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The 11th International Symposium on Adaptive Motion of Animals and Machines. Kobe University, Japan. 2023-06-06/09. Adaptive Motion of Animals and Machines Organizing Committee., Poster Session P49

New findings What is the central question of this study? Does Enterococcus faecium strain R30 (R30), a new lactic acid bacterial strain for supplementation, attenuate shifts in the typology of whole muscle fibres from slow- to fast-twitch by altering the autonomic nervous system in atrophied skeletal muscles? What is the main finding and its importance? R30 supplementation may attenuate the shifts in the typology of whole muscle fibres from slow- to fast-twitch fibres by upregulating peroxisome proliferator-activated receptor-γ coactivator-1α and activating the calcineurin-nuclear factor of activated T-cells signalling pathway, thus ameliorating the decrease in muscle endurance associated with disuse. Abstract Enterococcus faecium strain R30 (R30), a new lactic acid bacterial strain for supplementation, was hypothesized to attenuate shifts in the typology of whole muscle fibres from slow- to fast-twitch fibres in atrophied skeletal muscles. We further postulated that the prevention of slow-to-fast fibre shifts would suppress the decreased muscle endurance associated with atrophy. To evaluate the protective effects of R30, we analysed slow-to-fast fibre shifts and disuse-associated reduced muscle endurance. R30 was administered to rats with an acclimation period of 7 days before hindlimb unloading (HU) for 2 weeks. The composition ratio of the fibre type and the expression levels of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α), calcineurin and nuclear factor of activated T-cells (NFAT) were measured. Muscle endurance was evaluated at the end of the 2-week HU period in an in situ environment. R30 supplementation suppressed the slow-to-fast fibre switch and decreased the HU-induced expression of PGC-1α proteins and the deactivation of the calcineurin-NFAT pathway. Furthermore, R30 prevented a decrease in HU-associated muscle endurance in calf muscles. These results indicate that R30 supplementation may attenuate the shifts in the typology of whole muscle fibres from slow- to fast-twitch fibres via the upregulation of PGC-1α and the activation of the calcineurin-NFAT signalling pathway, thereby ameliorating the decrease in muscle endurance associated with disuse.

Protein-containing nutrients result in the efficient hypertrophy of muscles by increasing muscle protein synthesis. Soybean is often ingested by athletes or individuals who exercise; however, it takes very long to be absorbed. Lactic acid-fermented and enzyme-digested (LFED) soybean is absorbed faster than untreated soybean. This study aims at determining muscle protein synthesis after ingesting a single bolus of soybean or LFED soybean produced by lactic acid bacteria and protease digestion. Eight-week-old overnight-fasted ICR mice were administered powdered or LFED soybean. Mice were euthanized at 7, 15, 30, 60, 90, and 120 min after soybean intake. We have demonstrated that LFED soybean administration was quicker in stimulating muscle protein synthesis by activating mammalian target of rapamycin (mTOR) signaling than orally ingesting untreated soybean in the gastrocnemius muscle. These results suggested that LFED soybean is a more efficient source of nutrition for muscle hypertrophy than untreated soybean.

The biological effects of heat-killed Pediococcus acidilactici R037 (R037) were evaluated when orally administered in mice and rats. Oral R037 administration at a daily dose of 10 and 100 mg/kg for 3 wk dose-dependently reduced fasting and non-fasting serum triglyceride concentrations in KK-Ay/TaJcl mice, a model of type II diabetes, obesity, hypercholesterolemia, and hypertriglyceridemia. Serum levels of free fatty acids in the 100 mg/kg group tended to decrease (not statistically significant), and total cholesterol levels remained unchanged. Treatment with R037 resulted in a significant decrease in blood glucose (at 100 mg/kg) and liver weight (at 10 and 100 mg/kg), and a small body weight gain (at 100 mg/kg) as compared to those in control mice. In addition, oral R037 administration at 100, 200, and 400 mg/kg/d for 1 wk dose-dependently suppressed the increase in serum triglyceride levels in Wistar rats after oral fat loading. Moreover, intraduodenal injection of 120 mg of R037 in Wistar rats suppressed gastric vagal nerve activity (GVNA) indicating suppression of intestinal digestion and absorption of food, and suppression of appetite. The R037 injection potentiated epididymal white adipose tissue sympathetic nerve activity (WAT-SNA) and tended to potentiate pancreatic sympathetic nerve activity (PSNA), suggesting that R037 activated lipolysis. Taken together, these findings indicate that R037 lowers serum triglycerides, possibly through suppressing intestinal absorption and potentiating lipolytic pathways. R037 may be useful for primary prevention of coronary artery diseases in subjects with mild or borderline dyslipidemia in combination with lifestyle changes.

