Drinking deep-sea water restores mineral imbalance in atopic eczema/dermatitis syndrome.

OBJECTIVE: The effect of drinking deep-sea water on hair minerals was studied in patients with atopic eczema/dermatitis syndrome (AEDS). Study of hair minerals revealed an imbalance of essential minerals and an increase in toxic minerals in AEDS patients. DESIGN: After drinking deep-sea water (Amami no Mizu) for 6 months in AEDS patients, hair minerals (essential minerals and toxic minerals), clinical evaluation of the skin symptoms were compared before drinking with after drinking. SUBJECTS: After obtaining informed consent, 33 patients (mean age 26 y, range 1-50 y, 13 male and 20 female subjects) with mild to moderate AEDS were enrolled. RESULTS: After drinking deep-sea water, the levels of the essential mineral, potassium (K), were significantly decreased, while the levels of selenium (Se) increased. On the other hand, drinking deep-sea water significantly decreased the levels of the toxic minerals, mercury and lead. Moreover, after drinking deep-sea water, the skin symptoms were improved in 27 out of 33 patients. CONCLUSION: These results indicate that the mineral abnormalities/imbalance may be involved in the pathogenesis of AEDS, and that drinking deep-sea water may be useful in the treatment of AEDS.

Comparison of the isometric cervical extension strength and a cross-sectional area of neck extensor muscles in college wrestlers and judo athletes.

Increasing neck muscle strength can play an important role in preventing neck injuries in contact sports. The purpose of this study was to examine the actual conditions of the isometric cervical extension strength (ICES) and the cross-sectional area (CSA) of neck extensor muscles in male athletes participating in college wrestling and judo. The subjects comprised 18 wrestlers and 37 judo athletes from Nippon Sports Science University in Japan. The ICES was measured at eight angles (126 degrees, 108 degrees, 90 degrees, 72 degrees, 54 degrees, 36 degrees, 18 degrees, 0 degrees ). Transverse slices of 10 mm thickness were obtained at the position of each intervertebral disc between C2 and C3, C3 and C4, C4 and C5 and C5 and C6 using magnetic resonance imaging. The ICES of the wrestlers were significantly higher than those of the judo athletes. The ICES curve against the angle in wrestlers tended to differ from that of judo athletes. The CSA of neck extensor muscles in the wrestlers was significantly larger at all intervertebral levels examined than those of the judo athletes. A significant difference was observed in the CSA of the deepest area of neck extensor muscles between the groups although the difference was not significant in the superficial area. In this study, the ICES and the CSA in wrestlers were shown to be significantly higher and larger respectively than in the judo athletes, indicating a significant difference between these two sports.

Dermatomyositis with splenic and renal infarctions during corticosteroid therapy.

A 60-year-old woman was admitted to our hospital with complaints of muscle weakness and erythema on her extremities. Gottron's sign, heliotrope rash, elevation of serum myogenic enzymes, electromyography and magnetic resonance imaging findings established a diagnosis of dermatomyositis (DM). She was treated with 60 mg of daily prednisolone. One week later, she suddenly developed splenic and renal infarctions, which were considered to have resulted from vasculopathy associated with DM. Cyclophosphamide and anticoagulants along with increasing the dosage of corticosteroid were effective. This is the first report describing splenic and renal infarctions in a patient with adult-onset DM.

In vivo osteogenic capability of cultured allogeneic bone in porous hydroxyapatite: immunosuppressive and osteogenic potential of FK506 in vivo.

Fischer or ACI rat marrow cells were obtained from femoral shafts and were cultured to confluence in Eagle's minimal essential medium (EMEM) supplemented with 15% fetal bovine serum. After trypsinization, the cells were subcultured on porous hydroxyapatite (HA; Interpore 500) blocks in the presence of beta-glycerophosphate and 10 nM dexamethasone (Dex). After 2 weeks of subculture, a mineralized bone matrix with osteogenic cells developed on the HA pore surfaces. ACI or Fischer cultured bone tissue/HA constructs were implanted subcutaneously into the backs of Fischer rats and the immunosuppressant FK506 was given to the rats for 4 weeks. Implants were harvested 4 weeks and 8 weeks after insertion. At 4 weeks, the ACI constructs (allografts) showed high levels of osteogenic parameters (alkaline phosphatase [ALP] activity and osteocalcin content) and bone formation was observed together with active osteoblasts without obvious accumulation of inflammatory cells. At 8 weeks, active osteoblasts and progressive bone formation were still observed, while osteogenic parameters remained high and osteocalcin messenger RNA (mRNA) was detected. Without FK506 administration, the allografts showed neither bone formation nor osteocalcin mRNA and there were only trace levels of the osteogenic parameters. In the case of Fischer constructs (isografts), extensive bone formation was detected and all the osteogenic parameters were higher with FK506 than without FK506 at both 4 weeks and 8 weeks. These results indicate that cultured bone tissue/HA constructs possess a high osteogenic potential, even as allografts, and that FK506 not only has an immunosuppressive action, but also promotes bone formation.

