Effects of lysed Enterococcus faecalis FK-23 on allergen-induced serum antibody responses and active cutaneous anaphylaxis in mice.

BACKGROUND: Our previous studies have presented evidence that lysed Enterococcus faecalis FK-23 (LFK), a lysozyme and heat-treated probiotic product, can inhibit allergen-induced local accumulation of eosinophils in mice. OBJECTIVE: The purpose of this experimental study was to evaluate the influence of orally administrated LFK on the host immune responses. METHODS: BALB/c mice were sensitized subcutaneously, and challenged intraperitoneally by cedar pollen allergen. Blood and spleen samples were collected after oral administration of LFK 60 mg/day for 21 days. The serum levels of total and allergen-specific IgE and IgG2a antibodies and the production of IL-4, IL-5 and IFN-gamma generated by allergen-stimulated cultured splenocytes were determined. Additionally, the effect of LFK on active cutaneous anaphylaxis (ACA) induced by ovalbumin (OVA) challenge in mice was measured after 28 days LFK treatment. RESULTS: No significant differences in serum immunoglobulin levels, as well as in cytokine production of splenocytes were observed between LFK-treated and control mice (P>0.05). There was, however, an increasing tendency of allergen-specific IgG2a level in mice after LFK treatment for 21 days compared with controls (P=0.060). Furthermore, the serum ratio of specific IgE to IgG2a was found to be significantly decreased in the LFK group (P=0.005). In addition, a significant inhibition of OVA-induced ACA reaction was observed in mice that had been fed for 28 days with LFK compared with control mice (P=0.008). CONCLUSION: These results suggest that LFK shows an anti-inflammatory effect, which may be part of the mechanism for protection against IgE-mediated allergy.

Effect of lysed Enterococcus faecalis FK-23 (LFK) on allergen-induced peritoneal accumulation of eosinophils in mice.

BACKGROUND: The interest in anti-allergy immunoregulation by lactic acid bacteria has been growing for the last few decades. There is some evidence to suggest that lysed Enterococcus faecalis FK-23 (LFK) could relieve the clinical symptoms of pollinosis. However, the mechanism responsible for this phenomenon remains unknown. OBJECTIVE: To identify the effect of LFK, a lysozyme treated and heat-killed preparation from the lactic acid bacteria Enterococcus faecalis FK-23 strain, on allergen-induced eosinophil accumulation. METHODS: BALB/c mice were sensitized with ragweed pollen extract, and peritoneal accumulation of eosinophils was induced. A total of 60 mg (0.5 mL) LFK was orally administered to the experimental mouse every day during 21 days of the sensitization period. In addition, LFK 4 mg, 25 mg and 60 mg (each 0.5 mL) were also orally administered to a mouse of each group every day for 21 days. Saline was fed in a dose of 0.5 mL/mouse per day for the same duration as a control. RESULTS: Compared with control mice, LFK-treated mice exhibited decreased ragweed pollen allergen-induced peritoneal accumulation of eosinophils (P = 0.013), which showed a tendency to be in a dose-dependent fashion (P = 0.14). CONCLUSION: The results provide laboratory evidence of the role for LFK, a lactic acid bacteria preparation, in combating eosinophil accumulation.

Spasmolytic effect of peppermint oil in barium during double-contrast barium enema compared with Buscopan.

AIM: To evaluate the efficacy of peppermint oil in barium as a spasmolytic agent during a double-contrast barium enema (DCBE). MATERIALS AND METHODS: A total of 383 DCBEs with positive results from occult blood tests were assessed. Patients were assigned to one of four groups: peppermint in barium (n=91), peppermint in tube (n=90), Buscopan (n=105), or no treatment (n=97). After a screening sigmoidoscopy, the DCBEs were performed using air as a distending gas. In the Buscopan group, the DCBE was performed with an intramuscular injection of 20mg Buscopan at the start of the examination. Patients in the no-treatment group underwent DCBE without any spasmolytic agent. A peppermint oil preparation (30ml) was mixed in the barium solution for patients in the peppermint-in-barium group, and the same dose of peppermint oil was included in the enema tube in the peppermint-in-tube group. The presence of spasm on a series of spot films was evaluated without information about the type of spasmolytic agent used. RESULTS: The percentage of patients in the four groups (no treatment, Buscopan, peppermint in tube, and peppermint in barium) with absence of spasm in the entire colon on the series of spot films was 13.4, 38.1, 41.8, and 37.8%, respectively. In the group using peppermint oil or Buscopan, the rate of patients with non-spasm examination was higher than that in no-treatment group (p<0.0005). Peppermint oil had the same spasmolytic effect as the systemic administration of Buscopan in the transverse and descending colon. Peppermint oil had a stronger effect in the caecum and the ascending colon than a Buscopan injection (p<0.005). There was no advantage to placing peppermint oil in the enema tube over mixing it in the barium solution. A total of 157 polyps were found during the DCBE procedures, and no differences were observed in the number of lesions among the four groups. Peppermint oil did not impair image quality. CONCLUSION: Barium solution mixed with peppermint oil was safe and effective for the elimination of colonic spasm during the DCBE procedure, and it could be used instead of Buscopan.

