Effect of interleukin-1beta and dehydroepiandrosterone on the expression of lumican and fibromodulin in fibroblast-like synovial cells of the human temporomandibular joint.

Several epidemiological studies have reported that temporomandibular disorders (TMDs) are more prevalent in women than in men. It has recently been proposed that sex hormones such as estrogen, testosterone and dehydroepiandrosterone (DHEA) are involved with the pathogenesis of TMDs. Although studies have investigated the relationship between estrogen and testosterone and the restoration of TMDs, the relationship between DHEA and TMDs is unknown. The synovial tissue of the temporomandibular joint (TMJ) is made up of connective tissue with an extracellular matrix (ECM) composed of collagen and proteoglycan. One proteoglycan family, comprised of small leucine-rich repeat proteoglycans (SLRPs), was found to be involved in collagen fibril formation and interaction. In recent years, the participation of SLRPs such as lumican and fibromodulin in the internal derangement of TMJ has been suggested. Although these SLRPs may contribute to the restoration of the synovium, their effect is still unclear. The purpose of this study was to investigate the effect of DHEA, a sex hormone, on the expression of lumican and fibromodulin in human temporomandibular specimens and in cultured human TMJ fibroblast-like synovial cells in the presence or absence of the pro-inflammatory cytokine interleukin-1beta (IL-1beta). In the in vivo study, both normal and osteoarthritic (OA) human temporomandibular synovial tissues were immunohistochemically examined. In the in vitro study, five fibroblast-like synoviocyte (FLS) cell lines were established from human TMJ synovial tissue of patients with osteoarthritis. The subcultured cells were then incubated for 3, 6, 12 or 24 h with/without IL-1beta (1 ng/mL) in the presence or absence of DHEA (10 µM). The gene expression of lumican and fibromodulin was examined using the real-time polymerase chain reaction (PCR) and their protein expression was examined using immunofluorescent staining. We demonstrated that the expression of lumican significantly differs from that of fibromodulin in synovial tissue in OA and furthermore, that IL-1beta induced a significant increase in lumican mRNA and immunofluorescent staining in FLS compared to cells without IL-1beta. DHEA plus IL-1beta induced a significant increase in fibromodulin, but not in lumican mRNA, compared to DHEA alone, IL-1beta alone and in the absence of DHEA and IL-1beta. In immunofluorescent staining, weaker fibromodulin staining of FLS cells was observed in cells cultured in the absence of both DHEA and IL-1beta compared to fibromodulin staining of cells cultured with DHEA alone, with DHEA plus IL-1beta, or with IL-1beta alone. These results indicate that DHEA may have a protective effect on synovial tissue in TMJ by enhancing fibromodulin formation after IL-1beta induced inflammation. DHEA enhancement of fibromodulin expression may also exert a protective effect against the hyperplasia of fibrous tissue that TGF-beta1 induces. In addition lumican and fibromodulin are differentially expressed under different cell stimulation conditions and lumican and fibromodulin may promote regeneration of the TMJ after degeneration and deformation induced by IL-1beta.

Pilocarpine-induced status epilepticus causes acute interneuron loss and hyper-excitatory propagation in rat insular cortex.

