The purpose of this study was to examine the effect of ultrasound on inflammatory skeletal muscle
in vitro
and
in vivo
. C2C12 cells were cultured in medium with or without TNF-
α
or IL-1
β
. After ...stimulation with cytokines, the cells received ultrasound or sham exposure. Furthermore, the tibialis anterior (TA) muscle in C57BL/6 mice injured by cardiotoxin (CTX) were dissected after a series of ultrasound treatments and examined. Exposure of C2C12 cells to ultrasound resulted in down-regulation of cyclooxygenase-2 (COX-2) mRNA and protein expression induced by TNF-
α
or IL-1
β
, and up-regulated myogenin mRNA and protein depressed by TNF-
α
or IL-1
β
. In injured TA muscle induced by CTX, ultrasound caused increase of COX-2 mRNA at 1 day after ultrasound treatment, however, at day 5, reduction of COX-2 mRNA and protein. At day 5, ultrasound caused increase of myogenin mRNA and protein, increase of fast myosin protein, and increase of paired-box transcription factor 7 positive cells. At day 7, the cross-sectional area of myofibers in the ultrasound exposure side was significantly larger than that on the control side. In conclusion, ultrasound stimulation may be a better candidate as a medical remedy to promote myogenesis in inflammatory muscle states, such as muscle injury.
Low-intensity pulsed ultrasound (LIPUS) suppresses synovial hyperplasia and synovial cell proliferation characterized for rheumatoid arthritis, but the molecular mechanisms remain unknown. The ...purpose of this study was to examine the mechanotransduction pathway
via
the integrin/mitogen-activated protein kinase (MAPK) pathway in LIPUS exposure on the synovial membrane cells. Rabbit knee synovial membrane cell line, HIG-82, was cultured with or without FAK phosphorylation inhibitor, PF-573228. One hour after stimulation with PF-573228, the cells exposed to LIPUS for 20 min or sham exposure. A possible integrin/MAPK pathway was examined by immunofluorescence and Western blotting analysis with antibodies targeting specific phosphorylation sites on intracellular signaling proteins. LIPUS exposure increased phosphorylation of FAK, JNK, ERK, and p38, but the phosphorylation was inhibited by PF-573228. In conclusion, LIPUS exposure might be involved in cell apoptosis and survival of synovial membrane cells
via
integrin/FAK/MAPK pathway.
Glycyrrhizae Radix is the most frequently used crude drug in Japan and is prescribed in Kampo medicine for the treatment of a wide range of diseases. The major active ingredient of Glycyrrhizae ...Radix, glycyrrhizin (GL), has been shown to possess various pharmacological actions, but is also known to cause adverse effects such as pseudoaldosteronism. To avoid the adverse effects of GL, precautions have been indicated on the package inserts of Glycyrrhizae Radix-containing formulas depending on the amount of Glycyrrhizae Radix they contain. However, it remains unknown whether the extraction efficiency of GL from Glycyrrhizae Radix is constant throughout the different combinations of crude drugs in Glycyrrhizae Radix-containing formulas. To confirm the basis of the safety regulation, in this study we comprehensively determined the GL content of 25 major kinds of Kampo extracts compounding Glycyrrhizae Radix. We found that the GL content per daily dosage in all Kampo extracts are generally proportional to the compounding amount of Glycyrrhizae Radix, except in the case of shoseiryuto (Sho-seiryu-To). We also found that Schisandrae Fructus in Sho-seiryu-To decoction caused a lowered pH condition and drastically decreased the extraction efficacy of GL from Glycyrrhizae Radix. Moreover, we were able to confirm that the extraction efficiency of GL from Glycyrrhizae Radix is dependent on the pH value of the extraction solvent. The extraction efficiency of GL in the 25 kinds of Kampo extracts was not constant but it correlates significantly with the pH value of the decoction. Furthermore, the GL contents are well correlated with pseudoaldosteronism incidence data obtained from the Japanese Adverse Drug Event Report (JADER) database on the 25 kinds of Kampo extracts. This suggests that the GL content is a better index to consider to avoid the adverse effects of Glycyrrhizae Radix-containing Kampo formulas.
