Biomedical subjects
Publications and source records attributed to Y Nagao.
The expression of mRNA of leptin, the product of the obese gene, in bovine adipose tissue was analyzed by a lysate RNase protection assay. The mRNA level was significantly decreased by food deprivation for two days and partially recovered after 3 hr of refeeding, indicating that obese gene expression in the ruminant was regulated by feeding.
A subjective evaluation of beer drinkability and the degree of stomach fullness were found to correlate with the relaxed cross-sectional area of the pylorus antrum measured by real-time ultrasonography. Five kinds of beer with a different malt/adjuncts ratio and degree of attenuation were used. Each beer was given to 9 healthy volunteers at the rate of 3 ml/kg/15 min, and they each recorded the degree of stomach fullness, desire to drink and tastiness every 30 min. With increasing volume drunk, the degree of tastiness and desire to drink were lowered, and the degree of stomach fullness raised. The relaxed cross-sectional area of the pylorus antrum measured by an ultrasonic image analyzer every 30 min was highly correlated with the degree of stomach fullness, tastiness of the beer, and desire to drink (p = 0.0021, < 0.0001, 0.001). The beer giving the lowest degree of stomach fullness was appraised to be tasty and highly drinkable. These findings suggest that the rate of gastric emptying is one of the factors determining the drinkability of beer, and that measurement of the relaxed cross-sectional area of the pylorus antrum is useful to evaluate stomach fullness during beer drinking.
This study examined the timing of completion of meiosis I of bovine oocytes in which meiotic resumption had been inhibited by cycloheximide (CHX), and also determined the optimum interval of maturation in culture for subsequent fertilization and development. Most oocytes treated with CHX reached metaphase II at 16 hr in the maturation culture, while control oocytes did at 20 hr. CHX-treated oocytes cultured for 16 hr were normally fertilized but failed to develop into blastocysts. Maturation in culture for 20 hr resulted in comparable development for control oocytes. The results indicate that nuclear maturation of CHX-treated oocytes was completed 4 hr faster than for control oocytes, however the same interval of maturation as that of control oocytes is necessary for subsequent development to blastocysts.
To determine whether hepatitis C virus (HCV) replicates in bone marrow, we investigated positive- and negative-strand HCV RNA in bone marrow cells and fluids, and sera from patients with HCV infection. The study population consisted of 15 patients positive for antibodies to HCV (anti-HCV). Positive- and negative-strands HCV RNA were detected using highly strand-specific rTth reverse transcription-polymerase chain reaction (rTth RT-PCR) followed by Southern blotting analysis. Positive-strand HCV RNA was detected in 12 (80%) serum samples, in 13 (86.7%) bone marrow fluid specimens, and in 6 (40.0%) bone marrow cell samples. Negative-strand HCV RNA was detected in 9 (60.0%) serum samples, 11 (91.7%) fluid specimens, while it was not detected in bone marrow cells. The absence of negative-strand HCV RNA in bone marrow cells suggested that HCV does not replicate in these cells. Negative-strand HCV RNA detected in serum and bone marrow fluid samples may have been due to contamination with circulating HCV RNA from hepatocytes.
A 48-year-old woman with anti-epiligrin cicatricial pemphigoid (CP) who showed clinical features resembling epidermolysis bullosa acquisita was found to have adenocarcinoma of the stomach. Histological examination of lesional skin demonstrated a subepidermal blister. Direct immunofluorescence microscopy of perilesional skin revealed linear deposits of IgG and C3 at the basement membrane zone. The patient's serum contained IgG autoantibodies that bound to the dermal side of 1 mol/L NaCl-split normal human skin as determined by indirect immunofluorescence microscopy, and the lamina lucida as determined by indirect immunoelectron microscopy. The patient's serum immunoprecipitated laminin-5 from extracts and media of biosynthetically radiolabelled human keratinocytes. Immunoblot studies showed that the patient's autoantibodies specifically bound the alpha3 subunit of this laminin isoform. Fragility of the skin and bullous lesions disappeared after total gastrectomy, but soon reappeared possibly in association with metastatic disease in a lymph node. The possibility that anti-epiligrin CP may develop paraneoplastically in some patients is discussed.
