitates management of the cancer patient.

[CT in colon cancer].

CT pictures from 59 lesions of advanced colon cancer including rectal cancer were reviewed to evaluate a role of CT in preoperative staging diagnosis. CT findings were recorded following general rules for clinical and pathological studies on cancer of colon, rectum and anus, proposed by Japanese Society for Cancer of Colon and Rectum. 1) Tumors were detected in 90% of advanced colon cancers. 2) Sensitivity in local extension (S factor) was 58.0%. 3) Sensitivity in lymph node involvement (N factor) was 50.0%. 4) Sensitivity in final staging diagnosis, dividing colon cancer into two groups below st II and above st III, was 63.3%. 5) Further study should be necessitated to provide useful information for preoperative staging diagnosis of colon cancer.

Frequencies of HER-2/neu expression and gene amplification in patients with oesophageal squamous cell carcinoma.

The utilisation of antitumour T cells induced by cancer vaccination with HER-2 peptides or antibodies (Herceptin) against HER-2, as immunotherapy for oesophageal cancer, is a novel and attractive approach. It is important to clarify the frequencies of HER-2 expression and gene amplification in patients with oesophageal squamous cell carcinoma (SCC) and to evaluate the relationship between HER-2 status and HLA haplotype, since the candidates for HER-2 peptide-based vaccination are restricted to a certain HLA haplotype. We determined the frequency of HER-2 expression using the HercepTest for immunohistochemistry and HER-2 gene amplification by fluorescence in situ hybridisation (FISH) assay in oesophageal SCC (n=66). HER-2-positive tumours (1+/2+/3+) analysed by a HercepTest were observed in 30.3% of all the patients and HER-2 gene amplification evaluated by FISH was observed in 11.0% of all the patients, in which all HercepTest (3+) tumours were found to have gene amplification and three of six moderately positive (2+) tumours showed gene amplification. Furthermore, HER-2-positive cells were present more diffusely and were larger within each tumour in the patients who were HercepTest 3+ than those who were HercepTest 1+. Moreover, the survival rate in HER-2-positive group was significantly worse than that in HER-2-negative group. Also, the survival rate in the patients with HER-2 gene amplification was significantly worse than that without HER-2 gene amplification. In addition, oesophageal SCC patients with both HLA-A24-positive and HER-2-positive tumours (1+/2+/3+) accounted for 26% of these cases, and both HLA-A2- and HER-2-positive tumours accounted for 18% of them.

Pharmacokinetic study of the enterohepatic circulation of acetaminophen glucuronide in rats.

A pharmacokinetic model of the enterohepatic circulation of acetaminophen glucuronide was investigated in rats with particular attention to a lag time between biliary excretion and reabsorption. The plasma drug data obtained after acetaminophen glucuronide injection into the various sites of the gut confirmed that there is a lag time in the enterohepatic circulation and that the lag time is due to the intestinal transit period of the conjugate to the site of the hydrolysis. The value of the lag time was fairly close to that reported previously in the rat. Based on the result, a compartment model with periodic trigonometric function for the enterohepatic circulation was built up and the urinary excretion data were fitted to this model. Same parameters which are considered to be common to other glucuronide conjugates were in good agreement with those reported previously, indicating that the model and those values are useful to study the enterohepatic circulation for glucuronides of other compounds in the rat.

Production of recombinant mite allergen Der fI in insect cells and characterization of products--removal of pro-sequence is essential to IgE-binding activity.

Der fI is a major mite proteinaceous allergen found in house dust. We produced recombinant Der fI (reDer fI) in insect cells using a baculovirus expression system. Based on the molecular mass and N-terminal amino acid (aa) sequence analysis, reDer fI was found to be compromised of a mixture of two proteins, each of which includes a pro-sequence of different length. The reDer fI had IgE-binding activity at only 20% of that of native Der fI. The removal of the pro-sequence in an acidic solution drastically increased IgE-binding activity to almost the same as that of native Der fI, showing that the presence of the pro-sequence is inhibitory to the IgE-binding activity of Der fI.

The significance of the sweat test in chronic pancreatitis.

In order to study the disposition which is thought to be latent in chronic pancreatitis, we investigated the sweat chloride concentration of 95 normal subjects, 43 cases of chronic pancreatitis, 12 cases of cholelithiasis, 15 cases of peptic ulcers, 16 cases of hepatic diseases and 23 cases of diabetes mellitus with the sweat test, using the method of pilocarpine iontophoresis. We obtained the following results. (1) In normal subjects, the sweat chloride concentration was inclined to rise gradually with age from childhood to adulthood; the mean value of sweat chloride concentration was 30.0 mEq/liter in adults from 20 years old, and the upper limit was about 60 mEq/liter. (2) The mean value of sweat chloride concentration was 60.0 mEq/liter in chronic calcifying pancreatitis; this value was markedly higher than that of control subjects of the same age (p is less than 0.001). (3) The mean value of sweat chloride concentration in cholelithiasis, peptic ulcer and hepatic diseases did not differ significantly from control subjects. The mean value of sweat chloride concentration in diabetes mellitus was significantly higher than that of control subjects (p is less than 0.01), but was significantly lower than that in chronic pancreatitis (p is less than 0.01). (4) It was supposed that some cases of chronic pancreatitis have a congenital disposition toward abnormal secretion of sweat glands and epithelium in the pancreatic duct, resembling cystic fibrosis, and this disposition leads easily to pancreatic disorders when the individual is exposed to various external factors.

