Association of distinct clinical subsets with myositis-specific autoantibodies towards anti-155/140-kDa polypeptides, anti-140-kDa polypeptides, and anti-aminoacyl tRNA synthetases in Japanese patients with dermatomyositis: a single-centre, cross-sectional study.

OBJECTIVE: To determine the association of distinct clinical subsets with myositis-specific autoantibodies (MSAs) towards anti-155/140-kDa polypeptides [anti-155/140 antibodies (Abs)], anti-140-kDa polypeptides (anti-140 Abs), and anti-aminoacyl tRNA synthetases (ARS Abs) in Japanese patients with dermatomyositis (DM). METHODS: We compared the clinical features and short-term prognoses of 30 DM patients whose serological status included these MSAs. The MSAs were determined by immunoprecipitation. RESULTS: Anti-155/140 Abs (n = 5), anti-140 Abs (n = 8), and anti-ARS Abs (n = 7) did not overlap each other. All of the anti-155/140 Ab-positive patients (n = 5) were complicated by malignancies, as were all of the anti-140 Ab-positive patients (n = 8), who showed rapidly progressive interstitial lung disease (ILD). The survival rate at 6 months from the diagnosis of DM was significantly lower in the anti-140 Ab-positive patients than in the other patients. CONCLUSION: This is the first study to report, in a single cohort of DM patients, that distinct clinical subsets are distributed in an anti-155/140 Ab-positive group, an anti-140 Ab-positive group, or an anti-ARS Ab-positive group. Our data also confirm previous evidence that anti-155/140 Abs are involved in malignancies and that anti-140 Abs are involved in rapidly progressive ILD.

Scavenging effect of butylated hydroxytoluene on the production of free radicals by the reaction of hydrogen peroxide with N-methyl-N'-nitro-N-nitrosoguanidine.

The scavenging effects of the antioxidant butylated hydroxytoluene (BHT) on a hydroxyl free radical (.OH) produced in the reaction of H2O2 with N-methyl-N'-nitro-N-nitrosoguanidine [(MNNG) CAS: 70-25-7] and on free radicals derived from MNNG (.MNNG) were examined by electron spin resonance with the use of the spin-trapping agent 5,5-dimethyl-1-pyrroline-1-oxide (DMPO). BHT was added to the H2O2-MNNG-DMPO system in 50% acetonitrile and exposed to a tungsten-halogen lamp at an intensity of 0.3 mW/cm2 for 3 minutes at 3 degrees C for the first 2 minutes of irradiation. The amounts of .OH and .MNNG in the system decreased and reached constant levels with increase in BHT concentration. The spectrum of the H2O2-MNNG-BHT system showed the specific signal of BHT, whereas the spectra of the BHT solution and the H2O2-MNNG system did not show any signal. These findings indicate that BHT scavenged .OH and .MNNG, and in its reaction with them formed a stable free radical.

Production of hydroxyl-free radical by reaction of hydrogen peroxide with N-methyl-N'-nitro-N-nitrosoguanidine.

Production of a hydroxyl free radical (.OH) by reaction of hydrogen peroxide (H2O2) with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was examined by electron spin resonance using the X OH spin trapping agent 5,5-dimethyl-1-pyrroline-1-oxide (DMPO). The electron spin resonance spectra of the H2O2-MNNG-DMPO system after exposure to light at an intensity of 0.03 mW/cm2 for 5 min, and the DMPO- (.OH) spin adduct (2-hydroxy-5,5-dimethyl-1-pyrroline-1-oxide) generated by use of Fenton's reagent showed the same hyperfine structure and g-value. The signal of the DMPO adduct obtained in the H2O2-MNNG-DMPO system disappeared on addition of the .OH scavenger sodium benzoate. The addition of another .OH scavenger, ethanol, resulted in the appearance of a new signal due to trapping of the alpha-hydroxyethyl radical. These results show that .OH was formed in the H2O2-MNNG-DMPO system. The typical signal of the DMPO-(.OH) spin adduct was not observed in the system in the absence of light. The amount of DMPO-(.OH) spin adduct increased with increase in the concentration of H2O2 when the MNNG level was kept constant, and it changed with the concentration of MNNG at a constant H2O2 level, indicating that .OH was produced by the interaction of MNNG with H2O2. In the absence of H2O2, complicated trapped signals appeared in the spectrum of the MNNG-DMPO system in the light, but these signals were not observed when the system was kept in the dark. In the absence of MNNG, the H2O2-DMPO system did not show any signal, even in the light. These results indicate that interaction of free radicals derived from MNNG with H2O2 on exposure to light resulted in .OH production.

