Effects of sound stimulation on the central artery of the rabbit ear were studied as a somato-autonomic reflex. Vasoconstriction and dilatation, caused by metronome sound stimulation, were estimated from the temperature fluctuations in the central artery of the ear, measured by a thermistor. To enhance the detection of temperature rises, moderately high background levels of arterial tone were established by exposing the tips of the ears to water at a temperature of 10°C or 5°C, prior to sound stimulation. A fall in arterial temperature due to vasoconstriction was observed immediately after the start of the 1-min sound stimulation, with a subsequent temperature rise which overshot the original basal level due to vasodilatation. A positive correlation between the ear temperature before sound stimulation and the temperature fall (p<0.01), and a negative correlation between the ear temperature and the temperature rise (p<0.05) were obtained. The temperature fall was blocked by phenoxybenzamine (9 mg/kg, i.p., p<0.01). The subsequent rise was not influenced by atropine (3 mg/kg, i.p.) or phenoxybenzamine, however, it was attenuated by hexamethonium (6 mg/kg, i.p., p<0.05). The temperature fall at the beginning of sound stimulation was related to alpha-adrenergic mechanism. The subsequent temperature rise was thought to be related to parasympathetic mechanism, excluding cholinergic mechanism.
For the purpose of establishing a new in vitro model of adoptive immunotherapy, we synthesized two kinds of bispecific antibodies (BsAbs), i.e., (OK×L) BsAbs constructed with both OKT-3 (anti-CD3) and L-7-6 (anti-HCC), and (3G×L) BsAbs constructed with 3-G-8 (anti-CD16) and L-7-6 antibodies. These two BsAbs, having pairs of binding arms on their single molecule, showed similar binding to target cells as the parental monoclonal antibodies (OKT-3, 3-G-8 and L-7-6), when examined with FACS. Newly devised in vitro cytotoxicity tests revealed that LAK or PWM-stimulated LAK (PWM-LAK) cells did not show any significant cytotoxic activity to HCC cells, while both effector cells equally showed greatly enhanced cytotoxicity to HCC even at a low effector/target (0.3) in the presence of BsAbs (OK×L) for the efficient retargeting of the effector cells. Inasmuch as PWM-LAK cells proliferate in vitro 3-5 times faster than LAK cells, adoptive immunotherapy using PWM-LAK cells in combination with (OK×L) BsAbs should be very promising.
Carbon-11 labeled metabolites in human plasma were analyzed by high- performance liquid chromatography during positron emission tomography (PET) studies using the dopamine D2 ligand [11C]YM-09151-2 as well as the histamine H1, ligands [11C] doxepin and [11C] pyrilamine. For all the three tracers, blood clearance of the radioactivity was extremely rapid after an i.v. injection. The plasma protein-binding of [11C]YM-09151-2 and [11C] doxepin had protective effects upon the metabolic alteration of the ligands, whereas [11C] pyrilamine was free from the protein-binding and immediately degraded. The degradation of [11C] doxepin was more rapid in epileptic patients on medication than in normal subjects. These results indicate that analysis of metabolites in the plasma is necessary to determine the accurate arterial input function for quantitative PET measurement.
How 4β-phorbol 12-myristate 13-acetate (PMA) and ionomycin (Io), a calcium ionophore, affect on the atrial natriuretic peptide (ANP) stimulated cyclic-3', 5'-guanosine monophosphate (cGMP) production in cultured rat mesangial cells was examined. Cultured mesangial cells were prepared by isolated glomeruli from Sprague Dawley rats employing the sieving method and were used between the 3rd and 15th passage for experiments. cGMP and protein contents were measured by radioimmunoassay and Lowry method. Incubations with effectors were carried out either in the presence or absence of 0.5 mM 1-methyl-3-isobutyl-xanthine (MIX). The intracellular concentration of calcium ([Ca2+]i) was determined by using the Fura-2 method. Pretreatment with PMA, an activator of protein kinase C (PKC), attenuated ANP stimulated cGMP production in a time- and dose-dependent fashion, while αPDD (an inactive analog of PMA) did not inhibit cGMP production. PMA inhibition was reversed by addition of staurosporine, a protein kinase C inhibitor. Io attenuated ANP stimulated cGMP production in the absence but not in the presence of MIX. These findings suggested that PMA acts on ANP receptor or guanylate cyclase via activation of PKC in rat mesangial cells. Io may inhibit ANP stimulated cGMP production via activation of cyclic nucleotide phosphodiesterase.
The role of intestinal luminal bile on cholecystokinin (CCK) and gastric inhibitory polypeptide (GIP) release was examined in dogs with chronic external bile diversion. In 6 mongrel dogs, cholecysto-jejuno-cystostomy (C-J-C) was performed using a small segment of the middle small intestine interposed between the gallbladder and urinary bladder with section of the common bile duct. Butter dissolved in 30 ml of lukewarm water was orally ingested before surgery, and same amount of butter solution with or without graded volumes of canine bile were orally ingested between four to six weeks after C-J-C surgery. Increases in both plasma triglyceride and GIP levels after butter ingestion were almost completely abolished by C-J-C, but they were restored by oral bile feeding in a volume-dependent manner. Both basal and fat-stimulated CCK release were enhanced significantly by C-J-C, and oral bile feeding inhibited fat-stimulated CCK release by bile in a dose dependent manner. These data suggest that intraluminal bile regulates basal and fat-stimulated CCK release and fat-stimulated GIP release in dogs.
