Mirthful laughter differentially affects serum pro- and anti-inflammatory cytokine levels depending on the level of disease activity in patients with rheumatoid arthritis.

OBJECTIVES: To examine the effect of mirthful laughter in rheumatoid arthritis (RA), we evaluated the levels of serum cytokines before and after patients experienced mirthful laughter. METHODS: Forty-one patients with RA and 23 healthy subjects were enrolled. They listened to 'Rakugo', a traditional Japanese comic story, to induce mirthful laughter. We measured serum IL-6, IL-1beta, TNF-alpha, IL-4 and IL-1 receptor antagonist (IL-1Ra) concentrations before and after patients listened to the story. The RA subjects were divided into two groups. One was designated the 'difficult-to-control RA' group (CRP > or =1.0 mg/dl); The other group was regarded as the 'easily controlled RA' group (CRP <1.0 mg/dl). RESULTS: The basal levels of serum IL-6 and TNF-alpha in the RA patients were significantly higher than those in the healthy group. After experiencing mirthful laughter, the levels of serum IL-6 decreased significantly in the RA group but not in the healthy subjects. Interestingly, the level of serum TNF-alpha decreased only in the easily controlled RA group. Serum IL-4 concentration in the RA group was significantly higher than that in healthy subjects before the story. After the story, the level of serum IL-4 significantly decreased in the RA group, especially in the difficult-to-control RA group. In contrast, serum IL-1Ra concentration was statistically higher in the RA group than that in healthy subjects before the story, and a further increase was observed after the story, especially in the easily controlled RA group. CONCLUSIONS: Our findings suggest that mirthful laughter affects the levels of serum pro- and anti-inflammatory cytokines differentially, depending on the RA disease activity.

Effect of general anesthesia on the abnormal immune response in patients with rheumatoid arthritis.

OBJECTIVE: To evaluate the influence of mental stress on the neuroendocrine-immune system in patients with rheumatoid arthritis (RA). METHODS: Twenty-four patients with RA and 10 patients with osteoarthritis (OA) who underwent total knee or hip arthroplasty under general anesthesia were enrolled in this study. The blood levels of interleukin-6 (IL-6), IL-1 receptor antagonist (IL-1Ra), tumor necrosis factor-alpha (TNF-alpha), soluble TNF receptors (TNF-Rs) and other substances related to stress were measured just before administering anesthesia on the day of the operation when the patients lay on the operating table and roughly 30 min later when the patients were under general anesthesia without mental stress. These values were compared with those at the same time on the day before the operation, which were considered as controls. RESULTS: In patients with RA under general anesthesia, the levels of IL-6, TNF-alpha, and TNF-R1 and TNF-R2 in the peripheral blood were significantly decreased compared with the levels before anesthesia (p < 0.01). Before anesthesia the levels of IL-1Ra in the peripheral blood were significantly higher, and the level of IL-1Ra was enhanced after the administration of general anesthesia, when compared with the level on the day before the operation (p < 0.01). Such changes were not apparent in patients with OA. CONCLUSION: In patients with RA, excessive mental stress should be eliminated to modify the interaction between the stress-immune system and stress-endocrine system as a method to better control disease activity.

Uni-axial cyclic stretch induces Cbfa1 expression in spinal ligament cells derived from patients with ossification of the posterior longitudinal ligament.

Ossification of the posterior longitudinal ligament of the spine (OPLL) is characterized by ectopic bone formation in the spinal ligaments. Mechanical stress, which acts on the posterior ligaments, is thought to be an important factor in the progression of OPLL. To clarify this mechanism, we investigated the effects of in vitro cyclic stretch (120% peak to peak, at 0.5 Hz) on cultured spinal ligament cells derived from OPLL (OPLL cells) and non-OPLL (non-OPLL cells) patients. The mRNA expressions of Cbfa1 (an osteoblast-specific transcription factor), type I collagen, alkaline phosphatase (ALP), osteocalcin and integrin beta1 (a mechanotransducer) were increased by cyclic stretch in OPLL cells, whereas no change was observed in non-OPLL cells. The effects of cyclic stretch on the spinal ligament tissues derived from OPLL and non-OPLL patients were also analyzed by immunohistochemistry using an antibody against Cbfa1. The expression of Cbfa1 was increased by cyclic stretch at the center of the spinal ligament tissues of OPLL patients, whereas no change was observed in the tissues of non-OPLL patients. Furthermore, U0126, a specific inhibitor of MAPK kinase (MEK), suppressed the stretch-induced mRNA expressions of Cbfa1, ALP and type I collagen in OPLL cells. These results suggest that in OPLL cells, mechanical stress is converted by integrin beta1 into intracellular signaling and that Cbfa1 is activated through the MAP kinase pathway. Therefore, we propose that mechanical stress plays a key role in the progression of OPLL through an increase in Cbfa1 expression.