The purpose of this study was to examine the effect of low-intensity pulsed ultrasound (LIPUS) on the cell proliferation and growth of synovial membrane cells stimulated with inflammatory cytokines, ...and to evaluate the effectiveness of LIPUS treatment of synovitis in the knee joints of animal models for rheumatoid arthritis. The rabbit knee synovial membrane cell line, HIG-82, was cultured in medium with or without IL-1β or TNF-α. Four hours after stimulation with the cytokines, the cells received LIPUS or sham exposure. Cell proliferation and growth were then analyzed. Using MRL/
lpr
mice, the anti-inflammatory effects of LIPUS were also evaluated
in vivo
. Stimulation with proinflammatory cytokines significantly up-regulated cell proliferation which was significantly down-regulated by LIPUS exposure. In MRL/
lpr
mice, exposure of knee joints to LIPUS caused a significant reduction of histological damage compared to the control. Histological lesions were significantly reduced in the joints treated with LIPUS for 3 weeks. Cox-2-positive cells in the knee joints treated with LIPUS were markedly decreased compared to the control joints. Therefore, LIPUS stimulation may be a medical treatment for joint inflammatory diseases, such as synovitis.
Two known flavonoids were isolated from a tropical medicinal plant, Hyptis fasciculata (Labiatae), found in Brazil. Flavonoids were identified as cirsilineol (1) and cirsimaritin (2) by spectroscopic ...means and were exhibited potent antibacterial activity against Helicobacter pylori, and cirsilineol (1) had weak antibacterial activity against Escherichia coli and Salmonella enteritidis. Following up on the relationship between anti-H. pylori activity and flavonoids with methoxy groups, several methoxy flavonoids were evaluated for proliferation of H. pylori.
Besides various gastroduodenal diseases, Helicobacter pylori infection may be involved in autoimmune disorders like rheumatoid arthritis (RA) or idiopathic thrombocytopenic purpura. Such autoimmune ...disorders are often associated with autoreactive antibodies produced by B-1 cells, a subpopulation of B lymphocytes. These B-1 cells are mainly located in the pleural cavity or mucosal compartment. The existence of H. pylori urease-specific immunoglobulin A (IgA)-producing B cells in the mucosal compartment and of their specific IgM in the sera of acutely infected volunteers suggests the possibility that urease stimulates mucosal innate immune responses. Here, we show for the first time that purified H. pylori urease predominantly stimulates the B-1-cell population rather than B-2 cells, which produce antigen-specific conventional antibodies among splenic B220⁺ B cells. The fact that such stimulation of B-1 cells was not affected by the addition of polymyxin B indicates that the effect of purified H. pylori urease was not due to the contamination with bacterial lipopolysaccharide. Furthermore, the production of various B-1-cell-related autoreactive antibodies such as IgM-type rheumatoid factor, anti-single-stranded DNA antibody, and anti-phosphatidyl choline antibody was observed when the splenic B cells were stimulated with purified H. pylori urease in vitro. These findings suggest that H. pylori components, urease in particular, may be among the environmental triggars that initiate various autoimmune diseases via producing autoreactive antibodies through the activation of B-1 cells. The findings shown here offer important new insights into the pathogenesis of autoimmune disorders related to H. pylori infection.