The MAGE-4 gene, a member of the MAGE gene family, is expressed in various cancers, including head-and-neck squamous-cell carcinomas (HN-SCC), but is not expressed in any normal tissues except for the testis and placenta. The aim of this study was to determine whether serum MAGE-4 protein is a useful tumor marker for detection of HN-SCC. An enzyme-linked immunosorbent assay was used to measure serum level of MAGE-4 protein. The serum level of MAGE-4 in pre-operative HN-SCC patients was significantly higher than that in patients with non-malignant diseases (NMD) of the head and neck, volunteers undergoing cancer screening (VOL), or healthy donors (HD). When the cut-off level was determined at 1.15 ng/ml (mean plus 3 SD of HD), sera from 28 of 96 patients with HN-SCC (p < 0.0001 vs. the other groups), 7 of 82 patients with NMD, 2 of 92 with VOL, and 0 of 68 HD were positive for MAGE-4. Serum levels of MAGE-4 protein in all 7 HN-SCC patients whose sera were positive for MAGE-4 before operation decreased after operation, and, in one patient, a renewed rise in serum level was followed by recurrence. These results indicate that MAGE-4 protein is detectable in sera of a significant number of HN-SCC patients, and that serum MAGE-4 protein might be a useful tumor marker to monitor the recurrence of MAGE-4-positive HN-SCC.
Oral lichen planus (OLP) is frequently associated with hepatitis C virus (HCV) infection. We have previously reported that the pathogenesis of OLP arises from host rather than viral factors. In this study, we investigated the role of these factors in 41 patients with chronic hepatitis C: 22 with OLP (group 1) and 19 without OLP (group 2). All patients had antibodies to HCV (anti-HCV) and were serum HCV RNA-positive; none were HBsAg-positive. Immune abnormalities in serum were evaluated by testing for antinuclear antibody (ANA), rheumatoid factor (RF) activity, immunoglobulins (IgG, IgM, and IgA), and cryoglobulin. The rate for ANA positivity and IgM levels were significantly higher in group 1 than in group 2 (P < 0.05). Mean age in group 1 was significantly higher in group 2 (P = 0.0001). Of the factors tested, ANA, IgM, and age, logistic regression showed that age correlated independently with OLP (P = 0.003). In 5 patients in group 1, the infiltrating lymphocytic subsets of the OLP lesion were examined histopathologically. Predominant T cell infiltration was shown in all 5 patients. In addition to host factors, wer also examined viral factors in both groups of patients, measuring serum HCV RNA level and determining HCV genotype. There were no significant differences between the groups in these viral factors. This study suggested that host factors induced by the HCV infection are more important than viral factors in the pathogenesis of OLP associated with hepatitis C.
This study was conducted to assess the fertilizability and developmental capacity of bovine oocytes which had been maintained in meiotic arrest by either a protein synthesis inhibitor, cycloheximide (CHX), or an inhibitor of serine/threonine protein kinases, 6-dimethylaminopurine (6-DMAP). Both CHX and 6-DMAP reversibly prevented nuclear maturation of nearly all oocytes for 24 h. After the reversal of arrest, CHX-treated oocytes could be successfully matured and fertilized. They developed to the blastocyst stage at slightly lower rates than oocytes cultured without inhibition for 22 h prior to sperm addition but at higher rates than those incubated in a medium containing no inhibitors for 46 h prior to fertilization. Oocytes inhibited by CHX for 48 h matured and fertilized normally but failed to develop into blastocysts. Even though 6-DMAP-treated oocytes completed meiosis I after removal from the drug, the rates of fertilization and blastocyst formation were lower than for untreated oocytes or CHX-treated oocytes. Effects of adding FSH and/or estradiol-17 beta (E(2)) during CHX-inhibition for 24 h were also examined. Embryos from oocytes treated with CHX and E(2) or with CHX and FSH + E(2) developed into blastocysts at similar rates as the controls. Further development of inhibited oocytes was examined by transferring blastocysts derived from oocytes inhibited by CHX with FSH and E(2) for 24 h to recipient heifers. Two calves were obtained following transfer. These results indicate that CHX-inhibited oocytes retain developmental competence, while 6-DMAP-inhibited oocytes after the reversal of arrest have reduced capacities for fertilization and further development. The addition of FSH and E(2) during CHX-inhibition improves development to the blastocyst stage of the oocytes that are capable of initiating and maintaining pregnancy after embryo transfer to recipient animals.
We studied the incidence of GBV-C infection in patients with oral cancer or oral lichen planus (OLP). Subjects included 38 patients with oral cancer and 34 patients with OLP from whom serum was collected to detect GBV-C-RNA using reverse transcription (RT) "nested" PCR. GBV-C-RNA was positive in 1 (2.6%) oral cancer patient and 3 (8.8%) OLP patients. All 4 GBV-C-positive patients were also positive both for the antibody for hepatitis C virus (anti-HCV) and HCV-RNA, indicating simultaneous infection with GBV-C and HCV. In conclusion, GBV-C infection was found in 5.5% (4/72) of oral cancer or OLP patients and all patients infected with GBV-C also had HCV infection. This suggests that GBV-C infection is unlikely to play an important role in the pathogenesis of oral cancer and OLP.