Does being easily moved to tears as a response to psychological stress reflect response to treatment and the general prognosis in patients with rheumatoid arthritis?

OBJECTIVE: Psychological stress affects the condition of patients with rheumatoid arthritis (RA). We evaluated the neuroendocrine and immune responses (NEIRs) in the peripheral blood to psychological stress induced by deep emotion with tears in patients with RA. METHODS: We compared the levels of plasma cortisol and interleukin-6 (IL-6), the CD4/CD8 ratio, and natural killer (NK) cell activity in peripheral blood between the patients with easily controlled RA (CRP < 1.0 mg/dl) and those with difficult-to-control RA (CRP > or = 1.0 mg/dl) before and after the stress session. RESULTS: Psychological stress induced by deep emotion with tears had a greater influence on NEIRs in patients with difficult-to-control RA (CRP > or = 1.0 mg/dl) than in those with easily controlled RA (CRP < 1.0 mg/dl). The levels of plasma cortisol, IL-6, and the CD4/CD8 ratio were lower, while NK cell activity in the peripheral blood was higher in those who were not moved to tears than in those who were moved to tears. Patients who were moved to tears were apt to obtain good control of RA (CRP < 1.0 mg/dl) within one year. CONCLUSION: The patients with better RA control are easily moved to tears as an emotional expression; shedding tears is considered to suppress the influence of stress on the NEIRs, thus preventing the buildup of stress. Patients who were moved to tears had a more easily controlled RA compared with those who were emotionally affected but not moved to tears.

Roles of tyrosine kinase and protein kinase C in infarct size limitation by repetitive ischemic preconditioning in the rat.

In this study, we examined the possibility that infarct-size limitation by repetitive preconditioning (PC) is achieved by activation of both protein kinase C (PKC) and tyrosine kinase. In addition, we assessed whether such kinase activation is triggered by angiotensin II type 1 (AT1) and alpha1-adrenergic receptors and whether sarcolemmal and mitochondrial adenosine triphosphate (ATP)-sensitive potassium (K(ATP)) channels play roles as effectors of cardioprotection in the rat. Under pentobarbital anesthesia, myocardial infarction was induced by 20-min coronary occlusion and 3-h reperfusion in the rat. Infarct size was determined by tetrazolium and expressed as a percentage of area at risk (%IS/AR). PC with one cycle of 5-min ischemia/5-min reperfusion before 20-min ischemia significantly reduced %IS/AR from the control value of 49.4 +/- 2.0 to 35.4 +/- 2.8, and repetitive PC with two cycles of 5-min ischemia/5-min reperfusion further limited %IS/AR to 3.2 +/-0.9. Infarct-size limitation by single-cycle PC was completely abolished by a PKC inhibitor, staurosporine (100 microg/kg; %IS/ AR, 45.7 +/- 5.0). In contrast, the cardioprotection by repetitive PC was only partially blocked by staurosporine (%IS/AR, 19.8 +/- 2.4), another PKC inhibitor, polymyxin B (5 mg/kg; %IS/AR, 16.2 +/- 3.1), or a tyrosine kinase inhibitor, genistein (5 mg/kg; %IS/AR, 21.8 +/- 1.4). However, a combined injection of genistein and staurosporine additively inhibited protection of repetitive PC (%IS/AR, 36.4 +/- 1.7). Staurosporine, polymyxin B, or genistein alone did not modify %IS/AR in nonpreconditioned rat hearts. Infarct-size limitation by repetitive PC was not attenuated by pretreatment with a selective AT1-receptor blocker (CV11974, 10 mg/kg), prazosin (0.6 mg/kg; %IS/AR, 6.4 +/- 3.2 and 1.6 +/- 0.5, respectively). A selective blocker of mitochondrial K(ATP) channels, 5-hydroxydecanoate (3 mg/kg), completely abolished the cardioprotective effect (%IS/AR, 50.8 +/-3.5), but HMR1883 (3 mg/kg), a selective blocker of sarcolemmal K(ATP) channels, failed to inhibit the preconditioning effect (%IS/AR, 4.4 +/- 0.7). These findings suggest that repetition of PC provokes activation of both PKC and tyrosine kinase, leading to enhanced antiinfarct tolerance by opening of mitochondrial but not sarcolemmal K(ATP) channels. It is unlikely that activation of either AT1 or alpha1-adrenergic receptor alone is crucial to trigger preconditioning. Key Words: Tyrosine kinase-Genistein-Angiotensin II-alpha1-Adrenergic receptor-Sarcolemmal K(ATP) channel-Mitochondrial K(ATP) channel.