Recent clinical studies have shown that the insular cortex (IC) is involved in temporal lobe epilepsy and suggested that the IC mediates spreading of epileptic activity from the temporal lobe, including the hippocampus and amygdala, to the frontal cortex. However, little is known about anatomical and physiological features of the IC in models of temporal lobe epilepsy. The present study evaluated the distribution pattern of GABAergic interneurons, especially parvalbumin (PV)- and somatostatin (SS)-immunopositive neurons, and excitatory propagation pattern in the IC of rats 4-7 days and 2 months after pilocarpine-induced status epilepticus (4-7 d and 2 m post-SE rats, respectively). The number of PV-immunopositive neuron profiles in the agranular IC (AI) significantly decreased by 24.6% and 41.5% in 7 d and 2 m post-SE rats, respectively. The dysgranular and granular IC (DI+GI) exhibited only 5.2% loss of PV-immunopositive neurons in 7 d post-SE rats, while 2 m post-SE rats showed 30.4% loss of PV-immunopositive neurons. There was no significant change of the SS-immunopositive neuron profile numbers in the AI and DI+GI of 7 d and 2 m post-SE rats. The regions with decreased numbers of PV-immunopositive neuron profiles overlapped with those where many degenerating cells were detected by Fluoro-Jade B staining. The area of excitatory propagation responding to electrical stimulation of the caudal AI was expanded in 4-7 d post-SE rats, and excitation frequently propagated to the frontal cortex including the motor cortex. Optical signals in the AI of 4-7 d post-SE rats were larger in amplitude than those of controls. In contrast to the AI, the DI of 4-7 d post-SE rats showed similar excitatory propagation pattern and amplitude to that of controls. These results suggest that the region-specific loss of PV-immunopositive neurons occurred in the AI 4-7 d after pilocarpine-induced status epilepticus, which may play an important role in facilitating excitatory propagation in the IC.

Use of glazes on porcelain from near ground zero to measure Hiroshima neutron fluence.

Several porcelain samples from almost directly beneath the atomic explosion at Hiroshima on 6 August 1945, have been scanned for induced fission tracks, produced mostly by the thermal neutrons from the bomb due to interactions with trace uranium in their glass coatings. The ability to use porcelain opens a new and abundant material for retrospective dosimetry. Four different samples had thermal neutron fluences in 1945 of 1.0, 3.8, 4.1, and 8.9 x 10(12) cm(-2). The different values are not caused by track fading, but are likely to result from differing shielding at different nearby positions. Assuming that the three highest fluences, which have overlapping uncertainties, are at locations of minimum shielding, the statistically weighted thermal fluence in the air at ground level and ground zero was 4.8 x 10(12) cm(-2) with a statistical uncertainty of 15%. This value lies between the calculated value of 6.5 x 10(12) given in DS86 and the 3.7 x 10(12) inferred from induced radionuclides by Hoshi et al. (1998).

Transcriptional down-regulation of CXC chemokine receptor 4 induced by impaired association of transcription regulator YY1 with c-Myc in human herpesvirus 6-infected cells.

We have recently reported that down-regulation of CXC chemokine receptor (CXCR) 4 in CD4(+) T lymphocytes is induced by human herpesvirus (HHV) 6 infection. In this study, we further studied the mechanisms of HHV-6-induced CXCR4 down-regulation, focusing on the regulation of CXCR4 transcription. Down-regulation of CXCR4 transcription was detected in HHV-6A-infected JJHAN and HHV-6B-infected MT-4 cell lines, as we had previously reported for HHV-6-infected peripheral blood CD4(+) T lymphocytes. Luciferase assays revealed that a YY1-binding site around -320 relative to the transcription start site is important for down-regulation of CXCR4 transcription in HHV-6-infected cells. The binding activity of YY1, which is a repressor of CXCR4 transcription, to the CXCR4 promoter appeared to significantly increase in HHV-6-infected cells compared with the binding activity in mock-infected cells. Immunoprecipitation assays showed that in HHV-6-infected cells association of c-Myc with YY1 was decreased and that of Max with c-Myc was increased, whereas association of Mad with Max appeared to be decreased. The amounts of each of YY1, c-Myc, Max, and Mad proteins synthesized in cells were not altered by HHV-6 infection. These data indicate that the decreased association of YY1 with c-Myc that is caused by impaired interaction in the c-Myc/Max/Mad network results in increased binding activity of YY1 to the CXCR4 promoter, mediating down-regulation of CXCR4 production in HHV-6-infected cells.

Preparation of griseofulvin for topical application using N-methyl-2-pyrrolidone.

