Eudesmane-Type Sesquiterpene Lactones Inhibit Nuclear Translocation of the Nuclear Factor κB Subunit RelB in Response to a Lymphotoxin ß Stimulation.

The transcription factor nuclear factor κB (NF-κB) regulates various biological processes, including inflammatory responses. We previously reported that eudesmane-type sesquiterpene lactones inhibited multiple steps in the canonical NF-κB signaling pathway induced by tumor necrosis factor-α and interleukin-1α. In contrast, the biological activities of eudesmane-type sesquiterpene lactones on the non-canonical NF-κB signaling pathway remain unclear. In the present study, we found that (11S)-2α-bromo-3-oxoeudesmano-12,6α-lactone, designated santonin-related compound 2 (SRC2), inhibited NF-κB luciferase reporter activity induced by lymphotoxin ß (LTß) in human lung carcinoma A549 cells. Although SRC2 did not prevent the processing of the NF-κB subunit p100 induced by LTß, it inhibited the nuclear translocation of RelB and p52 in response to the LTß stimulation. In contrast to (-)-dehydroxymethylepoxyquinomicin, SRC2 inhibited the LTß-induced nuclear translocation of the RelB (C144S) mutant in a manner similar to wild-type RelB. While eudesmane derivatives possessing an α-bromoketone moiety or α,ß-unsaturated carbonyl moieties inhibited LTß-induced NF-κB luciferase reporter activity, eudesmane derivatives possessing an α-bromoketone moiety exhibited stronger inhibitory activity on the LTß-induced nuclear translocation of RelB than those possessing a single α-methylene-γ-lactone moiety. The results of the present study revealed that SRC2 inhibits the nuclear translocation of RelB in the non-canonical NF-κB signaling pathway induced by LTß.

Quinacrine Inhibits ICAM-1 Transcription by Blocking DNA Binding of the NF-κB Subunit p65 and Sensitizes Human Lung Adenocarcinoma A549 Cells to TNF-α and the Fas Ligand.

Quinacrine has been used for therapeutic drugs in some clinical settings. In the present study, we demonstrated that quinacrine decreased the expression of intercellular adhesion molecule-1 (ICAM-1) induced by tumor necrosis factor (TNF)-α and interleukin-1 (IL-1) α in human lung adenocarcinoma A549 cells. Quinacrine inhibited ICAM-1 mRNA expression and nuclear factor κB (NF-κB)-responsive luciferase reporter activity following a treatment with TNF-α and IL-1α. In the NF-κB signaling pathway, quinacrine did not markedly affect the TNF-α-induced degradation of the inhibitor of NF-κB or the TNF-α-induced phosphorylation of the NF-κB subunit, p65, at Ser-536 and its subsequent translocation to the nucleus. In contrast, a chromatin immunoprecipitation assay showed that quinacrine prevented the binding of p65 to the ICAM-1 promoter following TNF-α stimulation. Moreover, TNF-α and the Fas ligand effectively reduced the viability of A549 cells in the presence of quinacrine only. Quinacrine down-regulated the constitutive and TNF-α-induced expression of c-FLIP and Mcl-1 in A549 cells. These results revealed that quinacrine inhibits ICAM-1 transcription by blocking the DNA binding of p65 and sensitizes A549 cells to TNF-α and the Fas ligand.

CT findings of thoracic manifestations of primary Sjögren syndrome: radiologic-pathologic correlation.

Primary Sjögren syndrome is an immune-mediated exocrinopathy characterized by lymphoplasmacytic infiltration of the salivary and lacrimal glands. Various systemic extraglandular disorders are associated with primary Sjögren syndrome, and the thorax is commonly affected. The pulmonary manifestations of primary Sjögren syndrome may be categorized as airway abnormalities, interstitial pneumonias, and lymphoproliferative disorders; in each category, bronchiectasis or centrilobular nodules, nonspecific interstitial pneumonia, and lymphoid interstitial pneumonia are common. These manifestations do not usually occur in isolation; they are concomitantly seen with other types of lesions. Mucosa-associated lymphoid tissue (MALT) lymphoma and amyloidosis are key components of lymphoproliferative disorders, and MALT lymphoma should always be considered because its morphologic characteristics are similar to those of benign lymphoproliferative disorders. Amyloidosis is rare but important because it carries a risk for underlying MALT lymphoma or plasmacytoma, and it may lead to hemoptysis during biopsy. In addition, thin-walled air cysts are characteristic of primary Sjögren syndrome, irrespective of the main pulmonary manifestations. Lymphadenopathy and multilocular thymic cysts may be seen in the mediastinum. During the follow-up period, there is a risk for acute exacerbation of interstitial pneumonia and development of malignant lymphoma. Often, primary Sjögren syndrome is subclinical, but there are various underlying risks. Thus, imaging findings are important. In addition to the various types of interstitial pneumonia and airway abnormalities, air cysts and mediastinal manifestations may help diagnose primary Sjögren syndrome.

