Photodynamic therapy of experimental choroidal neovascularization with a hydrophilic photosensitizer: mono-L-aspartyl chlorin e6.

PURPOSE: To demonstrate the selective localization of the hydrophilic photosensitizer mono-L-aspartyl chlorin e6 (NPe6) in experimental choroidal neovascularization in nonhuman primate eyes. METHODS: Sixty-seven experimental choroidal neovascular lesions (CNV) were created in the fundi of Macaca monkeys using the modified Ryan's model and documented by fluorescein and indocyanine green angiography. To determine the biodistribution of NPe6 and the optimal timing of laser irradiation after dye administration, NPe6 angiography and fluorescence microscopy with NPe6 were performed. Photodynamic therapy (PDT) was performed at various dye doses (0.5-10.0 mg/kg) and laser fluences (7.5-225.0 J/cm2) on the CNV and on 10 areas of normal retina and choroid. Treatment outcomes were assessed by fluorescein and indocyanine green angiography and confirmed by light and electron microscopy. RESULTS: NPe6 fluorescence microscopy demonstrated intense fluorescence of CNV and retinal pigment epithelial cells. Choroidal vessel walls and outer retina adjacent to CNV fluoresced moderately; retinal vessel walls and microcapillaries had trace fluorescence. The fluorescence of CNV lesions on fluorescein angiography became stronger than that of retinal vessels 20-60 minutes after dye injection. Choroidal neovascular lesion closure was achieved with NPe6 PDT without significant damage to the sensory retina. Histology demonstrated necrosis of CNV endothelial cells with minimal damage to surrounding tissues. CONCLUSIONS: NPe6 PDT selectively localizes to experimental CNV in nonhuman primates, resulting in occlusion of CNV with sparing of the neurosensory retina.

[Prognosis of chronic hepatitis C with regard to the aim of treatment].

Epidemiology of hepatitis C virus(HCV) infection and clinical prognosis of chronic hepatitis C were presented here to reveal the object of treatment of Chronic Hepatitis C. Hepatitis C Virus is transmitted by blood and blood products. After acute HCV infection, about 70% developed persistent HCV infection, and the diagnosis is by finding viral RNA in the serum of patients with anti-HCV antibody. Persistent HCV infection causes chronic hepatitis, in which the natural clearance of HCV is almost impossible and there is almost no natural cure for chronic hepatitis caused by HCV. Chronic hepatitis C tends to develop gradually and to progress to liver cirrhosis, and is involved in the pathogenesis of hepatocellular carcinoma. In Japanese patients with chronic hepatitis C, 45% developed liver cirrhosis pass through a phase of chronic active hepatitis over a 15-year course after initial HCV infection, and 25% developed hepatocellular carcinoma over a 20-year course after the initial HCV infection. In addition the remaining patients may start to develop rapidly to chronic active hepatitis and to liver cirrhosis after 20 to 30 years duration of inactive phase. Thus, this type of chronic hepatitis reveals a poor long-term prognosis. For etiological treatment of chronic hepatitis C, eradication of persistent HCV infection is needed. If this is impossible, then preventing the development of liver cirrhosis and hepatocellular carcinoma is important.

[Epidemiological trend of chronic hepatitis C].

Though patients with chronic hepatitis C after blood transfusion hepatitis decrease dramatically during the past decade, patients with it after non-transfused acute hepatitis C are still in existence. This means that new patients with chronic hepatitis C are on the decrease but not diminish. To the contrary, the number of patients with chronic hepatitis C is increasing. This depends on mainly dig up the undiscovered patients. These result in increasing the patients with hepatocellular carcinoma.

Effect of decompression enlargement laminoplasty for posterior shifting of the spinal cord.

