Cytokine/chemokine profiles contribute to understanding the pathogenesis and diagnosis of primary Sjögren's syndrome.

To investigate the pathogenesis of localized autoimmune damage in Sjögren's syndrome (SS) by examining the expression patterns of cytokines, chemokines and chemokine receptors at sites of autoimmune damage. mRNA expression of these molecules in the labial salivary glands (LSGs) and peripheral blood mononuclear cells (PBMCs) from 36 SS patients was examined using a real-time polymerase chain reaction-based method. Subsets of the infiltrating lymphocytes and chemokines/chemokine receptors expression in the LSG specimens were examined by immunohistochemistry. Cytokines/chemokine concentrations in the saliva were analysed using flow cytometry or enzyme-linked immunosorbent assay. mRNA expression of T helper type 1 (Th1) cytokines, chemokines and chemokine receptors was higher in LSGs than in PBMCs. In contrast, mRNA expression of Th2 cytokines, chemokines [thymus and activation-regulated chemokine (TARC/CCL17), macrophage-derived chemokine (MDC/CCL22)] and chemokine receptor (CCR4) was associated closely with strong lymphocytic accumulation in LSGs. Furthermore, TARC and MDC were detected immunohistochemically in/around the ductal epithelial cells in LSGs, whereas CCR4 was detected on infiltrating lymphocytes. The concentrations of these cytokines/chemokines were significantly higher in the saliva from SS patients than those from controls, and the concentrations of Th2 cytokines/chemokines were associated closely with strong lymphocytic accumulation in LSGs. These results suggest that SS might be initiated and/or maintained by Th1 and Th17 cells and progress in association with Th2 cells via the interaction between particular chemokines/chemokine receptors. Furthermore, the measurement of cytokines/chemokines in saliva is suggested to be useful for diagnosis and also to reveal disease status.

Selective localization of T helper subsets in labial salivary glands from primary Sjögren's syndrome patients.

The aim of this study was to investigate the initiation and progression of autoimmune damage in the lesions of labial salivary glands (LSGs) from primary Sjögren's syndrome (SS) patients by examining the selective localization of T helper (Th) subsets such as Th1, Th2, Th17 regulatory T cells (T(regs)) and follicular T helper cells (Tfh). The expression of cytokines and transcription factors associated with these Th subsets in the LSGs from 54 SS patients and 16 healthy controls was examined using real-time polymerase chain reaction (PCR) and immunostaining. Additionally, infiltrating lymphocytes without germinal centre (GC(-)) and with GC (GC(+)) in the LSGs specimens from eight SS patients were extracted selectively by laser capture microdissection (LCM). The mRNA expression of these molecules was compared between the two sample groups of GC(-) and GC(+) by real-time PCR. The mRNA expression of cytokines and transcription factors of all T helper (Th) subsets in the LSGs from the SS patients was increased significantly in comparison with controls. In LSGs from the SS patients, Th2 and Tfh was associated closely with strong lymphocytic infiltration; however, Th1, Th17 and T(regs) was not. In the selectively extracted lesions of LSGs, Th1 and Th17-related molecules were detected strongly in the GC(-), while Th2 and Tfh-related molecules were detected in the GC(+). In contrast, no significant association with strong lymphocytic infiltration was observed in T(reg)-related molecules. These results indicate that SS has selective localization of Th subsets such as Th1, Th2, Th17 and Tfh in the LSGs, which is associated closely with disease severity and/or status. SS might be initiated by Th1 and Th17 cells, and then progressed by Th2 and Tfh cells via GC formation.

Close association between oral Candida species and oral mucosal disorders in patients with xerostomia.

