Omega filter installed in the 1MV microscope of Kyushu University.

A 1.25MV high-voltage electron microscope with a B-type omega filter has been successfully installed at Kyushu University. An image detection chamber has been set inside a concrete block below the ground level without changing the frame structure for anti-vibration. Nearly the same design as that for the 200kV microscope has been kept for the present omega filter except for its size. A new pre- and post-filter lens system with rotation-free imaging has been designed. Energy resolution, beam shape and stability of the filter have been measured. Some application data have been obtained to demonstrate the performance of the filter.

Genetic variability of respiratory syncytial virus subgroup a strain in 15 successive epidemics in one city.

The genetic variability of 125 respiratory syncytial virus (RSV) subgroup A isolates over 15 successive epidemics from 1980 to 1995 in an urban population of Japan was determined. Allocation of isolates into lineages was archived by reverse transcriptase-polymerase chain reaction amplification of selected regions of the nucleoprotein (NP) and attachment (G) protein gene followed by restriction fragment length polymorphism (RFLP) analysis. Three and seven distinct restriction patterns of the NP and G gene were observed, respectively. When the NP and G gene RFLP analyses were combined, ten different genetic lineages were identified in the 125 isolates. The strains with the same genotype were isolated in each epidemic and the dominant lineages were replaced by others after every one to three consecutive epidemics. Nucleotide and amino acid sequencing of the variable region of G gene of these predominant isolates revealed differences of 5--28% between strains. There was, however, no apparent accumulation of diversity with age to indicate progressive changes. The dominant strains were often closely related to those isolated in other parts of the world at a similar time. These observations suggest that dominant RSV strains are replaced frequently by others that have been co-circulating or have recently entered the community from a worldwide reservoir. The change of dominant strains may be influenced by the buildup of immunological resistance in the community to successive epidemics of the same strain.

Salmonella enteritidis FliC (flagella filament protein) induces human beta-defensin-2 mRNA production by Caco-2 cells.

Antimicrobial peptides are crucial for host defense at mucosal surfaces. Bacterial factors responsible for induction of human beta-defensin-2 (hBD-2) mRNA expression in Caco-2 human carcinoma cells were determined. Salmonella enteritidis, Salmonella typhimurium, Salmonella typhi, Salmonella dublin, and culture supernatants of these strains induced hBD-2 mRNA expression in Caco-2 human carcinoma cells. Using luciferase as a reporter gene for a approximately 2.1-kilobase pair hBD-2 promoter, the hBD-2-inducing factor in culture supernatant of S. enteritidis was isolated. The supernatant factor was heat-stable and proteinase-sensitive. After purification by anion exchange and gel filtration chromatography, the hBD-2-inducing factor was identified as a 53-kDa monomeric protein with the amino-terminal sequence AQVINTNSLSLLTQNNLNK, which is identical to that of the flagella filament structural protein (FliC) of S. enteritidis. Consistent with this finding, the 53-kDa protein reacted with anti-FliC antibody, which prevented its induction of hBD-2 mRNA in Caco-2 cells. In agreement, the hBD-2-inducing activity in culture supernatant was completely neutralized by anti-FliC antibody. In gel retardation analyses, FliC increased binding of NF-kappaB (p65 homodimer) to hBD-2 gene promoter sequences. We conclude that S. enteritidis FliC induces hBD-2 expression in Caco-2 cells via NF-kappaB activation and thus plays an important role in up-regulation of the innate immune response.

Hemodynamic changes in two patients with retinal circulatory disturbances shown by fluorescein angiography using a scanning laser ophthalmoscope.

PURPOSE: To assess hemodynamic changes in two patients with severely affected retinal circulation. METHODS: A 62-year-old man with central retinal artery occlusion and a 46-year-old woman with branch retinal vein occlusion were studied by fluorescein angiography with a scanning laser ophthalmoscope (SLO). Fluorescein angiography with SLO revealed hypofluorescent clumps of different sizes and hyperfluorescent dots in large retinal vessels. The velocities of the hypofluorescent clumps were calculated between two points on the same vessel, and movements of the hypofluorescent clumps and the hyperfluorescent dots were investigated. RESULTS: The velocities of the hypofluorescent clumps were slow and varied in the same vessel. The velocities of the hypofluorescent clumps increased at the sites with narrow calibers. The hypofluorescent clumps occasionally changed size in the bloodstream. The hypofluorescent clumps flowed along the walls of retinal vessels. Distance between consecutive hypofluorescent clumps was wide. Some vessels filled with hypofluorescent clumps were also detected. Rolling hyperfluorescent dots were seen in fluorescent plasma. CONCLUSIONS: The hypofluorescent clumps were concluded to be packed erythrocytes and the hyperfluorescent dots corresponded to leukocytes and platelets moving in the vessels. Fluorescein angiography with SLO is a useful method for evaluating hemodynamic changes using the hypofluorescent clumps in severely affected retinal circulation.