STUDY DESIGN: A study to measure the shifts of the spinal cords and the effects of decompression laminoplasty in 65 patients with cervical lesions who underwent computed tomographic myelography before and after laminoplasty. OBJECTIVES: To investigate limitations of the spinal cord posterior shift after laminoplasty and to clarify the optimal decompression areas to obtain effective posterior shifting. SUMMARY OF BACKGROUND: Although several types of laminoplasty have been performed, all procedures share the common purpose of posterior decompression. No previous studies have examined the limitations of posterior decompression or the optimal decompression range. METHODS: The distance from the posterior edge of each vertebral body or disc level to the posterior edge of the spinal cord was measured by computed tomographic myelography. After the posterior shift was determined by calculating the difference between pre- and postsurgical distances, the relations between posterior shift and neck alignment, clinical results, and the areas of decompression were analyzed. RESULTS: The spinal cord shift ranged from a maximum of 6.6 mm to a minimum of 0 mm. Clinically, spinal cord shifts greater than 3 mm were associated with good clinical outcomes. Upward or downward advanced laminoplasty was related to larger spinal cord shifts at the upper or lower cervical spine. CONCLUSIONS: A mean spinal cord shift of > 3 mm was associated with good clinical outcomes after laminoplasty. In cases with compressive lesions at the upper or lower cervical spine, extension of decompression one level above or one level below likely results in a greater posterior spinal cord shift at these lesions.

Malignant insulinoma presenting a non-functioning metastatic liver tumor 14 years after resection of the primary tumor.

A 58-year-old woman who had undergone resection of insulinoma 14 year earlier visited our clinic complaining of abdominal discomfort. Computerized tomographic scan showed multiple liver tumors, and a diagnosis of metastatic tumor of malignant islet cell tumor was confirmed histologically. No oversecretion of hormones or hypoglycemic episode was observed on readmission. Thus, the insulinoma seemed to have transformed to a clinically non-functioning tumor. The patient was treated with transcatheter arterial embolization, resulting in clinical improvement with marked reduction in tumor size. Features of interest in this case included; (1) transformation to non-functioning metastatic liver tumor 14 years after resection of insulinoma, (2) the usefulness of transcatheter arterial embolization for multiple metastatic tumor of malignant islet cell tumor.

Clinical application of hepatitis C virus core protein in early diagnosis of acute hepatitis C.

A fluorescence enzyme immunoassay (FEIA) for the quantitative measurement of hepatitis C virus (HCV) core protein has recently been developed. In this study, we studied the clinical usefulness of this measurement in patients with acute hepatitis C. Eighteen patients with post-transfusion acute hepatitis C were enrolled in the study; 5 patients showed resolution of hepatitis with disappearance of HCV viremia, while the remaining 13 patients did not. A second generation HCV antibody, HCV RNA, and HCV core protein were measured in serial serum samples taken within 1 month of the onset of acute hepatitis and 3, 6, 12, 24, and 36 months after onset. Within the first month after disease onset, the positivity rates of HCV RNA (100%; P = 0.0014) and HCV core protein (89%; P = 0.0300) were both significantly higher than that of HCV antibody (56%). Six months after disease onset, the positivity rate of HCV antibody had increased, to 100%, and the positivity rates of HCV RNA and HCV core protein began to decrease. HCV core protein levels did not differ between patients with resolved and unresolved disease in the first month after disease onset. These findings indicate that FEIA, a simple assay, for the measurement of HCV core protein was useful for the early diagnosis of acute hepatitis C.

Relationship of 31P MR spectroscopy to the histopathological grading of chronic hepatitis and response to therapy.

PURPOSE: In vivo phosphorus-31 MR spectroscopy (31P MRS) was performed in the human liver in order to investigate the relation between: the ratios of phosphorus metabolites in the liver; the histopathological grading of chronic hepatitis; and the response to therapy. MATERIAL AND METHODS: Hepatic 31P MRS using the DRESS method (depth-resolved surface-coil spectroscopy) was carried out in 45 patients with chronic viral hepatitis or autoimmune hepatitis, and in 16 control subjects. We measured the ratios of the peak areas of phosphomonoesters (PME), inorganic phosphate (Pi), or phosphodiesters (PDE) to the peak area of beta-adenosine triphosphate (ATP). RESULTS: The PDE/ATP ratio of patients with chronic hepatitis or liver cirrhosis was lower than that of control subjects (liver cirrhosis = 0.74; chronic active hepatitis = 1.13-1.21; normal = 1.43); only a small difference was found in the PME/ATP and Pi/ATP ratios. There was no correlation between the spectra and histopathological grading or response to therapy, but the response to therapy was poor when a reduced PDE/ATP ratio was present. CONCLUSION: The PDE/ATP ratio measured by 31P MRS makes it possible to identify the transition of chronic active hepatitis into liver cirrhosis with a poor response to therapy.

Analysis of bronchoalveolar lavage fluid in patients with chronic hepatitis C before and after treatment with interferon alpha.