OBJECTIVE: Heightened interest in oral health has lead to an increase in patients complaining of xerostomia, which is associated with various oral mucosal disorders. In this study, we investigated the relationship between Candida species and oral mucosal disorders in patients with xerostomia. SUBJECTS AND METHODS: We evaluated whole salivary flow rate and presence of oral mucosal disorders in 48 patients with xerostomia and 15 healthy controls. The number of Candida species was measured as colony-forming units after propagation on selective medium. Identification of Candida at the species level was carried out by polymerase chain reaction and restriction fragment length polymorphism analysis. We then examined the relationship between Candida species and oral mucosal symptoms. RESULTS: Compared with controls, patients with xerostomia exhibited significantly decreased whole salivary flow rate, increased rate of oral mucosal symptoms, and higher numbers of Candida. Salivary flow rate negatively correlated with the number Candida. Among patients with oral candidiasis, Candida albicans was isolated from the tongue mucosa and Candida glabrata was isolated from the angle of the mouth. CONCLUSION: These results suggest that particular Candida species are involved in the pathogenesis of oral mucosal disorders in patients with xerostomia.

[Intraoperative right-to-left shunting induced by supine position in simultaneous lung resection with repair of atrial septal defect].

This report describes the case of a 49-year-old woman, who was suffered severe hypoxemia attributable to right-to-left shunting through an atrial septal defect (ASD) during the combined surgery for lung cancer and ASD in supine position. Right-to-left shunting has been reported to occur after lung resection but not during it. According to our continuous measurement of pulmonary arterial pressure and oxygen saturation, changes in hemodynamics during lobectomy in supine position was supposed to differ from that in lateral position, which may contribute to right-to-left shunting. A combined lung resection with heart surgery was performed safely and resulted in preventing postoperative complications induced by ASD.

Pancreaticopleural fistula visualized by computed tomographic scan combined with endoscopic retrograde pancreatography.

We report a case with left pleural effusion caused by a pancreaticopleural fistula, which was formed between a small pseudocyst in the pancreatic body and the posterior mediastinal side of left costophrenic recess. The patient, a 66-year-old male with an alcohol drinking habit, had cough and chest pain but no symptoms or signs suggestive of abdominal pathology. In this case, computed tomographic scan performed immediately after endoscopic retrograde pancreatography was very useful not only in demonstrating the presence of the pancreatic internal fistula but in clarifying its anatomical relation to the surrounding organs.

Valvuloplasty for aortic valve regurgitation resulting from cusp prolapse.

Three adults, 2 with tricuspid aortic valve and 1 with bicuspid valve, underwent valvuloplasty for aortic valve regurgitation resulting from cusp prolapse. Surgical procedures consisted of combined cusp plication by triangular cusp resection and subcommissural annuloplasty. Doppler echocardiography revealed trivial aortic valve regurgitation intraoperatively and less than I/IV at discharge in all cases. After mean follow-up of 15 months, 2 tricuspid aortic valve patients remain I/IV regurgitation and II/IV in the bicuspid patient. Although long-term results remain unclear, our results show that this procedure is feasible and beneficial in patients with aortic valve regurgitation due to cusp prolapse.

[Association of cytochrome P-450 1A1 (CYP1A1) gene polymorphism to smoking status and hematologic findings].

Measures to control smoking are important in the field of preventive medicine. In order to clarify differences in susceptibility in individuals to lung cancer of genotypes of CYP1A1, which are considered to be related to lung carcinogenesis were evaluated in 391 healthy males to study relationship to smoking status and hematological findings. No correlation was observed between genotypes of CYP1A1 and smoking status. White blood cell counts in smokers with a Val allele were significantly higher than in those without a Val allele. Multiple regression analysis showed that the genotype of CYP1A1 and daily cigarette consumption had significant relationship with white blood cell count in smokers. However, red blood cell count, hemoglobin, hematocrit, MCV, MCH, or MCHC were not significantly associated with genotype of CYP1A1. There have been no previous reports on the relationship between the genotype of CYP1A1 and hematological findings. In consideration of the epidemiologic findings that many individuals with increased white blood cell counts have cancer or cardiac diseases, and reports that the increase in the white blood count was associated with poor respiratory function, white blood cell count may be candidate to for being a risk marker and thus contribute to prevention of these diseases.

