Force-induced IL-8 from periodontal ligament cells requires IL-1beta.

During orthodontic tooth movement, mechanical stresses induce inflammatory reactions in the periodontal ligament (PDL). We hypothesized that chemokines released from PDL cells under mechanical stress regulate osteoclastogenesis, and investigated the profiles and mechanisms of chemokine expression by human PDL cells in response to mechanical stress. In vitro, shear stress and pressure force rapidly increased the gene and protein expressions of IL-8/CXCL8 by PDL cells. Consistently, amounts of IL-8 in the gingival crevicular fluid of healthy individuals increased within 2 to 4 days of orthodontic force application. The PDL cells constitutively expressed low levels of IL-1beta, which were not further increased by mechanical stress. Interestingly, neutralization of IL-1beta abolished IL-8 induction by mechanical stresses, indicating that IL-1beta is essential for IL-8 induction, presumably though autocrine or paracrine mechanisms. Finally, experiments with signal-specific inhibitors indicated that MAP kinase activation is essential for IL-8 induction.

A radiographic study of the mandibular third molar root development in different ethnic groups.

BACKGROUND: The nature of differences in the timing of tooth formation between ethnic groups is important when estimating age. AIM: To calculate age of transition of the mandibular third (M3) molar tooth stages from archived dental radiographs from sub-Saharan Africa, Malaysia, Japan and two groups from London UK (Whites and Bangladeshi). MATERIALS AND METHODS: The number of radiographs was 4555 (2028 males, 2527 females) with an age range 10-25 years. The left M3 was staged into Moorrees stages. A probit model was fitted to calculate mean ages for transitions between stages for males and females and each ethnic group separately. The estimated age distributions given each M3 stage was calculated. To assess differences in timing of M3 between ethnic groups, three models were proposed: a separate model for each ethnic group, a joint model and a third model combining some aspects across groups. The best model fit was tested using Bayesian and Akaikes information criteria (BIC and AIC) and log likelihood ratio test. RESULTS: Differences in mean ages of M3 root stages were found between ethnic groups, however all groups showed large standard deviation values. The AIC and log likelihood ratio test indicated that a separate model for each ethnic group was best. Small differences were also noted between timing of M3 between males and females, with the exception of the Malaysian group. These findings suggests that features of a reference data set (wide age range and uniform age distribution) and a Bayesian statistical approach are more important than population specific convenience samples to estimate age of an individual using M3. CONCLUSION: Some group differences were evident in M3 timing, however, this has some impact on the confidence interval of estimated age in females and little impact in males because of the large variation in age.

Cloning and characterization of the gene encoding a mitochondrially localized DNA topoisomerase II in Dictyostelium discoideum. Western blot analysis.

We cloned a gene (topA) encoding DNA topoisomerase II from Dictyostelium discoideum nuclear DNA using oligo probes corresponding to the consensus amino acid sequences found in the gene in other eukaryotes. The gene encoding a predicted polypeptide of 1282 amino acids with M(r) of about 146 kDa, is a single copy that is expressed as a polyadenylated 4.5 kb RNA. The predicted amino acid sequence shares similarity with those of other eukaryotes with identity between 32 and 46%. The protein is 260-300 amino acids shorter in the C-terminal region and 50-80 longer in the N-terminal region than those of other eukaryotes. In TopA of D. discoideum, the N-terminal region with stretches of charged and hydrophilic amino acids is predicted to fold into an amphiphilic alpha-helix which is characteristic of a mitochondrial targeting signal presequence. Four independent polyclonal antibodies against bacterially expressed GST fusion proteins containing four portions of the polypeptide detected a single band on Western blots at about 135 kDa. Western blots analysis of subcellular fractions revealed that this protein is localized in mitochondria. The protein and the mRNA are present in growth phase and during development, although levels of both declined as development proceeded.

Improvement of coronary artery endothelial dysfunction with lipid-lowering therapy: heterogeneity of segmental response and correlation with plasma-oxidized low density lipoprotein.