We attempted to prepare a new griseofulvin formulation for topical application using N-methyl-2-pyrrolidone (NMP). Griseofulvin dissolves poorly in both water and oil, but dissolves in NMP to a concentration of about 100 mg/ml. A soybean oil-water emulsion with soybean lecithin and NMP as emulsifier and co-solvent, respectively, was prepared using a Microfluidizer, a high-pressure homogenizer. The size of the droplets in emulsion was about 200 nm, and the emulsion was stable for over 3 months. The skin permeation of griseofulvin through Yucatan micropig skin was studied in vitro using vertical type cells under donor phase open conditions. The permeation of griseofulvin from the NMP-water mixture (0-40%) into the skin tended to increase with increasing NMP concentration, although this finding was not statistically significant. Permeation from emulsion (oil phase, 20%; NMP 10-40%) was significantly higher than that from the water-NMP mixture. Permeation from the oil-NMP mixture was highest among the formulations investigated, and permeation from emulsion under donor phase closed conditions was significantly lower than that under open conditions. We believe that the evaporation of water from the emulsion after application to the skin was an important factor in skin permeation enhancement. When the emulsion containing 3% l-menthol was applied, a sufficient skin concentration (47 microg/cm3 in dermis) was obtained.

HLA class I-restricted lysis of leukemia cells by a CD8(+) cytotoxic T-lymphocyte clone specific for WT1 peptide.

The Wilms tumor (WT1) gene has been reported to be preferentially expressed in acute leukemia cells, regardless of leukemia subtype and chronic myelogenous leukemia cells in blast crisis, but not in normal cells. This finding suggests strongly that WT1 protein is a potential target of immunotherapy for human leukemia. In this study, we established a CD8(+) cytotoxic T-lymphocyte (CTL) clone directed against a WT1-derived peptide and examined its immunologic actions on leukemia cells. A CD8(+) CTL clone, designated TAK-1, which lysed autologous cells loaded with a WT1-derived 9-mer peptide consisting of the HLA-A24 (HLA-A*2402)-binding motifs was established by stimulating CD8(+) T lymphocytes from a healthy individual repeatedly with WT1 peptide-pulsed autologous dendritic cells. TAK-1 was cytotoxic to HLA-A24-positive leukemia cells expressing WT1, but not to HLA-A24-positive lymphoma cells that did not express WT1, HLA-A24-negative leukemia cells, or HLA-A24-positive normal cells. Treating leukemia cells with an antisense oligonucleotide complementary to the WT1 gene resulted in reduced TAK-1-mediated cytotoxicity, suggesting that target antigen of TAK-1 on leukemia cells is the naturally processed WT1 peptide in the context of HLA-A24. TAK-1 did not inhibit colony formation by normal bone marrow cells of HLA-A24-positive individuals. Because WT1 is overexpressed ubiquitously in various types of leukemia cells, but not in normal cells, immunotherapy using WT1 peptide-specific CTL clones should be an efficacious treatment for human leukemia. (Blood. 2000;95:286-293)

Auxiliary partial orthotopic living donor liver transplantation as an aid for small-for-size grafts in larger recipients.