Recent increase in the frequency of infant measles in Japan.

BACKGROUND: Because there has been no major outbreak of measles in Japan during the last decade, subclinical boosting of pre-existing immunity by exposure to epidemic measles is unlikely to have occurred. Consequently, the measles immunity of mothers of this generation is less well developed and they provide less effective passive immunity to their infants. An increase in the frequency of infant measles is therefore probable. METHOD AND RESULTS: A retrospective review was undertaken of the clinical records of all measles cases that occurred in the last 3 years in eight hospitals and two out-patient clinics in Hokkaido. Of 541 recorded cases, 96 (17.7%) were infants and of these, 66 patients (68.8%) were over 9 months of age. Only 20 children had received measles vaccination. CONCLUSION: There has been a recent increase in the frequency of infant measles, suggesting an increased population of infants susceptible to measles before the age of 1. Of the older patients, 95% had not been vaccinated against measles. Increasing the rate of measles vaccination may help suppress future measles outbreaks and assist in protecting infants until they pass their first birthday.

Respiratory syncytial virus infection of human respiratory epithelial cells enhances inducible nitric oxide synthase gene expression.

The induction kinetics of the mRNA of interferon regulatory factor 1 (IRF-1), inducible nitric oxide synthase (iNOS), and proinflammatory cytokines in respiratory syncytial virus (RSV)-infected human type 2 alveolar epithelial cells (A549 cells) were analyzed semiquantitatively by RT-PCR. RSV enhanced IRF-1 and iNOS mRNA expression as early as 4 h after RSV infection and this enhancement lasted several hours. No IFN-gamma gene expression was observed during the whole course of the infection. Expression of IFN-beta, IL-1beta, and TNF-alpha genes was observed slightly at 4 h and became marked 7 h after infection. Addition of neutralizing antibodies to these cytokines to the culture had no effect on the induction of iNOS mRNA. The iNOS transcriptional activity in RSV-infected cells was significantly enhanced by an exogenous cytokine mixture (IL-1beta, TNF-alpha, and IFN-gamma). An apparent nitric oxide (NO) production was identified only when cytokines were added together with RSV infection. A significant increase of iNOS gene expression was observed in nasopharyngeal exudate cells obtained from infants during the acute phase of RSV bronchiolitis. These observations suggest that RSV infection of human respiratory epithelial cells induces the iNOS gene both in vitro and in vivo; this induction may occur rather promptly and involves transcriptional activator IRF-1 induced by the RSV infection itself. The iNOS gene, which is initially induced by RSV infection, may be further enhanced in a paracrine fashion by proinflammatory cytokines released by infection-activated inflammatory cells.

Reduced passive measles immunity in infants of mothers who have not been exposed to measles outbreaks.

Natural measles infection usually confers life-long immunity which is transferred from mothers to their offspring, protecting them from natural measles until the age of about 12 months. Recently, however, natural measles has been observed with increased frequency in infants under the age of 12 months. Natural measles outbreaks in the city of Sapporo have been suppressed by widely applied measles vaccination. Passive measles immunity in 160 neonates (cord blood), born during the last 17 y in Sapporo, Japan was determined by a neutralization (NT) antibody test. The mothers of these infants had had natural measles infection during childhood. Geometric mean titres (GMTs) of cord blood NT antibodies gradually decreased after 1989 and the GMTs of the most recently born infants were significantly lower than those of infants born in the first few years of the study. These observations suggest that even in mothers who experienced natural measles in childhood, recurrent exposure to natural measles is necessary in order to maintain adequate antibody levels for effective passive immunity of their infants.

Immunochromatography test for rapid diagnosis of adenovirus respiratory tract infections: comparison with virus isolation in tissue culture.

The sensitivity and the specificity of a new commercial rapid 10-min adenovirus antigen immunochromatography (IC) test were determined by comparison with the sensitivity and specificity of virus isolation. Of 169 pharyngeal swabs from children with suspected adenovirus respiratory tract infections, 95 (56%) were culture positive for adenovirus. The IC test was sensitive (detecting 69 of these 95 infections [72.6%]) and completely specific (identifying 74 of 74 specimens [100%]) when it was compared with cell culture. The test detected adenovirus serotypes 1, 2, 3, 5, and 7 with almost equal sensitivities. This test is not only rapid and easy to perform but also sensitive and specific for adenovirus respiratory tract infections. The test is sufficiently rapid to be used at the bedside or in an outpatient clinic, with the result being available during a patient's first examination.