Evaluation of a surgical gamma probe for detection of 18F-FDG.

Because fluorine-18 fluorodeoxyglucose (18F-FDG) is an excellent tumor-localizing radio-pharmaceutical, a hand-held radiation detection probe capable of localizing an area with high 18F-FDG uptake would make radioguided surgery possible. In this laboratory study, we investigated the capability of a widely used intraoperative gamma probe with a cadmium zinc telluride (CdZnTe) detector for detection of 18F-FDG. For sensitivity tests, an 0.1- ml 18F-FDG preparation was made to act as a point source with radioactivities of 1.0, 2.0, and 3.0micro Ci (37 kBq, 74 kBq, 111 kBq). Relative transmission across the side wall of the probe and sensitivity at each source-to-probe distance were measured. For simulation studies, 2 l of 18F-FDG solution (0.02micro Ci, 0.74 kBq/ml) served as normal background. One ml of 18F-FDG was prepared to simulate tumors with radioactivities of 0.05, 0.1, 0.2, and 0.4micro Ci (1.85 kBq, 3.7 kBq, 17.4 kBq, and 14.8 kBq). The ratios of the radioactive concentration of tumor to that of the background were 2.5, 5, 10, and 20:1, respectively. The tested gamma probe was shown to be sensitive to 18F. The high-energy annihilation radiation was detected from the side wall of the probe despite application of a supplementary collimator. The count rate decreased markedly as the source-to-probe distance increased, owing to the effects of the inverse-square law. In the simulation studies, the probe detected a considerable amount of background activity. However, the measured count rate increased with the increasing source-to-background ratio. In our setting, the probe was capable of distinguishing the 18F-FDG source from the background when the source-to-background ratio was no less than 5:1. To make a surgical application feasible, however, collimation or shielding against high background radiation is necessary.

Elevation of serum hepatic aminotransferases during treatment of rheumatoid arthritis with low-dose methotrexate. Risk factors and response to folic acid.

Sixty-six rheumatoid arthritis (RA) patients were analyzed retrospectively to assess the incidence and risk factors for elevation of serum hepatic aminotransferases during methotrexate (MTX) therapy. The effect of folate supplementation on serum ALT and RA activity was evaluated prospectively in 14 patients who showed a sustained high serum level of ALT. The frequency of elevation of serum AST or ALT was 4-5 times greater than in patients taking other DMARDs. Multivariate linear regression analysis demonstrated that elevation of ALT was independently associated with sex (female), obesity, baseline ALT, MTX dose, and gastrointestinal side effects. Folate supplementation caused ALT levels to decrease in all patients within 3 months. Eleven patients showed no change of RA activity, but 3 patients dropped out of the study because of the exacerbation of RA. These results suggest that careful monitoring of serum hepatic aminotransferases is necessary in patients with predisposing factors, especially those receiving more than 0.15 mg/kg of MTX weekly. Folate supplementation can reverse the sustained elevation of ALT, but might cause exacerbation of RA in some patients.

Treatment of ulcerative colitis with germinated barley foodstuff feeding: a pilot study.

BACKGROUND: Germinated barley foodstuff (GBF) has been shown to attenuate intestinal injury in animal models, largely by increasing luminal short-chain fatty acid production. AIM: To investigate the safety and efficacy of GBF in the treatment of ulcerative colitis (UC). METHODS: Ten patients with active UC received 30 g of GBF daily for 4 weeks in an open-label treatment protocol while the baseline anti-inflammatory therapy was continued. The response to treatment was evaluated clinically and endoscopically. Pre- and post-treatment stool concentrations of short-chain fatty acids were measured by gas-liquid chromatography. RESULTS: Patients showed improvement in their clinical activity index scores, with a significant decrease in the score from 6.9+/-1.4 to 2.8+/-1.5 (mean+/-S.E.M., P < 0.05). The endoscopic index score fell from 6.1+/-2.3 to 3.8+/-2.3 (P < 0.0001). Patients showed an increase in stool butyrate concentrations after GBF treatment (P < 0.05). No side-effects were observed. CONCLUSIONS: Oral GBF therapy may have a place in management of ulcerative colitis, but controlled studies are needed to demonstrate its efficacy in the treatment of this disorder.