In vivo osteogenic durability of cultured bone in porous ceramics: a novel method for autogenous bone graft substitution.

BACKGROUND: Bone marrow cells differentiate into bone-forming osteoblasts when cultured in medium supplemented with 15% fetal bovine serum, ascorbic acid, beta-glycerophosphate, and dexamethasone. METHODS: To investigate in vivo osteoblastic activity and bone matrix formation by cultured bone marrow cells, Fischer rat marrow cells were cultured for 2 weeks in porous hydroxyapatite (HA) and then subcutaneously implanted into 7-week-old male syngeneic rats. The implants were harvested after 8 and 52 weeks for biochemical and histological analyses. RESULTS: At both times, formation of lamellar bone accompanied by regeneration of marrow were seen in many of the HA pores. When a fluorochrome (calcein) was administered at 50 weeks after implantation, it was detected in the pores of implants harvested at 52 weeks. Osteoclastic resorption followed by new bone formation was seen in some pores at 52 weeks, indicating that bone remodeling was continuing. The alkaline phosphatase activity of implants harvested at 52 weeks was comparable to that at 8 weeks, whereas the osteocalcin content of the implants harvested at 52 weeks was about twice that at 8 weeks. CONCLUSION: These results demonstrated that there was persistent in vivo osteogenic and hematopoietic activity in the prefabricated bone/HA constructs, and indicated that normal bone tissue was regenerated after grafting of the constructs, which were brittle before implantation. Tissue engineering using HA and cultured marrow cells culture may provide an alternative method of bone transplantation for patients with skeletal disorders, although further in vivo and in vitro experiments are needed.

Al2O3 doped apatite-wollastonite containing glass ceramic provokes osteogenic differentiation of marrow stromal stem cells.

Fresh marrow cells were obtained from femora of Fischer rats and cultured in a medium containing 15% fetal calf serum (FCS) until confluence. After trypsinization, cells were subcultured at a cell density of 100 x 10(3)/35-mm well in the presence of FCS, beta-glycerophosphate, and ascorbic acid phosphate on four different culture substrata. The period of subculture was 2 weeks; the substrata used were the culture dish, apatite-wollastonite containing glass ceramic (AW), hydroxyapatite coated AW (HA/AW), and Al2O3 doped AW (Al/AW). The HA coating was attained by the incubation of AW in simulated physiological solution. The glass matrix of AW and HA/AW contained MgO, CaO, P2O5, and SiO2; Al/AW contained Al2O3 in addition to these components. The subculture on Al/AW substratum showed many alkaline phosphatase (ALP) positive nodules and the highest ALP activity. On a Northern blot analysis the housekeeping gene of beta-actin mRNA was evenly detected from the cells cultured on all substrata; however, bone-specific osteocalcin mRNA was only detected from the cells on Al/AW. These results indicate that Al/AW provokes the osteoblastic differentiation of marrow stromal stem cells.

Molecular cloning, expression, and characterization of a novel mouse liver SULT1B1 sulfotransferase.

A mouse liver homogenate was shown to contain enzymatic activities catalyzing the sulfation of 3,4-dihydroxyphenylalanine (Dopa) and tyrosine isomers with a pH optimum of 8.25. Western blot analysis revealed a 34 kDa protein exhibiting immunologic cross-reactivity to antiserum against rat liver SULT1B1 sulfotransferase. By employing the reverse transcriptase-polymerase chain reaction (RT-PCR) technique, a 910-base pair product encoding the putative mouse liver SULT1B1 sulfotransferase was obtained. Using this PCR product as a probe, a cDNA containing the entire open reading frame of the mouse liver SULT1B1 sulfotransferase was cloned from a mouse liver Lambda ZAP cDNA library. The nucleotide sequence indicated it is a new enzyme. The deduced amino acid sequence exhibited 87.6, 72.3, 55.9, 54.2, 52.8, 51.1, and 49.4% identity to the amino acid sequences of the rat liver SULT1B1 sulfotransferase, human thyroid hormone sulfotransferase, mouse phenol sulfotransferase, rat liver phenol sulfotransferase, rat liver hydroxyarylamine sulfotransferase, mouse estrogen sulfotransferase, and rat estrogen sulfotransferase. Upon transfection of COS-7 cells with an expression vector (pcDNA3) harboring the cDNA encoding this new enzyme, a 34 kDa protein exhibiting immunologic cross-reactivity to antiserum against the rat liver SULT1B1 sulfotransferase was expressed. The recombinant sulfotransferase exhibited enzymatic activities toward Dopa and tyrosine isomers, as well as dopamine and 3,3',5-triiodo-L-thyronine. Northern blot analyses indicated the SULT1B1 sulfotransferase was predominantly expressed in liver, but not in the other ten mouse organs examined. Furthermore, the enzyme was found to be expressed in a developmental stage-dependent manner, being at a very low level in liver samples from 1-day-old mice and then gradually increasing to the maximum level in liver samples from 4-week-old mice.

