Effect of Lactobacillus delbrueckii LAB4 on hepatic and pancreatic sympathetic nerve activity and blood glucose levels in rats.

Various lactobacilli have been suggested to exert beneficial effects in humans. In this study, we examined the effects of intraduodenal (ID) administration of heat-killed Lactobacillus delbrueckii LAB4 (LAB4) on activities of efferent sympathetic nerves innervating the liver and pancreas. Consequently, it was observed that ID administration of LAB4 significantly reduced either the efferent hepatic sympathetic nerve activity (hepatic-SNA) or pancreatic sympathetic nerve activity (pancreatic-SNA) in urethane-anaesthetised rats. Moreover, the effect of acute and chronic administration of LAB4 (1×109 cells/ml) on hyperglycaemia induced by intracranial injection of 2-deoxy-D-glucose (2DG) were examined in conscious rats. We found that LAB4 significantly inhibited 2DG-induced hyperglycaemia. These findings suggest that ID administration of heat-killed LAB4 might lower plasma glucose level via changes in the autonomic nervous system in rats.

Effect of heat-killed Lactobacillus brevis SBC8803 on cutaneous arterial sympathetic nerve activity, cutaneous blood flow and transepidermal water loss in rats.

AIM: To evaluate the efficacy of the effects of heat-killed Lactobacillus brevis SBC8803 (HK-SBC8803) on the standard physiological markers of skin health of cutaneous arterial sympathetic nerve activity (CASNA), cutaneous blood flow and transepidermal water loss (TEWL) and to determine whether SBC8803 targets serotonin 5-HT3 receptors in rats. METHODS AND RESULTS: A set of three experiments were conducted to examine the effects of SBC8803 on CASNA, cutaneous blood flow and TEWL using Wistar and hairless rats. Two additional experiments further attempted to determine whether HK-SBC8803 was targeting the serotonin 5-HT3 receptors by pretreatment with the 5-HT3 antagonist granisetron. Administration of HK-SBC8803 in the first three experiments caused marked inhibition of CASNA and significant elevation of cutaneous blood flow under urethane anaesthesia as well as significant decrease in TEWL on the dorsal skin of conscious hairless rats. Pretreatment with granisetron decreased the effects of HK-SBC8803 on CASNA and cutaneous blood flow. CONCLUSIONS: These findings suggest that HK-SBC8803 reduces CASNA, increases cutaneous blood flow and decreases TEWL and that 5-HT3 receptors may be involved in CASNA and cutaneous blood flow responses. SIGNIFICANCE AND IMPACT OF THE STUDY: HK-SBC8803 could be a useful substance in the treatment/prevention of skin problems, specifically chapped or dry skin.

Pharmacological characterization of cyclosporine A-induced kaolin intake in rats.

Kaolin intake behavior of rats is known to be one of the useful animal models to evaluate the emetic and antiemetic actions of drugs. The present study was aimed at elucidating the pharmacological characterization of cyclosporine A (CsA)-induced kaolin intake in rats. Subchronic treatment (once a day for 3 days) with CsA produced a dose- and time-dependent increase in kaolin intake. Scopolamine (muscarinic antagonist), mepyramine (selective histamine H(1) antagonist) and diphenhydramine (H(1) and muscarinic antagonist) but neither domperidone (dopamine D(2) antagonist) nor ondansetron (serotonin 5-HT(3) antagonist) significantly inhibited CsA-induced kaolin intake. These findings suggest that an activation of central muscarinic and H(1) receptor is closely associated with CsA-induced kaolin intake in rats. Use of scopolamine and/or diphenhydramine may be possible regimens to alleviate and avoid nausea and vomiting in patients with CsA therapy.

Residue aspartate-147 from the third transmembrane region of Na(+)/H(+) antiporter NhaB of Vibrio alginolyticus plays a role in its activity.

NhaB is a bacterial Na(+)/H(+) antiporter with unique topology. The pH dependence of NhaB from Vibrio alginolyticus differs from that of the Escherichia coli NhaB homolog. Replacement of Asp-147 with Glu made high H(+) concentrations a requirement for the NhaB activity. Replacement of Asp-147 with neutral amino acids inactivated NhaB.

[Proposed mechanism of action of metalloendopeptidase-F in the treatment of patients with chronic hepatitis B or C infection].