Short-term low dose methotrexate ameliorates abnormal bone metabolism and bone loss in adjuvant induced arthritis.

OBJECTIVE: To clarify whether short term weekly methotrexate (MTX) therapy aggravates bone abnormalities in adjuvant induced arthritis (AIA) or improves them through its antiarthritic effect. METHODS: Bone metabolism and bone mineral density (BMD) were studied in 6 groups of Lewis rats: (1) normal controls, (2) rats given MTX 0.3 mg/kg weekly, (3) rats given MTX 3 mg/kg weekly, (4) AIA rats, (5) AIA rats given MTX 0.3 mg/kg weekly, and (6) AIA rats given MTX 3 mg/kg weekly. Osteogenic activity was determined from serum osteocalcin levels and number of marrow fibroblast colony forming units (osteogenic precursor cells). Bone resorptive activity was assayed by detecting osteoclast-like cells and pit formation in bone marrow cultures. RESULTS: In control rats, MTX (3 mg/kg weekly) suppressed osteogenic activity, as shown by low serum osteocalcin levels and decreased growth of marrow fibroblast colony forming units. Osteoclast-like cells and pit formation in bone marrow cultures from control rats were increased by MTX, but BMD was unchanged. In rats with AIA, MTX (3 mg/kg) suppressed arthritis and restored the decreased osteogenic activity of bone marrow cells, and reduced their increased bone resorptive activity. These changes resulted in a significant increase of periarticular BMD in the femur. CONCLUSION: Low dose weekly MTX therapy had a favorable effect on abnormal bone metabolism and osteopenia in rats with AIA.

Periarticular osteopenia in adjuvant induced arthritis: role of interleukin-1 in decreased osteogenic and increased resorptive potential of bone marrow cells.

OBJECTIVE: To clarify the local osteogenic and bone resorptive potential of periarticular bone in adjuvant induced arthritis (AIA). METHODS: Formation of fibroblast colony forming units (FCFU; osteogenic precursor cells) and osteoclast-like cells in bone marrow culture was studied in AIA rats. Osteoclast-inducing activity in the AIA rat bone marrow was assayed by the addition of the marrow supernatant from rats with AIA to control cultures. Bone mineral density was determined by dual x ray absorptiometry. RESULTS: Marrow from AIA rats and that from animals receiving recombinant human interleukin-1 (IL-1) beta for seven days grew significantly fewer FCFU than control marrow. Formation of osteoclast-like cells was increased in bone marrow cultures from rats with AIA, especially when bone marrow cells were cultured in the presence of marrow supernatant. Formation of resorption lacunae on ivory slices was increased in the marrow cultures from rats with AIA, especially from the right (adjuvant inoculated) tibia. AIA rat marrow supernatant promoted osteoclast-like cell formation in control culture, and this was significantly suppressed by an anti-IL-1 antibody. Rats with AIA showed a significant decrease in the bone mineral density of the periarticular regions of the tibia and femur. CONCLUSION: An uncoupled state in bone resorption-formation linkage, possibly mediated through an increase of IL-1 in the bone marrow, may contribute to the development of periarticular osteopenia in inflammatory arthritis.

Solitary pyogenic thalamic abscess--two case reports.

We report two patients with solitary thalamic abscesses, occurring among 91 consecutive patients (2.2%) with computed tomography (CT)-diagnosed and surgically-verified brain abscess experienced in our college during 1975 to 1991. A 9-year-old girl with congenital heart disease experienced frequent vomiting followed by left hemiparesis and deterioration of consciousness. CT demonstrated a right thalamic ring-enhanced lesion. Purulent material was aspirated via a burr hole. She died of heart failure on the 5th postoperative day. Autopsy disclosed diffuse brain swelling and an encapsulated abscess in the right thalamus, which had ruptured into the third ventricle. A 30-year-old female experienced headache, nausea, and vomiting, which progressed to somnolence and right hemiparesis. CT demonstrated a left thalamic ring-enhanced lesion. Purulent material was aspirated by stereotactic procedures. All symptoms had resolved by the end of the 2nd postoperative week.

Giant negative U waves in a patient with uncontrolled hypertension and severe hypokalemia.