Free neutral
d-amino acids have previously been detected in human plasma, usually accounting for less than 2% of the total free amino acid concentration (
d-amino acid ratio) Nagata, Y., Masui, R., ...Akino, T., 1992a. The presence of free
d-serine,
d-alanine and
d-proline in human plasma. Experientia 48, 986–988. Nagata, Y., Yamamoto, K., Shimojo, T., 1992b. Determination of
d- and
l-amino acids in mouse kidney by high-performance liquid chromatography. Journal of Chromatography 575, 147–152. Nagata, Y., Yamamoto, K., Shimojo, T., Konno, R., Yasumura, Y., Akino, T., 1992c. The presence of free
d-alanine,
d-proline and
d-serine in mice. Biochimca et Biiophysica Acta 1115, 208–211. In the present study to search for the source of free
d-amino acids,
d- and
l-enantiomers of the major non-essential amino acids, i.e., the free form of serine, alanine, proline, aspartate and glutamate were analyzed by HPLC in human saliva, submandibular glands and oral epithelial cells. The
d-enantiomer ratios to total of free alanine or proline were 35% and 20%, respectively, in saliva. The ratios of the other
d-amino acids were less than 11%. The effect of ingested food and oral bacteria on the saliva amino acid levels was suggested to be insignificant.
d-Alanine and
d-aspartate were also detected in the submandibular gland in ratios up to 5%, and
d-alanine and
d-proline were found in oral epithelial cells in ratios of 18% and 5%, respectively. The submandibular gland and oral epithelial cells are suggested to be possible sources of the saliva
d-alanine and
d-aspartate.
ABSTRACT
U. urealyticum, a member of the family Mycoplasmataceae, is often detected in the vagina of pregnant women. In this study, the possible association of ureaplasmal infection with preterm ...delivery was examined, as was the capacity of ureaplasmal LP to stimulate monocytes in vitro to produce pro‐inflammatory cytokines relevant to preterm delivery. A hundred cases of normal delivery and 45 cases of preterm delivery were randomly selected. A mAb against U. urealyticum urease, that selectively and positively stained it in vaginal secretions of infected women but not in those of uninfected women, was generated. The preterm delivery group showed a significantly higher incidence of vaginal infection with this bacteria than the normal delivery group. Since the LP of Mycoplasma has potent biological activity, ureaplasmal LP was extracted. THP‐1 cells, and human monocytic cells, produced IL‐8, a potent pro‐inflammatory cytokine associated with preterm delivery, and showed apoptotic cell death in response to the LP in vitro. These results suggest that U. urealyticum infection might play a causative role in preterm delivery via LP‐induced IL‐8 production and apoptosis.
Background: Triple therapy with amoxicillin, clarithromycin, and a proton‐pump inhibitor is a common therapeutic strategy for the eradication of Helicobacter pylori (H. pylori). However, frequent ...appearance of clarithromycin‐resistant strains is a therapeutic challenge. While various quinones are known to specifically inhibit the growth of H. pylori, the quinone 1,4‐dihydroxy‐2‐naphthoic acid (DHNA) produced by Propionibacterium has strong stimulating effect on Bifidobacterium. We were interested to see whether DHNA could inhibit the growth of H. pylori in in vitro or in vivo experimental setting.
Materials and Methods: The minimum inhibitory concentration (MIC) of DHNA was determined by the agar dilution method. The inhibitory action of DHNA on the respiratory activity was measured by using an oxygen electrode. Germ‐free mice infected with H. pylori were given DHNA in free drinking water containing 100 μg/mL for 7 days.
Results: DHNA inhibited H. pylori growth at low MIC values, 1.6–3.2 μg/mL. Likewise, DHNA inhibited clinical isolates of H. pylori, resistant to clarithromycin. However, DHNA did not inhibit other Gram negative or anaerobic bacteria in the normal flora of the human intestine. Both H. pylori cellular respiration and adenosine 5′‐triphosphate (ATP) generation were dose‐dependently inhibited by DHNA. Similarly, the culture filtrates of propionibacterial strains inhibited the growth of H. pylori, and oral administration of DHNA could eradicate H. pylori in the infected germ‐free mice.
Conclusions: The bifidogenic growth stimulator DHNA specifically inhibited the growth of H. pylori including clarithromycin‐resistant strains in vitro and its colonization activity in vivo. The bactericidal activity of DHNA was via inhibition of cellular respiration. These actions of DHNA may have clinical relevance in the eradication of H. pylori.
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