OBJECTIVES: This study assessed coronary artery endothelial function in patients with hypercholesterolemia before and after lipid lowering, using quantitative angiography to examine the acetylcholine (Ach) response along the entire analyzable vessel. BACKGROUND: Lipid lowering reverses endothelial dysfunction, but whether improvement occurs only in some segments and not others has not been established. Statistical correlation of improvement with specific lipid moieties remains undefined. METHODS: Quantitative angiography was performed after Ach (10(-6), 10(-5), 10(-4) M) in 29 patients with coronary atherosclerosis before and 18 +/- 5.2 months after lipid-lowering treatment (statins, bile sequestrant resins). Standard lipid moieties and markers of oxidized low density lipoprotein (LDL) (immunoglobulin G and M autoantibody titers to malondialdehyde-LDL, E06 epitope) were measured serially. RESULTS: Pre-treatment of the vessel diameters at control and with 10(-6)M, 10(-5) M and 10(-4) M Ach were 2.108 +/- 0.085, 2.086 +/- 0.087, 2.069 +/- 0.084 and 1.963 +/- 0.097 mm (M +/- SE), respectively, and increased at follow-up to 2.139 +/- 0.094, 2.119 +/- 0.086, 2.127 +/- 0.084 and 2.080 +/- 0.085 mm (p < 0.0001). Improvement in the most constricted and modest declination in the more dilated segments were observed. Change in the E06 and Apolipoprotein A-1 titers correlated with improved vasomotion (p = 0.027 and 0.005, respectively). The pre- and post-treatment levels of the E06 epitope, as well as the post-treatment IgM autoantibody titer to MDA-low density lipoprotein, also correlated (p < 0.028, < 0.001 and p < 0.004, respectively). CONCLUSIONS: Drug treatment reverses endothelial dysfunction, but the effect is heterogeneous. Most coronary segments show enhancement, while others show declination of dilation, underscoring the importance of assessing the entire analyzable artery. Improvement in vasomotion correlates most significantly with markers of plasma-oxidized low-density lipoprotein.

A neuropathological study of a case of lupus erythematosus with chorea.

Neuropathological examination in a 48-year-women of SLE with chorea was performed. Histological examination of the putamen showed a widespread neuronal loss associated with reactive astrocyte proliferation and neuropil rarefaction at both sides. Disappearance of large neurons was more prominent than that of small ones. Although a few old and fresh microinfarcts were scattered in the same area, there was no significant pathological abnormality in small vessels. The caudate nuclei also showed a few irregular microinfarcts and spotty loss of neurons associated with reactive astrocytosis. These neuropathological changes might be related to the appearance of choreatic movement in patients with SLE.

A case of protein-losing enteropathy in idiopathic thrombocytopenic purpura with decreased IgA.

A young woman presented with high fever and edema in January, 1984, and was diagnosed as having systemic lupus erythematosus. Prednisolone administration failed to improve her symptoms. In May she was admitted to hospital because of elevated erythrocyte sedimentation rate (ESR), hypoproteinemia, hypogammaglobulinemia, hypocomplementemia, positive antinuclear antibody, elevated immune complex level, and diarrhea. Edema disappeared following administration of diuretics and albumin, although the pathogenesis was still undetermined. In September, she was referred to our institution because of severe watery diarrhea and hypoproteinemia. Endoscopic examination showed a diffuse inflammatory lesion in the duodenum and the colon. Radioisotopic 51Cr-albumin study results were compatible with protein-losing enteropathy. Hypoproteinemia and inflammatory changes of the intestine were improved by antibiotics, suggesting that the inflammatory lesion was caused by bacterial infection. Despite the improvements in clinical symptoms and laboratory findings, the serum IgA level was still low and the thrombocytopenia remained. The morphological characteristics of the megakaryocytes were consistent with idiopathic thrombocytopenic purpura. In May, 1986, the thrombocytopenia deteriorated, causing purpura. Prednisolone was administered again, and this resulted in normalization of the platelet count, although the IgA level remained low. Finally the prednisolone was stopped, and the IgA level gradually recovered, with the improvement of the enterocolitis. The exact pathogenesis of the whole picture in this case is unclear, but an 8-year-long clinical course suggests that the protein-losing was caused by an infectious enterocolitis superimposed on IgA deficiency.

Rabbit granulomatous enterocolitis induced by injection of muramyl dipeptide emulsified with Freund's incomplete adjuvant.