The biological effects of heat-killed Pediococcus acidilactici R037 (R037) were evaluated when orally administered in mice and rats. Oral R037 administration at a daily dose of 10 and 100 mg/kg for 3 wk dose-dependently reduced fasting and non-fasting serum triglyceride concentrations in KK-A

Objective A chronic decrease in neuromuscular activity results in atrophy and capillary regression in skeletal muscles. The purposes of the present study were to determine the effects of enterococcus faecium strain R30 (R30) administration on 1) the hemodynamics of the rat soleus muscle and 2) the capillary regression normally associated with hindlimb unloading (HU). Methods Experiment 1: the velocity of red blood cells (VRBC) was measured for up to one hour after administration of R30 with or without the β-blocker propranolol. Experiment 2: R30 was administered daily to control and HU rats for two weeks. Mean capillary luminal diameter, volume, and the levels of endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) protein were measured. Results Experiment 1: VRBC was faster 20, 40, and 60 min after than before the administration of R30: this effect was suppressed by propranolol administration. Experiment 2: R30 administration during HU increased capillary luminal diameter and volume and eNOS and VEGF protein levels in the soleus of HU rats. Conclusions The results suggest that R30 increased VRBC in the soleus muscle via muscle sympathetic nerve activity (Experiment 1) and that R30 supplementation lessens the capillary regression normally associated with HU via the eNOS/VEGF pathway (Experiment 2). This article is protected by copyright. All rights reserved.

Objectives: The present study sought to evaluate the effect of electrical stimulation (ES) by using kilohertz frequency on muscle atrophy induced by sepsis. Methods: Seventeen male ICR mice were randomly divided into 3 groups: control, lipopolysaccharide (LPS)-injected for 4 days, LPS plus ES (LPS+ES). Sepsis was induced by 4 days of an intraperitoneal LPS injection (10 µg/g body weight/day). LPS+ES animals received the LPS injections and ES twice a day for 4 days. ELISA and western blot analysis determined the plasma levels of inflammatory cytokines and ubiquitinated proteins, while the tibialis anterior muscles were weighed and muscle fiber cross-sectional area (CSA) were measured to assess muscle atrophy, which were analyzed by Student’s t-test and ANOVA. Results: LPS induced increased plasma levels of inflammatory cytokines, significant muscle mass loss (LPS:-29.0%, LPS+ES:-23.1%), decreased fiber cross-sectional area, and an up-regulation of atrogin-1 and ubiquitinated proteins in the tibialis anterior muscle compared with the control. ES attenuated the sepsis-induced loss of muscle mass and decreased fiber CSA, as well as attenuated the atrogin-1 and ubiquitinated protein up-regulation. Conclusions: Electrical stimulation may prevent sepsis-induced muscle atrophy through ubiquitin-proteasome pathway inhibition.

We found a facile one‐step synthetic method for water‐soluble fullerenols bearing 8–16 nitrogen‐containing substituents as well as more than 16 hydroxyl groups from commercially available C60 as a starting material. The water solubility of one of the products showed the same as the highest one reported (>200 mg/mL). Other fullerenols bearing hydrogen or diethylamino group on the same fullerene cage were also synthesized by the modified method using hydrogen peroxide as a simple hydroxylation reagent.