An increase in ALS incidence on the Kii Peninsula, 1960-2009: a possible link to change in drinking water source.

We investigated changes in the incidence of amyotrophic lateral sclerosis (ALS) in the Koza/Kozagawa/Kushimoto area (K. area) in the Kii Peninsula, Japan in 1960-2009. Probable and definite ALS cases diagnosed using El Escorial criteria were collected during a five-decade period: period I-V, 1960-2009. Forty-three ALS patients matched the selection criteria in the overall K. area, including three patients on Oshima, a small island opposite the mainland K. area. The age- and gender-adjusted incidence of ALS in the overall K. area (standardized for the 2005 Japanese population) decreased from 5.47/100,000 (95% CI 1.86-9.08) in period I to 0.61/100,000 (95% CI-0.28-1.50) in period III, and then increased to 4.39/100,000 (95% CI 1.70-7.07) in period V. On Oshima, the age- and gender-adjusted incidence of ALS was 9.45/100,000 (95% CI-7.39-26.29) in period V. The present research indicates an increase of ALS incidence in the K. area, especially on Oshima. A limitation of this study was the small population.

Seed dormancy in Trillium camschatcense (Melanthiaceae) and the possible roles of light and temperature requirements for seed germination in forests.

PREMISE OF THE STUDY: Seeds of some temperate plants require multiple treatments to break complex forms of dormancy, such as deep simple double morphophysiological dormancy (MPD), but embryo growth and radicle and cotyledon emergence have not been studied in depth for this level of MPD. Here we studied Trillium camschatcense, a species that is purported to have this MPD and that is sensitive to habitat fragmentation with decreased recruitment at forest edges. METHODS: Trillium camschatcense seeds were collected and experiments conducted in Hokkaido, Japan over 5 years. Growth of embryos and emergence of radicles and cotyledons were tracked in relation to field temperature to determine specific temperature and light requirements for these stages under laboratory conditions. KEY RESULTS: Low (winter) temperatures overcame dormancy in the underdeveloped embryos, and embryo growth and radicle emergence occurred between July and September, ca. 1 year after seed dispersal. Radicles emerged optimally over a narrow temperature range (20-25°C), in darkness, and at constant temperatures. Roots developed during the second autumn. Cotyledons needed a second low temperature (second winter) to emerge from seeds with roots, doing so in April, slightly over 1.5 years after dispersal. CONCLUSIONS: Seeds of T. camschatcense have deep simple double MPD and requirements for radicle emergence: darkness and constant temperatures. Ecologically, edges of forests may be deleterious for germination of the species due to increased light and to higher temperature fluctuations as compared to the interiors of forests. Thus, these specific requirements may play an important role in reducing seed germination of this plant at forest edges.

Pleuroparenchymal fibroelastosis (PPFE)-like finding on CT in daily practice -prevalence and serial changes.