Proliferating cell nuclear antigen in normal and regenerating rat livers.

Immunohistochemical analysis using proliferating cell nuclear antigen (PCNA) antibody shows a negligible number of cells stained in normal liver, but much higher numbers in regenerating liver 24 and 48 h after surgery. We also verified different results by biochemical analysis. Two forms of PCNA, L type (eluted at low concentrations of KCl from a phosphocellulose column) and H type (eluted at high KCl concentrations), were observed in the nucleoplasm of regenerating livers 24 and 48 h after surgery. Treatment of the H type fraction with nuclease caused the H type to disappear and the amount of L type to increase. PCNAs in the cytoplasm are P type (eluted in the pass through fraction) and L type. Surprisingly, the total amounts of P type and L type in cytoplasmic extracts are comparable to those of L type and H type in the nucleoplasm. These results suggest that newly synthesized PCNA is immediately converted into the P and L complex forms. The P type and some of the L type that lacks a nuclear localization signal remain in the cytoplasm; the rest of the L type with a nuclear localization signal is transferred into the nuclei. Then, some of the L type in the nucleoplasm forms the H type, which binds to DNA. These three types of PCNA are also found in significant amounts in the nucleoplasm and cytoplasm of normal rat liver despite its nonproliferating state.

Characterization of the cytotoxic activity of nitric oxide generating N-nitroso compounds.

The NO-generating abilities of aromatic N-nitroso compounds (nitrosoureas, nitrosamides and nitrosamines), and N-acetyl-S-nitroso-DL-penicillamine at ambient temperature were compared by employing the Griess reaction. 3,3-Dibenzyl-1-(4-tolyl)-1-nitrosourea showed the greatest NO-generating ability among the tested compounds. The NO-generating ability of the aromatic N-nitrosoureas and N-nitrosamides was greater than that of the N-nitrosamines, presumably reflecting differences in electrostatic repulsion between the carbonyl oxygen and nitroso oxygen in these compounds. In addition, a conjugative effect between the aromatic ring carbon and neighboring nitrogen influences the NO-generating ability; the conjugative effect in the case of N-nitrosoureas and N-nitrosamides having an ortho-alkyl substituted aromatic ring, or N-nitrosamines having a bulky N-group, such as tert-butyl, is decreased by an increase in steric hindrance around the nitroso group. The N-NO bond then becomes more stable. The NO-generating ability was related to the reciprocal of the ID50 value for growth inhibition of cultured L-5178 Y cells by the aromatic N-nitroso compounds. On the other hand, NO production from the aliphatic N-nitroso compounds was not observed under our conditions, and these N-nitroso compounds did not show effective cytotoxic activity.

Clinical evaluation of a new continuous intraarterial blood gas monitoring system in the intensive care setting.

