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A sharp rise in the number of patients with infectious gastroenteritis was observed in the 25th week of year 1996 in the Takahashi-Ashin district by researchers with the Infectious Disease Surveillance Program for tuberculosis and other infectious diseases in the Okayama Prefecture. This sharp rise occurred coincidentally with an outbreak of enterohemorrhagic Escherichia coli O157:H7 (EHEC O157) infection in Niimi City of the Takahashi-Ashin district. However, this phenomenon of coincidental outbreaks was not observed during the outbreak of EHEC O157 infection in Oku Cho. By reviewing outpatients' charts in a sentinel hospital in Niimi City for the Infectious Disease Surveillance Program, it was noted that patients with acute gastrointestinal infection visiting the hospital during the increased incidence of infectious gastroenteritis may have been included as misclassified cases of EHEC O157 infection. On the other hand, the exponential probability plotting of symptomatic patients with EHEC O157 infection in Niimi City revealed a breaking point which suggested a dual exposure to contaminated food or an overlap with other acute gastrointestinal infections. The latter possibility was discounted, because stool culture-positive patients with EHEC O157 infection also exhibited a similar breaking point, and furthermore, the coincidental increase in infectious gastroenteritis in the same area was attributable to the EHEC O157 infection. The present study demonstrates the association between the sharp rise in gastroenteritis and the outbreak of EHEC O157 in the Takahashi-Ashin district. A careful analysis of the cases of infectious gastroenteritis by the Infectious Disease Surveillance Program would have predicted the outbreak of EHEC O157.

The rat model is ideal for investigating various reactions to small intestinal transplantation (SIT). The conventional surgical model (hand-suture method), however, requires microsurgical techniques and remains difficult for beginners to perform at a high success rate. We have employed the SIT model using the cuff method, by which the vessels are anastomosed without sutures. All of the fellows who used the hand-suture models needed over 8 +/- 5.8 months until they achieved a 70% success rate. In contrast, the fellows employing the cuff method mastered SIT models after 6 weeks' practice. The cuff technique is a simplified and quickly mastered alternative to the hand-suture method that may be desirable for researchers who wish to apply the method to SIT experiments and whose primary purpose is not microsurgery.

We have developed a reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus (HCV) RNA using in vitro synthesized Renilla luciferase (Rluc) RNA as an internal control. Using this method, the 5'-noncoding region of HCV RNA (144 nucleotides) and Rluc RNA (276 nucleotides) were efficiently amplified in a single tube, and the sensitivity and specificity of this method were comparable to standard RT-nested PCR. This method was successfully performed on RNA specimens obtained from in vitro HCV-infected human hepatocyte PH5CH8 cells, which support HCV replication. In addition, we demonstrated that this method was useful for the evaluation of antiviral reagents by confirming the anti-HCV activity of bovine lactoferrin, which we previously found to be a new inhibitor of HCV infection. Therefore, this method may be useful for the studies of not only HCV but also of other viruses.

The accessory ascending cervical artery (Murakami et al., 1996), which arises from the subclavian artery and ascends between the scalenus anterior and medius muscles, was studied in 87 Japanese adult cadavers (174 sides), with special attention being given to its origin, distribution, and relationship to other arteries at the cervical or scalenus region. In 154 sides (88.5%), the accessory ascending cervical artery was found to originate from the subclavian artery behind the scalenus anterior muscle, and to branch out to the scalenus anterior and medius muscles as well as those entering the 5th and 6th intervertebral foramens along the 6th and 7th cervical nerves. This artery arose independently in 105 sides. The accessory ascending cervical artery issued off or formed a common trunk with the transverse cervical artery and/or costocervical trunk in 49 sides. In cases lacking the accessory ascending cervical artery, it was usually compensated for by the costocervial trunk and/or transverse cervical artery (18 sides). Common trunk formation with the vertebral, internal thoracic, or suprascapular arteries was not observed. The authors suggest that the accessory ascending cervical artery, the transverse cervical artery, and the costocervical trunk should be grouped into one arterial system, a system that may be a remnant of the precostal longitudinal anastomoses of intersegmental arteries of the dorsal aorta behind the scalenus anterior muscle.