OBJECTIVE: To characterize the prevalence of PPFE (pleuroparenchymal fibroelastosis)-like finding on CT in daily practice and to identify the risk factors for its progression. MATELIALS & METHODS: 2416 consecutive daily CT examinations were screened for PPFE-like finding. CT images with PPFE-like finding were retrospectively reviewed for the extent, maximum height, presence or absence of intraalveolar fibroelastosis (IAFE), emphysema, interstitial lung disease (ILD), suprasternal depression at the level of clavicle end, and bronchiectasis in upper lobe, and anterior-posterior/transverse diameter ratio (AT ratio) of thoracic cage. Serial CT scans more than 3 years before the baseline scan were also reviewed and compared when available. RESULTS: 380 patients (median age of 65, M:F = 153:227) were recognized as having PPFE-like finding. The lowest level of PPFE-like finding was T5 in median, horizontal extent was 0-25 % in 229/380, 25-50 % in 66 and >50 % in 24. Median height was 8.0 mm. IAFE was seen in 75, emphysema in 71, ILD in 59, chronic bronchial disorder in 71, suprasternal depression in 148, upper-lobe bronchial dilatation in 124. Mean AT ratio was 0.63. Serial images were available in 131 patients. 11 showed significant progression, 57 showed mild progression. Existing ILD lesion and >25 % horizontal extent were significantly related to significant progression. CONCLUSIONS: PPFE-like finding was seen relatively in high proportion. Extensive lesion in axial plane and co-existing ILD might be risk factors of progression.

4-O-Methylascochlorin inhibits the prolyl hydroxylation of hypoxia-inducible factor-1α, which is attenuated by ascorbate.

4-O-Methylascochlorin (MAC), a methylated derivative of ascochlorin, was previously shown to promote the accumulation of hypoxia-inducible factor (HIF)-1α in human breast adenocarcinoma MCF-7 cells. In the present study, we further investigated the effects of MAC on the expression and function of HIF-1α in human fibrosarcoma HT-1080 cells. MAC promoted the accumulation of the HIF-1α protein without affecting its constitutive mRNA expression and augmented the transcriptional activation of HIF target genes. Ascorbate, but not N-acetylcysteine, attenuated MAC-mediated HIF-1α accumulation. MAC-induced increases in HIF-1α transcriptional activity were also attenuated by ascorbate. MAC inhibited the hydroxylation of HIF-1α at the proline 564 residue, while it was reversed by ascorbate. MAC slightly decreased the intracellular concentration of ascorbate. The present results demonstrated that MAC promoted the accumulation of HIF-1α by preventing prolyl hydroxylation, and ascorbate attenuated the MAC-mediated inhibition of HIF-1α prolyl hydroxylation.

Brain cytokines and the 5-HT system during poly I:C-induced fatigue.

Fatigue is evoked not only by peripheral factors, such as muscle fatigue, but also by the central nervous system (CNS). For example, it is generally known that the feeling of fatigue is greatly influenced by psychological aspects, such as motivation. However, little is known about the central mechanisms of fatigue. The clinical symptoms of chronic fatigue syndrome (CFS) are shown to include disorders in neuroendocrine, autonomic, and immune systems. On the other hand, it has been demonstrated that cytokines produced in the brain play significant roles in neural-immune interactions through their various central actions, including hypothalamo-pituitary and sympathetic activation, as well as immunosuppression. In this article, using the immunologically induced fatigue model, which was achieved by intraperitoneal (i.p.) injection of synthetic double-stranded RNAs, polyriboinosinic: polyribocytidylic acid (poly I:C) in rats, we show an involvement of brain interferon-alpha (IFN-alpha) and serotonin (5-HT) transporter (5-HTT) in the central mechanisms of fatigue. In the poly I:C-induced fatigue rats, expression of IFN-alpha and 5-HTT increased, while extracellular concentration of 5-HT in the medial prefrontal cortex decreased, probably on account of the enhanced expression of 5-HTT. Since the poly I:C-induced reduction of the running wheel activity was attenuated by a 5-HT(1A) receptor agonist, but not by 5-HT(2), 5-HT(3), or dopamine D(3) receptor agonists, it is suggested that the decrease in 5-HT actions on 5-HT(1A) receptors may at least partly contribute to the poly I:C-induced fatigue.

Endotoxin inhibitor blocks heat exposure-induced expression of brain cytokine mRNA in aged rats.

