Chemoprevention of heterocyclic amine-induced mammary carcinogenesis in rats

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is one of the most prevalent carcinogenic heterocyclic amines in the environment, targeting the colon, prostate, pancreas, and mammary gland in rodents. Chemopreventive effects of synthetic and naturally occurring compounds on PhIP-induced rat mammary carcinogenesis were investigated in a series of experiments. In a PhIP feeding model, groups of 20-21 female F344 rats each, were treated with 0.02% PhIP alone or PhIP plus 0.5% 1-O-hexyl-2,3,5-trimethylhydroquinone (HTHQ), 1% green tea catechins, 1% alpha-tocopherol, 0.1% ellagic acid, or 1% chlorophyllin, each in the diet, or 0.1% caffeine in drinking water for 52 weeks. To assess the mechanism of HTHQ and caffeine inhibition of PhIP-induced carcinogenesis, effects of these compound on the in vitro metabolic activation of PhIP were examined in the presence of S9 mix. In the next series of experiments, the PhIP intragastric dose model was applied to allow separate investigation of the effects of chemicals during the initiation and postinitiation periods. In these experiments, female Sprague-Dawley rats were given eight intragastric doses of 100 mg/kg body weight during the first 4-8 weeks for initiation. Either during initiation or after initiation, or only after initiation, animals were treated with either corn or perilla oil at doses of 5 and 20%, conjugated fatty acid derived from safflower oil (CFA-S) or perilla oil (CFA-P) at a dose of 1%, arctiin at doses of 0.02 and 0.2% in the diet, or sodium nitrite (NaNO(2)) at a dose of 0.2% in drinking water. In the PhIP feeding model, administration of PhIP alone for 52 weeks induced adenocarcinomas in 40% of rats, but the incidence was remarkably reduced to 5% by the simultaneous treatment with 0.5% HTHQ, a strong lipophilic phenolic antioxidant, or to 10% by 0.1% caffeine. Administration of 1% chlorophyllin exerted similar, albeit weaker, effects. alpha-Tocopherol at a dose of 0.5% only reduced the multiplicity of carcinomas, and 1% green tea catechins only the mean size of mammary tumors. In a metabolic activation study of PhIP, HTHQ and caffeine clearly inhibited the formation of metabolites. In the PhIP gastric dose model, among the naturally occurring compounds examined, a plant lignan arctiin, perilla oil, which contains a large amount of n-6 alpha-linolenic acid, and CFA-S or CFA-P inhibited mammary tumor development, particularly in the postinitiation period, although a clear dose response was not observed. Treatment with 0.2% NaNO(2) in the initiation period was found to lower the volume of mammary tumors. The present results indicate that a number of compounds may be candidate chemopreventive agents against PhIP-induced mammary carcinogenesis, acting through different mechanisms and depending on the stage of carcinogenesis.     

In vivo characteristics of low molecular weight copoly (D,L-lactic acid) formulations with controlled release of LH-RH agonist

Amorphous and crystalline copolymers with a relatively low molecular weight of 1800 were synthesized by direct copolycondensation of D-lactic acid and L-lactic acid in the absence of a catalyst, to evaluate their in vivo capabilities as biodegradable carriers for drug delivery systems. A luteinizing hormone-releasing hormone agonist, des-Gly10-(D-Leu6)-LH-RH ethylamide, was incorporated in a fine cylindrical copolymer formulation, under melt-pressing technique, a mild heat-pressure condition. This formulation was implanted subcutaneously in the back of male rats. The rate of in vivo degradation of amorphous copolymer was much faster than that of crystalline copolymer. Contrary to this tendency, the in vivo release of the drug from this amorphous formulation was held constant over a longer period, compared with the crystalline formulation. This can be closely related to the difference in dispersion of the drug in the formulation.     

Expression of EGF, EGF-receptor, p53, v-erb B and ras p21 in colorectal neoplasms by immunostaining paraffin-embedded tissues