We induced granulomatous enterocolitis in rabbits by injecting them with muramyl dipeptide (MDP), a subunit of the peptidoglycan polymers that endow the bacterial cell wall with structural rigidity, emulsified with Freund's incomplete adjuvant (FIA). Injections of 0.1 ml of a water-in-oil emulsion of MDP and FIA were given submucosally at six sites in the rectum and colon, 10 cm proximal to the anus, using a flexible endoscope. Four rabbits each were sacrificed 1, 2, and 4 weeks after a single injection of the emulsion. Another 4 rabbits each were injected six times at 1- and 2-week intervals, and were sacrificed 1 and 2 weeks after the last injection of the emulsion, respectively. In all 20 rabbits, injected with the MDP emulsion, histological findings of the colon consisted of cellular infiltrations of plasma cells and lymphocytes, granulomatous lesions, and granulomas, although the findings differed in degree. Cellular infiltration in hyperplastic villi and denuded epithelia of the small intestine were seen in 2 of 8 rabbits repeated that received MDP emulsion injections. The histological changes in this animal model may be useful for studying the pathogenesis of inflammatory bowel disease in humans.

Extraintestinal manifestations of granulomatous enterocolitis induced in rabbits by long-term submucosal administration of muramyl dipeptide emulsified with Freund's incomplete adjuvant.

We examined whether extraintestinal manifestations of granulomatous enterocolitis in rabbits might be produced by the long-term administration of muramyl dipeptide which represents the basic fragment of the bacterial cell wall, emulsified with Freund's incomplete adjuvant. Muramyl dipeptide emulsion was injected submucosally at six sites in the rectum and colon, 10 cm proximal to the anus, each time with a flexible endoscope. Seven rabbits were injected nine times or more every month, and all were sacrificed 1 month after the last injection. The histological changes in the colon in the seven rabbits were mononuclear cell infiltration, epithelioid granulomas, granulomatous lesion, and denuded and regenerative epithelia, although the changes differed in degree. In five of the seven rabbits, histological examination of the liver showed pericholangitis and periductal fibrosis, findings analogous to sclerosing cholangitis in patients with inflammatory bowel disease. In four of the seven rabbits, fibrosis bridging mainly between portal and portal veins, and, in places, between portal and central veins, was seen. Two of the seven rabbits developed polyarthritis. The histological changes in our model suggest that continuous stimulation with bacterial cell wall fragments may be involved in the extraintestinal manifestations of chronic intestinal inflammation such as that seen in inflammatory bowel disease.

Pericholangitis in a rabbit colitis model induced by injection of muramyl dipeptide emulsified with a long-chain fatty acid.

Rabbit colitis has been induced by injection of muramyl dipeptide emulsified with a long-chain fatty acid. The muramyl dipeptide emulsion was injected submucosally at six portions of the rectum and colon, 10 cm proximal to the anus, using a flexible endoscope. Six rabbits were injected six times every 2 weeks and subsequently killed 2 weeks after the last injection. The histological changes of the colon that occurred in all 6 rabbits were mononuclear cell and histiocyte infiltration with sporadic eosinophils, transmural infiltration, and well-maintained goblet cell populations. These changes were different in degree. In 4 of 6 rabbits histological examination of the liver showed pericholangitis and periductal fibrosis mimicking the pericholangitis frequently seen in patients with inflammatory bowel disease. Fibrosis bridging between the portal and portal veins occurred in 2 rabbits, and noncaseating granuloma was seen in 1 rabbit. These histological changes in our model have led to the suggestion that continuous stimulation with bacterial cell wall fragments may be involved in chronic intestinal inflammation and extraintestinal manifestations such as pericholangitis.

[A randomized controlled study on imipenem/cilastatin sodium in comparison to aztreonam + lincomycin in treating severe infections in patients with malignant tumors or hematological diseases].

A randomized controlled study was conducted to compare effects of imipenem (IPM) (1.0-1.5 g/day) with those of aztreonam (AZT) (4 g/day) +lincomycin (LCM) (1,200-2,400 mg/day) in patients with malignant tumors or hematological diseases and severe infections. A total of 95 patients entered the study between October 1989 and March 1991. Forty-seven patients were treated with IPM and the remaining 48 patients were given AZT+LCM. No statistically significant differences existed in age, sex or underlying diseases between the 2 groups. Overall, the clinical cure rate of the IPM group was 53%; This was significantly higher than the 31% cure rate obtained in the AZT+LCM group (P less than 0.05). The difference was significant in patients whose granulocyte counts were less than 1,000/microliters, but not in those whose granulocyte counts were 1,000/microliters or higher. Side effects were observed in 5 patients given IPM and one given AZT+LCM. In conclusion, no significant differences appeared to exist regarding clinical efficacy and safety between the 2 treatment regimens.