Non-A, non-B hepatitis in experimentally infected chimpanzees with the NIH strains F and H.

The NIH strains F and H of non-A, non-B hepatitis were studied by transmission to chimpanzees. They developed biochemical and ultrastructural evidences of non-A, non-B hepatitis. Chimpanzee's plasma containing the strain F was passaged further three times in the present study, and the results were identical to the original report made by SHIMIZU et al. In contrast to the original report, however, the prototype strain H inoculum or human plasma supposedly containing only the strain H, induced both SHIMIZU's cytoplasmic and nuclear changes in one chimpanzee. This result may pour further oil on the flame of recent controversy regarding the sequestration of the strains H and F, but we believe at present that the strains H and F are separate infectious agents. This is based on the fact that the strain F never induced SHIMIZU's nuclear changes, and our detection of an additional infectious agent of non-A, non-B hepatitis identical to the NIH strain H. We suspect the NIH strain H inoculum provided to us was contaminated by the strain F. The conclusion of controversy regarding the strain H should await for further studies including a further cross challenge study between the strain H and the strain F. Ultrastructural changes of the chimpanzee's liver associated with non-A, non-B hepatitis are described.

Further studies of 48-1 antigen in serial liver biopsies from chimpanzees infected with hepatitis delta virus.

The 48-1 antibody, initially reported to react specifically with non-A, non-B infected liver tissue, has been found to react also with liver specimens from chimpanzees infected with hepatitis delta virus (HDV). To clarify further the relation between HDV and appearance of the antigen reacting with the 48-1 antibody (48-1 Ag), immunoperoxidase studies were carried out on serial liver specimens from chimpanzees infected with HDV. Immunohistochemical and serological findings suggested that the appearance of 48-1 Ag paralleled that of HDV. Double immunoperoxidase staining revealed HDAg in the nucleus and 48-1 Ag in the cytoplasm of the same hepatocytes as well as in different hepatocytes separately. The course of appearance of microtubular aggregates paralleled that of 48-1 Ag. The present results suggested that expression of 48-1 Ag was related to infection with HDV, probably because expression of this antigen is induced from the host genome.

Time-related changes in the distribution of 45Ca in the developing enamel of rat incisors as revealed by radioautography.

The distribution and movement of calcium through the enamel organ and into the forming enamel was examined by means of quantitative 45Ca radioautography in rat incisors. Dislocation of radiocalcium in the specimen was minimized during histologic and radioautographic processing by using rapid freeze/freeze-substitution and dry emulsion coating methods. At 30 sec. after the 45Ca injection, distinct peaks of radioactivity occurred in the connective tissue immediately adjacent to the enamel organ and the infranuclear compartment of secretory ameloblasts. An intense labeling also occurred in the superficial layers of the forming enamel extending 15 microns below the surface. The grain density in the distal cytoplasm of secretory ameloblasts increased at later time periods, whereas all other regions of the enamel organ showed a considerable decrease in radioactivity. The radioactivity in the infranuclear compartment of ameloblasts with numerous mitochondria remained relatively high at 2 min. but was abolished by 10 min. after the injection. The grain density in the enamel matrix became much stronger but the labeled regions only extended to 20, 30, and 40 microns below the surface at 2, 10, and 60 min. after the injection, respectively. The application of wet emulsion over similarly prepared sections caused a severe dislocation of radiocalcium in the specimens. These data confirmed the rapid penetration of systemically administered radiocalcium into the surface layers of forming enamel and its slow diffusion to the deeper layers. The time-related changes in relative grain densities at various regions of the ameloblasts support the coexistence of a relatively slow transcellular pathway for calcium through the secretory ameloblast layer.

Production of antibodies directed against microtubular aggregates in hepatocytes of chimpanzees with non-A, non-B hepatitis.