A 66-year-old woman with a long history of hypertension had an electrocardiogram with giant negative U waves in left precordial leads despite hypokalemia. This seems to be the first report of giant negative U waves induced by uncontrolled hypertension with hypokalemia. The occurrence of negative U waves in the presence of profound hypokalemia is an important observation because it masks the electrocardiographic manifestation of hypokalemia.

Long-term effects of intense sound on hair cells of Corti's organ and endocochlear DC potential.

Cochleograms of guinea pig ears were made 30-40 days after exposure to intense pure tones of 300 Hz, 500 Hz, 2 kHz, or 4 kHz at 130-150 dB SPL for 4-24 h. At 4 kHz, hair cells in the basal turn disappeared totally, in the second turn moderately, and were relatively undamaged in the third and apical turns. At 500 Hz, hair cells in the second and third turns were almost completely injured and at 300 Hz moderately damaged in the third and apical turns although the basal turn remained undamaged. At 2 kHz for 9 h, hair cells were almost completely injured in all turns. Negative endocochlear DC potential (negative EP) induced by furosemide was observed in the basal turn but not in the third turn of animals exposed to 300 Hz. Contrarily, negative EP was observed in the third turn but not in the basal turn of animals exposed to 4 kHz. We conclude that the hair cells of Corti's organ play an essential role in the production of negative EP.

Appearance of neurons with glucocorticoid receptors and neurovascular links in the embryonal rat hypothalamus grafted in the third ventricle.

We have investigated the appearance of the transmitter phenotypes of hypothalamic neurons in grafts transplanted into the third ventricle of adult female rats. The grafts were the mediobasal hypothalamus and the preoptic area of 12.5-day-old rat embryos, and were examined 40-100 days later. Wheat germ agglutinin (WGA) was injected into the jugular vein of several animals for the examination of the existence of neurovascular associations. Three days after the injection, WGA appeared to have been incorporated into the neurons in the paraventricular, periventricular, and arcuate nuclei of the host animals. In the grafts, WGA was also seen incorporated in certain neurons which were found immunoreactive for tyrosine hydroxylase (TH), rat corticotropin-releasing factor (rCRF), substance P (SP), or somatostatin (SRIH). Neurons immunoreactive for neuropeptide Y (NPY) and ACTH did not seem to incorporate WGA. These findings suggest that the neurons containing TH, rCRF, SP, or SRIH link with fenestrated capillaries developed in the grafts. The immunoreactivity for glucocorticoid receptor (GR) was detected mainly in the nucleus of certain neurons and glial cells in the grafts as well as in the host hypothalamic neurons. In the grafts, strong GR immunoreactivity was detected in the cells immunoreactive for TH, NPY, and rCRF as in the host animals. It is concluded that the undifferentiated hypothalamic neurons differentiate to synthesize GR as well as definitive peptides and TH in the grafts.

Hypothalamic neurons from a developmental aspect.

The development of hypothalamic neurons was examined in vivo and in transplanted grafts in rats. The neurons appeared in vivo in distinctive chronotopical schedules showing such morphological characteristics as synaptocrine or hemocrine neurons. These phenotypical properties seemed to be primed already by day 12.5 of gestation in rats, because the grafted hypothalamic primordia from 12.5-day-old embryonal rats differentiated neurons, which express these neuronal properties in the third ventricle of adult female rats. The synaptocrine neurons projected to other neurons, suggesting the establishment of synaptic contacts, and the hemocrine neurons projected to vasculatures developed in the grafts, suggesting the accomplishment of neurovascular associations.

Effect of intermittent sound stimulation on cochlear microphonics and the possible preventive effect of coenzyme Q10.

Observations have been made on the effect of intermittent intense sound stimuli, using the CM as an index. Experiments were conducted regarding the effectiveness of the drug CoQ10 in ameliorating the acoustic injury due to this traumatization. The CM was measured by the differential electrode method from the basal turn of the cochlea. The individual differences between animals were monitored using intensity functions. Pure tones at a frequency of 4 kHz were used both for the traumatizing signal and the CM generating signal. Acoustic traumatizing signals lasted 1 min with a 5 min silent interval, and were repeated 10 times. Both the chronic depression and the transient depression of CM were measured after each traumatization. In control animals, the chronic depression progressed in an exponential manner and the transient depression was constant after each stimulation. In CoQ10 treated animals, the chronic depression was milder than that in the control animals. The transient depression was equivalent in the two groups. The effectiveness of CoQ10 on the acoustic injury was discussed on the basis of the results obtained with the present experiment.