BACKGROUND: Previous studies have shown that patients with idiopathic pulmonary fibrosis (IPF) were more likely to be seropositive for hepatitis C virus (HCV) than normal controls, and that patients with chronic hepatitis C treated with interferon alpha (IFN-alpha) sometimes developed pulmonary fibrosis. The possibility that HCV infection and/or treatment with IFN-alpha are involved in the pathogenesis of pulmonary fibrosis or alveolitis was investigated. METHODS: A prospective non-randomised study was performed in 13 healthy controls and in patients with chronic hepatitis C before (n = 13) and after (n = 10) treatment with IFN-alpha. Bronchoalveolar lavage (BAL) fluid cell counts, ratios and T cell subsets, and the concentrations of interleukin (IL)-1 beta, tumour necrosis factor(TNF)-alpha, and hepatocyte growth factor (HGF) were measured. RESULTS: Lymphocyte counts in the BAL fluid were significantly increased in both groups of patients (median (range) values: before treatment, 36.8 (1.5-226.0); after treatment, 16.2 (4.5-97.6)) compared with the normal controls (3.3 (0.5-32.3)). In the pretreatment group the activated T cell (HLA-Dr positive) count was also increased (51 (40-74)) compared with that in the normal controls (27 (4-52)), but after treatment it was decreased (40 (0-76)) compared with the pretreatment count. Administration of IFN-alpha did not affect these parameters. IL-1 beta, TNF-alpha, and HGF were not detected. CONCLUSIONS: These findings suggest that HCV infection is associated with increased counts of lymphocytes and neutrophils in BAL fluid and that treatment with IFN-alpha appears to alter lymphocyte surface markers.

Serum thrombopoietin (c-Mpl ligand) levels in patients with liver cirrhosis.

To clarify the role of c-Mpl ligand (thrombopoietin: TPO) in liver cirrhosis (LC), we examined serum TPO levels (sTPO) in patients with LC (N = 44), chronic hepatitis (CH; N = 13) and healthy controls (N = 41) by an enzyme-linked immunosorbent assay. Although platelet counts of all LC patients (89 +/- 59 x 10(9)/l; mean +/- SD) were lower than those of controls and CH patients, sTPO levels in LC patients (1.23 +/- 0.51 fmol/ml) were the same as those in controls (1.22 +/- 0.37) and CH patients (1.18 +/- 0.36). Platelet counts were significantly higher in splenectomized patients than in unsplenectomized patients, but the sTPO level did not differ between these two groups. In LC patients, the sTPO level was not correlated with the platelet count, but was correlated with prothrombin time, activated partial thromboplastin time, and total bilirubin, indicating that production of TPO in the liver decreases slightly with the development of liver dysfunction. Our findings suggest that production of TPO is maintained in LC patients and their thrombocytopenia is not due to a defect in platelet production.

Responses of periodontal nerve terminals to experimentally induced occlusal trauma in rat molars: an immunohistochemical study using PGP 9.5 antibody.

The response of periodontal nerves to experimentally induced occlusal trauma in rat molars was assessed by immunohistochemistry for protein gene product 9.5 (PGP 9.5) at light and electron microscopic levels, and by computerized image analysis. The occlusal surface on the left upper first molar of 8-wk-old male Wistar rats was raised approximately 1 mm under ether anaesthesia. The rats were perfusion-fixed on d 1, 2, 3, 4, and 7 after bite-raising and then decalcified for 2-3 wk. Frozen sagittal cryostat sections were stained by the avidin-biotin complex method. By the second day after bite-raising many Ruffini endings were swollen and their outline unclear at the light microscopic level. Transmission electron microscopy disclosed PGP 9.5 reaction products within Ruffini endings that had unusually long cytoplasmic projections extending through enlarged slits of the Schwann sheaths and also diffuse extracellular PGP 9.5-immunoreactivity near the Ruffini endings. From d 2 to 4, thin nerve fibres on the pressure side of the periodontal ligament were orientated irregularly and had a prominent beaded appearance. An increase in beaded nerve terminals occurred at d 2-4 post elevation, and decreased later. These results suggest that occlusal trauma indices specific changes in the distribution and shape of nerve terminals in the periodontal ligament.

The natural course of chronic hepatitis C: a comparison between patients with genotypes 1 and 2 hepatitis C viruses.