Whether or not prophylactic excision of the extrahepatic bile duct is appropriate for patients with pancreaticobiliary maljunction without bile duct dilatation.

BACKGROUND/AIMS: The standard treatment for patients with a pancreaticobiliary maljunction (PBM) without bile duct dilatation remains controversial. METHODOLOGY: We followed up 29 patients with such PBM who mainly underwent a cholecystectomy alone. The ages of the patients ranged from 3 to 76 years (average age 47.3 years) and the ratio of males to females was 8 vs. 21. When the diameter of the common bile duct was less than 10mm, such bile ducts were diagnosed to have no dilatation. The main clinical indications for surgery were cholecystolithiasis in 15 patients, choledocholithiasis in 3, cholecystocholedocholithiasis in 2, gallbladder polyp in 2, adenomyomatosis in 2, cholecystitis in 2, and protein plug in 1. RESULTS: The amylase levels of gallbladder bile in 20 patients ranged from 115 to 460,200 IU/mL (a mean of 191,698 IU/mL). One patient died of gastric cancer 182 months after surgery and two patients died of other diseases 153, 171 months after surgeries, respectively. The remaining 26 patients have all been doing well for 36 months to 326 months after surgery (a median follow-up period, 160.5 months). The 10- and 15-year survival rates were 100% and 89.7%. CONCLUSIONS: In conclusion, a prophylactic resection of the extrahepatic bile duct and biliary diversion could be unnecessary for patients with PBM without bile duct dilatation.

Thermostable peroxidase activity with a recombinant antibody L chain-porphyrin Fe(III) complex.

In order to engineer a new type of catalytic antibody, we attempt to use a monoclonal antibody L chain as a host protein for a porphyrin. TCPP (meso-tetrakis(4-carboxyphenyl)porphyine) was chemically synthesized and Balb/c mice were immunized using TCPP as a hapten. Two hybridoma cells (03-1, 13-1), that produce monoclonal antibody against TCPP, were obtained. Genes for both H and L chains of monoclonal antibodies were cloned, sequenced and overexpressed using E. coli as a host. ELISA and fluorescence quenching method show that the independent antibody L chains from both Mab03-1 and Mab13-1 have specific interaction with TCPP. Furthermore, the recombinant antibody L chain from Mab13-1 exhibits much higher peroxidase activity than TCPP Fe(III) alone. The enzyme activity was detectable with pyrogallol and ABTS (2,2-azinobis-3-ethylbenzthiazolin-6-sulfonic acid) but not with catechol. This new catalytic antibody was extremely thermostable. Optimum temperature of the peroxidase reaction by the complex of 13-1L chain and TCPP Fe(III) was 90 degrees C, while that the TCPP Fe(III) alone was 60 degrees C.

The Noh mask effect: vertical viewpoint dependence of facial expression perception.

Full-face masks, worn by skilled actors in the Noh tradition, can induce a variety of perceived expressions with changes in head orientation. Out-of-plane rotation of the head changes the two-dimensional image characteristics of the face which viewers may misinterpret as non-rigid changes due to muscle action. Three experiments with Japanese and British viewers explored this effect. Experiment 1 confirmed a systematic relationship between vertical angle of view of a Noh mask and judged affect. A forward tilted mask was more often judged happy, and one backward tilted more often judged sad. This effect was moderated by culture. Japanese viewers ascribed happiness to the mask at greater degrees of backward tilt with a reversal towards sadness at extreme forward angles. Cropping the facial image of chin and upper head contour reduced the forward-tilt reversal. Finally, the relationship between head tilt and affect was replicated with a laser-scanned human face image, but with no cultural effect. Vertical orientation of the head changes the apparent disposition of facial features and viewers respond systematically to these changes. Culture moderates this effect, and we discuss how perceptual strategies for ascribing expression to familiar and unfamiliar images may account for the differences.

Control of photoinduced electron transfer from zinc-porphyrin to methyl viologen by supramolecular formation between monoclonal antibody and zinc-porphyrin.