We examined the effects of FK506 and its non-immunosuppressive derivative, GPI1046, on H2O2-induced reduction of cell viability and apoptotic cell death in Neuro 2A cells. Our results suggest that the protective properties of GPI1046 against H2O2-induced reduction of cell viability are equipotent with those of FK506 and may be mediated by increased intracellular concentrations of glutathione (GSH). In addition, both FK506 and GPI1046 prevented apoptotic cell death in Neuro 2A cells, although the antiapoptotic effect of FK506 was somewhat stronger than that of GPI1046. These findings suggest that non-immunosuppressive immunophilin ligands such as GPI1046 might be potentially useful in treatment of neurodegenerative diseases without serious side effects such as immune deficiency.

The applications of ultraviolet (UV) light (336 nm) on the upper oesophageal strips of frog elicited relaxant responses in the presence of NaNO2 (50 microM). The tissues were mounted under the tension 0.5 g in an organ bath containing Ringer solution, maintained at 25 degrees C and gassed with 100% O2. The responses were recorded on a kymograph via an isotonic lever. Antimegaloblastic agents, including hydroxocobalamin (1, 10, and 100 microM), cyanocobalamin (1, 10, 25, and 100 microM), and folic acid (1, 10, 50, 100, and 200 microM), significantly attenuated the relaxation response to UV light. Folinic acid (1, 10, 25, and 100 microM), however, enhanced the relaxation. Pyrogallol (50 microM), hydroquinone (50 microM), and diethyldithiocarbamic acid (8 mM) were found ineffective for attenuation, though FeSO4 (200, 400, and 500 microM) and hemoglobin (50 microM), respectively, exerted significant inhibition. L-arginine methylester (500 microM) did not impair UV-induced relaxation. Based on these results, we concluded that a mechanism involving undefined action(s) of antimegaloblastic drugs may cause alterations in the UV light-induced relaxation of the tissue used.

Although the rehabilitation of patients with chronic obstructive pulmonary disease (COPD) improves both exercise capacity and quality of life, a standard protocol for COPD patients has not been established. To clarify whether physiologic and quality-of-life improvements can be achieved by an inpatient pulmonary rehabilitation program 5 days per week for 3 weeks, 18 patients with COPD were enrolled in a rehabilitation program. The physical exercise training regimen consisted of respiratory muscle stretch gymnastics and cycle ergometer exercise training. Pulmonary function tests, an incremental ergometer exercise test, a 6-min walking test, and a quality of life assessment by the Chronic Respiratory Questionnaire were administered before and after the program. The peak VO2, an indicator of maximal exercise capacity, did not increase, although the 6-min walking distance, an indicator of functional exercise capacity, increased significantly after rehabilitation. There was a significant improvement in the quality of life in terms of dyspnea, fatigue, and emotional state. These findings suggest that even a 3-week program may be beneficial for COPD patients. Increases in functional exercise capacity, even without an increase in maximal exercise capacity, are helpful for reducing dyspnea and improving quality of life parameters in patients with COPD.

1. In the normal rabbits the intravenous application of adrenaline cause hyperglycaemie, but in rabbits which have been previously splanchniectomized on both sides, the adrenaline application always betrayed hyperglycaemia. 2. In the normal rabbits ergotoxine had no influence on the blood sugar content. Contrary to the normal animals, a rabbit which received previously an intravenous injection of ergotoxine by which the vasoconstriction of the blood vessels was made impossible, sugar puncture was not effected in the blood sugar content. 3. Contrary to the normal animals, in the rabbits which were treated previously with intravenous application of ergotoxine, the electrical stimulation of the basis of the brain did not cause hyperglycaemia. 4. By the perfusion of the liver of toad with Ringer solution, on adding of adrenaline in the perfusing liquid, discharge of the sugar in the perfusate was increased in percentage, but the absolute quantity was reduced on account of the reduction of the amount of perfusate. 5. The discharge of sugar from the perfusing liver of the toad, which was previously perfused sufficiently with ergotoxine-Ringer solution, on adding of adrenaline in the perfusing liquid showed no change compared with these without ergotoxine and adrenaline application. 6. The piqure-hyperglycaemia may be assumed as adrenaline-hyperglycaemia. The origin of adrenaline-hyperglycaemia is rather mechanical, since the torrent of blood stream is increased by the strong contraction of blood vessels of skin and muscles than those of liver, washing out the reserved sugar in liver.

