Evaluation of electron radiation damage to green fluorescent protein.

Green fluorescent protein (GFP) emits light when irradiated by not only light but also electrons. This electron-induced light emission called cathodoluminescence (CL) can be used to realize a high-resolution light emission microscopy based on the irradiation of a very narrow electron beam. To implement CL mapping in life sciences the investigation of the damage resistance of GFP to electron irradiation needs to be clarified. In this study, we investigated the electron radiation damage to GFP by analyzing the change in the CL intensity during electron beam irradiation. Since some of the CL spectra changed in shape during electron irradiation, the change in the intensity between 585 and 605 nm were measured. The characteristic doses at different electron current densities and electron energies were investigated. The characteristic dose of EGFP is much larger than that of coronene, which is one of the stable organic molecules against the electron beam irradiation.

Development of a surface conductivity measurement system for ultrahigh vacuum transmission electron microscope.

The surface conductivity measurement system using a micro-four-point probe (M4PP) had been developed for the ultrahigh vacuum transmission electron microscope (UHV-TEM). Since the current distribution in the sample crystals during the current voltage measurement by the M4PP is localized within the depth of several micrometers from the surface, the system is sensitive to the surface conductivity, which is related with the surface superstructure. It was installed in the main chamber of the TEM and the surface conductivity can be measured in situ. The surface structures were observed by reflection electron microscopy and diffraction (REM-RHEED). REM-RHEED enables us to observe the surface superstructures and their structure defects such as surface atomic steps and domain boundaries of the surface superstructure. Thus the effects of the defects on the surface conductivity can be investigated. In the present paper we present the surface conductivity measurement system and its application to the Si(111)-square root(3) x square root(3)-Ag surface prepared on the Si(111) vicinal surfaces. The result clearly showed that the surface conductivity was affected by step configuration.

Typing of Herpes Simplex Virus in Patients with Uveitis.

Aim: To examine the type of herpes simplex virus (HSV) in cases with uveitis.Materials and Methods: Intraocular fluid specimens obtained from 3 cases with herpetic iridocyclitis and 6 cases with acute retinal necrosis (ARN) were examined by polymerase chain reaction (PCR). HSV typing was performed by the restriction patterns of the PCR products. Serum samples obtained from these cases and 33 cases with uveitis were examined by neutralization test (NT) for the availability of the typing of HSV.Results: The restriction patterns of the PCR products amplified from 3 specimens of iridocyclitis revealed HSV type 1 DNA. HSV type 2 DNA was identified in 5 of 6 cases of ARN and HSV type 1 DNA was found in only one case. The results of serum NT titers correlated with the typing of the amplicons.Conclusions: In the cases studied, HSV type 1 was the dominant etiological agent in herpetic iridocyclitis, while HSV type 2 plated a similar role in HSV-associated ARN. The examination of the serum NT may be helpful for the identification of the etiological types of HSV in patients with uveitis.

[Typing of herpes simplex virus in patients with uveitis].

AIM: To examine the type of herpes simplex virus (HSV) in cases with uveitis. MATERIALS AND METHODS: Intraocular fluid specimens obtained from 3 cases with herpetic iridocyclitis and 6 cases with acute retinal necrosis (ARN) were examined by polymerase chain reaction (PCR). HSV typing was performed by the restriction patterns of the PCR products. Serum samples obtained from these cases and 33 cases with uveitis were examined by neutralization test (NT) for the availability of the typing of HSV. RESULTS: The restriction patterns of the PCR products amplified from 3 specimens of iridocyclitis revealed HSV type 1 DNA. HSV type 2 DNA was identified in 5 of 6 cases of ARN and HSV type 1 DNA was found in only one case. The results of serum NT titers correlated with the typing of the amplicons. CONCLUSIONS: In the cases studied, HSV type 1 was the dominant etiological agent in herpetic iridocyclitis, while HSV type 2 played a similar role in HSV-associated ARN. The examination of the serum NT may be helpful for the identification of the etiological types of HSV in patients with uveitis.

Progressive outer retinal necrosis caused by herpes simplex virus type 1 in a patient with acquired immunodeficiency syndrome.

OBJECTIVE/BACKGROUND: To identify the etiologic agent of rapidly progressive outer retinal necrosis (PORN) in a 32-year-old man with acquired immunodeficiency syndrome (AIDS), who had retinitis developed from cytomegalovirus (CMV). Multiple yellowish spots appeared in the deep retina without evidence of intraocular inflammation or retinal vasculitis, diagnosed clinically as PORN. Death occurred after failure of multiple organs. DESIGN: Case report. METHODS: Both globes were taken at autopsy, fixed in formalin, and examined histopathologically and immunohistochemically to identify causative agents in the retinal lesions. MAIN OUTCOME MEASURE: Immunohistochemistry. RESULTS: All layers of the retina were severely damaged and contained focal calcification. Cytomegalic inclusion bodies were found in cells in the damaged retina of the right eye. Immunohistochemical studies for herpesviruses revealed the presence of CMV antigens in the right retina at the posterior pole and herpes simplex virus type 1 (HSV-1)-specific antigen in the periphery of both retinas. No varicella-zoster virus (VZV) antigen was detected in either retina. CONCLUSIONS: PORN has been described as a variant of necrotizing herpetic retinopathy, occurring particularly in patients with AIDS. Although the etiologic agent has been reported to be VZV, HSV-1 can be an etiologic agent.