Modulation of bone mass, strength, and turnover by a new benzamide compound, DU-6712, in adjuvant-induced arthritic rats.

To assess the reductions in bone mass, strength, and turnover in adjuvant-induced arthritic rats and to examine the effect of a new benzamide compound, DU-6712, on these parameters, a 28-day dosing experiment was performed using 8-week-old female Lewis rats. Arthritis was induced by injecting the adjuvant into the hind paw. The age-dependent increases in the body weight, lumbar bone mineral content and density (BMC and BMD) and compressive strength were disturbed in the arthritic rats. At 14 days, the histomorphometric parameters of bone formation (BFR/BS and BFR/BV) and the serum osteocalcin levels were significantly reduced compared with the baseline controls. However, the BMC values corrected for body weight did not differ significantly between the arthritic and normal rats, and the bone minerals were not reduced compared with the baseline controls. At 28 days, the parameters of bone minerals and strength of the lumbar body in the arthritic rats, both with and without correction for body weight, were significantly reduced compared with the baseline controls. The trabecular mineralizing surface remained significantly reduced and the osteoclast numbers were increased. DU-6712 at the doses of 5, 15, and 45 mg/kg, orally administered daily from the start of the experiment, significantly prevented the development of the chronic paw edema at 28 days. The reductions in the parameters of bone minerals, strength, and trabecular bone formation, and the increase in osteoclast number were alleviated by this agent. Age-dependent increases in the lumbar height, disturbed by the adjuvant injection, were also maintained. These data indicated that a 28-day period is necessary to obtain sufficient reductions in the bone mass and strength of the lumbar body concerning the model of secondary osteoporosis in adjuvant arthritic rats. DU-6712 was able to prevent these reductions by modulating the bone turnover in this arthritis model.

Dose rate effectiveness and potentially lethal damage repair in normal and double-strand break repair deficient murine cells by gamma-rays and 5-fluorouracil.

SCID (severe combined immunodeficiency) fibroblasts established from C.B 17-scid/scid embryos showed higher sensitivity to high (1.105 Gy/min) and low (0.00069 Gy/min) dose rate gamma-rays and also to 5-fluorouracil, a cancer sedative producing double-strand breaks, than wildtype cells from C.B17- +/+ embryos. Furthermore, SCID cells were deficient in repairing DNA damage induced by high dose rate gamma-rays even after dose fractionation and after 24 h recovery periods, while wildtype cells showed an apparent repair ability on DNA damage after these gamma-ray exposures. This is the first report to prove that SCID cells lack the repair of gamma-ray-induced potentially lethal damage and also of 5-fluorouracil-induced double-strand breaks. However, SCID cells showed a significantly higher survival rate by low dose rate exposure than by high dose rate exposure as in the case of wildtype cells, indicating that SCID cells have a deficiency in DNA repair for high dose rate gamma-rays, but not for low dose rate exposure. This suggests an important finding that the dose rate effect (diminution of cell killing by low dose rate exposure) is caused not only by the repair of double-strand breaks induced by gamma-rays but in most parts by less yields of double-strand breaks due to dispersed or low intensity ionization in the cell.

Enhancement of prostaglandin E2 production by epidermal growth factor requires the coordinate activation of cytosolic phospholipase A2 and cyclooxygenase 2 in human squamous carcinoma A431 cells.