Chronic hepatitis B and C virus infections have been characterized by the pathophysiological features with a high incidence of progression to cirrhosis and development of hepatocellular carcinoma. The viral persistence produced by escape mutations from virus-specific cytotoxic T lymphocytes (CTL) response may lead to upregulation of delayed-type hypersensitivity immune response, which causes hepatic tissue damage through non specific macrophage activation and CTL response and promotes pathogenesis of hepatic fibrosis. In a preliminary clinical study, a novel metalloendopeptidase-F (MEP-F) has been shown to be effective in the treatment of patients with either chronic hepatitis B or C infection. Oral administration of MEP-F resulted in a significant reduction of the serum levels of HBs antigen and HCV RNA and improvement in the liver function abnormalities. However, the mechanism of action of MEP-F is not yet well understood. There are accumulating evidences showing an important role of alpha 2-macroglobulin-proteinase complexes in regulatory mechanisms of immune response and repairing within impaired and inflammatory tissues. In this article, reviewing the pharmacological and biological properties of alpha 2-macroglobulin-proteinase complexes, the mechanism of anti-viral effect of MEP-F is examined based on the clinical findings. It is indicated that alpha 2-macroglobulin-MEP-F complexes may induce macrophage/Kuppfer cell activation and proliferation through binding their receptors on the cells and activating signaling cascades, which enhance both anti-viral specific and nonspecific immune responses. alpha 2-Macroglobulin-MEP-F complexes may also augment cellular immunity and hepatic regeneration by neutralizing the immunosuppressive and fibrogenic activities of transforming growth factor-beta.

Prefrontal and striatal dopamine metabolism during enhanced rebound hyperphagia induced by space restriction--a rat model of binge eating.

BACKGROUND: Several lines of evidence indicate that abnormalities in brain dopamine and serotonin metabolism may play an important role in bulimia nervosa. However, the regional neurochemical mechanism of the binge eating is poorly understood. Our purpose was to elucidate brain neurochemical mechanisms of binge eating using a rat model. METHODS: The dopamine release and metabolism in the prefrontal cortex (PFC) and in the ventrolateral striatum (VLS) of rats were studied using microdialysis during enhanced rebound hyperphagia induced by space restriction (an animal model of binge eating). RESULTS: The rats showed rebound hyperphagic state when they were released from scheduled feeding (2 hours/day feeding for 7 days). The hyperphagia was further enhanced when they were put in a space-restricted cage where their mobility was restricted. Dopamine release and metabolism were increased both in the PFC and in the VLS during the enhanced rebound hyperphagia. CONCLUSIONS: These results tentatively suggest that increased dopamine release and metabolism in the PFC and in the VLS may be related to space restriction and to activation of motor function involved in feeding behavior, respectively. The enhanced rebound hyperphagia induced by space restriction may be useful as an animal model of binge eating.

Bromocriptine enhances feeding behavior without changing dopamine metabolism.

Bromocriptine is an ergot derivative and has been thought to act as a selective D2 receptor agonist, but its effects on dopamine release in vivo have not been confirmed. We administered bromocriptine into the striatum of rats and studied the effects on feeding behavior and dopamine release. Bromocriptine was perfused via a microdialysis probe into the ventrolateral striatum of rats fasted for 22 h, and the rats were then allowed to feed freely for 6 h. Bromocriptine perfusion increased food intake in a dose-dependent manner, whereas the extracellular concentrations of dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) did not change. Perfusion of (-) sulpiride, a selective D2 receptor antagonist, decreased food intake, but increased dopamine release and the levels of DOPAC and HVA. Pretreatment with (-)sulpiride perfusion for 1 h prior to bromocriptine perfusion inhibited the increase of food intake induced by bromocriptine, and it increased dopamine release and the levels of DOPAC and HVA. These findings suggest that bromocriptine directly perfused into the ventrolateral striatum acts selectively on postsynaptic D2 receptors and enhances feeding behavior.

Effects of fluvoxamine on food intake during rebound hyperphagia in rats.