Photoinduced electron transfer from tetrakis(4-carboxy-phenyl porphyrin)-zinc complex (Zn-TCPP) to an acceptor molecule (methyl viologen; MV2+) has been found to be controlled by the complex formation of monoclonal antibody 03-1 for the porphyrin (TCPP) and Zn-TCPP. Although there are no ground-state interactions between Zn-TCPP and MV2+ for a 2:1 complex of antibody 03-1 and Zn-TCPP, the fluorescence of Zn-TCPP is quenched by the addition of MV2+. The Stern-Volmer plots and emission lifetime studies show that there is a long-range electron transfer through the antibody 03-1.

Effects of nitric oxide on matrix metalloproteinase-2 production by rheumatoid synovial cells.

Nitric oxide (NO) is a multifunctional messenger molecule generated from L-arginine by a family of enzymes, including nitric oxide synthase (NOS). This study was performed to examine whether NO modulates the production of matrix metalloproteinases (MMPs), which degrade all components of extracellular matrix (ECM), in rheumatoid synovial cells. We investigated the effects of exogenously generated NO by a NO donor, S-nitroso-N-acetyl-DL-penicillamine (SNAP), on the MMPs production by rheumatoid synovial cells. Culture media conditioned by SNAP-treated synovial cells were examined by gelatin zymography and immunoblot analysis. Incubation of synovial cells with SNAP resulted in gelatinase A production in a dose-dependent fashion. Furthermore, RT-PCR analysis demonstrated that MMP-2 mRNA expression was induced in SNAP-treated synovial cells. In contrast, SNAP did not influence the production of TIMP-1 and TIMP-2, which preferentially inhibit MMP-2, by rheumatoid synovial cells. Our data indicate that NO could modulate MMP production by rheumatoid synovial cells and therefore contribute to ECM degradation of articular components in RA.

Mucous gland adenoma of the trachea resected with an endoscopic neodymium: yttrium aluminum garnet laser.

We report a case of mucous gland adenoma of the trachea in a 73-year-old male revealed by bronchoscopy. The tumor was resected with a contact neodymium: yttrium aluminum garnet (Nd-YAG) laser after five years of observation. The tumor was histologically peculiar because it presented numerous cystically dilated, or irregularly shaped mucus-filled glands lined with cuboidal or tall columnar cells. In some parts, the lining cells of the tumor showed papillary proliferation. We diagnosed this tumor as a mucous gland adenoma of the trachea. We review the clinical features of this rare tumor and discuss the usefulness of the laser in the diagnosis and the therapy.

Immunocytochemical presentation of alpha-fetoprotein-producing gastric cancer in ascitic fluid: a case study.

A case of primary gastric cancer without hepatic metastasis showing extremely high alpha-fetoprotein (AFP) levels is reported. This case illustrates the application of the immuno-peroxidase technique to ascitic fluid cytology. Papanicolaou-stained smears of the ascites permitted the diagnosis of a metastatic carcinoma. A positive reaction to AFP was demonstrated in the tumor cells in the ascitic fluid cellular samples as well as in the paraffin-embedded tissue section of the primary gastric carcinoma. Rising AFP levels were also detected in ascitic fluid. AFP fractionation using lectin-affinity-crossed-line immunoelectrophoresis showed the hepatic rather than yolk sac type. Reports of such occurrences are few; no study, to the best of our knowledge, has previously documented cytological and immunocytochemical diagnosis in ascitic fluid. AFP-producing gastric cancer should be considered in the differential diagnosis.

Immunocytochemical demonstration of carcinoembryonic antigen in cerebrospinal fluid with carcinomatous meningitis from rectal cancer.

The case presented illustrates the application of the immunoperoxidase technique to cerebrospinal fluid (CSF) cytology. The cytologic findings in a Papanicolaou-stained slide of the CSF permitted the diagnosis of a metastatic carcinoma. Positive reactions to carcinoembryonic antigen (CEA) were demonstrated in the tumor cells in the CSF sample as well as in the paraffin-embedded section of the primary rectal cancer. Rising CEA levels were also detected in both CSF and serum. The determination of cancer-associated antigens, such as CEA in the CSF specimen, may be useful in establishing the presence of metastatic tumor in the CSF.

Cytologic demonstration of Entamoeba histolytica using immunoperoxidase techniques. Report of two cases.

An indirect immunoperoxidase technique to detect Entamoeba histolytica in cell samples from patients suspected to have amebiasis is described. Using a rat antiserum specific for E. histolytica, the organism was clearly identified both in smears and in cell blocks. This immunoperoxidase technique seems to offer great possibilities for a specific, accurate and rapid identification of amebic infestation in diagnostic cytology.