[Acquired pulmonary stenosis caused by mediastinal mature teratoma: a case report].

A 13-year-old girl was admitted with a sudden onset of chest oppression. Mediastinal teratoma was suspected on chest X-ray and CT scan. A grade 2/6 systolic murmur was heard at the upper left sternal area and cardiac catheterization showed mild pulmonary stenosis. After resection of the tumor, the murmur disappeared and histopathological diagnosis was mature teratoma. This is a rare case report of pulmonary stenosis caused by mediastinal mature teratoma.

[A case of lung metastasis from Meibomian gland carcinoma of eyelid with effective chemotherapy].

A 47-year-old man was admitted to Shinshu University Hospital in 1992 because of cough and chest abnormal shadow. In 1987, he had an operation for Meibomian gland carcinoma of his right lower eyelid, and also received radiotherapy. His chest X-ray film on admission showed tumor shadows in the bilateral upper lung fields. The chest CT scan showed tumors in the right S1 and the left S3. Bronchofiberscopic findings demonstrated stenosis of the right upper bronchus and an endobronchial mass occluding the left B3. A biopsy specimen of the endobronchial mass revealed sebaceous carcinoma, which was identical with that of the resected eyelid tumor. He received two courses of chemotherapy in a combination of CDDP 75 mg/m2 and ADM 50 mg/m2. The tumors on chest X-ray and CT scan became small by the chemotherapy. Bronchofiberscopy after the chemotherapy also revealed that the stenosis of the right upper bronchus improved and the endobronchial mass in the left B3 had disappeared. He died 8 months after initial chemotherapy with a response duration of 7 months.

Site-specific effect of estradiol on gene expression in the adipose tissue of ob/ob mice.

Sex hormones are likely to be involved in sex differences in adipose tissue distribution. To test whether estrogen regulates genes expressed in the adipose tissue in a site-specific manner, we studied the effect of exogenous estradiol on the gene expression in visceral and subcutaneous adipose tissues of male ob/ob mice. We screened genes expressed in a site- and sex-specific manner, and genes that were affected by exogenous estradiol by DNA chip analysis. They were verified by real-time PCR. Myosin heavy chain 2B (Myh4) and phosphoglycerate mutase muscle-specific subunit (Pgam) were expressed specifically in the subcutaneous adipose tissue, and uroplakin IIIb (UP3) was expressed specifically in the visceral adipose tissue. DEAD-box Y RNA helicase (DBY) and eukaryotic initiation factor 2 gamma Y (eIF2gamma Y) were expressed only in male adipose tissue. X-chromosome inactive specific transcript (Xist) was expressed only in female adipose tissue. When estradiol was subcutaneously administrated to male mice, the expression of monocyte-chemoattractant protein-1 (MCP-1) and androgen receptor (AR) genes was regulated in a site-specific manner. The difference in the amount of estrogen receptor did not account for the site-specific effect of estrogen. Our findings show that estrogen affects the expression of some adipocyte genes in a site-specific manner.

Pulmonary hemodynamics and lung function during chronic paraquat poisoning in sheep. Possible role of reactive oxygen species.

The purpose of this study was to establish a chronic model of paraquat-induced lung injury. To examine the role of reactive oxygen species in this form of lung injury, we measured malondialdehyde (MDA) and superoxide dismutase (SOD) activity in the lungs. Paraquat (5 mg/kg intramuscularly) caused a significant decrease in dynamic lung compliance from 128.5 +/- 9.2 to 63.3 +/- 11.8 ml/cm H2O (p less than 0.05), with a significant increase in AaPO2 3 wk after paraquat. Histologic findings in the lungs showed a gradual increase in the number of granulocytes and alveolar wall thickening with proliferation of reticular fibers and were coincident with the changes in physiology. A transient decrease in pressor responses to hypoxia was observed 1 wk after paraquat, although pulmonary hemodynamics did not change. The amount of lung MDA 3 wk after paraquat increased from the baseline value of 0.73 +/- 0.04 to 1.12 +/- 0.10 nmol/mg protein (p less than 0.05). SOD activity in the lung tissue significantly decreased from 6.47 +/- 0.20 to 4.82 +/- 0.25 U/mg protein (p less than 0.05) 1 wk after paraquat and remained at low levels for 3 wk. These findings suggest that a small dose of paraquat causes chronic lung injury characterized by granulocyte infiltration and lung fibrosis. Reactive oxygen species may play an important role in this chronic lung injury, and the inability to increase antioxidant defense may contribute to the reaction.