We examined the effects of fluvoxamine on food intake during rebound hyperphagia induced by a time-restricted feeding schedule in rats. Rats were allowed access to food for only 2 h daily for 7 days, and then had free access to food for 7 consecutive days. The daily food intake of the rats was dramatically increased, by 42.5% (rebound hyperphagia), for 7 days of the free-feeding period. Intraperitoneal injection of fluvoxamine decreased food intake significantly in a dose-dependent manner for the first 3 h of feeding during 7 days. When rats were allowed access to one of the standard, carbohydrate-, fat-, or protein-rich diets in the free-feeding period following the time-restricted feeding schedule, fluvoxamine significantly decreased food intakes of standard, carbohydrate- and fat-rich diets on all days, and the protein-rich diet after the 2nd day of the free-feeding period. These results indicate that fluvoxamine, irrespective of the diet composition, suppresses rebound hyperphagia induced by a time-restricted feeding schedule, but that its effect is short-lived.

Immunoblastic lymphadenopathy-like T-cell lymphoma displaying rearrangement of both IgH and TCR beta genes after 4-year follow-up of idiopathic eosinophilia.

Immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma (IBL-T) occurred in a 60-year-old female after a 4-year follow-up of idiopathic eosinophilia and upper pharyngeal inflammatory tumor with infiltration of mature eosinophils. Gene analysis of tumor cells revealed rearrangement of both IgH and TCR beta genes. The patient died of lymphoma seven months after the onset of the illness, in spite of chemotherapy against lymphoma. The relationship between eosinophilia and the pathogenesis of IBL-T, as well as the significance of the rearrangement of both IgH and TCR beta genes are discussed.

D2 receptors in the ventrolateral striatum are involved in feeding behavior in rats.

To study the role of dopamine D1 and D2 receptors in the ventrolateral striatum in feeding behavior, a D1 receptor agonist (CY 208-243), a D1 receptor antagonist (SCH 23390), a D2 receptor agonist (quinpirole), and a D2 receptor antagonist [(-)-sulpiride] were perfused via a microdialysis probe into the ventrolateral striatum of rats fasted for 22 h. Then the rats were allowed to feed freely for 6 h. Sulpiride perfusion at a high concentration suppressed food and water intake significantly, whereas dopamine release and the levels of DOPAC and HVA were increased at all concentrations. In contrast, quinpirole perfusion at a high concentration increased food intake by 41%. Dopamine release and the levels of DOPAC and HVA were decreased at all concentrations. On the other hand, neither CY 208-243 nor SCH 23390 changed food intake or dopamine release, but both drugs decreased water intake. These results suggest that D2 receptors in the ventrolateral striatum have a more important role than D1 receptors in the feeding behavior of rats.

[NEO-MACOP-B chemotherapy for the treatment of advanced-stage aggressive non-Hodgkin's lymphoma].

We conducted a new chemotherapy, NEO-MAC OP-B (addition of etoposide and mitoxantrone to MACOP-B with half dose of methotrexate and half administration of doxorubicin), to reduce severe mucositis, which is a major toxic effect of MACOP-B, and to increase its effect with etoposide and mitoxantrone as new non-cross resistant drugs. Between Jan. 1989 and Mar. 1993, 12 patients with previously untreated advanced aggressive non-Hodgkin's lymphoma (NHL), 2 patients with adult T cell lymphoma, and 3 patients with relapsed NHL, were treated with NEO-MACOP-B. After termination of NEO-MACOP-B therapy, 83.3% of 12 patients with previously untreated NHL were in complete remission (CR). After median follow-up of 22 months, Kaplan-Meier estimates showed that overall survival of 12 previously untreated patients was 71.4%, and relapse-free survival of complete responder was 83.3%. Toxic effects on all 17 patients were moderate with a lower incidence of severe mucositis (only one patient with relatively severe stomatitis, WHO Grade 3). No treatment related deaths were observed. Thus, NEO-MACOP-B is an effective and safe treatment for advanced stage aggressive NHL.

Dot-blot hybridization using digoxigenin-labeled cDNA probe complementary to the S1 gene of avian infectious bronchitis virus permits discrimination between virus strains.

