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Background: Our previous studies have presented evidence that lysed Enterococcus jaecalis 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-γ 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. [ABSTRACT FROM AUTHOR]

Various fatty acids are attracting considerable interest for their anticancer effects. Among them, fatty acids containing conjugated double bonds show one of the most potent cytotoxic effects on cancer cells. Here, we focused on the cancer cell killing activity of jacaranda seed oil. The seed oil of jacaranda harvested from Miyazaki in Japan contained 30.9% cis-8, trans-10, cis-12 octadecatrienoic acid, called jacaric acid (JA). Fatty acid prepared from this oil (JFA) and JA strongly induced cell death in human leukemia HL-60 cells. On the other hand, linoleic acid and trans-10, cis-12 conjugated linoleic acid (<10 μM) did not affect cell proliferation and viability. An increase in the sub-G₁ population and internucleosomal fragmentation of DNA was observed in JA- and JFA-treated cells, indicating induction of apoptotic cell death. Finally, the cytotoxic effects of JA and JFA were completely abolished by α-tocopherol. Taken together, these data suggest that jacaranda seed oil has potent apoptotic activity in HL-60 cells through induction of oxidative stress.

We have shown previously that jacaric acid (JA; 8c,10t,12c-18:3), which has a conjugated triene system, has a strong anti-tumor effect. However, the characteristics of absorption and metabolism of JA have yet to be determined in vivo, and the details of absorption and metabolism of JA in the small intestine are particularly unclear. This information is required for effective use of JA in humans. Therefore, in this study we examined absorption and metabolism of JA using cannulation of the thoracic duct in rats. Emulsions of two test oils, jacaranda seed oil and tung oil, which contain JA and α-eleostearic acid (α-ESA; 9c,11t,13t-18:3), respectively, were administered to rats and lymph from the thoracic duct was collected over 24 h. We examined the rate of absorption of JA and possible conversion to a conjugated linoleic acid (CLA)containing a conjugated diene system. The positional isomerism of the CLA produced by JA metabolism was determined using gas chromatography-electron impact/mass spectrometry. The rate of absorption and percentage conversion of JA were compared with those of α-ESA. We found that JA is rapidly absorbed and converted to a CLA in rats and that the percentage conversion of JA was lower than that of α-ESA. This is the first report on the absorption and metabolism of JA and this information may be important for application of JA as a functional food.

Kampo is a traditional Japanese medicine originating from ancient Chinese medicine which included the administration of herbal prescription, lifestyle advice and acupuncture. Orally administered Kampo prescriptions are believed to be influenced by diet and intestinal microbiota. However, reports on the Kampo administration effects are still limited. Shoseiryuto (TJ-19), which has anti-allergic and anti-inflammatory properties, is a Kampo prescription used clinically for the treatment of allergic bronchial asthma. We examined whether Shoseiryuto administration is affected by a probiotic product, lysed Enterococcus faecalis FK-23 (LFK). BALB/c mice were sensitized with cedar pollen allergen, and the peritoneal accumulation of eosinophils was induced. During a sensitization period of 21 days, varying amounts of Shoseiryuto (and saline as a control) were administered to the mice. The accumulation of eosinophils was significantly reduced by 30 mg/day doses of Shoseiryuto but not by 3 or 9 mg/day doses. Similarly, 3 mg/day Shoseiryuto, 30 mg/day LFK, 3 mg/day of Shoseiryuto co-administered with 30 mg/day of LFK, and saline control were compared. A significant reduction in the accumulation of eosinophils was observed at 3 mg/day Shoseiryuto co-administered with 30 mg/day of LFK. These results suggest that Shoseiryuto-mediated anti-allergic effects are enhanced by the probiotic (LFK). Although not significant statistically, serum allergen-specific and total IgE levels in the treatment group exposed to the mixed agent (ie. Shoseiryuto and LFK) were generally lower than those receiving either one alone. The results indicate a synergistic effect of a Kampo medicine (Shoseiryuto, Xiao-Qing-Long-Tang in Chinese) and lysed Enterococcus faecalis FK-23 on allergic responses in mice.