Columnar cell carcinoma of the thyroid.

A case of columnar cell carcinoma of the thyroid occurring in a 50-year-old female is described. Histologically, the 2 cm tumor showed a prominent papillary architecture with thin fibrous cores covered by columnar cells and marked nuclear stratification. It also showed microfollicular, glandular, and solid patterns. The nuclear features were different to those of conventional papillary carcinoma and similar to those of follicular tumors. The tumor was principally encapsulated with vascular and minimal capsular invasion. The tumor cells were positive for thyroglobulin. The tumor was DNA diploid with a low S phase traction as determined by flow cytometry. The patient had no lymph node or distant metastasis. The patient was well and without disease 9 months after surgery. The possibility that the neoplasm is one of poorly differentiated thyroid carcinomas rises.

Bax is downregulated in inflamed colonic mucosa of ulcerative colitis.

BACKGROUND AND AIMS: One form of epithelial cell injury in inflamed colonic mucosa in ulcerative colitis (UC) is reported to involve apoptosis of these cells. Bcl-2 family proteins Bax and Bcl-2 are the major regulators of apoptosis. The aim of this study was to elucidate the involvement of the Bax/Bcl-2 system in induction of apoptosis of the inflamed colonic epithelium in UC. METHODS: Colonic epithelium was isolated from colonic biopsy specimens. Expression of CD95, Bax, Bcl-xL, and Bcl-2 proteins was determined by western blotting. Bax gene expression was assessed by both reverse transcription-polymerase chain reaction (RT-PCR) followed by Southern hybridisation and a real time PCR assay. RESULTS: Equal levels of expression of CD95, Bcl-xL, and Bcl-2 proteins were noted in normal and UC colonic epithelia. Equal levels of expression of Bax protein and mRNA were noted in epithelia of normal colon and inactive UC. Levels of expression of Bax protein and mRNA were markedly reduced in inflamed UC colonic epithelium. CONCLUSIONS: Our study showed for the first time downregulation of Bax in inflamed colonic epithelium of UC. The Bax/Bcl-2 system did not seem to be involved in induction of apoptosis of epithelial cells in the inflamed colonic mucosa of UC.

Granulocyte colony-stimulating factor does not exacerbate endotoxin-induced lung injury in sheep.

Because it has been suggested that neutrophils play a role in endotoxin-induced lung injury, we examined the effects of increased peripheral blood-neutrophils induced by recombinant human granulocyte-colony-stimulating factor (rhG-CSF) on the lung dysfunction when induced by endotoxin in awake sheep. We prepared chronically instrumented awake sheep with lung lymph fistulae and catheters for hemodynamic monitoring. We compared alterations of pulmonary hemodynamics, lung lymph balance, circulating leukocytes, plasma and lung lymph concentrations of thromboxane B2 and 6-keto-prostaglandin F1 alpha, and arterial blood gases in sheep that received Escherichia coli endotoxin infusion intravenously (1 microgram/kg) in the following two groups: Group 1 (n = 6) received endotoxin alone, and Group 2 (n = 7) received rhG-CSF (125 micrograms/day) intravenously for 3 days before endotoxin was administered in the same manner as in Group 1. The rhG-CSF remarkably increased circulating neutrophils to 24,600 +/- 2,500/microliters in Group 2 sheep at baseline when compared with the level in Group 1 (5,700 +/- 550 microliters). After endotoxin infusion, we observed significant decreases in circulating neutrophils in both groups. There were no differences between the two groups in the time course of changes in pulmonary hemodynamics, arterial blood gases, or arachidonate products after endotoxemia. However, lung lymph flow and lung lymph protein clearance during the late phase were lower in rhG-CSF-treated sheep than in those that received only endotoxin.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of continuous infusion of nitroglycerin on pulmonary hemodynamics, lung lymph balance, and prostanoid products in the response to endotoxin in awake sheep.