The formation of the swim-bladder gases of some sea and fresh water fishes were investigated and the results may be summarized as follows : 1. As a rule, oxygen content in the swim-bladder is higher in a fish living at greater depth than at shallow, and sea water fishes, than fresh water ones. 2. Oxygen content in the swim-bladder of the fish living at great depth decreases after 1-2 days stay in the aquarium. 3. Carbon dioxide content in the swim-bladder of all fishes examined is very small. 4. Through the poisoning of carbon monoxide, the swim-bladder gas decreases in its oxygen content and increases slightly in its carbon dioxide. 5. Corresponding to the artificial increase or decrease of the external pressures influencing the body surface of the fish, oxygen content of the swim-bladder gases increases or decreases respectively. 6. After the evacuation of the swim-bladder gases, newly formed gases always contain high oxygen percentage. 7. When oxygen or carbon dioxide of high concentration are injected in the swim-bladder, these gases diffuse out easily through the wall of the swim-bladder during 1-2 days. 8. Oxygen dissociation curve of carp blood is remarkably steep compared with the human blood, and influenced very much with the presence of carbon dioxide so as to decrease the affinity of the blood to oxygen. 9. Histological examination of the swim-bladder of Sebastiscus marmoratus and Carassius auratus indicate characteristic structure of the blood capillaries which distributed in the internal layer of membranes and sinus-like dilated. 10. From the above experimental results, some considerations on the gas formation in the swim-bladder were offered.

1. Veratrine greatly sensitizes the action of potassium ions on frog's rectus muscle. Quinine inhibits the sensitizing action of veratrine. 2. Perfusate of the preparation from the stimulated veratrinized hind limbs evokes a contraction of the veratrinized test muscle. 3. The other purfusates of the preparations perfused with Ringer's solution or with veratrine-Ringer's solution do not affect the test muscle. But a similar contraction is observable if potassium chloride of a certain strength, is added to the perfusates. 4. It can be shown that the chemical agent causing the muscular contraction is potassium ions, 5. The above experimental results may be wholly explained by the assumption that veratrine increases the permeability of muscle cell membrane to potassium ions.

A patient with a diffuse, small cleaved cell, non-Hodgkin's lymphoma associated with marked hypecalcemia was described. Antibody to the adult T-cell leukemia-lymphoma virus was absent. Although bone marrow was infiltrated by lymphoma cells, destructive or lytic bone lesions could not be detected. The serum level of immunoreactive parathyroid hormone C-terminal (PTH-C) was normal. The serum level of 1, 25-dihydroxyvitamin D was lower than normal. This case suggests that other humoral substances produced by lymphoma cells may be responsible for hypercalcemia.</P>

The cardioprotective effect of calmodulin antagonists, trifluoperazine (TFP) and N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide (W-7) was examined on the isolated rat heart exposed to hypothermic and ischemic conditions by measuring distribution of lysosomal enzymes in myocardial cells, and leakage of creatine kinase (CK) during reperfusion and postischemic recovery in myocardial systolic function. Experimental hearts were infused with 20 degrees C Krebs-Henseleit bicarbonate buffer (KHB) or KHB containing TFP or W-7 for 2min every 30min during hypothermic ischemia. After ischemia for 120min at 20 degrees C, rat hearts were reperfused at 37 degrees C for 30min. TFP and W-7 improved functional recovery and prevented CK release. In TFP treated hearts, leakage of lysosomal enzymes was reduced significantly, whereas stabilization of lysosomes by W-7 did not occur. These results suggest that calcium-calmodulin dependent enzymes may play an important role in the development of cellular damage of the myocardium during hypothermic ischemia, although levels of leakage of lysosomal enzymes may be unreliable predictors of functional recovery after hypothermic ischemia.</P>