We demonstrated the effect of epidermal growth factor (EGF) on the production of PGE2 in human squamous carcinoma A431 cells. The production of PGE2 was increased by stimulating the cells with EGF for 2 h and reached a maximum for 10 h. EGF was also found to augment the release of arachidonic acid (AA) following the increase in phospholipase A2 (PLA2) activity (1.7-fold). The induced PLA2 activity was diminished by 4-bromophenacyl bromide, but not by dithiothreitol, indicating that the EGF-induced release of AA was due to the increase in the activity of cytosolic PLA2 (cPLA2). On the other hand, cyclooxygenase (COX) activity was increased (1.6-fold) within 2 h after the EGF-treatment and the induced activity was inhibited by cycloheximide. In addition, Northern blot analysis showed that the level of COX-2 mRNA was increased by the EGF-treatment, whereas no COX-2 mRNA was detected in the untreated cells, indicating that the EGF-induced COX activity was resulted from the increase in the production of COX-2. These results suggest that EGF augments the production of PGE2 by increasing not only the activity of cPLA2 but also the production of COX-2 in A431 cells.

Bilirubin oxidation provoked by endotoxin treatment is suppressed by feeding ascorbic acid in a rat mutant unable to synthesize ascorbic acid.

We examined the possibility that bilirubin oxidation is provoked in vivo by using scurvy-prone ODS-od/od rats treated with endotoxin (lipopolysaccharide). Recently, bilirubin oxidative metabolites were isolated from human urine and named biotripyrrin-a and biotripyrrin-b. In ODS-od/od rats fed an ascorbic-acid-free diet, the concentration of bilirubin metabolites in urine was increased 7.0-fold at 3 h after injection of lipopolysaccharide and 4.4-fold at 10 h compared to the control rats injected with saline. The dietary supplement of ascorbic acid, the physiological antioxidant, suppressed the increase in bilirubin metabolites in urine after lipopolysaccharide injection: concentrations of biotripyrrin-a and biotripyrrin-b in urine collected 6.5-10 h after the injection were lower in rats fed an ascorbic-acid-supplemented diet than in rats fed an ascorbic-acid-free diet. Moreover, feeding of ascorbic acid suppressed the hepatic mRNA level of heme oxygenase-1, the rate-limiting enzyme of bilirubin biosynthesis, in rats injected with lipopolysaccharide. These findings indicate that bilirubin oxidation is markedly stimulated in lipopolysaccharide-treated rats and suggest that bilirubin and ascorbic acid have physiologically protective effects against oxidative stress.

Abdominal actinomycosis: barium enema and computed tomography findings.

A case of abdominal actinomycosis is described in a woman with recurrent right lower abdominal pain and low-grade fever without history of appendectomy. Past history included the use of an intrauterine device (IUD) until 10 years before manifestation of these symptoms. We followed up the patient, via diagnostic imaging, for 7 months. On initial barium enema, a polypoid lesion was visualized at the bottom of the cecum and there was constriction of the sigmoid colon; the appendix was not seen. Seven months later, poor extension at the cecum, severe constriction in the sigmoid colon, and narrowing of the terminal ileum were also visualized. On computed tomography (CT), the lesion was initially localized only in the ileocecal region adjacent to the sigmoid colon. After 7 months, the lesion had infiltrated adjacent anatomic components and showed direct infiltration of the pelvic space. Differential diagnosis was difficult, as it was not obvious whether this was a pelvic abscess due to inflammation or appendiceal carcinoma. Laparotomy was performed. Macroscopically, the lesion was not limited to the ileocecal region, but involved the right ureter, tubes the Fallopian and ovary, bladder, psoas muscle, and abdominal wall. Pathology findings showed, chronic inflammatory tissue with evidence of actinomycosis. Although previous reports have described a lack of specific findings in this disease. When actinomycosis is suspected, CT is recommended to define its extent.

[Quantitative analysis of organophosphorous pesticide residues in Chinese drugs].

This paper reports the GC determination of 13 organophosphorous pesticides in Chinese drugs Flos Ionicerae and Moluodan etc by the present method of Japan for determining pesticide residues. The results suggest that the determined samples do not contain the above said 13 organophosphorous pesticides.

[Quantitative analysis of organochlorine pesticide residues in Chinese drugs].

This paper reports the GC determination of 20 organochlorine pesticides in Chinese drugs Flos Ionicerae and Moluodan etc by the present method of Japan for determinaing pesticide residues. The results suggest that except Folium Isatidis, Radix Codonopsis and Sanqi Pian all accord with the for provisions pesticide residues in Japanese foodstuffs.

Bilirubin is oxidized in rats treated with endotoxin and acts as a physiological antioxidant synergistically with ascorbic acid in vivo.