We examined the effects of continuous intravenous infusion of nitroglycerin (NTG) on lung dysfunction induced by endotoxemia in awake sheep chronically instrumented with lung lymph fistula. We measured the responses of hemodynamics, lung lymph balance, and thromboxane (Tx)B2 and 6-keto-prostaglandin (PG) F1 alpha levels in plasma and lung lymph to endotoxin administration (1 microgram/kg, intravenously [IV], over 30 min) with and without continuous infusion of NTG (1 microgram/kg/min). Continuous infusion of NTG alone (n = 5) over 5 hr did not significantly alter systemic, pulmonary hemodynamics, and/or lung lymph fluid filtration. Infusion of endotoxin alone (n = 7) caused remarkable increases in pulmonary artery pressure (Ppa) and lung lymph flow (Qlym) in the early phase. Continuous infusion of NTG (n = 6) significantly prevented the early increases in Ppa and Qlym after endotoxin. The increased values of TxB2 and 6-keto-PGF1 alpha in both plasma and lung lymph after endotoxemia showed the same increases in groups with and without NTG. These findings suggest that the reduction of pulmonary artery pressure induced by NTG decreased the filtration of fluid into the lungs associated with endotoxemia in sheep, and that the mechanism of vasodilating action of NTG is not due to modifications of constrictive-dilated cyclo-oxygenase products of arachidonate, such as TxA2 and PGI2.

Influence of pepsinogen gene polymorphisms on serum pepsinogen.

We identified pepsinogen C (PGC) gene polymorphisms by means of PCR, which amplified DNA in the region within the intron between exons 7 and 8, and by 6% polyacrylamide gel electrophoresis. Six alleles were found in a Japanese population. The frequencies of these alleles in 408 unrelated Japanese individuals were 0.074, 0.026, 0.335, 0.237, 0.016 and 0.314, respectively. The serum pepsinogen II level significantly decreased in the order of the allele 6 homozygote, the allele 6 heterozygote and the other genotypes (chi 2 = 7.850, D.F. = 2, p = 0.020). These findings indicated that the genetic background of serum pepsinogen should be considered when screening for stomach cancer by this procedure.

[Arachidonic acid metabolites on peripheral blood plasma in patients with bronchial asthma].

To assess the role of arachidonic acid metabolites in pathogenesis of bronchial asthma, we measured thromboxane B2 (TxB2), 6-keto PGF1 alpha, leukotriene (LT) C4, and LTB4 in venous blood plasma in patients with bronchial asthma and in controls. The level of TxB2 was significantly higher in 18 asthmatics with attacks than that in 11 controls (77.3 +/- 48.8 pg/ml vs 48.8 +/- 9.4 pgml). The levels of 6-keto PGF1 alpha in both asthmatics with attacks (17.8 +/- 6.7 pg/ml) and without an attack (16.4 +/- 9.7) were significantly higher than that in controls (11.6 +/- 3.9 pg/ml). The levels of LTC4 in asthmatics with attacks (0.84 +/- 0.11 ng/ml, n = 11) were significantly higher than that in controls, furthermore the level of LTC4 in asthmatics without an attack. The level of LTB4 was significantly higher in asthmatics with attacks (295.0 +/- 160.7 pg/ml, n = 26) than that in controls (161.7 +/- 25.3 pg/ml, n = 12) and asthmatics without an attack (182.4 +/- 97.9 pg/ml, n = 22). These findings suggest that the arachidonic acid metabolites are related to the asthma attack and are associated with the pathogenesis of bronchial asthma.