Metabolic and hormonal mechanisms of mineral metabolic adaptation to induced hypokinetics in rats.

During simulated hypokinetics, many biological functions have a tendency to stabilize at different levels with the passing of time. In this experiment, hypokinetic exposure was induced in albino rats by the suspension method for 5 weeks. A new plateau in urinary K+ excretion was attained during the third week, after a transient decrease during the first week. Since the formation of this steady plateau is considered a manifestation of the adaptation of this function, the hypokinetic exposure period was divided into a prestabilized phase and a stabilized phase. To ascertain the biochemical mechanisms of this adaptation process, the effects of aldosterone and anti-aldosterone substances (e.g., tetracycline) on K+ and Na+ excretion patterns in each phase were investigated. Decreasing aldosterone activity in the prestabilized phase and increasing activity in the stabilized phase are recognized as one of the main factors in the renal adaptation process to hypokinetics.

[Pathogenesis-screening tests for liver dysfunction in the asymptomatic patients with elevated ALT values and their diagnostic efficacies in primary care medicine].

We have evaluated the diagnostic efficacies of ultrasonography and hepatitis C virus (HCV) antibody measurement to differentiate pathogenesis of liver dysfunction in the asymptomatic adults with elevated ALT value. Among 4256 visitors to PL Tokyo Health Control Center for their health examination, 463 cases (11%) showed abnormal liver function including elevation of ALT value. Ultrasonography and HCV antibody measurement using the second generation reagent had been applied to 362 cases in order to screen the etiology of liver dysfunction. The ultrasonography succeeded to establish the diagnosis of fatty liver in 137 cases (38%) and 41 cases (11%) demonstrated positive HCV antibody. There were 4 cases with positive HBs antigen, however, it was found that their abnormal liver function was attributed to other etiology such as fatty liver and alcoholic liver dysfunction rather than chronic type B hepatitis. HCV antibody-positive cases showed higher levels of total protein, ZTT, AST, ALT, and lower levels of albumin, A/G, total cholesterol, triglyceride, gamma-GT and cholinesterase value than other cases. HCV antibody titers were not correlated to hepatic parenchymal damage estimated by ALT or cholinesterase value. Only a little correlation was observed between HCV antibody titers and HCV-RNA amounts determined by the competitive reverse transcription-polymerase chain reaction (RT-PCR) method. These results indicate sufficient diagnostic efficacies of ultrasonography and HCV antibody measurement for a pathogenesis differentiation in the asymptomatic patients with liver dysfunction, and these examinations should be employed as the first-step screening tests for the etiology determination of liver diseases in the primary care medicine.

[A clinical application of laser scanning cytometer--the significance in the cytology as an additional diagnostic technique].

Laser scanning cytometer (LSC) is a new machine similar to flow cytometer but with advantages for certain clinical and research applications. LSC is a microscope based and measures cells on a slide with the position of each cell on the slide. This new technique of LSC can be utilized on extremely small specimens and enables direct correlation of all of the measured fluorescent parameters with light microscopic cytologic morphology. To date, LSC has been successfully used to perform DNA content analysis of numerous specimen types and automated analysis of fluorescence in situ hybridization specimens. In this report, we describe characteristics of LSC comparison with flow cytometry and a clinical application of LSC focused on DNA content analysis in clinical specimens with pulmonary disorders. LSC provides a number of benefits that may make it more suitable for clinical laboratories than FCM.

[Clinical evaluation of hepatitis C virus RNA quantification by competitive reverse-transcription PCR].

We have quantified the hepatitis C virus (HCV) RNA by competitive reverse-transcription PCR method (Amplicor) HCV quantification monitor kit) in sera with positive HCV antibody measured by means of the second generation HCV antibody assay. Among the visitors to PL Tokyo Health Control Center for their health examination and the patients to National Defense Medical College Hospital, 123 HCV antibody-positive cases were examined. A positive but low correlation between the amount of HCV-RNA and the titer of HCV antibody (r = 0.508, p value < 0.0001) was obtained. HCV-RNA was not detectable in 19 HCV antibody-positive cases. Among them, 10 cases showed normal ALT values. In the cases with more than 1000 copies/ml of HCV-RNA, the greater the amount of HCV-RNA the higher ALT values were observed, while the titer of HCV antibody was not correlated to ALT. This study demonstrated the dissociation and low correlation between HCV-RNA amount and antibody-titers in some patients, which may recommend direct quantification of RNA for clinical evaluation of the patients with positive-HCV antibody. The quantification of HCV-RNA by such rapid and simple methods can be applicable for the determination of HCV-RNA amount in routine laboratory works.