本研究の代表者らは、《まちの保健室》事業の趣旨に基づき看護系大学で展開される「専門相談」の一環として、精神看護学担当教員による『こころの健康相談』を立ち上げ、平成15年度より実践している。これまでの活動実績と、来談者のニーズおよび相談の成果に関する調査結果から、当活動は地域の関連専門機関との補完的な役割を担い得るという実践的意義が示唆された。そこで、今回は、当活動独自のアプローチと役割の探求を目的として、相談記録の質的帰納的分析ならびに地域の関連機関の専門職への面接調査を実施した。平成17・18年度の全相談件数65件に関する相談記録を分析した結果、、、、、、、、などのアプローチの要素が抽出され、来談者のニーズに応じて組み合わされていることが明らかになった。また、地域の関連機関に所属する専門職のうち研究協力に同意の得られた10名(医師5名、精神保健福祉士2名、臨床心理士2名、保健師1名)に対する面接調査の結果、、、といった当活動への認識、、、などの役割期待が語られるとともに、活動頻度を増やすことや連携体制についての提案が得られた。以上の結果および考察を踏まえて、大学を拠点とした看護師による『こころの健康相談』の実践活動について、概念モデルの試案を作成した。今後は妥当性の検討および精錬を重ね、活動の普及と人材育成に貢献し得る実践モデルを開発することが課題である。, As a part of the specialized consultations developed at a nursing-affiliated university based on the concepts of the Town Healthcare Room project, the researchers in this study have been undertaking mental health nursing consultations since 2003. The needs and the benefits of the consultees were examined by ongoing interview surveys and the written questionnaire surveys of the consulties. As a result, it is suggested that these activities can cover complementary roles with related specialized organizations. Then with the goal of finding an original approach to these activities, we undertook a qualitative analysis of the approach offered by the consultation process and conducted studies via related regional organizations on the awareness of and role expectations toward the activities. The following findings were obtained. A qualitative analysis of the records of 65 consultations in 2005 and 2006 revealed the elements of the approach, including the recipience of worries, the recognition of one's own strengths, a review of the stress situation, an increase in coping competence, advice on psycho-education, the provision of specialized knowledge regarding illness and treatment, an assessment of mental state, and the provision of information on social resources, as well as a variety of techniques combined to address the needs of the consultees. Of specialists who belonged to regional organizations, ten persons agreed to cooperate in the research (five physicians, two mental health welfare workers, two clinical psychologists, and one public health nurse). As a result of the interview surveys, we heard about awareness of the activities, including their "ease of use," "feeling safe to talk about issues," and "nurse's own role," as well as their coverage of areas that specialized organizations can not cover, regional contributions that can only be made by a university, the specialized nature of nursing, and other role expectations, and at the same time we received suggestions to step up the frequency of the activities and proposals regarding the liaison system. In view of the above findings and considerations, we prepared a proposal for a concept model of mental health nursing consultation activities based at a university.

Using a novel hydrogen peroxide heating method, we synthesized milky white, water-soluble polyhydroxylated fullerenes (fullerenols) with 36-40 hydroxyl groups (estimated average) along with 8-9 secondary bound water molecules. The fullerenols exhibited high water solubility up to 58.9 mg/mL in a neutral (pH = 7) condition. Dynamic light scattering analysis showed a high dispersion property, to give a narrow particle size distribution within 0.7-2.0 nm.

本研究の代表者らは、《まちの保健室》事業の趣旨に基づき看護大学教員が展開する『専門相談』の一環として、精神看護学担当教員による『こころの健康相談』を立ち上げ、平成15年度より実践している。1年間の活動実績から、来談者にとっての、精神神経科や心療内科に求めるものとは異なるニーズが示唆された。そこで、2年目の活動実績の検討ならびに継続来談者への面接調査を行い、来談者のニーズおよび相談の効果について検討した。その結果、相談件数は1年目の約1.8倍に増加しており、来談理由については、精神的問題をもつ子どもの親としての悩みや家族関係のストレスが過半数を占めていた。自分自身の健康問題では抑うつ状態が最も多く、その半数は神経科に通院服薬中であり、その他に通院中の心療内科から紹介された人もいた。これらのことから、来談者の多くは精神科・神経科、心療内科などの医療機関の活用もしくは補完的な機能に関するニーズを有しているものと考えられた。継続来談者への面接調査の結果をも含めて検討した結果、(1)専門的な判断や助言、(2)じっくりと話を聴いてもらうこと、(3)適切な社会資源の情報、などに関するニーズが明らかになった。相談による効果については、継続来談者には、(1)心理的な安定が得られた、(2)問題解決への動機づけが高まった、(3)専門的な情報や助言を主体的に活用するようになった、(4)状況の改善や自己効力感の高まりが得られた、(5)悩みはありながらも安定した生活を営めるようになった、などの前向きな変化がみられた。, In accordance with the purpose of the "Community Healthcare Room" project, in 2003 the representatives of this research set up the Mental Health Consultation Room with a base at the College of Nursing Art and Science, University of Hyogo. Since then, mental health nursing consultation has been provided by faculty members teaching mental health and psychiatric nursing. This paper reports the results of a study that examined the needs of clients and effects of the consultations, through a review of activities during the second year of the project's operation and interviews with those who continue to visit the consultation office. The number of consultation cases in the second year increased to 1. 8 times the number seen in the first year. As for the reasons for visiting the mental health consultation room, the majority of reasons cited were worries as a parent who has a child with mental health problems or stress from family relationships. Among visitors who had their own health problems, the largest number of people complained about depressive moods, half of whom see a psychiatric doctor regularly and take medication. There were some people who visited the consultation room on the recommendation of their psychiatric doctors. From these findings, it is suggested that many visitors expect the consultation room to perform complementary functions with a medical institution such as a psychiatric clinic and psychosomatic medicine clinic. As a result of examining the results of interviews with those continuing to go to the consultation room, in addition to findings from past activities, the following needs of visitors have been identified : (1) professional judgment and advice ; (2) active listening ; and (3) information on and referral to appropriate social resources. Regarding the effects of consultation, the following positive changes were seen in visitors (1) increased mental stability ; (2) increased motivation for solving problems ; (3) more proactivity in seeking professional information and advice ; (4) improved conditions and increased selfefficacy ; and (5) becoming able to have a stable life despite someworries.