Copolymers (IABb) composed of N-isopropylacrylamide (I), N-(3-dimethylaminopropyl)-acrylamide (A), 3-acrylamidophenylboronic acid (B), and a hydrophobic comonomer, n-butyl methacrylate (b), were synthesized as cell culture substrata since we previously learned that bovine aortic endothelial cells (BAECs) cultured on the phenylboronic acid-containing copolymer (IAB) differentiated into capillary structures after 26 days culture. The synthesized IABb copo ymers contained higher boron and amine moieties than the IAB copolymer. The results of the dynamic contact angle measurement revealed that IABb copolymer-coated surfaces showed a relatively hydrophobic nature, changing to hydrophilic in response to the aqueous environment. BAECs cultured on the copolymer substrata developed into capillary networks after 7 days. This is probably due to the enrichment of boron and amine segments in the vicinity of the hydrophilic copolymer surface, enhancing more pronounced interaction of boronates with cell membrane glycocalyx. The introduction of n-butyl methacrylate into the polymers might enhance the diffusion of the hydrophobic segments to the bulk polymers and the concentration of relatively hydrophilic segments at the outermost polymer surfaces by contact with water. A copolymer (IAP) without boronic acid groups was also prepared using N-phenylacrylamide (P) as a comonomer instead of 3-acrylamidophenylboronic acid in the IAB copolymer to investigate the effect of boronic acid moieties on the capillary formation of the cultured cells. Although the endothelial cells seeded on the copolymer without boronic acid groups adhered during the early culture period, these cells showed neither proliferation nor differentiation and detached from the surface after 13 days. These results strongly support the opinion that the phenylboronic acid groups in the copolymers are responsible for the specific induction of tissue formation of BAECs through the interaction with glycoconjugates on the cell membranes.
A 49-year-old man was treated with cisplatin and pirarubicin for tongue cancer. After the second course of chemotherapy, partial seizures including transient motor aphasia, tonic finger movement, and loss of consciousness were observed. The EEG showed frequent diffuse (multiple) spike and slow wave discharges. Following the administration of carbamazepine and diazepam, no seizures occurred and no paroxysmal discharges were observed or EEGs. We conclude that carbamazepine and diazepam administration was effective.
We examined the effect of ulinastatin in comparison with prednisolone (PSL) and salazosulfapyridine (SASP), well-known drugs for ulcerative colitis (UC), on two experimental UC models induced by dextran sulfate sodium (DSS) or trinitrobenzene sulfonic acid (TNB) in rats. Ulinastatin at the doses of 3000 approximately 10,000 units/kg/day (i.v.) significantly ameliorated the formation of erosion and infiltration of inflammatory cells in colonic mucosa in DSS-induced rat UC models. Moreover, ulinastatin at the dose of 10,000 units/kg/day (i.v.) significantly suppressed inflammation with ulcer in the colonic mucosa in TNB-induced rat UC models. PSL at the dose of 1 mg/kg/day (p.o.) also was as effective as ulinastatin on the above two UC models, while SASP at the dose of 100 mg/kg/day (p.o.) was less effective than ulinastatin and PSL. In addition, ulinastatin inhibited the activities of elastase and cathepsin G from human leukocytes, and it suppressed TNF alpha, IL-8 and superoxide production by rat macrophages and rabbit leukocytes in vitro. These results suggest that the suppression of inflammatory mediators produced by leukocytes is involved in the mechanism of the anti-UC action of ulinastatin.
The effects of BRL35135A ((R*,R*)-(+/-)-methyl-4-[2-[2-hydroxy-2 -(3-chlorophenyl)ethylamino]propyl]-phenoxyacetate hydrobromide), a beta 3-adrenoceptor agonist, on visceral and subcutaneous fat weight and metabolic disorders were studied in genetically obese C57BL/KsJ db/db mice and Zucker fa/fa rats. In db/db mice, four weeks of oral administration of BRL35135A (0.5 and 5 mg/kg/day) decreased body weight gain and reduced white fat weight. The rates of reduction of white fat weight were in the order mesenteric fat > retroperitoneal fat > subcutaneous fat. In fa/fa rats, daily administration of BRL35135A (0.05 mg/kg/day)) for 6 weeks reduced the visceral white fat weight/total energy intake ratio, particularly for mesenteric fat, without any clear effect on body weight gain. This tendency of the compound to exert effects on visceral fat was consistent with the findings that the effect of BRL37344 ((R*,R*)-(+/-) -methyl-4-[2-[2-hydroxy-2-(3-chlorophenyl)ethylamino]propyl]-phenoxyacet ic acid), an active metabolite of BRL35135A, on the lipolytic activity of isolated adipocytes and the tissue concentration of [14C]BRL37344 in male Wistar rats were each greater in visceral fat than in subcutaneous fat. Moreover, BRL35135A at 0.05 mg/kg/day elevated serum insulin levels and improved hyperglycemia in db/db mice without reducing body weight gain, whereas at doses of 0.5 and 5 mg/kg/day it ameliorated hyperglycemia and hyperlipidemia, and tended to decrease serum insulin levels. In fa/fa rats, BRL35135A (0.005 mg/kg/day) was also effective in improving hyperinsulinemia, glucose intolerance, and hypertriglyceridemia without any effect on body weight gain or fat distribution. These findings suggest that the improvement of metabolic disorders by BRL35135A may be due to improvement in insulin resistance as well as reduction of visceral fat weight.