[High inducibility of Epstein-Barr virus replication in B lymphocytes in Vogt-Koyanagi-Harada disease].

PURPOSE: The role of Epstein-Barr virus (EBV) reactivation in the pathogenesis of Vogt-Koyanagi-Harada (VKH) disease was examined. MATERIAL AND METHODS: Using B lymphocytes obtained from 8 patients with VKH disease and 10 patients with other types of uveitis, immortarized lymphoblast lines were established and infected with EBV. The degree of EBV activation in each lymphoblast line, in the presence and absence of various stimuli, was assessed by measuring the expression of 3 different antigens involved in replication by immunofluorescent staining and western blot analysis. Quantification of EBV DNA in cell culture supernatants was done by polymerase chain reaction. RESULT: Cell lines established from VKH patients expressed more viral antigens that those established from patients with other types of uveitis. There were greater amounts of EBV DNA in the VKH cell lines. CONCLUSION: B lymphocytes from VKH patients may be more susceptible to EBV activation, and the reactivation of EBV may be involved in the pathogenesis of VKH.

[A study of 44 patients with Kirisawa type uveitis].

We studied 50 eyes of 44 patients with acute retinal necrosis, Kirisawa type uveitis (KU), in order to examine clinical symptoms, pathogenic viruses, clinical grading, therapy and prognosis for this disease. Varicella-zoster virus (VZV) was the pathogenic organism in 37 eyes of 31 patients, while herpes simplex virus (HSV) was responsible in 13 eyes of 13 patients. There were more elderly patients in the VZV-KU group than in the HSV-KU group. In addition, mutton fat keratic precipitates and retinal exudates were more common in VZV-KU than in HSV-KU. We divided KU eyes into 3 clinical grades: severe, serious, and mild. Using statistical analysis, we found that the VZV-KU group had a significantly greater number of severe and serious cases than the HSV-KU group. Furthermore, some HLA antigens were found to be statistically more common in the VZV-KU group, although no associations were found in the HSV-KU group. 32% of VZV-KU and 67% of HSV-KU eyes had a final visual acuity (fVA) of greater than 0.5. When eyes with an fVA of greater than 0.1 were compared to eyes with an fVA of less than 0.1, we found that combined therapy using acyclovir, interferon beta, and prednisolone was especially effective for VZV-KU, although no significant difference was found for HSV-KU. Thus, it is essential to determine the pathogenic virus causing KU, in order to understand the disease pathogenesis as well as to select appropriate treatment.

[Cytomegalovirus retinitis and Pneumocystis carinii choroidopathy in a patient with AIDS].

A case of Pneumocystis carinii (P. carinii) choroidopathy is reported. The patient was a 17-year-old man with acquired immunodeficiency syndrome (AIDS) who developed bilateral, multifocal, yellow-white, round, flat choroidal lesions ranging in size from 1/8 to 1/6 of the disc diameter while undergoing treatment for cytomegalovirus retinitis. He had had P. carinii pneumonia 43 months previously, and received inhaled pentamidine as suppressive therapy. Over a 4-week period of observation, the choroidal lesions appeared to enlarge slowly and increased in number in the posterior pole, with no clinical evidence of intraocular inflammation and retinal involvement. He was diagnosed as having P. carinii choroidopathy and treated with intravenous pentamidine. Three months after systemic pentamidine therapy was begun the choroidal lesions disappeared. Despite the fact that P. carinii choroidopathy is a rare opportunistic ocular infection, ophthalmic manifestations may be an important initial marker of extrapulmonary disseminated infection in some patients. Therefore we recommend ophthalmologic examinations in patients with AIDS who receive long-term aerosolized pentamidine prophylaxis for pneumonia.

Polymerase chain reaction for diagnosis of herpetic intraocular inflammation.