New yellow-green pigments based on cerium orthophosphate were synthesized by the solid-state reaction in order to develop ecological and environmentally-friendly specific pigments. The pigments were obtained by doping alkaline earth metal ions into a cerium orthophosphate lattice to form their solid solutions. The products were characterized by using X-ray powder diffraction, UV-Vis spectroscopy, Fourier-transform infrared spectroscopy (FT-IR), Raman spectroscopy, and colorimetric assessment. From these results, it has been evidenced that the formation of solid solution is necessary to have color coordinates in the synthesized pigment samples. Since the color of the pigments was almost the same yellow-green in all solid solution samples, the origin of the coloration was attributed to the appearance of the principal broad absorption band corresponding to the 4f-5d electron transition of Ce 3 + and the additional O 2 p Ce 4 f charge transfer transition of Ce 4 + by the doping of alkaline earth ion into CePO 4 lattice.

Our research has focused on the molecular design of immunotherapeutic vaccines and the advancement of mite-allergy diagnosis. Here, we describe the research history of the major group 1 and group 2 allergens, immunoelectrophoretic analyses covering the complete spectrum of mite allergens, our results on allergens with distinctive characteristics (a conjunctival congestion-eliciting antigen [LM2], an immunotherapeutic antigen [HM2] with high efficacy and without definite adverse reactions, and a potent T-cell stimulatory antigen [HM1] with secretion of IFN-γ), the full spectrum and immunochemical properties of the major and other important mite allergens (including our newly described allergens: a pan-allergen [tropomyosin, group 10], a potent T-cell stimulatory allergen [M-177, apolipophorin, group 14] and its peptide fragments Mag1 and Mag3, a moderate IgE-binding allergen [gelsolin/villin, group 16], an EF-hand Ca2+-binding allergen [group 17], and a less IgE-binding allergen [heat shock protein 70]), and prospects for the development of immunotherapeutic and diagnostic agents.

Background: We have previously demonstrated that high-molecular mite antigen (HM1) from Dermatophagoides farinae feces is an allergen which binds to mite-allergic patients IgE. HM1 also induced a proliferative response in lymph node cells from mite-immunized mice as well as nonimmunized mice. In the present study, we demonstrated that HM1 induced T cell proliferation and investigated the HM1-stimulated T cell proliferative pathways using nonallergic human peripheral blood mononuclear cells (PMBC). Methods: Blood samples were obtained from 10 healthy donors. Using primary culture, T cell response stimulated with HM1 was performed on purified T cells, CD19+ cell-depleted PBMC and CD11b+ cell-depleted PBMC. In addition, interleukin (IL)-5 and interferon (IFN)-γ produced by mite-allergic and healthy donors stimulated with HM1 were estimated by enzyme immunoassay. Results: T cell proliferation was detected only in CD19+ cell-depleted PBMC. When T cells were cocultured with CD11b+ cells they recovered their proliferative response to HM1. In addition, the pathway of HM1-stimulated T cell proliferation did not involve HLA class II restriction. Both activated CD11b+ cells and their conditioned media were needed to induce HM1-stimulated T cell proliferation. Furthermore, HM1 induced IFN-γ production in both healthy and allergic donors. Conclusion: The high-molecular mite antigen, HM1, induced a proliferative response of T cells in healthy as well as allergic donors, without HLA class II restriction. Our results suggest that further investigation of HM1 could constitute a valuable avenue of research into complex allergic diseases.