A monoclonal antibody (MAb), OPT1, reactive with T cells in formalin-fixed, paraffin-embedded tissue sections, has been identified through immunization with activated T cells from peripheral blood lymphocytes (PBL). The antibody is an IgG1 antibody as demonstrated by the Ouchterlony technique. By cytofluorometric analysis, almost all CD3+ lymphocytes and only a few CD20+ lymphocytes of peripheral blood expressed the OPT1 antigen. Nonhematolymphoid cell lines were negative for OPT1 by the immunoperoxidase staining using acetone-fixed cell lines. On the contrary, peripheral T cells, cells of two T cell lines out of four and a part of the cells of one B cell line out of two were positive for OPT1. The immunoperoxidase staining of paraffin-embedded tissue sections revealed that most of lymphocytes in T cell areas of lymph nodes expressed OPT1 antigen. Some lymphocytes in both cortex and medulla of the thymus and erythroid precursors of the bone marrow were OPT1+. In the malignant lymphoma series, approximately 90% of T cell lymphomas and 6% of B cell lymphomas reacted with OPT1. None of the Reed-Sternberg cells nor Hodgkin cells in Hodgkin's disease were positive. Consequently, OPT1 may be useful for the diagnosis and study of malignant lymphomas and other related lesions.</P>

In order to clarify difference of the mucosal immunity in various sites of normal large and small intestines, we studied the population of lymphocyte subsets and immunoglobulin (Ig)-containing cells in situ in biopsy specimens taken from various sites (ascending colon, sigmoid colon and rectum) of the large intestine and from the duodenum using an immunohistochemical method. Monoclonal antibodies against pan-T (Leu 1), cytotoxic/suppressor T (Leu2a), helper/inducer T (Leu3a), suppressor T (Leu15) and natural killer/K (Leu7) cells, and polyclonal antibodies to human IgG, IgA and IgM were used. In the duodenum, intraepithelial lymphocytes (IELs) were more prominent than in the large intestine. Immunoelectron microscopic observation revealed that some Leu2a+ IELs possessed pseudopods extending into intestinal epithelial cells, indicating that some IELs belong to the cytotoxic T cell subset. Leu7+ IELs were scarcely observed and Leu7+/Leu1+ ratio was higher in the large intestine than in the duodenum. Furthermore, the number of Leu7+ cells were more in the distal than the proximal colon. In the lamina propria Ig-containing cells tended to be fewer in the rectum than in the duodenum and the proximal colon. Our findings may suggest the variation of local immune responses and the difference of assigned immunological functions among the various sites of the intestines.

To gain further insight into the central nervous system (CNS)-action of beta-adrenergic blocking agents (beta-blockers), we examined the effects of various kinds of beta-blockers on opioid receptors (Op-Rs) using radiolabeled receptor assay (RRA). We demonstrated that beta-blockers are competitively bound to Op-Rs in the CNS. Sodium index of beta-blockers in [3H]naloxone binding study indicated that beta-blockers had the mixed agonist-antagonist activity of opiates. The relative potency of beta-blockers in opioid RRA was negatively correlated with their membrane stabilizing activity. Neither beta-blocking activity nor intrinsic sympathomimetic activity was correlated with IC50 values of beta-blockers in opioid RRA. While it is widely accepted that beta-blockers have a tranquilizing activity, a part of the tranquilizing action of beta-blockers may be mediated through Op-Rs in the CNS. Although beta-blockers may have effects on their own receptors (beta-receptors) in the CNS, the more precise mechanisms of central action of these drugs must be further investigated.

Effects of the mesenteric nerve stimulation (MNS) on the twitch contraction induced by field stimulation were investigated regarding the relationship between myenteric neurons and extrinsic cholinergic nerves in the guinea-pig mesenteric nerve-ileal preparation. The twitch contraction was inhibited after MNS. The inhibition of the twitch contraction after MNS was induced twice, just after MNS (1st inhibition) and 2-3 min later (2nd inhibition) (type I), or once, just after MNS (1st inhibition) (type II), in recovery course of twitch contraction for 6-8 min. The 1st inhibition was slightly decreased by guanethidine and hexamethonium. The inhibitory response (1st inhibition) in both types I and II was recovered to the control level by pretreatment with naloxone (recovered twitch contraction), but the late inhibitory response (2nd inhibition) was markedly observed after 2-3 min in types I and II. Either the 1st or the 2nd inhibition was not altered by capsaicin, desensitization to calcitonin gene-related polypeptide (CGRP), vasoactive intestinal polypeptide (VIP), somatostatin, or galanin. The recovered twitch contraction in types I and II was decreased by CGRP-desensitization, or capsaicin. These results suggest that the first inhibitory response was induced by enteric opioid neurons connected with extrinsic cholinergic nerves, but the 2nd inhibition was induced by unknown substances other than CGRP, VIP, somatostatin, and galanin. The twitch contraction may partly be induced by endogenous neurokinin-like substances. And, some CGRP containing neurons, which connect with extrinsic cholinergic nerves, probably activate the intrinsic excitatory neurons.