The relevance of TH1 and TH2 cells in immediate and nonimmediate reactions to gelatin-containing vaccine.

BACKGROUND: The immune mechanism of gelatin allergy, especially the participation of TH1 and TH2 cells and their cytokine secretion, has not been investigated. OBJECTIVE: We investigated the characteristics of T lymphocytes from patients allergic to gelatin-containing vaccine by secondary in vitro stimulation of circulating mononuclear cells with gelatin. METHODS: We studied 8 children with a history of immediate-type reactions and 8 with nonimmediate-type reactions after inoculation of gelatin-containing vaccine. The expression of IFN-gamma (TH1 ), IL-2 (TH1 ), IL-4 (TH2 ), and IL-13 (TH2 ) mRNA was examined semiquantitatively by using a reverse transcriptase PCR. IgE antibody to bovine gelatin was measured with the fluorometric ELISA system, and gelatin-specific T-cell responses were detected by an in vitro lymphocyte proliferation assay. RESULTS: Patients with an immediate reaction all had gelatin-specific IgE antibody, whereas others did not. However, all patients exhibited positive T-lymphocyte responses specific to gelatin. Lymphocytes from subjects with nonimmediate-type reactions generally expressed very weak or sometimes no IFN-gamma, IL-2, or IL-13 genes and especially no IL-4 gene. On the other hand, the lymphocytes of subjects with immediate-type reactions significantly expressed not only IL-4 and IL-13 but also IFN-gamma and IL-2 mRNA. CONCLUSION: Our observations suggest that both gelatin-specific TH2 and TH1 responses are involved in the pathogenesis of the immediate reaction to gelatin. The gelatin-specific IL-4 and/or IL-13 responses consistently observed in patients with an immediate reaction may be associated with the production of gelatin-specific IgE antibody.

Respiratory syncytial virus infection of human respiratory epithelial cells enhances both muscarinic and beta2-adrenergic receptor gene expression.

The possible changes in transcriptional activities of the M1 muscarinic acetylcholine receptor (mAChR) and beta2-adrenergic receptor (AR) genes in respiratory syncytial virus (RSV)-infected human type 2 alveolar epithelial cells (A549 cells) were analyzed semiquantitatively by reverse transcription-polymerase chain reaction (RT-PCR). RSV enhanced M1 mAChR gene expression significantly at 4 hrs post infection (p.i.), and this enhancement persisted until 10 hrs, after peaking at 7 hrs. Beta2-AR gene expression also increased significantly as early as at 1 hr p.i. and persisted for more than 10 hrs. These transcriptional enhancements were observed in cells treated with live but not with inactivated virus. Our observations suggest that RSV infection of human respiratory epithelial cells may enhance the expression of both parasympathetic and sympathetic receptors. The upregulated M1 mAChR gene in virus-infected cells may correlate with temporal airway hyperresponsiveness in subjects with RSV or other respiratory virus infection. The enhanced beta2-AR gene expression in peripheral lungs might explain the excessive mucus secretion observed during viral pneumonitis.

[Resection and reconstruction of sternum].

In case of sternal resection, it is necessary to preserve bone material indispensable for the stability of the anterior chest wall and air tightness of the thoracic cavity, and the support of the chest wall integrity must be restored by some means. Various techniques have been applied to the reconstruction of the chest wall following resection. During the last 10 years, we have performed reconstructive operation for 6 cases of the chest wall following resection of the sternum in recurrent cases of breast cancer or invaded case of primary breast cancer. In these patients, the chest wall was reconstructed using a rib-latissimus dorsi osteomyocutaneolus flap or a latissimus dorsi myocutaneous flap. The sternum was totally resected in 3 cases, and in all 3 cases, reconstructed using a rib-latissimus dorsi osteomyocutaneous flap. Although postoperative pulmonary function decreased, all cases could be relieved from endotracheal intubation within 17 hours after operation, and had no problems in activities of daily living or occurrence of chest flailing or paradoxical movement of the chest. An artificial material (expanded polytetrafluoroethlene patch) was used in only one patient for the reconstruction of the osseous thorax, but this case developed infection during postoperative chemotherapy. After this experience, we used only biological materials for the reconstruction of the chest wall and postoperatively performed radiotherapy and/or chemotherapy on all cases. We have observed no flap infection or detachment since then. One characteristic of using the latissimus dorsi myocutaneous flap is that it is easily elevated and rarely causes serious postoperative esthetic or functional problems. The flap is also easily utilized to reinforce the osseous thorax because ribs immediately below the latissimus dorsi muscle are readily mobilized as a pedicle graft. Reconstruction of the chest wall following resection of the sternum, described in this report, allowed us to perform radiotherapy and/or chemotherapy without serious postoperative complications on the cases relapsing after treatment of breast cancer. The 2-year survival rate is 50% and one of these cases survived up to 10 years after resection of the sternum. Thus we prefer to perform resection of the sternum for sternal recurrence of breast cancer if there are no metastatic lesions in other organs.