Background: The house dust mite Dermatophagoides farinae is the most common aeroallergen causing human allergic asthma. Previously, we demonstrated that a high-molecular-weight allergenic fraction (HM1), which was abundant in D. farinae extracts, induced a proliferative response of T cells from healthy donors. The induction was mediated through the activation of macrophages without MHC class II restriction. In this study, we investigate whether HM1 influences the development of airway inflammation in murine models of asthma. Methods: BALB/c mice were injected twice intraperitoneally with D. farinae fecal extract (Dff) at an interval of 5 days. They were exposed daily to aerosolized antigen (group 1: Dff, group 2: HM1, group 3: HM1-depleted Dff and group 4: PBS) for 10 days. The effect of HM1 on their airway inflammation was evaluated by measuring acetylcholine-induced airway hyperresponsiveness and inflammatory cell infiltration in lung tissue. Results: The inhalation of the whole fecal extract or the HM1 fraction induced airway hyperresponsiveness which was detectable after 24 h and was maintained for as long as 120 h. The inhalation of extract depleted of the HM1 fraction induced hyperresponsiveness measured at 24 h but this was not maintained for 120 h. Macrophage infiltration was significantly prolonged in mice inhaling the whole extract and the HM1 fraction compared to the HM1-depleted extract. Conclusion: The inhalation of the high-molecular-weight HM1 fraction of D. farinae prolonged airway hyperresponsiveness and macrophage inflammation in a mouse model of hypersensitivity. The results indicate that the HM1 fraction which can induce T cell proliferation through macrophage activation may play a role in the duration of airway responsiveness.

We investigated the suppressive effect of lactic acid bacteria (LAB) isolated from traditional South Asian fermented milk `dahi' on the development of atopic dermatitis (AD) using NC/Nga AD model mice. In the initial evaluation, we confirmed the effect of LAB on serum total IgE using ovalbumin (OVA)-induced type 1 allergy model mice. Forty-one bacterial strains isolated from dahi were evaluated for their ability to induce interleukin (IL)-12 production and suppress IL-4 production in splenocytes obtained from OVA-sensitized mice. Of the 41 strains tested, Lactobacillus delbrueckii subsp. lactis R-037 exhibited the greatest IL-12 induction, suggesting that it is a potent Th1 inducer. Oral administration of heat-treated R-037 significantly suppressed the elevation of serum total IgE in OVA-induced type 1 allergy model mice. In NC/Nga AD model mice, oral administration of heat-treated R-037 reduced inflammatory auricular thickness and alleviated the AD clinical score although the effect on serum total IgE level was unclear. Histopathological findings showed a tendency toward improvement of inflammation. Hyperkeratosis in particular showed improvement in dermatitis skin lesions. These results suggest that oral administration of R-037 may alleviate AD.

Mouse embryonic stem (ES) cells can be maintained in an undifferentiated state in the presence of leukemia inhibitory factor (LIF), a member of the interleukin-6 cytokine family. In other mammals, this is not possible with LIF alone. Chicken ES-like cells (blastodermal cells) have only been cultured with mouse LIF because chicken LIF was not available. However the culture system is imperfect and chicken ES-like cells equivalent to mouse ES cells were not observed. In the present study, we cloned the cDNA-encoding chicken LIF using mRNA subtraction and RACE methodology. The chicken LIF cDNA encodes a protein with approximately 40% sequence identity to mouse LIF. It has 211 amino acids including a putative N-terminal signal peptide of 24 residues. Chicken blastodermal cells were cultured in the presence of bacterially expressed chicken LIF or mouse LIF. The expression of alkaline phosphatase and embryonal carcinoma cell monoclonal antibody-1 and stage-specific embryonic antigen-1 and the activation of STAT3 were examined, all of which are indices of the undifferentiated state. Exposure in the blastodermal cells to recombinant chicken LIF but not to mouse LIF maintained the expression of these various markers. After 9 days of incubation, the blastodermal cells formed cystic embryoid bodies in the presence of mouse LIF but not in the presence of recombinant chicken LIF. We conclude that chicken LIF is able to maintain chicken ES cell cultures in the undifferentiated state.