Immunohistochemical studies were performed to clarify the significance of the expression or overexpression of epidermal growth factor (EGF), EGF-receptor (EGFR), p53, v- erb B, ras p21 in 23 cases each of tubular adenoma and adenocarcinoma. The expression of EGF, EGFR, p53, v- erb B, and ras p21 in paraffin-embedded tissues, from 46 patients with colorectal tumors (adenoma: 23 cases; 14 mild dysplasia, six moderate dysplasia, three severe dysplasia, adenocarcinoma: 23 cases; 17 well differentiated, two moderately differentiated, three poorly differentiated, one mucinous carcinoma was analyzed immunohistochemically using anti-EGF, EGFR, p53, v- erb B and ras p21 antibodies. The EGF and ras p21 tended to express more strongly in carcinoma cases than in the adenoma cases, and in severe and moderate dysplasia than in mild dysplasia (EGF: stained positive in five adenomas [21.74%] and 17 adenocarcinomas [73.91%]; ras p21: stained positive in six adenomas [26.09%] and 14 adenocarcinomas [60.87%]. The EGFR stained positive in two adenomas (8.70%) and two adenocarcinomas (8.70%). The p53 and v- erb B showed positive staining only in the carcinoma cases (p53: stained positive in four cases [17.39%]; v- erb B: stained positive in eight cases [34.78%]). This study suggests that these factors seem to have some role in the progression of colon neoplasms. It suggests that genetic alteration is not always equal to the overexpression of protein products, but that it reflects them well, and that the staining makes some contribution to differential diagnosis in colorectal neoplasms.     

Effects of rotation and distortion on perceptual priming

We conducted two experiments to specify the properties of the representation underlying perceptual priming. We transformed a particular property of stimuli between study and test. We used novel stimuli to eliminate contamination by semantic memory. They were rotated by 0 degree, 90 degrees, or 180 degrees in Experiment 1 and were distorted horizontally in Experiment 2. These transformations were chosen for topological structures of stimuli to be kept unchanged. In Experiment 1 the priming effect of the rotated stimuli was smaller than that of the identical ones. The rotational angles had no effect. More surprisingly, the horizontally distorted stimuli showed the same amount of priming as the identical ones in Experiment 2. In contrast, recognition performance of the rotated or horizontally distorted stimuli was lower than that of the identical ones. Statistical independence between priming and recognition was found in both experiments. Thus, recognition was statistically and functionally independent of priming. These results suggest that the priming representation encodes both orientation-independent and orientation-dependent information, and that the global topological structure is critical for priming.     

Synthesis and ion-coupling reactions of telechelic polystyrene having cyclic onium salt groups

A series of mono- and bifunctional polystyrenes ( 1 c and 1 ′ c ) having 1-methylpyrrolidinium salt end groups were prepared through sequential derivatization, i.e., tosylation and quaternization reactions, of prepolymers having 3-hydroxypropyl groups ( 1 a and 1 ′ a ) produced by end-capping reactions of the relevant living polymers. The 1-methylpyrrolidinium salt end group was found to undergo a selective ring-opening reaction at 100°C by nucleophilic attack of a benzoate counter anion introduced by ion-exchange reaction. The ion-coupling reaction of 1 c and 1 ′ c with poly(styrene-co-acrylate salt) was found to take place upon coprecipitation of an equimolar mixture into methanol to afford ionically linked pseudo-graft and network products, respectively. The subsequent heat treatment, converting the ionic bond into a covalent one, results in branched and crosslinked polystyrene with predetermined structural parameters.     

Immunogenetic background of patients with autoimmune fatigue syndrome

We have previously reported that approximately 50% of children with chronic nonspecific complaints were positive for antinuclear antibodies (ANA), and that a novel autoantibody to a 62 kD protein (anti-Sa) was found in 40% of these ANA-positive patients. Therefore, we proposed a distinct disease entity termed autoimmune fatigue syndrome (AIFS). We hypothesized that if autoimmune mechanisms did play an important role in the pathogenesis of AIFS, it is possible that it is immunogenetically regulated as observed in other autoimmune disorders. In order to examine the immunogenetic background of AIFS patients, HLA-A, -B, -C, and -DR loci were analyzed serologically in 61 AIFS patients. AIFS was found to be positively associated with the class I antigen HLA-B61 and with the class II antigen HLA-DR9, with odds ratios of 2.77 (p = 0.015, Pcorr = 0.48) and 2.60 (p= 0.012, Pcorr = 0.17), respectively. A negative association was also found between AIFS and HLA-DR2 with odds ratio of 0.25 (p = 0.029, Pcorr = 0.041). When comparing anti-Sa positive AIFS patients with healthy controls, the odds ratios associated with HLA-B61, DR9, and DR2 were 3.42 (p = 0.021, Pcorr = 0.22), 3.96 (p = 0.0011, Pcorr = 0.015), and 0.16 (p = 0.0022, Porr = 0.031), respectively. Thus, the HLA associations observed in this study suggested that immunogenetic background might play a role in AIFS.     