[Specific quantification of gamma-seminoprotein-alpha 1 antichymotrypsin complex in serum by monoclonal antibody-based enzyme immunoassay].

Gamma-seminoprotein (gamma-Sm) is one of the serine proteinases in seminal plasma, and is well known as a tumor marker of prostate cancer. In the blood, a major portion of gamma-Sm combines with alpha 1 antichymotrypsin (ACT), a serine proteinase inhibitor. We developed a sensitive enzyme immunoassay (EIA) for the gamma-Sm-ACT complex using two different monoclonal antibodies (MoAbs). One MoAb, prepared against gamma-Sm, is used for the capture, and the other, prepared against ACT, is conjugated with horseradish peroxidase and used for detection. The detectable range of this assay in clinical applications was from 0.2 to 50 units/ml. The intra-assay coefficients of variation (CV%) obtained from ten repeated assays of three sera were 3.6 to 5.5%. The mean gamma-Sm-ACT complex concentration in the sera of normal individuals was determined to be 0.86 +/- 0.11 units/ml for males (n = 50) and 0.11 +/- 0.08 units/ml for females (n = 54). There was no significant increase in the level of the complex with increasing age in males. The ratio of the gamma-Sm-ACT complex to free gamma-Sm tended to be significantly higher in patients with prostate cancer than in those with benign prostatic hyperplasia.

[Mitral valve replacement and closure of ventricular septal defect in the first two months of life].

A 2-month-old female infant weighing 4.2 kg was admitted with severe congestive heart failure and respiratory distress. The patient was operated upon under a diagnosis of severe congenital mitral regurgitation and ventricular septal defect. There was thickening of the anterior leaflet and short thickened chordae. The valve was judged not amenable to repair and it was replaced with a 17 mm St. Jude Medical valve. She was extubated on the 5th postoperative day. Post operative anti-coagulant therapy was initiated with warfarin potassium, dipyridamole and ticlopidine hydrochloride. The postoperative course was uneventful and the patient is doing well 9 months after the operation.

[Long-term results after the total cavopulmonary connection].

Between July, 1988 and November, 2002, 108 patients underwent total cavopulmonary connection (TCPC) at Kobe Children's Hospital. The primary malformation was univentricular heart in 40 tricuspid atresia in 21, mitral atresia in 16, and other complex cardiac defects in the remaining 31. Fenestrated TCPC, staged TCPC, and off-pump TCPC were performed in 39, 26, and 15 high risk patients, respectively. Nitric oxide inhalation was administered in 46 patients. The mean follow-up period was 4.3 years (range, 1 month to 14 years). There were 10 early deaths due to low cardiac output syndrome in 4, thrombosis in 3, tracheal bleeding in 2, and disseminated intravascular coagulation in 1. There were 5 late deaths due to congestive heart failure in 2 patients, arrhythmia in 1, cerebral infarction in 1, and subarachnoid hemorrhage in 1. Late complications included arrhythmia in 17 patients, systemic desaturation caused by abnormal systemic venous channels in 10, pleural or pericardial effusion in 3, chylothorax in 1, and aortic valve incompetence in 1.

[Concomitant mitral valvuloplasty and fenestrated total cavopulmonary connection: a case report].

A seven-year-old boy with tricuspid atresia successfully underwent a fenestrated total cavopulmonary connection and mitral valvuloplasty. Preoperative cardiac catheterization showed a mean pulmonary artery pressure of 16 mmHg. Pulmonary arteriography showed poor development of the branches (PA index: 180). Echocardiography revealed mild to moderate mitral valve incompetence due to prolapse of anterior leaflet. Mitral valve was exposed through the trans-septal approach. The excess chorda length was tucked into a longitudinal split in the top of the posterior papillary muscle. Then wedge resection of the redundant segment of the anterior leaflet and bilateral annuloplasty were performed. Finally, a total extracardiac cavopulmonary anastomosis with a 6 mm fenestration was completed. Postoperative clinical course was uneventful, and he is doing well with no recurrence of mitral incompetence 1 year after the operation.

[Midterm results of bilateral enlargement of the aortic valve ring for valve replacement].