Our research has focused on the molecular design of immunotherapeutic vaccines and the advancement of mite-allergy diagnosis. Here, we describe the research history of the major group 1 and group 2 allergens, immunoelectrophoretic analyses covering the complete spectrum of mite allergens, our results on allergens with distinctive characteristics (a conjunctival congestion-eliciting antigen [LM2], an immunotherapeutic antigen [HM2] with high efficacy and without definite adverse reactions, and a potent T-cell stimulatory antigen [HM1] with secretion of IFN-gamma), the full spectrum and immunochemical properties of the major and other important mite allergens (including our newly described allergens: a pan-allergen [tropomyosin, group 10], a potent T-cell stimulatory allergen [M-177, apolipophorin, group 14] and its peptide fragments Mag1 and Mag3, a moderate IgE-binding allergen [gelsolin/villin, group 16], an EF-hand Ca2+-binding allergen [group 17], and a less IgE-binding allergen [heat shock protein 70]), and prospects for the development of immunotherapeutic and diagnostic agents.

We have previously demonstrated that high-molecular mite antigen (HM1) from Dermatophagoides farinae feces is an allergen which binds to mite-allergic patients IgE. HM1 also induced a proliferative response in lymph node cells from mite-immunized mice as well as nonimmunized mice. In the present study, we demonstrated that HM1 induced T cell proliferation and investigated the HM1-stimulated T cell proliferative pathways using nonallergic human peripheral blood mononuclear cells (PMBC).Blood samples were obtained from 10 healthy donors. Using primary culture, T cell response stimulated with HM1 was performed on purified T cells, CD19+ cell-depleted PBMC and CD11b+ cell-depleted PBMC. In addition, interleukin (IL)-5 and interferon (IFN)-gamma produced by mite-allergic and healthy donors stimulated with HM1 were estimated by enzyme immunoassay.T cell proliferation was detected only in CD19+ cell-depleted PBMC. When T cells were cocultured with CD11b+ cells they recovered their proliferative response to HM1. In addition, the pathway of HM1-stimulated T cell proliferation did not involve HLA class II restriction. Both activated CD11b+ cells and their conditioned media were needed to induce HM1-stimulated T cell proliferation. Furthermore, HM1 induced IFN-gamma production in both healthy and allergic donors.The high-molecular mite antigen, HM1, induced a proliferative response of T cells in healthy as well as allergic donors, without HLA class II restriction. Our results suggest that further investigation of HM1 could constitute a valuable avenue of research into complex allergic diseases.

Here we describe the detection of T-cell epitope region on the house dust mite allergen Mag 3, which has been shown to trigger T-cell proliferation in mite-allergic asthmatic patients. We first examined murine T-cell epitope using T-cell fraction prepared from recombinant Mag 3 (r-Mag 3)-primed H-2k mice. Initial proliferation assay with truncated r-Mag 3 indicated that N-terminal 113 amino acid region was required for triggering T-cell activation. Subsequent epitope scanning with synthetic overlapping peptides revealed that T-cell reactive region was assigned within amino acid range 56-75. We also explored human T-cell determinant using specific T-cells from mite-allergic patients. Intriguingly, we found that amino acid range 56-85, a portion partially overlapping with that identified in r-Mag 3-primed mice, was exclusively recognized by T-cells from different patients. Further investigation of unique T-cell epitope region found in this study would provide insight into the development of animal therapeutic model and/or peptide vaccine for asthma.

Background High-molecular size mite antigen fraction (HM1) induced macrophage activation and prolonged airway inflammation from mite-sensitized mice. In the present study, we investigated the inflammatory factors in the HM1-activated macrophage in both non-immunized splenocytes and bronchoalveolar lavage (BAL) from mite-immunized and HM1-exposed mice. Methods Dermatophagoides farinae feces (Dff) extract was divided into HM1 and HM1-depleted fraction (DH) by size-exclusion chromatography. Transcriptional gene induction of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-12 and cytokine-inducible nitric oxide synthase (iNOS) in splenic macrophages stimulated with HM1 or DH was analyzed by using semiquantitative reverse transcription–polymerase chain reaction. Nitric oxide (NO) production was measured by using diaminofluoresceins (DAF), fluorescence indicators. Results Gene expression of TNF-α, IL-12 p40 and iNOS was observed with HM1 stimulation of splenic macrophages from non-immunized mice. In addition, the release of NO induced by HM1-stimulated splenic macrophages increased in a dose-dependent manner. However, splenic macrophages stimulated with DH induced TNF-α and IL-12 p40, but not iNOS, gene expression. Similarly, significant iNOS mRNA expression was detected in alveolar macrophages recovered from HM1-exposed mice, but not from DH-exposed mice. Conclusion In the present study, HM1 induced iNOS mRNA expression and NO production both in vitro and in vivo . The present results suggest that the ability of HM1 to induce NO production may be one of the causes aggravating airway inflammation in HM1-exposed mice.