41 students took either docosahexaenoic acid (DHA)-rich oil capsules containing 1.5-1.8 grams DHA/day (17 females and 5 males) or control oil capsules containing 97% soybean oil plus 3% fish oil (12 females and 7 males) for 3 mo in a double-blind fashion. They took a psychological test (P-F Study) and Stroop and dementia-detecting tests at the start and end of the study. The present study started at the end of summer vacation and ended in the middle of mental stress such as final exams. In the control group extraggression (aggression against others) in P-F Study was significantly increased at the end of the study as compared with that measured at the start (delta = +8.9%, P = 0.0022), whereas it was not significantly changed in the DHA group (delta = -1.0%). The 95% CI of differences between the DHA and control groups were -16.8 to -3.0%. DHA supplementation did not affect the Stroop and dementia-detecting tests. Thus, DHA intake prevented extraggression from increasing at times of mental stress. This finding might help understand how fish oils prevent disease like coronary heart disease.
A case with recurrent hyperparathyroidism secondary to chronic renal insufficiency is reported. The patient had undergone total parathyroidectomy and autotransplantation of parathyroid tissue five years ago. Bone scintigraphy clearly demonstrated skeletal involvement of secondary hyperparathyroidism and 99mTc-methoxyisobutylisonitrile scintigraphy clearly demonstrated a hyperfunctioning parathyroid autograft.
A novel series of pivaloyloxy benzene derivatives has been identified as potent and selective human neutrophil elastase (HNE) inhibitors. Convergent syntheses were developed in order to identify the inhibitors which are intravenously effective in an animal model. A compound of particular interest is the sulfonanilide-containing analogues. Structure-activity relationships are discussed. Structural requirements for metabolic stabilization are also discussed.
Lichen planus (LP) is a common oral disorder that may represent a mucosal reaction to a variety of factors, including hepatitis C virus (HCV). To determine whether viral factors play a role in oral lichen planus (OLP) pathogenesis, we measured serum HCV RNA and determined HCV genotype in patients with chronic hepatitis C accompanied by OLP. The subjects included 43 patients with chronic hepatitis C: 23 with OLP (group 1) and 20 without OLP (group 2). Serum was collected from all subjects and used to quantify HCV RNA by the branched DNA signal amplification assay; HCV genotypes were classified by the reverse transcription-polymerase chain reaction (RT-PCR) method into types I, II, III and IV. Comparison of patient characteristics disclosed that the mean age of group 1, 60.7 years, was significantly higher (P = 0.001) than that of group 2 (46.4 years). No significant differences were seen between sexes in values of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), total protein (T protein), albumin and gamma-globulin. There were also no significant differences in HCV RNA levels or HCV genotypes between groups. The findings suggested that OLP pathogenesis was a result of host factors rather than viral factors.
Oral lichen planus (OLP) is frequently seen in patients with hepatitis C virus (HCV) infection. To clarify the role of HCV in OLP pathogenesis, we investigated the occurrence and progression of oral lesions in chronic hepatitis C patients treated with interferon. Oral surgeons examined 24 hepatitis C patients (15 men, nine women; mean age 48.1 years) for oral lesions before, during and after interferon (IFN) treatment. OLP was observed in 16.7% (4/24). Two patients had OLP before treatment, one during and one after treatment. Those who developed OLP during or after treatment had neither improvement nor disappearance of OLP even when serum HCV RNA became negative. Leucoplakia was seen in four patients before treatment and oral cancer in one patient 6 months after completing treatment. OLP can occur, exacerbate and persist during IFN treatment for hepatitis C, even when serum HCV RNA becomes negative. The present study suggested that OLP pathogenesis in hepatitis C is due to host factors induced by HCV infection rather than direct HCV participation. Treating physicians should be aware of OLP occurrence or exacerbation by IFN treatment with hepatitis C patients, but IFN therapy is not necessarily contraindicated in these patients.