To investigate the age-related changes in the expression of interleukin-1beta (IL-1beta) and its related substances in the brain during heat stress, we measured amounts of mRNAs for IL-1beta, cyclooxygenase-2 (COX-2), and an inhibitor of nuclear factor (NF)-kappaB-beta (IkappaB-beta) that is known to reflect an activation of NF-kappaB, in the cortex, cerebellum, and hippocampus using a quantitative real-time capillary PCR method. The basal levels of IL-1beta mRNA in aged rats (108-110 weeks old) was significantly higher than those in young animals (10-11 weeks old) in these brain regions. Heat exposure (33 degrees C) for 1 h enhanced the expression of IL-1beta and COX-2 mRNAs in aged rats but not in young ones. The amount of lipopolysaccharide (LPS) assessed by its bioactivity in the cortex increased by heat exposure only in aged rats. To further examine an involvement of LPS in the increase in mRNAs, an endotoxin inhibitor (EI), a synthetic peptide that detoxifies LPS by binding to the toxic component of LPS, lipid A, was intraperitoneally injected before heat exposure in aged rats. An intraperitoneal injection of EI significantly attenuated the heat exposure-induced increases in mRNAs for IL-1beta, COX-2, IkappaB-beta, and the LPS activity. Administration of EI also debilitated the heat exposure-induced hyperthermia and responses of plasma ACTH and catecholamines. These findings, taken together, suggest that the bacterial translocation is involved in the mechanisms of the responses to heat exposure in aged rats including the increased expression of mRNAs for IL-1beta and its related substances in the brain.

Purification and characterization of formate oxidase from a formaldehyde-resistant fungus.

A formate oxidase activity was found in the crude extract of a formaldehyde-resistant fungus isolated from soil. The fungus was classified and designated as Aspergillus nomius IRI013, which could grow on a medium containing up to 0.45% formaldehyde and consumed formaldehyde completely. The specific activity of formate oxidase in the extract of the fungus grown on formaldehyde was found to be considerably higher than that in the extracts of the fungus grown on formate and methanol. Formate oxidase from the fungus grown on formaldehyde was purified to homogeneity. The enzyme had a relative molecular mass of 100000 and was composed of two apparently identical subunits that had a relative molecular mass of 59000. The enzyme showed the highest activity using formate as substrate. Hydrogen peroxide was formed during the oxidation of formate. The Michaelis constant for formate was 15.9 mM; highest enzyme activity was found at pH 4.5-5.0. The enzyme activity was strongly inhibited by NaN(3), p-chloromercuribenzoate and HgCl(2).

Stem cell factor modulates paired-pulse facilitation and long-term potentiation in the hippocampal mossy fiber-CA3 pathway in mice.

The effects of recombinant mouse stem cell factor (rmSCF) on paired-pulse facilitation (PPF) and long-term potentiation (LTP) in the mossy fiber (MF)-CA3 pathway were examined in mouse hippocampal slices by recording field EPSPs. When PPF was measured before and 30 min after tetanic stimulation, the initial PPF positively correlated with the amplitude of LTP and negatively correlated with changes in PPF (PPF after LTP minus initial PPF), indicating a presynaptic component in MF-CA3 LTP. Bath application of rmSCF for 30 min also produced negative correlation between initial PPF and changes in PPF after rmSCF, suggesting common mechanisms of the LTP- and rmSCF-induced modulation of PPF. The rmSCF-induced negative correlation was abolished by simultaneous perfusion with K252a, a receptor tyrosine kinase inhibitor, and by wortmannin, a phosphatidylinositol-3'-kinase inhibitor. Although SCF activates phospholipase A(2) (PLA(2)) and diacylglycerol (DAG) lipase to produce arachidonic acid (AA) in mast cells, mepacrine, a PLA(2) inhibitor, but not RHC80267, a DAG lipase inhibitor, abolished the negative correlation. The induction of LTP was prevented by perfusion with rmSCF started 30 min before tetanus, while preincubation of slices with antibody for SCF receptor, c-kit, blocked LTP, suggesting that the intrinsic SCF is involved in the induction of LTP and the blockade of LTP by rmSCF might be due to an occlusion of SCF/c-kit signaling. In addition, since c-kit is expressed on the postsynaptic CA3 neurons but not on the MF terminals in mice, effects of rmSCF on PPF may be mediated by the PLA(2)-induced AA acting as a retrograde messenger.

Somatic symptoms most often associated with depression in an urban hospital medical setting in Japan.