Digoxigenin-dUTP-labeled DNA probe was prepared from a cDNA clone complementary to the gene encoding S1 region of the spike protein of infectious bronchitis coronavirus (IBV) strain M41. The probe exclusively reacted with four strains at 56 degrees C which were grouped to the same serotype as the strain used for the probe. In contrast, at 68 degrees C, the probe reacted only with the homologous strain and did not react even with the strains belonging to the same serotype. The dot-blot hybridization thus appeared serotype-specific at 56 degrees C and strain-specific at 68 degrees C. In addition, it was revealed that the S1 gene has some nucleotide sequence variation even among strains in the same serotype. This technique should be applied to determining serotypes of the virus isolates and to differentiating field isolates from the vaccine strain.

Scheduled feeding caused activation of dopamine metabolism in the striatum of rats.

The effects of a time-restricted feeding schedule on dopamine (DA) release and its metabolites output in the striatum of freely moving rats were studied. Rats had access to food for only 2 h daily for 7 successive days. On the 1st or 7th day, the extracellular concentrations of DA, 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA) in the ventrolateral striatum were measured by in vivo brain microdialysis during 2 h of exposure to food-related stimuli followed by 2 h of access to food. Extracellular concentrations of DA and its metabolites did not change during the period of exposure to food-related stimuli or during feeding on the 1st day. On the 7th day, extracellular DOPAC and HVA concentrations increased significantly during 2 h of feeding, but not during exposure to food-related stimuli, compared with basal levels. Extracellular DA concentration did not change significantly. These results indicate that scheduled feeding caused activation of DA metabolism in the ventrolateral striatum and facilitate feeding-related motor activity in feeding behavior.

[Successful treatment of prolonged anemia after major ABO incompatible bone marrow transplantation for a case of myelodysplastic syndrome with recombinant erythropoietin].

A delay in red cell recovery after ABO-incompatible bone marrow transplantation (BMT) is often observed. The authors experienced a case of prolonged anemia after a major ABO incompatible BMT for myelodysplastic syndrome which was successfully treated with recombinant human erythropoietin (Epo). Effects of Epo were confirmed by the recurrence of anemia after withdrawal of Epo as well as the rapid reincrease in reticulocytes on readministration. The patient received a dose of Epo which was similar to the amount used for renal anemia, however serum concentration of Epo after administration exceeded endogenous Epo levels. Epo may have a beneficial role in the treatment of prolonged anemia after BMT.

[Adequate requirements of energy and protein in postoperative total parenteral nutrition].

Adequate requirements of energy and protein in post operative total parenteral nutrition (TPN) were determined by studying the effects of energy and protein dose on nitrogen retention at varying levels of surgical stress assessed by urinary excretion of total catecholamines. Fifty-two patients received esophagectomy (severe stress), and gastric or colorectal operations (moderate stress) fed exclusively by TPN perioperatively were divided into 6 groups according to the dose level of protein and energy; Group I: 40 kcal, 1.0 g.protein/kg.day (9 patients), Group II: 40 kcal, 1.5 g.protein/kg.day (11 patients), Group III: 40 kcal 2.0 g.protein/kg.day (8 patients), Group IV: 40 kcal, 3.0 g.protein/kg.day (7 patients), Group V: 30 kcal, 2.0 g.protein/kg.day (11 patients), and Group VI: 50 kcal, 2.0 g.protein/kg.day (6 patients). Daily nitrogen balance (NB) and urinary excretion of total catecholamines (U-CA) were determined everyday pre- and post-operatively. Significantly negative correlations between U-CA and NB were observed. Statistically significant differences among the correlations of all groups were recognized. In moderate stress, increasing dose of protein and energy improved NB, and positive NB was achieved when 1.5-2.0 g.protein/kg.day with 35-40 kcal/kg.day was provided. Protein dose exceeding 3.0 g/kg.day caused the rise of BUN. In severe stress, such as following esophagectomy, NB was not improved by increasing dose of nutrients.

Immunosuppressive effect of infectious bursal disease virus strains of variable virulence for chickens.

Infection of chicks with attenuated Lp and Sp clones of the RF-1 strain of infectious bursal disease virus was shown to exert no immunosuppressive effect, whereas the parent strain RF-1tc and the original virulent strain RF-1wt were immunosuppressive. One-day-old chicks infected with Lp and Sp clones showed no suppression of immunological response to live Newcastle disease vaccines B1 and TCND, and to bivalent infectious coryza vaccine. On the other hand, infection with RF-1tc or RF-1wt strains was immunosuppressive for these vaccines. The immunosuppressive effect of RF-1tc and RF-1wt strains was more pronounced for infectious coryza vaccine and B1 vaccine than for TCND vaccine. The immunosuppressive effect of RF-1tc and RF-1wt strains was lower when chicks were infected with these strains at the age of 21 days than when they were infected at one day of age.