Frequency of steroid sulfatase deficiency in Hiroshima.

A retrospective survey was performed between 1983 and 1995 to determine the frequency of steroid sulfatase (STS) deficiency in Hiroshima. Males with ichthyosis were diagnosed enzymatically. During 1979-95 in Hiroshima Prefecture, 275,943 males were born and 28 had STS deficiency. The observed frequency of STS deficiency was 1 per 9855 males. Therefore, STS deficiency is fairly prevalent in Japan.

Allergic disease as an association of steroid sulphatase deficiency.

Ten of 31 patients with steroid sulphatase (STS) deficiency were found to have an allergic disease (bronchial asthma, allergic rhinitis, or atopic dermatitis). STS deficiency may predispose patients to allergic disease.

Enzymatic diagnosis of steroid sulfatase deficiency by high performance liquid chromatography.

We established a reversed phase high performance liquid chromatographic (HPLC) method of assaying lymphocyte steroid sulfatase activity using estrone sulfate as the substrate. Application of this method for diagnosis of 8 patients allowed us to clearly distinguish the patients from the normal controls. This method is simpler and less expensive than the method previously reported, since neither radioisotope labeled substrates nor radioisotope facilities are required. We consider it to be easily used and widely available in most clinical laboratories.

Anatomical investigations of the cutaneous branches of the circumflex scapular artery and their communications.

The vascular anatomy of the cutaneous branches of the circumflex scapular artery and their communications with perforators supplying adjacent territories are described, based on dissections of 20 scapular regions. The cutaneous branches of the circumflex scapular artery are distributed in three areas by the ascending, horizontal and descending branches. The existence of the ascending branch was constant, confirming the reliability of the ascending scapular flap.

Pedunculated club-shaped swelling in the nostril.

This paper presents a skin mass found in the nostril of a boy who had no other anomalies. The skin mass was similar to the pedunculated masses with median cleft that have been reported previously. The literature is reviewed, and the relationship between skin masses associated with true median cleft and the skin mass in our patient is discussed.

[Three infants with severe myocardial ischemia diagnosed by echocardiography: investigation of the cause of transient myocardial dysfunction].

We examined 3 infants with persistent pulmonary hypertension. They also showed signs of heart failure. The signs of cardiac dysfunction noted in the acute phase were resolved after treatment with catecholamines and vasodilators. Contrary to the general concept that transient myocardial dysfunction is secondarily caused by persistent pulmonary hypertension, left myocardial dysfunction accompanied pulmonary hypertension and was followed by right myocardial dysfunction. High blood concentration of thromboxane B2 was reported in 2 neonates with persistent pulmonary hypertension. We considered thromboxane A2 as a possible cause of coronary spasm, resulting in myocardial ischemia.

Effects of periodic administration of Nocardia rubra cell-wall skeleton on immunoglobulin production and B-cell-stimulatory factor activity in vitro in workers at a poison gas factory.

The former workers at the Okunojima poison gas factory (poison gas workers) are a high-risk group for malignant neoplasms and show abnormalities in cellular immunity. At the same time, poison gas workers often have chronic respiratory diseases, such as chronic bronchitis, and are highly susceptible to respiratory infections. To explore the possibility of immunological cancer prevention, we have periodically administered 200 micrograms Nocardia rubra cell-wall skeleton (N-CWS) to poison gas workers once every 3 months since December 1978. During this period, we noted a significantly lower incidence of influenza among poison gas workers receiving N-CWS than in those not receiving the drug during the influenza epidemic. This finding suggested that the administration of N-CWS enhanced the resistance of these workers to infections. Therefore, periodical administration of N-CWS to poison gas workers was considered to enhance the reduced T-cell function of normalizing antibody production by stimulating the production of B-cell-stimulatory factor (BSF). In the present study, to clarify the mechanism of immunosuppression in the poison gas workers and to examine the effects of continual administration of N-CWS on this condition, we compared the immunoglobulin production and the proliferative and differentiative activities of B-cell-stimulatory factor (BSF) of peripheral blood mononuclear cells (PBMC), in poison gas workers treated or not treated with N-CWS. Comparisons were also made with age-matched healthy controls. In the untreated poison gas workers, immunoglobulin and BSF production of PBMC were reduced as compared with the control group. On the other hand, in the poison gas workers receiving N-CWS, immunoglobulin and BSF production of PBMC were restored nearly to the control level. These results show that in vitro antibody production in the poison gas workers was reduced and that a reduction in BSF production of T cells was one of its causes.