The authors present a polymerase chain reaction method for rapid and direct diagnosis of herpetic intraocular infections using small volume samples of intraocular fluid from 29 patients with various intraocular inflammatory diseases and 24 controls with senile cataract. Of six patients with early acute retinal necrosis from whom aqueous humor was tested, four were found to be positive for the presence of varicella-zoster (VZV) DNA while the other two were positive for the presence of herpes simplex virus (HSV) DNA. One of the patients with HSV DNA had been tested at an extremely early stage, at which time the aqueous humor viral antibody ratio did not predict a specific viral infection. Among four patients with acute retinal necrosis in relatively late stages following treatment with acyclovir from whom vitreous was obtained and tested, only one was found to have the presence of any viral DNA (VZV). On the other hand, the vitreous viral antibody ratio was found to be predictive of VZV infection in all four cases. VZV DNA was also detected in aqueous humor samples from four patients with suspected herpes zoster anterior uveitis, while HSV DNA was found in the aqueous humor of one patient with nonspecific keratouveitis. Neither human cytomegalovirus DNA nor human herpesvirus-6 DNA was detected in any sample included in this study. Finally, Epstein-Barr virus DNA was detected in the aqueous humor of the majority of patients studied and identified in cataract patients as well, suggesting either low specificity of the authors' assay for this virus or ubiquity of this virus in human eyes. In summary, the PCR method proved to be a very useful tool in establishing an etiological diagnosis in patients in the early stages of acute retinal necrosis, and in patients with anterior uveitis due to suspected HSV or VZV infection.

[A case of psittacosis].

A 67-year-old Japanese housewife, who had been attended the out patient department of medicine, Fukuoka Dental College (FDC) Hospital for paroxysmal atrial fibrillation, was admitted to FDC Hospital because of high fever, exhaustion, anorexia, myalgia and mild stupor. Her ECG finding revealed atrial fibrillation and roentgenologic examination of the chest showed diffuse opacities in the left lung field (S10) without pleural effusion. As she had told her physician that her pet parakeet had been dead recently, she was diagnosed immediately as psittacosis. She was instantly treated with minocycline orally and deslanoside intravenously. Laboratory findings on admission disclosed the following results: Complement-fixing antibodies against Chlamydia psittaci were 1:64, and liver dysfunction (GOT 253, GPT 86, LDH 846) was shown. The white blood cell count was 4,700 associated with shift to the left, C-reactive protein was 6 plus and the erythrocyte sedimentation rate was 109 mm in 1 hour. The course in the hospital was satisfactory and after 38 hospital days she was discharged with complete recovery from the psittacosis. It is emphasized the importance of that the question about the history of contact with psittacine birds or other avian species is essential to diagnose psittacosis.

[A study of proteinuria. (12) The incidence of the patients with proteinuria, microscopic hematuria and glycosuria in Fukuoka Dental College students in 1987 and 1988].

An investigation was made on the incidence of patients with proteinuria, microscopic hematuria and glycosuria in 757 (in 1987) and in 681 (in 1988) students of Fukuoka Dental College; and the results were as follows. 1) The incidence of patients with proteinuria was 6.3% (48/757) in 1987, and 6.9% (47/681) in 1988. 2) The incidence of patients with microscopic hematuria was 4.0% (30/757) in 1987, and 5.0% (34/681) in 1988. 3) The incidence of patients with glycosuria was 0.5% (4/757) in 1987, and 0.1% (1/681) in 1988. 4) Further medical examinations were made on 33 students in 1987, and 37 students in 1988. a) 27 students in 1987 and 34 students in 1988 were diagnosed as normal respectively. b) In three cases, clinical diagnoses could not be established without long-term observation and two cases were diagnosed as urinary tract infection and a case was diagnosed as possible chronic glomerulonephritis in 1987. c) In 1988, a case was diagnosed as urinary tract infection, a case was urethritis and a case should be needed a longterm observation.

[Tooth extraction in a case of thrombasthenia].

Thrombasthenia is a congenital platelet disorder characterized by the normal platelet count, prolonged bleedingtime, absence of platelet aggregation and defective clot retraction. Clinically, purpura, epistaxis, gingival bleeding and excessive bleeding after minor injuries or operations are the main manifestations. We experienced a case of a 15-year-old boy suffering from thrombasthenia required extraction of 6/6. The problems of nature of the disease and its familial character are described and the problems of management are outlined.

[Statistical analysis of new out-patients in the Department of Medicine, Fukuoka Dental College Hospital. 10: From January 1 to December 31, 1987].

To clarify the common diseases of new out-patients, a retrospective study was made in 1.523 out-patients, who visited our department from January 1 to December 31, 1987. Of 1,523 out-patients, 3,513 diseases were diagnosed in 1,350 patients and the others were normal subjects who visited the department for obtaining health certificates. Of these 3,513 diseases, 1,220 (34.7%) were respiratory diseases including upper respiratory infections, 471 (13.4%) were gastrointestinal diseases, 396 (11.3%) were renal and urinary tract diseases, 371 (10.6%) were liver and biliary tract diseases including pancreas diseases, 354 (10.1%) were cardiovascular diseases, 257 (7.3%) were allergic, muscle, bone and connective tissue diseases, 206 (5.9%) were neurologic diseases, 153 (4.4%) were metabolic diseases, 46 (1.3%) were endocrine diseases and 39 (1.1%) were hematologic diseases. Of these 1,523 out-patients 20 (1.5%) patients were positive for HBs antigen.