From 1989, 4 patients underwent bilateral enlargement of the aortic valve ring for valve replacement. Age at the operation ranged from 2 to 8 (mean 6) years; body weight ranged from 14.9 to 25.4 (mean 19.0) kg. This procedure enabled us to implant a prosthesis 3 to 4 sizes larger (19 to 23 mm) than that measured with the native aortic annulus (13 to 17 mm). There was no late death and no cardiac event over a mean follow-up period of 6.2 years. Pressure gradient across the prosthesis measured by echocardiography was 40 mmHg in 1 patient who underwent aortic valve replacement with the use of 19 mm St. Jude Medical valve at 2 years of age. There was no significant pressure gradient in other 3 patients. All patients showed normal left ventricular function. We conclude that bilateral enlargement of the aortic valve ring for valve replacement has provided good midterm results with no mortality and no cardiac event.

[Reconstruction of the pulmonary outflow tract without external conduit].

Between October 1987 and December 2000, 50 patients underwent reconstruction of the pulmonary outflow tract without external conduit. The primary malformation was tetralogy of Fallot with pulmonary atresia in 37, double outlet of right ventricle in 4, corrected transposition of the great arteries in 4, transposition of the great arteries with ventricular septal defect and pulmonary stenosis in 4, and double outlet of left ventricle in 2. Mean age at operation was 7.2 years, and mean body weight was 18.3 kg. To reconstruct posterior wall of the pulmonary outflow tract, interposition of autologous pericardium was performed in 24, direct anastomosis between pulmonary trunk and ventriculotomy in 13, longitudinal incision from ventriculotomy through pulmonary trunk in 12, and interposition of left atrial appendage in 1. Anterior wall was reconstructed with monocusp valved outflow patch (MVOP). There was one hospital death and no late death. At 10 years, the freedom from reoperation for pulmonary outflow tract obstruction was 100%, and freedom from reoperation for any cause was 86.6%. Transcatheter stenting for peripheral pulmonary stenosis was performed in 6 patients 2 to 10 months after operation.

[Arterial infusion of anticancer agent and lipiodol using a totally implanted drug delivery system].

We attempted arterial infusion of anticancer agent using a totally implanted drug delivery system in 52 patients who had inoperable liver cancer or the scheduled adjuvant chemotherapy after hepatic resection. The response rate of the cases using lipiodol was 38%, while that of the cases using only ADM and MMC was 0%. We studied changes in serum concentration of ADM and MMC. The results indicated that using 60% Urographin to make ADM, MMC-lipiodol emulsion was effective for targetting and control release.

[Study of complications after endoscopic sclerotherapy for esophageal varices--particularly 2 cases that changed portal circulation after sclerotherapy].

In 116 patients with esophageal varix due to liver cirrhosis who received the endoscopic sclerotherapy, the following complications were observed after the sclerotherapy: 38 ulcer formation; 19 chest pain; 16 hypotension; 15 fever; 12 pleural effusion; 6 esophageal stenosis and so on. Furthermore, two cases with marked changes of portal circulation were experienced after the sclerotherapy. The first case had severe bleeding out of duodenal varix four months after the sclerotherapy and died because of massive bleeding. The second case had hepatic encephalopathy six months after the sclerotherapy. In both cases, angiography revealed the development of collateral veins after the sclerotherapy. Because the abrupt intercept of the esophageal varix by the successful sclerotherapy causes the increase of the other collateral blood flow, the changes of portal circulation must be watched carefully afterward.

[Nucleotide metabolism in remnant rat liver after major hepatic resection].

Major hepatic resection is restricted in the presence of cirrhosis, because the cirrhotic liver is less able to regenerate. In the present study, to clarify what factors play a major role as inhibitors of the regeneration process, we focused on the changes in liver purine nucleotides and their catabolite levels in rats with cirrhosis. Decreases in adenine nucleotides and guanine nucleotides were observed after resection both in normal and thioacetamide-induced cirrhotic livers. These were prolonged in rats with cirrhosis. Tissue levels of hypoxanthine and xanthine increased both in the control group and in the cirrhotic group. The increase in xanthine level was remarkable in the cirrhotic group compared with the control group. These results indicate that xanthine oxidase activity is increased in cirrhotic liver after major hepatic resection. Xanthine oxidase catalyzes the oxidation of hypoxanthine to xanthine and xanthine to uric acid. Xanthine and uric acid are end metabolites of purine nucleotides. On the other hand, superoxide is generated in association with this reaction. Therefore, the disturbance in the energy metabolism and the increase in superoxide formation which are caused by the activation of xanthine oxidase might be involved in the regeneration process as inhibitory factors in cirrhotic rat liver.