Flow-dependent water permeability of the rabbit descending limb of Henle's loop

Because completely opposite results have been reported on the water permeability of the rabbit descending limbs of Henle's loop (DLH), we rigorously examined water permeability of the upper portion of the descending limb of the rabbit long-looped nephron. Even when the double-cannulation method was used in an attempt to reduce the resistance of tubular outflow, the collected fluid-to-perfusate inulin ratio was equal to or very close to the bathing fluid-to-perfusate osmolality ratio, indicating that osmotic equilibration occurred along the tubule by absorption of water. When perfusion rates were controlled by varying the height of the fluid reservoir connected to the perfusion pipette, osmotic (Pf) as well as diffusional (Pdw) water permeability was shown to be correlated with perfusion rate and/or perfusion pressure. Pf and Pdw at zero perfusion rate as determined from the values of the intercept of regression lines were 253 X 10(-3) and 4.54 X 10(-3) cm X s-1, respectively. The maximal values for Pf and Pdw were 737-1,098 X 10(-3) and 18.3 X 10(-3) cm X s-1, respectively. By changing the resistance to perfusion at the tubular outflow, it was shown that changes in Pf paralleled changes in perfusion rate rather than changes in perfusion pressure. Under stop-flow conditions the luminal fluid volume rapidly decreased after the osmolality of the bathing fluid was increased, suggesting that the segment is highly permeable to water even at zero flow rate. Reflection coefficients for urea and NaCl were 1.01 and 0.82, respectively. These data support the view that this segment is highly permeable to water and that increases in osmolality along the DLH in vivo may be accounted for mainly by abstraction of water rather than addition of solutes.     

A Morphological and Immunohistochemical Study of Lymphoid Germinal Centers in Synovial and Lymph Node Tissues from Rheumatoid Arthritis Patients with Special Reference to Complement Components and Their Receptors

Rheumatoid arthritis (RA) is one of the immune complex (IC) diseases in which lymphoid germinal centers (GCs) are found in the synovial tissue. Simultaneously, patients with RA often show swelling of lymph nodes. The morphology and function of the lymph node GCs in patients with RA is not clear. The aim of this study was to evaluate the differences in morphology and immunoreactions to complement (C) components, their receptors, and lgM-rheumatoid factor (RF) between synovial GCs and lymph-node GCs in RA. Furthermore, the relationship between these immuno-reactive substances and follicular dendritic cells (FDCs) in GCs was investigated. The tissues examined were 41 RA synovial specimens, seven RA lymph nodes with massive lymphadenopathy, and 10 non-RA lymph nodes. The number of synovial GCs was relatively decreased in comparison with lymph-node GCs in RA, and the diameter of each synovial GC was smaller than that of each lymph-node GC. The synovial GCs were edematous and less cellular, and moreover, those from RF-seronegative cases were smaller than those from RF seropositive cases. On the other hand, the lymph-node GCs in RA were larger, more cellular and hyperplastic, but contained more tingible-body macro-phages (TBMs) and neutrophils. In the GCs of both synovial tissues and lymph nodes in RA, early C components (C1q, C4, C3c, C3d), IgM RF, and C3b receptor (C3bR) and C3d receptor (C3dR) were expressed as a lacy network by light microscopy, and were demonstrated on the surfaces of FDCs and lymphocytes, and in the intercellular spaces by electron microscopy. Furthermore, immuno-staining for dendritic reticulum cells (DRC, DAKO DRC1) was observed in a lacy pattern by light microscopy and on the cell surface of FDCs by electron microscopy. In the GCs of non-RA lymph nodes, early C components, C3bR, C3dR, and DRC showed a similar reaction pattern, but IgMRF did not. Consequently, no marked difference in immunoreactions in the GCs, except for the immunoreactions of late C components, was found between synovial tissues and lymph nodes in RA. On the basis of these findings, we discuss the possibility of the presence of a RF-IC.     

Eosinophil response in mast cell-deficient W/WV mice

The combination of cyclophosphamide treatment and Toxocara canis infection is known as an effective way of causing a high level of eosinophilia in mice. When this treatment was applied to congenitally anemic, mast cell-deficient W/WV mice, eosinophil response was far less than that of their normal littermate +/+ mice. The degree of the defective eosinophil response in the peripheral blood of W/WV mice was severer than that in the bone marrow. The defective eosinophil response of W/WV mice was completely restored by bone marrow grafting 8 weeks prior to cyclophosphamide treatment and T. canis infection. The kinetics of the recovery of eosinophil response in the bone marrow of W/WV mice after bone marrow grafting was faster than that in the peripheral blood. Chemotactic reactivity of eosinophils obtained from bone marrow or peritoneal cavity of W/WV mice was essentially comparable to that of +/+ mice. These results suggest that, in addition to the production of eosinophils in the bone marrow, mast cell-derived factors play an important role in the mediation of peripheral blood eosinophilia.