BACKGROUND: In countries where living donors are the only source of liver grafts, restrictions on graft size are a serious obstacle for the expansion of indications for adult recipients. To overcome this problem, auxiliary partial orthotopic liver transplants (APOLT*) was performed on the basis of the concept that the residual native liver would support the graft function until the graft had grown enough to function by itself. METHODS: APOLT as an aid for small-for-size (SFS) grafts was reviewed retrospectively to evaluate its feasibility. Between April 1995 and March 1998, 20 recipients underwent APOLT, which was indicated because of a SFS graft in 15 of them. The indication was based on the estimated graft/recipient's body weight ratio (GRWR). If the ratio was <0.8%, APOLT was performed. The other 5 patients had a graft with a GRWR >0.8% and underwent APOLT on the basis of the residual native liver supporting the graft function temporarily, 4 for supplementation of the defective enzyme in metabolic liver diseases and one for leaving the potential of the regeneration of the native liver in fulminant hepatic failure. The recipients who underwent APOLT because of a SFS graft were categorized as the SFS group, and the others were the second group. RESULTS: In the SFS group, the age of the recipients ranged from 13 to 48 (median 23). The original indications of this group were fulminant hepatic failure in 2 recipients, acute deterioration of chronic liver diseases in 3, Wilson's disease in 2, biliary atresia in 4, primary biliary cirrhosis in 3, and primary sclerosing cholangitis (PSC) in one. The actual GRWR ranged from 0.45 to 0.72 (median 0.55). The graft was implanted after resection of the left lateral segment of the native liver. Except in the first two patients, the portal vein to the residual native liver was completely transected so that all of the portal blood drained into the graft liver. This procedure was successful in 9 patients. The cause of death in the other 6 was mainly infection. The mortality rate among the recipients with signs of advanced liver failure, such as massive ascites or hepatic coma, was higher, even though APOLT was used to support the SFS graft. In the second group, in the other five recipients who underwent APOLT for other indications, one recipient with fulminant hepatic failure died of sepsis caused by the dehiscence of bilio-enteric anastomosis. CONCLUSIONS: APOLT as an aid for a SFS graft is technically viable. This procedure can thus expand the indication of living donor liver transplants for adult recipients when the native liver retains some functional capability to support the grafted liver during the immediate postoperative period.

Effects of blood perfusion rate on the optimization of RF-capacitive hyperthermia.

The effects of the blood perfusion rate on the optimization of heating conditions in radio-frequency capacitive hyperthermia were examined using numerical simulations. When the blood perfusion rate in the tumor was smaller than approximately one-half that of normal tissues, optimal selective heating of the tumor was obtained.

[Comparison of therapeutic effects of SMANCS (+TAE) and Non-SMANCS/LpTAE].

Therapeutic effects of SMANCS and LpTAE were evaluated for hepatocellular carcinoma (HCC). Since June 1995, SMANCS has been used in 59 patients for their first treatment. LpTAE had been performed for HCC before introduction of SMANCS in our hospital, and 71 patients treated after 1992 were chosen for comparison with the therapeutic effect of SMANCS. Among the patients treated with SMANCS, complete and partial responses (CR and PR) were obtained in 24 cases (41%) and 17 cases (33%), respectively. SMANCS accompanied by TAE was more effective than SMANCS alone. The effects did not depend on the level of the hepatic arterial branch at which SMANCS was administered. In patients treated with LpTAE, CR and PR were obtained in 12 cases (17%) and 18 cases (25%), respectively. SMANCS was significantly more effective than LpTAE. Because of our short experience with SMANCS, we could only show a two year survival rate. The one- and two-year survival rates for SMANCS were 71% and 57%, respectively. They were not significantly different from those for LpTAE, at 80% and 60%. Despite good results of treatment for HCC, a better prognosis could not be expected by SMANCS in this study. These results may be explained as follows. The evaluating the cause of death within two years after first treatment, hepatic failure was more common in patients treated with SMANCS. After treatment by SMANCS, 11 patients (55%) died from hepatic failure. On the other hand, 4 patients (15%) died from hepatic failure after LpTAE. Although there is no significant difference of Child Pugh score, this may indicate that SMANCS has been used for patients with lesser hepatic reserve and this leads to early deaths in patients treated with SMANCS. However, because of the short experience in this study, further observation is necessary for precise evaluation of clinical efficacy of SMANCS.

NF00659A1, A2, A3, B1 and B2, novel antitumor antibiotics produced by Aspergillus sp. NF 00659. I. Taxonomy, fermentation, isolation and biological activities.

Five novel cytotoxic antibiotics, NF00659A1 (1), A2 (2), A3 (3), B1 (4) and B2 (5) were discovered. They were isolated from a culture mycelium of Aspergillus sp. These compounds were proved to have 4,5-seco-tricyclic diterpene alpha-pyrone structure by spectroscopic analyses. They showed potent antitumor activities against human ovarian carcinoma A2780 and human colorectal adenocarcinoma SW480 cells, but did not show any antimicrobial activities at 1,000 micrograms/ml against Gram-positive and Gram-negative bacteria, yeasts and fungi.