Since the amounts of hepatogenous enzymes discharged into the intestinal tract remain unknown, this study was initiated to evaluate the amounts of the enzymes in the intestinal tract. Whole gut lavage fluid (polyethyleneglycol electrolyte solution) was administered orally to 42 subjects, consisting of 5 patients with hepatoma, 10 with chronic hepatitis, 10 with colon polyps, and 17 control subjects without liver disease. Two hr after the large intestinal lavage, the digestive tract juice was aspirated by colonoscopy, and the bilirubin (Bil), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) in the aspirates were measured. A positive correlation between the AST and LDH values was found, and a significant difference in these values between the hepatic disorders and the normal controls was noticed. A significant positive correlation between the ALP and Bil values was found, and a statistical difference in these values between the group of colon polyps and the controls and other groups was observed. This lavage fluid technique enables to estimate the amounts of hepatic enzymes discharged into the intestinal tract, thereby opening a new avenue for future enzyme research.
Cigarette smoking is considered to be a risk factor of bladder cancer. We investigated the relationship between bladder cancer mortality rates and cigarette smoking prevalence in various countries, and examined whether only the difference of smoking prevalence could explain the difference of mortality rates of bladder cancer in each country or not. The age-adjusted mortality rates from bladder cancer were high in European countries and low in Asian countries. The sex ratios ranged between 2 to 6 in most of the countries. For 20 countries, the mortality rate from bladder cancer was not correlated with the cigarette smoking prevalence. However, when the analysis was restricted to 16 countries in Europe, North America and Oceania (Caucasians), the correlation coefficients between them were high, being 0.68 (p<0.01) in males and 0.49 (0.05<p<0.1) in females. In both Japan and the United States, cigarette smoking prevalence declined while the incidence of bladder cancer increased in males, both being stable in females. Race might be considered to be a strong risk factor of bladder cancer, followed by the prevalence of cigarette smoking. It is necessary to follow the changes of incidence rates of bladder cancer in the future to evaluate the influence of cigarette smoking.
To examine which of the faster or slower large myelinated nerve fibers (alpha fiber group) are affected earlier by diabetes mellitus (DM), the distribution of nerve conduction velocities (DCV) and conventional sensory nerve conduction velocities (SCV) in the median nerve were measured in 10 male DM patients aged 35-58 (mean 50) years. The patients had suffered from DM for 0-30 (mean 11) years; their fasting blood sugar (FBS) levels were 105-363 (mean 219) mg/100 ml. The DCV was expressed by the conduction velocities below which 10, 20, ……, 80 and 90% of active fibers lie (V10, V20, ……, V80 and V90 velocities, respectively). In DM patients, the V30 to V90 velocities of the DCV as well as the SCV were significantly smaller than those of age-matched male controls. The results of multiple regression analysis indicated that FBS was inversely related to V50 to V90 velocities; duration of the disease was also significantly related to V80 and V90 velocities. These data suggest that faster nerve fibers are more susceptible to DM than slower fibers are; changes in conduction velocities of the faster fibers are closely associated with FBS and duration of DM.
After immunizing mice with a human megakaryoblastic leukemia cell line, M-MOK, we obtained two monoclonal antibodies which recognize the human c-kit receptor. The monoclonal antibodies, designated MTK1 and MTK2, were found to specifically recognize Balb/3T3 cells transfected with human c-kit cDNA and not parent Balb/3T3 cells while showing different immunological, biochemical and biological behaviors. Both allowed visualization of the 140 kDa c-kit protein by Western blot analysis, but MTK1 detected only positive band with non-reducing conditions for sodium dodecyl sulfatepolyacrylamide gel electrophoresis. MTK1 partially inhibited the stem cell factor (SCF) induced proliferation of M-MOK cells, whereas, MTK2 was without effect. MTK1 also inhibited the bone marrow derived colony forming unit granulocyte/ macrophage (CFU-GM) formed by granulocyte-macrophage colony stimulating factor (GM-CSF) and SCF. Not only anti-CD34 antibodies (HPCA-1) but also MTK1 could be shown to concentrate bone marrow CFU-GM and burst forming unit erythroid (BFU-E) effectively. The presently described monoclonal antibodies may therefore be useful for functional analysis of the ligand binding domain of the human c-kit receptor, as well as for further classification of hematopoietic stem cells in addition to the CD34 positive cells.
Samples of abnormal endometrium, which were composed of 5 cases of atypical hyperplasia (ATH), 12 cases of well differentiated adenocarcinoma (G1), 17 cases of moderately differentiated adenocarcinoma (G2), 11 cases of poorly differentiated adenocarcinoma (G3) and 2 cases of serous papillary adenocarcinoma (SPA), were analyzed by flow cytometry. With respect to the ploidy level, all cases of both ATH and G1 showed diploidy, and the proliferative index (PI) was 14.32 and 16.33, respectively. In contrast, 3 of G2 cases, 3 of G3 cases and all of SPA cases showed aneuploid patterns, and the PI was 21.06, 27.91 and 31.52, respectively. There were statistical differences between the former group and the latter group as to both the ploidy and the PI. It was found that ATH and G1 had similar biological behavior.