Somatic symptoms are known to be a prominent characteristic in patients with depression. The aim of the present study was to investigate the major somatic and psychiatric symptoms associated with depression in each sex. Patients with a DSM-IV diagnosis of depressive disorders (n=335) and comparison patients without depression (n=423) among new outpatients in an urban hospital medical setting were eligible for study. The relationship between the the most distressing subjective symptoms at first visit, confirmed by the patient's description on a health questionnaire and the admitting physician's interview, and depression was investigated in each sex. Most (73.5%) of the complaints in patients with depression were somatic. In a simple logistic regression analysis, diarrhea, excessive sweating and psychomotor retardation in men and headache, depressed mood and grief in women were associated with depression. In multiple logistic regression analysis, diarrhea, excessive sweating and weight loss in men, and headache, dysesthesia and grief in women, as well as sleep disturbance, loss of appetite, general fatigue loss on interest and agitation in both sexes, were significantly associated with depression. Fever in men was also associated with depression by Fisher's exact test.

Somatic symptoms most often associated with depression in an urban hospital medical setting in Japan.

Somatic symptoms are known to be the major manifestation in patients with depression. The aim of the present study was to investigate the major somatic and psychiatric symptoms associated with depression in each sex. Patients with a DSM-IV diagnosis of depressive disorders (n=335) and comparison patients without depression (n=425) among new outpatients in an urban hospital medical setting were eligible for study. The relationship between the three most distressing subjective symptoms at the first visit, confirmed by the patient's description on a health questionnaire and the admitting physician's interview, and depression was investigated in each sex. Most (77.4%) of the complaints in patients with depression were somatic. In a simple logistic regression analysis, diarrhea, excessive sweating and psychomotor retardation in men, and headache, depressed mood and grief in women were associated with depression. In multiple logistic regression analysis, diarrhea, excessive sweating and weight loss in men, and headache, dysesthesia and grief in women, as well as sleep disturbance, loss of appetite, general fatigue, loss of interest and agitation in both sexes, were statistically significantly associated with depression. Fever in men was also associated with depression by Fisher's exact test.

Acupuncture and moxibustion for stress-related disorders.

Acupuncture and moxibustion, which medical doctors are licensed by the government of Japan to perform, can improve the psychological relationship between doctors and patients, especially when it is disturbed by a "game", a dysfunctional interpersonal interaction that is repeated unintentionally. This advantage is due to the essential properties of acupuncture and moxibustion. Acupuncture and moxibustion are helpful in treating somatoform disorders, especially musculoskeletal symptoms. In Japan, a holistic acupuncture and moxibustion therapy called Sawada-style has been developed. This is based on fundamental meridian points that are considered to have effects on central, autonomic nervous, immune, metabolic, and endocrine systems to regulate the whole body balance. In addition, some of the fundamental points have effects on Qi, blood, and water patterns associated with major depression, generalized anxiety disorder, eating disorders, and somatoform disorders. The fixed protocol of Sawada-style would be suitable for large-scale, randomized, controlled studies in the future. Recent systematic reviews indicate that electroacupuncture would be a useful addition to antidepressant therapy for some symptoms accompanying fibromyalgia. Acupuncture and moxibustion are also recommended for irritable bowel syndrome, instead of Western drug therapy. Surprisingly, the dorsal prefrontal cerebral cortex, which is associated with a method of scalp acupuncture applied for gastrointestinal disorders, has been found to be activated in patients with irritable bowel syndrome. It is quite possible that regulation of this cortical area is related to the effect of scalp acupuncture. This acupuncture method can be effective not only for irritable bowel syndrome but also for other stress-related gastrointestinal disorders.

Seed germination and seedling development ecology in world-wide populations of a circumboreal Tertiary relict.