The objective of the present study was to assess the effects of Enterococcus sp. strain TN-3 isolated from deep seawater on inhibition of eosinophil accumulation, IgE production and active cutaneous anaphylaxis (ACA). We investigated the effects of viable and non-viable TN-3 on allergen-induced peritoneal eosinophil accumulation in mice. Viable (5.4 x 1010 colony-forming units per 60 mg) or non-viable TN-3 (60 mg) was orally administered to BALB/c mice that had been sensitised with the cedar pollen (Cryptomeria japonica) allergen. Oral administration of non-viable TN-3 was effective in suppressing eosinophil accumulation while viable TN-3 was ineffective. We also examined the dose-response relationship for non-viable TN-3 in regard to eosinophil accumulation, IgE production and ACA in allergen-primed mice. Non-viable TN-3 was orally administered at doses of 15 mg (low dose), 30 mg (medium dose) and 60 mg (high dose) to BALB/c mice that had been sensitised with cedar pollen allergen. The anti-allergic effects expressed as inhibition of eosinophil accumulation, IgE production and ACA were found at the low and high doses, but not at the medium dose. These results suggest that non-viable TN-3 exhibited anti-allergic effects at doses of 15 and 60 mg.

Recent clinical trials have shown the possibility of probiotics in prevention and treatment of allergic diseases. The purpose of this experimental study was to assess the influence of lysed Enterococcus faecalis FK-23 (LFK) on allergic responses in different mouse strains. We performed a comparative study on the effects of LFK for allergen-induced peritoneal accumulation of eosinophils and serum total IgE concentration by using BALB/c, C57BL/6, C3H/HeN and C3H/HeJ mice. There was no significant difference in total number of peritoneal accumulated cells induced by cedar pollen allergen between the control and LFK groups in any strain of mice (p > 0.05); however, the ratio of eosinophils to total accumulated cells was significantly decreased in LFK-treated mice of BALB/c (p = 0.016), C3H/HeN (p = 0.010) and C3H/HeJ (p = 0.004), but not C57BL/6 (p > 0.05). No significant difference in serum total IgE concentration was found between the control and LFK groups of different mouse strains (p > 0.05). These results reveal a different effect of LFK on suppressing allergen-induced local eosinophila in inbred strains of mice, suggesting the effectiveness of probiotics on limiting allergy might be under the influence of individual genetic background.

Background: Recent epidemiological studies have indicated that early life receipt of antibiotics may be associated with an increased risk of developing atopic disorder. Lysed Enterococcus faecalis FK-23 (LFK), a probiotic product of E faecalis, has been shown to have inhibitory effects on allergen-induced immune responses in mice., Objective: The purpose of this study was to evaluate the effects of LFK on immune responses and intestinal microflora in antibiotic-treated, and allergen-sensitized weaning mice., Methods: Three-week-old BALB/c mice were sensitized with cedar pollen allergen to establish the experimental model. The allergen-induced peritoneal accumulation of eosinophils, serum levels of total and allergen-specific immunoglobulin (Ig) E and IgG2a, and the intestinal bacterial flora were determined in the control, antibiotic, LFK and antibiotic-LFK groups (n = 7 in all groups). Orally administered erythromycin, one kind of macrolide antibiotic, was used for the experiments., Results: There was no significant difference in the allergen-induced peritoneal accumulation of eosinophils and serum specific IgE and IgG2a levels in erythromycin-treated mice compared to a control group. However, the ratio of serum total IgE to IgG2a levels was significantly increased in erythromycin-treated mice relative to that found either in LFK-treated mice or in erythromycin-treated mice with LFK supplementation. The total aerobes, total anaerobes and Enterococcus species of intestinal microflora were not significantly different among all groups. Lactobacillus species were distinctly eliminated in the mice exposed to erythromycin on day 7 and totally recovered in erythromycin-treated mice with LFK intervention on day 28, but could not be recovered in the erythromycin-treated mice without LFK intervention., Conclusions: Our results suggest that LFK may improve the intestinal ecosystem disturbed by antibiotic use, and thereby prevent subsequent development of atopy. However, whether different antibiotics have different effects on immune responses needs to be addressed further.

We examined the effects of a fermented vegetable product (FVP), fermented by yeast as well as lactic acid bacteria, on body weight and fat deposition after ovariectomy in 10-wk-old Sprague-Dawley rats. The rats were divided into the following 5 groups: (1) ovari-ectomized rats fed as usual (OVX); (2) ones receiving FVP at a daily dose of 100 mg/kg (LOW); (3) ones receiving FVP at a daily dose of 1000 mg/kg (HIGH); (4) ones receiving a calcium-free diet (Ca-FREE); (5) sham-operated rats (SHAM). A slight increase in body weight was observed in the LOW group compared with the OVA group. The rate of increase in total fat content was lowest in the LOW group, but both bone weight and strength were similar to those in the other ovariectomized groups. In conclusion, the low dose of FVP turned out to reduce fat content without affecting bone weight and strength.