[A case of non-small-cell lung cancer with intramedullary spinal cord metastasis diagnosed pre-mortem].

The patient was a 54-year-old man who in May 1999 received a diagnosis of squamous cell carcinoma, T4 N2 M1, stage IV. Systemic chemotherapy and stereotactic radiosurgery were performed only to result in further progression of the disease. In August 1999, he experienced gait disturbance due to lumbar pain. Rehabilitation improved the gait disturbance and he was discharged. In October, since the pain reappeared and there was numbness in the right leg, he was readmitted. Brain MRI revealed multiple brain metastasis and whole brain irradiation was performed. But his symptoms deteriorated, and palsy of the right leg ensued. Later, bladder dysfunction also developed. Since spinal cord MRI revealed intramedullary metastasis at Th 12 and L1 levels, we performed radiotherapy for the lumbar medullary lesion, together with systemic chemotherapy. After chemoradiotherapy the tumor size decreased and the pain improved. Cases of lung cancer with intramedullary metastasis are rare, especially those diagnosed before death.

[Microspectrophotometric analysis of nuclear DNA contents related to the presence of alpha-fetoprotein in human hepatocellular carcinoma].

A total of 28 surgically resected hepatocellular carcinomas, including both fresh-smeared specimens and paraffin-embedded tissues, was stained with alpha-fetoprotein (AFP) antibody. In all cases we measured the DNA contents using microspectrophotometry. In 5 cases, analyses of the nuclear DNA histogram of both the AFP-positive and negative cells from the same specimen were successfully performed. The ratio of the S-G2.M phase of the AFP-positive cells was 2.24 times more than that of the AFP-negative cells. In addition, not only the ratio of S-G2.M phase cells, but also the incidence of aneuploidy pattern of AFP positive stained cases was higher than the negative stained cases. Histologically, the AFP-positive cases contained various differentiated tissues more than the negative cases. From these results, we confirmed that AFP production was related to the growth of HCC, and the results suggest that AFP producing HCC have a higher potential for growth than AFP non-producing HCC.

[Cytofluorometric and histopathological studies on non-cancerous liver lesions after lipiodol-transcatheter arterial embolization].

We have investigated the effects of preoperative Lipiodol-Transcatheter arterial embolization (Lp-TAE) on both the nuclear DNA content of noncancerous liver cells by DNA-cytofluorometry (using NIKON SPM-RF1-D), and the histopathological findings of liver lesions. Lp-TAE through the hepatic artery was performed on ten primary hepatocellular carcinoma patients using Gelfoam in combination with adriamycin (40 mg) or mitomycin C (20 mg), emulsified in Lipiodol (10 ml) and 60% Urografin (2 ml). And hepatic resection was carried out after one or two months later in each case. Histologically, primary hepatocellular carcinoma lesions showed various patterns of necrosis, while the non-cancerous liver cells did not show hepatocellular infarction in the liver cirrhosis. The results of the nuclear DNA content showed an increase in the fractions of poliploid cells in the non-cancerous lesions treated with Lp-TAE compared with that in the normal age-matched controls. But we found no remarkable differences in the ploidy patterns between Lp-TAE treated patients and untreated patients with liver cirrhosis. In conclusion, preoperative Lp-TAE does not induce any remarkable histological hepatocellular damage of non-cancerous lesion and does not affect nuclear DNA content of non-cancerous liver cells.

[Analysis of nuclear DNA heterogeneity of the hepatocellular carcinoma (HCC)].

To evaluate the objective proliferative activity in HCC nuclear DNA contents were measured by means of microspectrophotometry and at the same time the immunohistochemical technique using anti-PCNA antibody was employed. Surgically resected 26 HCCs were analyzed in terms of cell proliferative activity and regional heterogeneity. The analysis was performed by immunohistochemical demonstration of PCNA and pathologic histochemical study in formalin-fixed, paraffin-embedded specimens and cytophotometric measurements of nuclear DNA contents in fresh specimens. The results were as follows. 1) Nine HCCs showed regional ploidy heterogeneity. 2) PCNA labeling index and histological grade of the marginal area was much higher than that of the central area. From these results, we concluded that in the process of the HCC progression proliferative activity was decreased in the central area and was not decreased in the marginal area.