BACKGROUND AND AIMS: Temperate forests are disjunct in the Northern Hemisphere, having become fragmented from the earlier widespread (Tertiary) boreotropical forest. We asked 'What are the contemporary patterns of population variation in ecological traits of a Tertiary relict in a macroecological context?'. This issue underpins our understanding of variation in populations occurring in the same biome but on different continents. METHODOLOGY: We examined characters associated with root and shoot emergences among populations of Viburnum opulus in temperate forests of Asia, North America and Europe. This species has complex seedling emergence extending over several years and requiring various temperature cues. PRINCIPAL RESULTS: Populations varied in germination responses and clustered into groups that were only partly related to varietal status. Whereas roots (at warm temperatures) and shoots (following a cold period) simultaneously emerged from seeds of all populations when simulated dispersal occurred in winter, they were delayed in some populations when dispersal occurred in summer. CONCLUSIONS: Viburnum opulus populations, some separated by 10 300 km, showed high similarity in seedling development and in germination phenology, and we suggest that stabilizing selection has played a key role in maintaining similar dormancy mechanisms. Nevertheless, there was some degree of variation in other germination characters, suggesting local adaptation.

Genotypic and phenotypic diversity of rhizobia isolated from Lathyrus japonicus indigenous to Japan.

Sixty-one rhizobial strains from Lathyrus japonicus nodules growing on the seashore in Japan were characterized and compared to two strains from Canada. The PCR-based method was used to identify test strains with novel taxonomic markers that were designed to discriminate between all known Lathyrus rhizobia. Three genomic groups (I, II, and III) were finally identified using RAPD, RFLP, and phylogenetic analyses. Strains in genomic group I (related to Rhizobium leguminosarum) were divided into two subgroups (Ia and Ib) and subgroup Ia was related to biovar viciae. Strains in subgroup Ib, which were all isolated from Japanese sea pea, belonged to a distinct group from other rhizobial groups in the recA phylogeny and PCR-based grouping, and were more tolerant to salt than the isolate from an inland legume. Test strains in genomic groups II and III belonged to a single clade with the reference strains of R. pisi, R. etli, and R. phaseoli in the 16S rRNA phylogeny. The PCR-based method and phylogenetic analysis of recA revealed that genomic group II was related to R. pisi. The analyses also showed that genomic group III harbored a mixed chromosomal sequence of different genomic groups, suggesting a recent horizontal gene transfer between diverse rhizobia. Although two Canadian strains belonged to subgroup Ia, molecular and physiological analyses showed the divergence between Canadian and Japanese strains. Phylogenetic analysis of nod genes divided the rhizobial strains into several groups that reflected the host range of rhizobia. Symbiosis between dispersing legumes and rhizobia at seashore is discussed.

Temperature requirements differ for the two stages of seed dormancy break in Aegopodium podagraria (Apiaceae), a species with deep complex morphophysiological dormancy.

Only a few studies have considered the possibility that low temperature requirements may vary among stages of dormancy break in seeds with morphophysiological dormancy (MPD). We show that this lack of consideration in previous studies on seed dormancy and germination of Aegopodium podagraria might explain the low germination percentages and/or the relatively long periods of incubation needed for germination. Under natural temperatures, embryos began to grow in September and were fully elongated by late December; most growth occurred when the average daily mean temperature was about 10°C. Radicles emerged under snow in late winter, and cotyledons emerged after snowmelt in early spring. In laboratory experiments, 100% of the embryos grew to full length at both 0 and 5°C, whereas 0°C was much more effective than 5°C in overcoming the physiological dormancy in seeds after embryos were fully elongated. Following radicle emergence, cotyledons emerged readily in a wide range of temperatures ≥5°C. GA(3) did not substitute for the low temperature requirement for dormancy break. Seed dormancy in A. podagraria fits Nikolaeva's formula for deep complex MPD, i.e., C(3)B-C(3). Better germination of seeds pretreated at 0° than at 5°C has practical implications for cultivating this species.

Mental and somatic symptoms related to suicidal ideation in patients visiting a psychosomatic clinic in Japan.

Patients with suicidal ideation (SI) have various mental or somatic symptoms. A questionnaire-based interview elicited details concerning mental and somatic symptoms in patients visiting a psychosomatic clinic in Japan. Univariate logistic regression analyses followed by multiple regression models using a stepwise method were selected for identifying the candidate symptoms. Overall, symptoms related to depression were associated with SI in both sexes. Although women showed more various somatic symptoms associated with SI than men, many of those associations were diminished once severity of the depression was controlled. The current results suggest that a variety of self-reported symptoms, mainly related to depression, might reveal suicidal risk in outpatients with an urban hospital clinical setting.