NF1 gene mutations in Japanese with neurofibromatosis 1 (NF1).

Neurofibromatosis 1 (NF1) is an autosomal dominant disease characterized by abnormalities in multiple tissues derived from the neural crest. We analysed 50 unrelated Japanese patients for NF1 mutations by using polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) analysis for exons 28 to 36. Here, we demonstrate a single base pair (bp) insertional mutations in exon 31 in one patient (5843insA/5844insA/5845insA/5846insA) and a single adenine to guanine transitional mutation 4 bp upstream from the 3' end of intron 31 in two unrelated cases. The insertional mutation in exon 31 was novel and resulted in premature termination of the transcript. The other intron 31 mutations resulted in 4 bp insertions of cDNA between exon 31 and exon 32 with premature termination of the transcript, indicating that those transitions of intron 31 caused aberrant splice acceptor sites upstream from the 5' end of exon 32. However, as the same mutation of intron 31 has been reported previously in two cases of unrelated Caucasians, the splice junction mutation of intron 31 is thought to be common among different ethnic groups.

Some aspects of occupational safety and health in green tea workers.

The objectives of this study were to evaluate the health and safety conditions of 36 male and 27 female green tea workers who were aged 40-69 years and had worked for at least 5 years in green tea production procedures. The Mean +/- SD of age was 57.1 +/- 5.8 years in males and 54.4 +/- 6.4 years in females. The mean working career was 26.1 +/- 9.5 years and 24.3 +/- 9.1 years, respectively. The most commonly subjective complaints developed while at work was pain in the lower back at a rate of 72.2% in males and 63.0% in females. Regarding persistent subjective complaints, female workers had significantly (P < 0.05) higher prevalence rate for breath shortness (25.9%) compared to that of male workers (5.6%). It was observed that the male workers whose occupational career was equal to or more than 28 years had significantly higher prevalence rates for stiffness in the neck and pain in the arms, compared to those of male workers whose occupational career was less than 28 years. The prevalence of nasal allergy was 11.1% either in the male or female workers. The total rate of accidents during working hours in the male workers was 22.2%, and that in the female subjects was 18.5%. The mean frequency weighted vibration magnitude produced by using tea-leaf plucker was in the range of 2.4-3.5 m/s2. The mean equivalent A-weighted noise level while using the same equipment was 100.6 +/- 5.0 dB(A). The need for occupational safety and health programs in these small farm settings are discussed.

[Whole head measurement of auditory evoked magnetic field using a MR-linked helmet shaped MEG system].

Authors compared MR anatomy and magneto-encephalographic (MEG) functional methods in locating auditory cortex in 20 normal volunteers and 56 patients with intracranial structural lesions. Auditory evoked response was measured over the entire head, using a helmet shaped 64 channel MEG system. In each subject, three dimensional MR image was obtained with identical coordinate system of MEG. Best fit sphere to each subject's head was calculated from MRI data. Signal source of auditory evoked MEG was estimated using a two dipole model in the best fit sphere. Position and orientation of dipoles were superimposed on each subject's MRI. Both left and right dipolar patterns appeared about 100 ms after unilateral auditory stimuli (N100m) in 19 normal subjects and 47 patients. These dipolar patterns overlap each other across the midline. A two dipole model successfully localized bilateral N100m dipoles in the upper surface of superior temporal planes. N100m response was missing unilaterally in 1 normal subjects and 9 patients inspite of normal audiometry findings, suggesting sub-clinical auditory dysfunction. The MR-linked whole head MEG is a non-invasive tool to localize source of auditory evoked N100m response even in the case with intracranial structural lesions.

Acetic acid intoxication by rectal administration.