The present study was designed to evaluate a new continuous intraarterial blood gas monitoring system under routine clinical intensive care conditions. Nine mechanically ventilated adult patients were enrolled in this study. A multiparameter intravascular sensor was inserted into the radial or dorsalis pedis artery through a 20-gauge cannula in each patient. The accuracy of the sensor for pH, Pco2, and Po2 values was evaluated by comparing the data simultaneously obtained from the monitoring system and from conventional blood gas analysis. Measurements were performed for 3 days for each sensor. A total of 62 blood samples were obtained for comparison. The ranges of measured variables were: pH 7.185-7.602, Pco2, 28.8-68.5 mmHg, and Po2 45.2-542.4 mmHg. The overall bias ±precision values were 0.002±0.018 for pH units, 0.53±2.04mmHg for Pco2, and -1.62±20.00 mmHg for Po2. In clinically important ranges of Po2, less than 200 mmHg in particular, the bias and precision values were -2.25±6.48 mmHg in the range of less than 100mmHg, and 0.98±14.38 mmHg in the range of 100-200 mmHg. Variations of sensor accuracy as a function of elapsed time were within the clinically acceptable range throughout the study period. These findings suggest that this new device is sufficiently useful for routine clinical settings.

Juvenile hyaline fibromatosis. Case report with five years' follow-up.

Juvenile hyaline fibromatosis (JHF) is a rare hereditary disorder named by Drescher et al. in 1969. As recently as 1985, only 30 cases had been reported worldwide. We report the case of a 9-year-old girl who was diagnosed with JHF at age 3 and has been closely followed since. She initially had slowly growing multiple soft tumors over her entire body as well as hypertrophic gingiva and mild bone deformities. She was originally misdiagnosed with infantile myofibromatosis at age 3. However, at age 6, because of the light and electron microscopic findings of the tumors, she was diagnosed as having JHF. Currently, at age 9, she has nodular lesions developing over her body as well as bone changes that are progressing with no evidence of regression.

Efficacy of interferon alfa therapy in chronic hepatitis C patients depends primarily on hepatitis C virus RNA level.

To clarify the viral factors that may predict the therapeutic effect of interferon (IFN) in chronic hepatitis C (CHC) patients, we investigated the quantitative serum hepatitis C virus (HCV) RNA level, genotype, and liver biopsy histological features in 60 patients who were treated with 360 x 10(6) U of natural IFN-alpha for 36 to 48 weeks and for more than 12 months after therapy. A branched DNA (bDNA) assay was used to measure HCV RNA levels. All responders, defined as those individuals with normal alanine transaminase (ALT) levels at 48 weeks after therapy, had less than 2 x 10(6) HCV RNA Eq/mL before administration of IFN. Of 39 patients with RNA levels (less than 2 x 10(6) Eq/L) 23 (59.0%) were responders. The genotype was determined for each patient using type-specific polymerase chain reaction (PCR) primers. There was a significant difference in rate of response between subtype 1b and subtypes 2a and 2b (P < .0002); however, all responders had less than 2 x 10(6) Eq/L independent of genotype. In a multivariate analysis, RNA level was the most statistically significant factor affecting response to IFN. Although disease severity, as defined by histological features, was not statistically correlated with nonresponse, patients that responded to IFN tended to have less severe disease.

Malignant hypercalcemia due to gastric endocrine cell carcinoma.

A 62-year-old man was admitted to our Neurology Unit due to consciousness disturbance. Laboratory data showed marked hypercalcemia and azotemia. Serum parathyroid hormone-related protein (PTHrP) level was extremely high. We performed intensive hemodialysis for renal failure, but his condition deteriorated rapidly. On day 10, he died of multiple organ failure. The autopsy revealed gastric undifferentiated adenocarcinoma with systemic dissemination. Immunohistological study showed positive PTHrP staining in carcinoid-like parts of the tumor. This is the first reported case of malignant hypercalcemia due to PTHrP-producing carcinoid or endocrine cell carcinoma of the stomach.

[Radiation therapy of carcinoma of the esophagus in the aged--its results and problems].