We studied the use of high-performance liquid chromatography (HPLC), using a solid phase extraction column (Bond Elut cartridge column), for the simple, rapid and sensitive determination of serum clonazepam levels in epileptic patients. Extracted aliquots were analyzed by HPLC, using a reverse phase ODS column (mu-Bondapak C18). The analytical mean recovery of clonazepam added to the blank serum averaged 99.9%. The detection limit was as high as approximately 2 ng/ml in the serum. The reproducibilities were 2.3-8.6 CV % in the within-day assay and 6.5 CV % in the between-day assay, indicating that the analysis method was effective in the determination of clonazepam serum levels. Accordingly, we suggest that the present method, using a solid phase extraction column, may be useful for the routine monitoring of clonazepam serum levels in epileptic patients.</P>

Hematoxylin and eosin (H-E) stained liver sections of 47 autopsy cases of hepatic malignancies were examined. There were 43 cases of hepatocellular carcinoma (subtypes of 30 trabecular, 7 solid, 5 pseudoglandular, and one scirrhous carcinoma), 3 of cholangiocellular carcinoma and one of mixed carcinoma. After immunohistochemical staining, benign hepatocytes reacted positively with anti-epithelial membrane antigen (EMA). Hepatocellular carcinoma cells reacted more weakly than benign hepatocytes. It was noted that the microtubular structure, which could not be demonstrated even by alcian blue or cationic ferric hydroxide colloid stabilized with cacodylate (Fe-CaC), was clearly detected with anti-EMA. The EMA-positive microtubular structures may indicate terminal cholangiolar differentiation. Based on EMA, seven more cases formerly classified as hepatocellular carcinoma by H-E were reclassified as mixed carcinoma, totaling eight (17.0%). The histologic classification of "mixed carcinoma" has been 1.5 to 2.0% of primary liver cancers in Japan, but we suggest there may be more cases of "mixed carcinoma" identified in the future. In conclusion, we emphasize that EMA staining is useful for more accurate classification of hepatic tumors.

Proportional changes of lymphocyte subsets in the peripheral blood were monitored by two-color flow-cytometry in seven leukemia patients who had received allogenic bone marrow transplantation (BMT). Lymphocyte counts, and proportions of T and B-cells returned to normal ranges between the 2nd and 12th months after BMT. Activated T-cells prominently increased after BMT, and the values gradually returned toward normal. As to lymphocyte subsets, the proportions of CD 4+ cells had remained low, while those of CD 8+ cells high for a whole observation period after BMT. The changes of CD 4+ cells were caused by the decrease of suppressor-inducer T-cells (CD 4+ Leu 8+). High proportion of CD 8+ cells was mainly associated with increased suppressor T-cells (CD 8+ CD 11+). Among natural killer (NK) cells, highly active NK cells (CD 16+ CD 57-) markedly increased shortly after BMT, and gradually returned to normal. CD 16 -CD 57+ NK cells increased beyond normal ranges after the 2nd month. The incidence or degree of acute and chronic graft-versus-host diseases (GVHD) did not correlate with the changes of any lymphocyte subsets. The present results suggest that the increase of activated T-cells shortly after BMT reflects lymphocyte reconstitution. The prolonged immune deficiency after BMT might be related to either deficient expression of homing receptor (Leu 8 antigen) on CD 4+ cells or increased suppressor T-cells (CD 8+ CD 11+). In addition, the early increase of NK cells after BMT may compensate for the immune deficiency in BMT patients.