We have previously used Epstein-Barr virus transformation to established two clonal lymphoblastoid cell lines (48-1 and S-1) producing monoclonal antibodies against microtubular aggregates that appear in the hepatocytes of chimpanzees with non-A, non-B hepatitis (NANBH). To obtain additional antibodies directed against the same structure, the mouse hybridoma method was employed. Partially purified microtubular aggregates were prepared from liver homogenates of a chimpanzee with NANBH and used as the immunogen. Hybridoma cultures were first screened by radioimmunoassay against the partially purified antigen and secondly by immunofluorescence (IF) using liver sections from a chimpanzee with NANBH. Twenty-seven cultures exhibited positive IF reactions similar to those observed with the original antibodies, 48-1 and S-1, and were cloned by limiting dilution. The specificities of the monoclonal antibodies were tested by IF on liver biopsy specimens from chimpanzees with hepatitis A, B, D or NANBH and from normal chimpanzees. All the antibodies proved to be IgG. Immunoelectron microscopy revealed that all 27 antibodies bound to the same structure, the microtubular aggregates, in hepatocytes of chimpanzees with NANBH. To determine the size of the antigen polypeptide recognized by these antibodies, polyacrylamide gel electrophoresis and Western blot assays were performed. Nine of the 27 antibodies specifically reacted with a single polypeptide of Mr 44K (p44). The remaining 18 antibodies detected no antigen polypeptide on the filters. The anti-p44 antibodies were then tested using cross-competition assays with 125I-labelled antibodies, and were found to be classifiable into three groups. In addition, the results indicate that at least three distinct epitopes are located on p44: epitope A recognized by group 1, epitope B recognized by group 2 and epitope C recognized by group 3.

Superconductivity and magnetic fluctuations in Cd(2))Re(2)O(7) via Cd nuclear magnetic resonance and re nuclear quadrupole resonance.

We report Cd nuclear magnetic resonance (NMR) and Re nuclear quadrupole resonance (NQR) studies on Cd(2)Re(2)O(7), the first superconductor among pyrochlore oxides (T(c) approximately 1 K). The Re NQR spectrum at zero magnetic field below 100 K rules out any magnetic or charge order. The spin-lattice relaxation rate below T(c) exhibits a pronounced coherence peak and follows the weak-coupling BCS theory with nearly isotropic energy gap. The results of Cd NMR point to a moderate ferromagnetic enhancement at high temperatures followed by a rapid decrease of the density of states below the structural transition temperature of 200 K.

Secretin-like bioactivity in the duodenal mucosa in patients with peptic ulcer and chronic pancreatitis.

Biopsy specimens of the duodenal mucosa were assayed to determine their secretin-like activity in 9 controls, 9 patients with gastric ulcer, 19 patients with duodenal ulcer, 4 patients with gastric and duodenal ulcer, and 13 patients with chronic pancreatitis. The bioassay of secretin was done on the pancreatic secretion in anesthetized rats. The sensitivity was in the orcer of 0.0625 CHR unit/rat (4 ng/rat). In the range between 0.0625 and 0.5 CHR units a satisfactory dose dependency was recognized. The following results were obtained. 1) The level of duodenal mucosal secretin-like activity in patients with gastric ulcer was the same as that in the controls, but was elevated in 32% of the patients with duodenal ulcer, 50% of those with gastric and duodenal ulcer, and 8% of those with chronic pancreatitis. 2) The high level of secretin-like activity noted in patients with duodenal ulcer was suspected to be related to the hypersecretion of gastric acid which is characteristic of this disease, but there was no correlation between gastric acid secretion and secretin-like activity in the duodenal mucosa.

Reserve capacity of bicarbonate secretion in chronic pancreatitis.

When secretin was given by continuous intravenous infusion in the control subjects, the dose of secretin inducing maximal bicarbonate output was found to be around 6.0 CHR U/kg/hr. Then the pancreatic exocrine secretory response to sequential standard (1.2 CHR U/kg/hr) and augmented (6.0 CHR U/kg/hr) dose of secretin was studied in the controls, in patients with chronic pancreatitis and in its suspected cases. This new method for exocrine pancreatic function did not offer advantage for the diagnosis of well established chronic pancreatitis. But from the results obtained in suspected chronic pancreatitis it was supposed that the decline of increasing rate of bicarbonate output with augmentation of dose and the decrease of response to augmented dose of secretin might be one of functional disorders occurred in the early stage of chronic pancreatitis.

Muon spin relaxation and susceptibility studies of the pure and diluted spin 1/2 kagomé-like lattice system (CuxZn(1-x))3V2O7(OH2) 2H2O.

Muon spin relaxation and magnetic susceptibility measurements have been performed on the pure and diluted spin 1/2 kagomé system (CuxZn(1-x))3V2O7(OH)2 2H2O. In the pure x=1 system we found a slowing down of Cu spin fluctuations with decreasing temperature towards T approximately 1 K, followed by slow and nearly temperature-independent spin fluctuations persisting down to T=50 mK, indicative of quantum fluctuations. No indication of static spin freezing was detected in either of the pure (x=1.0) or diluted samples. The observed magnitude of fluctuating fields indicates that the slow spin fluctuations represent an intrinsic property of kagomé network rather than impurity spins.