Clinical records of 128 non-selected patients with esophageal cancer treated by radiation were reviewed to investigate reasons why treatment had to be discontinued, relationship between age and survival, and factors influencing prognosis. Radiation therapy was completed in 77 patients but was discontinued in 27 patients. Preoperative radiation was attempted in 24 patients. An overall median survival of 128 patients was 6.4 months, with 8.5 and 3.3 percent surviving 3 and 5 years. Median survivals of incomplete RT, completed Rt and RT + Surgery were 1.3, 7.7 and 11.3 months. By Kaplan-Meier analysis significant difference was observed in survival rate between incomplete RT and the other two groups, but not between RT and RT + surgery. T1 tumor cases with incomplete RT were characterized by a higher C-Score (higher incidence of comorbidity and complications), lower albumin concentration and poor performance status. Median survivals of 60 years, 70 and 80 were 12, 5.4 and 6.2 months, respectively. Performance status and C-score were significantly different between the 60 yr and 80 yr groups. Survival rates were also apparently affected by the size of the primary tumor and metastasis. Thus important factors influencing prognosis were performance status, albumin concentration, and comorbidity and complications, in addition to stage of tumor itself. Although performance status and albumin concentration were considered directly related to tumor stage, old age may have an adverse effect on these factors through increase of comorbidity and complication. The data may be useful for decision making for treatment of esophageal cancer of the aged.

Immunohistochemical localization of ganglioside components in hepatocellular carcinoma and liver cirrhosis using monoclonal antibody.

BACKGROUND: In our previous studies (1, 2) a remarkable difference in the patterns of gangliosides was demonstrated between normal human liver and hepatocellular carcinoma (HCC) on thin layer chromatography, consisting of a marked increased in GM2 and the unidentified gangliosides. The proliferation of the unidentified gangliosides was also revealed in certain types of liver cirrhosis. The precise chemical structure of the unidentified components still remains to be elucidated, and there are no comprehensive data on these components investigated in liver tissue from patients with liver cirrhosis, HCC, or malignancies of the biliary and gastrointestinal tracts. In view of this, we studied the immunohistochemical localization of these ganglioside components in these tissues. EXPERIMENTAL DESIGN: Two unidentified gangliosides were extracted and monoclonal antibody were prepared. Localization of these gangliosides on tissue sections of liver obtained from HCC and liver cirrhosis, and malignancies of the biliary and gastrointestinal tracts was investigated by performing immunohistochemical staining using monoclonal antibody. The carbohydrate composition and sialic acid species of these components were determined by gas-liquid chromatography analysis. The sialic acid linkage to the terminal moiety of these components was also investigated by sialidase treatment. RESULTS: Staining with monoclonal antibody against specific gangliosides revealed dense brown granules in HCC tissue sections, whereas normal liver, kidney, and spleen sections were negative. Tissue sections of cirrhotic livers generally stained weakly except one case that showed slight to moderate staining. These antigens were immunohistochemically negligible in sections from one case each of gastric, colon, and common bile duct cancer. The ganglioside components identified in HCC were confirmed to be sialosylparagloboside with an alpha 2-6galactosyl terminus as determined by gas-liquid chromatography analysis and enzymatic hydrolysis with different sialidases. CONCLUSIONS: These components may be a useful marker in the detection of HCC and for subclassification of HCC from the standpoint of ganglioside metabolism.

A non-radioisotopic reverse transcriptase assay using biotin-11-deoxyuridinetriphosphate on primer-immobilized microtiter plates.

We developed a non-radioisotopic (non-RI) reverse transcriptase assay (RTA). The reverse transcriptase (RT) incorporates biotin-11-deoxyuridine-triphosphate (bio-dUTP) using a poly(rA) template hybridized with oligo(dT) primer that is immobilized on the surface of a 96-well microtiter plate. This assay is thus semi-automated by adapting it to an ELISA testing format. The incorporation of bio-dUTP was enhanced by adding cold dTTP to the reaction mixture, optimally in a molar ratio 4:1 (dTTP:bio-dUTP). This non-RI RTA is more sensitive than the conventional RI assay for the detection of purified Rous-associated virus 2 (RAV-2) and of human immunodeficiency virus type 1 (HIV-1) lysate. Because of its simple procedure, higher sensitivity and non-use of RI materials, the assay can be utilized not only for virological studies but also for routine safety screening of biological products for retroviral contamination.