We examined the possibility that bilirubin physiologically acts as an antioxidant by using scurvy-prone ODS-od/od rats treated with endotoxin (lipopolysaccharide: LPS). Recently, bilirubin oxidative metabolites were isolated from human urine and named biotripyrrin-a and biotripyrrin-b. The LPS injection markedly increased bilirubin oxidative metabolites in urine of rats fed an ascorbic acid-free diet. This increase was supressed by feeding an adequate amount of ascorbic acid, a physiological antioxidant. the concentrations of biotripyrrin-a and -b in urine collected 6.5-10 h after the LPS injection were lower in rats fed an ascorbic acid-supplemented diet than in rats fed an ascorbic acid-free diet. Moreover, feeding with ascorbic acid suppressed the elevation of hepatic mRNA level of heme oxygenase-1, the rate-limiting enzyme of bilirubin biosynthesis, in rats injected with LPS. These findings suggest that bilirubin is oxidized in rats treated with LPS and acts as a physiological antioxidant synergistically with ascorbic acid in vivo.

Hyperbaric oxygenation reduces the cytostatic activity and transcription of nitric oxide synthetase gene of mouse peritoneal macrophages.

The effects of hyperbaric oxygenation (HBO) and hyperbaric air (HBA) on the cytostatic activity, peroxynitrite synthesis and transcription of the inducible nitric oxide synthetase (iNOS) gene of murine peritoneal macrophages were studied in vitro. Exposure of mice to HBO or HBA significantly reduced the cytostatic activity, peroxynitrite synthesis and transcription of iNOS mRNA of their macrophages stimulated with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). These results indicate that HBO and HBA treatments of mice reduce the cytostatic activity of peritoneal macrophages by reducing iNOS mRNA synthesis.

Compound heterozygous mutations affecting both hepatic and erythrocyte isozymes of pyruvate kinase.

Novel erythrocyte pyruvate kinase gene defects were found in a patient without a family history of consanguinity. The polymerase chain reaction products of the R-type pyruvate kinase cDNA from the propositus contained two point mutations of Ser80 (TCC)-->Pro (CCC) and Arg490 (CGG)-->Trp (TGG). Allele-specific polymerase chain reaction of the genomic DNA revealed that this patient was a compound heterozygote. The mobilities of the patient's L- and R-type pyruvate kinase by thin-layer polyacrylamide gel electrophoresis were abnormal. The results are consistent with the fact that these mutations are within exons common to the hepatic and erythrocyte isozymes.

Involvement of the pineal body in the Bi-Digital O-Ring Test.

As an interesting exception to the general clinical application of the Bi-Digital O-Ring Test (BDORT), the test could not be performed successfully in diagnosing a patient who suffered from cancer of the pineal body. The following examinations were attempted using the BDORT: 1) The thymus representation area, 2) the distal forearm during compression of the upper arm, 3) surface representations of diseased areas that had previously been identified by the indirect method with slide preparations of pineal body or lung cancer tissue, 4) holding of oncogene c-fos Ab2 and integrin alpha 5 beta 1. Results suggest that the failure of the BDORT might be due to the absence of pineal body function. It is suggested that the sensor for the BDORT might exist in the pineal body since 1) BDORT cannot be successfully performed when the eyes are closed, and the pineal body is sensitive to light; 2) electromagnetic resonance might stimulate this sensor; and 3) N-acetyltransferase, the enzyme that converts serotonin to methylserotonin in the pineal body, might be inhibited by activation of this sensor. As a result, serotonin levels might be increased by activation of this sensor, which might then inhibit finger flexor muscle contraction needed to maintain an O-ring in BDORT.

[Long-term administration of carmofur as a post-operative adjuvant chemotherapy for cervical adenocarcinoma. Cervical Adenocarcinoma Cooperative Research Association].

Cervical adenocarcinoma and adenosquamous cell carcinoma are low in radiation sensitivity, and the prognosis is said to be inferior to that of squamous cell carcinoma. Eleven facilities nationwide participated in the historical control study on the recurrence prevention effect of carmofur (HCFU) administered at 300 mg/day for more than two years postoperatively as an adjuvant chemotherapy to patients with cervical adenocarcinoma or adenosquamous cell carcinoma for which radical operation is possible. Registered for the study was a total of 252 patients: 77 patients for whom administration of carmofur was begun during the period from January 1987 and March 1989, 71 patients (control 1) who were treated for the cancer after January 1982 but did not receive adjuvant chemotherapy, and 104 patients who received adjuvant chemotherapies with other than carmofur (control 2). In analyses with the Kaplan-Meier method, the carmofur administration group demonstrated a better cumulative survival rate and disease free rate than control 1 (no adjuvant chemotherapy), suggesting carmofur was effective in preventing the recurrence of cervical adenocarcinoma and adenosquamous cell carcinoma.