The enhanced toxicity of acid instilled directly into the rectum, without benefit of dilution and neutralization in the upper intestine, is evident in a case of acetic acid intoxication by accidental rectal administration of 50 mL of 9% acetic acid to a 5-year-old boy. The complications included necrosis of the colon, acute renal failure, acute liver dysfunction, disseminated intravascular coagulopathy (DIC) and sepsis.

Splenic abscess associated with colon cancer: a case report.

The present report describes a case of colon cancer which presented with a rare complication of splenic abscess. A 52-year-old Japanese man with diarrhea, fever and chills was admitted to our hospital. He complained of fever, with chills at night, and abdominal pain occurring during the last month. The origin of the fever was investigated, and Escherichia coli grew from a blood culture. Multilocular splenic abscesses and wall thickening of the descending colon were revealed by CT scan, magnetic resonance imaging and ultrasound. A cancer of the descending colon was found by barium enema and colonoscopy. A curative resection was performed and the pathological report revealed the splenic abscess to have developed from a direct extension of, and perforation by, the carcinoma of the descending colon.

[Hemodilutional and blood salvaging autotransfusions in a case of aplastic anemia].

A 28-year-old female, weighing 46 kg, 155 cm in height, with aplastic anemia underwent implantation of iliac bone to the head of the femur. A combined technique of hemodilution and intraoperative blood salvaging was applied to supplement the blood loss during the operation. Intraoperative monitoring included continuous arterial pressure, heart rate, electrocardiograph, SVO2, bleeding time, prothrombin time (PT), activated thromboplastin time (APTT), and thromboelastography. A total of 900 ml of blood was drawn and the same volume of 5% albumin solution was infused over half an hour before the beginning of the surgery. During the operation, 2100 ml of blood was lost, and 1260 ml of autologous blood, 400 ml of homogeneous red blood cells and 5 units of fresh platelet were infused. The values of PT, APTT, bleeding time were within normal ranges after the surgery. Only 3 units of fresh platelet was infused in 2 weeks after the surgery. It was suggested that hemodilution and salvaging autotransfusion is safely performed and beneficial to minimize homogeneous blood transfusion even in a case of aplastic anemia.

Usefulness of 31P-MRS as a method of evaluating the viability of preserved and transplanted rat liver.

We used phosphorus-31 magnetic resonance spectroscopy (31P-MRS) to evaluate the viability of transplanted rat liver. Wistar rats were used as donors and recipients. The donor livers were preserved in saline (group 1), Euro-Collins solution (group 2), or in University of Wisconsin (UW) solution (group 3) for 3 and 6 h in groups 1, 2 and 3 and for 9 h in groups 2 and 3. Thereafter the livers were orthotopically transplanted. 31P-MRS spectra were measured after portal reperfusion. Finally, all the recipients were divided into survivors and non-survivors. Survival rates were better in group 3 than in groups 1 and 2. In the 9-h-preserved livers, the livers in group 3 showed a significantly higher beta-ATP/Pi ratio than those in group 2. Comparing survivors and non-survivors in the 6-h-preserved livers in group 2, survivors' livers showed significantly higher beta-ATP/Pi ratio than those of non-survivors. We concluded that 31P-MRS is a useful method for assessing viability of rat liver grafts.

[Miscellaneous contributions to the essential oils of plants from various territories. LI. On the components of essential oils of Torilis japonica (Houtt.) DC].

The essential oils of Torilis japonica (Houtt.) DC. (Japanese name, Yabujirami) of the family Umbelliferae were prepared from fresh whole herbs and fruits, and investigated by using gas chromatographic and mass spectrometric analyses. The fruits of this plant had been used as a substitute in Japan for a Chinese crude drug Zya-syo-si: Cnidii Monnieri Fructus. Fifty three components were positively identified in the essential oils. The main components of the essential oil were germacrene-D (57.9-71.8% in the oil), alpha-humulene (2.4-13.2%), bicyclogermacrene (1.9-5.4%), beta-caryophyllene (1.5-4.6%), and delta-cadinene (1.0-1.9%).