Characteristics and management of patients with fetal neuroblastoma.

At 35 weeks 6 days of gestational age, ultrasound evaluation of the fetal abdomen showed a mixed cystic mass in the superior pole of the left kidney. The mass was suspected to be an adrenal hemorrhage or neuroblastoma. The diagnosis was fetal neuroblastoma. Differential diagnosis enabled the fetal neuroblastoma to be distinguished from adrenal hemorrhage. The parameters of diagnosis of fetal neuroblastoma include no specific ultrasonographic pattern, lack of palpability, and no tumor markers. However, certain features do characterize fetal neuroblastoma, such as little metastases, complete resection at operation, and excellent prognosis. In cases of suspected neuroblastoma, a laparotomy performed as soon as possible is generally regarded as the best course of treatment. Nonetheless, biological analyses of the tumor may prove in the future to be necessary for determining whether or not laparotomy is the best treatment.

Tubular anal duplication--experiences with two cases.

Anal duplication is a very rare abnormality, especially in infants. Two cases of tubular anal duplication of the infant and neonate are reported in this paper. In all cases, removal of the duplicated anus through the perineal approach was accomplished without difficulty. The histology revealed a squamous epithelium with smooth muscle component around the cavity, combined with collumnar or transitional epithelium. There was no evidence of inflammation. The postoperative courses were uneventful with satisfactory anal function. The definite aetiology of this condition is still unknown, although several hypotheses have been proposed.

Abnormal hepatobiliary clearance of 99mTc-N-(2,6-diethylphenylcarbamoylmethyl)iminodiacetic acid in the altered state of thyroid--by an analysis of deconvolution method.

Hepatobiliary clearance of 99mTc-EHIDA was investigated in cases with altered thyroid function by deconvolution method. The results indicated that mean hepatic transit time of all control subjects revealed less than 10 minutes. On the other hand, mean hepatic transit time of cases with altered thyroid function revealed prolonged more than 13 minutes. Cases especially showing an elevated serum concentration of TSH compared with normal range (4.6 microU/ml) had a tendency of a high incidence of markedly prolonged mean hepatic transit time. These results suggest that thyroid hormone may influence on the hepatic metabolism of hepatobiliary radiopharmaceuticals. This phenomenon also could partly explain the cause of liver dysfunction seen in subjects with altered states.

Ganglioside variations in human liver cirrhosis and hepatocellular carcinoma as shown by two-dimensional thin-layer chromatography.

Gangliosides isolated from 5 cases of normal liver tissues, 11 cases of liver cirrhosis and 5 cases of hepatocellular carcinoma were compared in their concentrations and compositions. Quantitative analysis revealed no significant change of ganglioside levels between normal and cirrhotic liver tissues or hepatocellular carcinoma. There was also no significant difference (p greater than 0.05) between cirrhotic liver tissues and hepatocellular carcinoma. Two dimensional thin-layer chromatography of the total ganglioside preparations of liver tissues from both liver cirrhosis and hepatocellular carcinoma showed proliferation of GM2, GD3, GD1 and at least two unidentified components, named provisionally spots Nos. 1 and 2 in the present report, and loss of GM3. Sialidase treatment and thin-layer chromatography showed the components of these spots to be sialidase-labile monosialogangliosides and distinctly different from GD3 which was described elsewhere.

Localization in situ of c-myc mRNA and c-myc protein in adult mouse testis.

It has been suggested that c-myc, one of the proto-oncogenes, plays a role in normal somatic cell proliferation and differentiation. To define whether c-myc is only expressed during somatic cell division or is also expressed during meiotic cell division, the production of c-myc mRNA and protein were investigated in the mouse testis by using in situ hybridization with non-radioactive DNA probes and enzyme immunohistochemistry respectively. For in situ hybridization, T-T dimerized DNA probes were used and DNAs hybridized in situ were detected immunohistochemically using specific antibody against T-T dimer. The results indicate that c-myc mRNA and protein are expressed in a cell-cycle-dependent manner only in spermatogonia and not in spermatocytes and spermatids.