This study was conducted to clarify if the long-term histological outcome among patients with chronic hepatitis C differs according to whether they are infected with genotype 1 or 2 hepatitis C virus (HCV). We examined 140 patients with chronic hepatitis C. The HCV genotype was determined by the enzyme-linked immunosorbent assay (ELISA) based on genotypes 1 and 2 specific recombinant proteins; genotype 1 was found in 100 patients (96 were 1b and 4 were indeterminate) and genotype 2 in 36. The two groups showed no significant difference for any clinical background features. Deterioration of the grade of liver histology during the follow-up period was seen in 68.0 percent of the patients with genotype 1 as compared with 41.7 percent of those with genotype 2 (P < .01). Similarly, the deterioration of the stage of liver histology was more common in the former group than in the latter (63.0 percent and 38.9 percent respectively; P < .05). The mean serum HCV-RNA titer was significantly higher in the patients with genotype 1 than in those with genotype 2 (P < .001), and multivariate analysis showed the titer was one of the independent factors of the deterioration of the stage (P = .0044). This phenomenon may be related in part to the difference in pathogenicity between the two HCV genotypes. In conclusion, our results suggest that more severe progression of chronic hepatitis C is seen in patients showing genotype 1b compared with those with genotype 2.

Bronchoalveolar lavage fluid findings in patients with chronic hepatitis C virus infection.

BACKGROUND: Hepatitis C virus (HCV) infection has recently been incriminated as an aetiological agent in idiopathic pulmonary fibrosis. This study was performed to determine the cellularity and lymphocyte phenotypes of bronchoalveolar lavage (BAL) fluid in patients with chronic hepatitis C. METHODS: BAL fluid and lavage lymphocyte subsets from 13 patients (10 men) with active chronic hepatitis C, diagnosed by sustained elevated serum glutamic pyruvic transaminase and typical histological findings in the liver, were analysed. Lavage findings in these patients were compared with those from 13 healthy volunteers (eight men) as controls. RESULTS: There was no difference in total cell counts in lavage fluid between the two groups. Lavage lymphocyte and eosinophil numbers were increased in patients with chronic hepatitis C. Surface marker analysis of the lymphocyte populations showed increases in CD2, CD3, CD4, and HLA-DR. CD4/CD8 ratios were not different. CONCLUSIONS: The numbers of lymphocytes and eosinophils in BAL fluid are increased in patients with chronic hepatitis C. These findings suggest that HCV infection may trigger alveolitis.

Acute hepatitis C transmitted by needlestick accident despite short duration interferon treatment.

Hepatitis C virus (HCV) transmission by needlestick accidents involving hospital employees has become an important problem. The present report is of a case of acute hepatitis C that developed after a needlestick injury, despite short duration interferon treatment performed just after the accident in a trial effort to prevent HCV transmission. Nosocomial infection of HCV in medical employees is reviewed, and the current prospects for protecting them from HCV transmission after needlestick accident are discussed.

Usefulness of simple assays for serum concentration of hepatitis C virus RNA and HCV genotype in predicting the response of patients with chronic hepatitis C to interferon alpha 2a therapy.

The use of two new assays was evaluated for predicting the response to interferon (IFN) therapy in patients with chronic hepatitis C. The genotype of hepatitis C virus (HCV) was established by an enzyme-linked immunosorbent assay based on genotype-specific recombinant peptides of the NS4 region (genotyping ELISA). The concentration of HCV RNA was measured by a branched DNA assay (bDNA assay). Seventy-eight patients received the same regimen of IFN alpha 2a. Of the 74 patients assessed who completed the program, 38 (51.4%) were responders; i.e., their serum aminotransferase levels remained normal for 6 months or longer after stopping IFN, while 36 (48.6%) were nonresponders. The results of the HCV genotype determined by the genotyping ELISA and by the polymerase chain reaction (PCR) assay based on genotype-specific primers were similar. The serum concentrations of HCV RNA as measured by the bDNA assay and by the competitive PCR assay correlated closely and significantly (r = 0.82, P < 0.001). Multiple logistic regression analysis showed that the serum concentration of HCV RNA determined by the bDNA assay, the HCV genotype determined by the genotyping ELISA, and the histology activity index (HAI) of the liver were independently associated with IFN efficacy. By using these three variables in combination, a predictive rate of 82.4% was obtained. A lower level of HCV RNA, genotype 2 and a lower HAI score for liver histology were predictive of a favorable response to IFN. Thus, the genotyping ELSIA and the bDNA assay appear to be useful for clinical management of patients receiving IFN therapy.