Polymyositis in mice experimentally inoculated with Getah virus.

Mice inoculated intracerebrally with parent, large-plaque (LP) and small-plaque (SP) strains of Kanagawa strain of Getah virus showed clinically recumbency and paralysis. The LP strain caused recumbency more rapidly and killed mice more early after inoculation than the parent and SP strains. Microscopically, skeletal muscles of the whole body were involved showing degenerative or inflammatory changes. In mice inoculated with the parent or SP strains, there were degeneration and necrosis of the muscle fibers with inflammatory cell infiltration and regenerative reaction. The lesions were particularly conspicuous in muscles of the hind legs. In mice inoculated with the LP strain, most of the muscle fibers revealed degeneration and necrosis, but reactive changes were poor. In addition, the periosteum and muscular connective tissue were thickened with karyorrhexis. Electron microscopically, virus particles were recognized mainly in cisternae of sarcoplasmic reticulum in skeletal muscle fibers of mice inoculated with the LP strain, while they were rare in those of animals injected with the parent and SP strains. From these finding, it was suggested that Kanagawa strain of Getah virus has the virulence to skeletal muscles of mice.

The relation between pathogenicity and plaque size of Getah virus Kanagawa strain in suckling mice.

The relation among biological properties, particularly pathogenicity for suckling mice, and plaque size was studied in four virus strains: Getah virus strain Kanagawa; two strains obtained by plaque cloning of the Kanagawa strain, Getah Kanagawa SP (G-K-SP) strain which forms small plaques (SP) only and strain G-K-LP which forms large plaques (LP) only; and strain Haruna which forms SP only. There were no marked differences among the four strains in serological properties, growth curves and sensitivity to pH, trypsin and temperature. Strain G-K-LP showed higher pathogenicity for suckling mice than strain G-K-SP. However, the pathogenicity of strain Haruna, which forms SP only, was as high as that of strain G-K-LP. Some of the clones in SP of strain Kanagawa kill all mice in 5 to 6 days after inoculation while the others required 9 to 11 days or longer before causing death. The present study showed that the pathogenicity of Getah viruses shortly after being isolated from the field, such as the Kanagawa strain, is different between large and small plaques, and even among small plaques, at least in suckling mice, and that the pathogenicity has no relation to plaque size.

The production of transforming growth factor-beta in acute megakaryoblastic leukemia and its possible implications in myelofibrosis.

Acute myelofibrosis is often associated with acute megakaryoblastic leukemia (AMKBL). Although the exact mechanism for the progression of myelofibrosis in AMKBL is unclear, certain humoral factors from megakaryoblastic cells, the precursors of platelets, may be involved in the enhancement of collagen synthesis by bone marrow fibroblasts. The present study, therefore, is an investigation of the possible pathogenic role of transforming growth factor-beta (TGF-beta), known to be a very potent collagen-stimulating factor found in platelets in the myelofibrosis of AMKBL. The results obtained were as follows: (1) Conditioned media from peripheral megakaryoblasts taken from an AMKBL patient and from established megakaryoblast cell lines (MEG-01) had much greater stimulatory effects on collagen synthesis in bone marrow fibroblasts than conditioned media from other leukemic cell types. (2) Based on an assessment of soft agar colony formation, there was greater TGF-beta activity in media that had been conditioned from megakaryoblasts than in media from other leukemic cell types. (3) When compared with other leukemic-cell types, megakaryoblasts showed substantially greater expression of TGF-beta mRNA that was hybridized at 2.5 kb with a TGF-beta cDNA probe, and TGF-beta polypeptides were detected at 13 Kd with anti-TGF-beta antibodies. (4) The addition of the anti-TGF-beta antibody inhibited the stimulatory effects of the megakaryoblast conditioned medium on collagen synthesis in bone marrow fibroblasts. These results clearly suggest that megakaryoblasts produce and secrete an active form of TGF-beta and stimulate collagen synthesis in bone marrow fibroblasts in a paracrine manner.