Predominant Dissemination of PVL-Negative CC89 MRSA with SCCmec Type II in Children with Impetigo in Japan.

Background. The ratio of CA-MRSA in children with impetigo has been increasing in Japan. Methods. Antimicrobial susceptibilities of 136 S. aureus isolates from children with impetigo were studied. Furthermore, molecular epidemiological analysis and virulence gene analysis were performed. Results. Of the 136 S. aureus isolates, 122 (89.7%) were MSSA and 14 (10.3%) were MRSA. Of the 14 MRSA strains, 11 belonged to CC89 (ST89, ST91, and ST2117) and carried diverse types of SCCmec: type II (IIb: 3 strains; unknown subtype: 4 strains), type IVa (2 strains), and unknown type (2 strains). The remaining three strains exhibited CC8 (ST-8)-SCCmec type VIa, CC121 (ST121)-SCCmec type V, and CC5 (ST5)-nontypeable SCCmec element, respectively. None were lukS-PV-lukF-PV gene positive. Gentamicin- and clarithromycin-resistant strains were frequently found in both MRSA and MSSA. Conclusions. PVL-negative CC89-SCCmec type II strains are the most predominant strains among the CA-MRSA strains circulating in the community in Japan.

A monoclonal antibody that recognizes Epstein-Barr virus nuclear antigen 2 (EBNA 2) amino acids 1-58 does not react with EBNA 2 in native form, consistent with the self-association of EBNA 2 through the amino-terminus.

We have generated a mouse IgG1 monoclonal antibody (mAb) that recognizes amino acids 1-58 of Epstein-Barr virus (EBV) nuclear antigen 2 (EBNA 2) of type 1 EBV strain B95-8. mAb Y101 also reacted with EBNA 2 of EBV type 2 strains MISP and Jijoye in immunoblots, whereas Jijoye EBNA 2 was not detected by the widely used mAb PE2. mAb Y101, in contrast to mAb PE2, reacted with faster migrated, hypophosphorylated proteins of type 1 EBNA 2 as intensely as slower migrated, hyperphosphorylated ones. mAb Y101 did not react in fixed-cell immunostaining or cell extract immunoprecipitation. The results implicate that the amino-terminal epitope is not exposed in a native form, consistent with the previously reported idea of self-association of EBNA 2 through the amino-terminus. mAb Y101 is the first mAb to the EBNA 2 amino-terminus and will be useful for further analyses of the structure and function of EBNA 2.

Induction of interferon-inducible protein-10 and monokine induced by interferon-gamma from human endothelial cells infected with Influenza A virus.

Primary human umbilical vein endothelial cells (HUVECs) were infected with Influenza virus A/Aichi/2/68 (H3N2) in order to determine the role of endothelial cells in mediating inflammation induced upon virus infection. Structural proteins of the virus and mRNA of the M2 protein were detected in the infected cells, indicating that virus infection had occurred in HUVECs. The Influenza A virus-infected HUVECs showed elevated levels of gene expression of interferon (IFN)-inducible protein (IP)-10 and monokine induced by IFN-gamma (Mig), while heat-, formalin- and diethyl ether-inactivated viruses did not enhance the IP-10 and Mig gene expression. The results thus indicate that infection of live Influenza A virus is responsible for elevation of IP-10 and Mig gene expression. The elevation of IP-10 and Mig gene expression in infected HUVECs was not accompanied by the elevation of IFN-gamma gene expression, indicating that the elevation of IP-10 and Mig gene expression was independent of the IFN-gamma pathway.

Two cases with transient lipoprotein lipase (LPL) activity impairment: evidence for the possible involvement of an LPL inhibitor.

UNLABELLED: Two independent severe hypertriglyceridemic infants with transiently impaired lipoprotein lipase (LPL) activity were observed and the causes were explored. Both infants were female, born prematurely with low birth weight and developed hypertriglyceridemia (Fredrickson type V hyperlipidemia: high VLDL and low LDL/HDL) a few months after birth. While mass levels of their post-heparin plasma LPL and apoprotein C-II (apo C-II), a physiological activator of LPL, were normal, their post-heparin plasma LPL activities were remarkably impaired. Both of their mothers' post-heparin plasma LPL activities were slightly or moderately impaired as well, without a decrease in the LPL mass level. No mutations in the genes for LPL and apo C-II were detected in either patient. In an in vitro study with their serum at onset, we could not detect any distinct circulating inhibitors for LPL. There was no data supporting infection or autoimmune diseases, which might have an impact on LPL activity, during the follow-up period. Levels of their plasma triglyceride (TG) and total cholesterol (TC) were decreased quickly by a dietary intervention with medium-chain triglyceride (MCT) milk and kept normal even after stopping the intervention at around age 1 year. However, their low post-heparin LPL activity persisted and returned to normal at around age 2 years. Their low HDL cholesterol levels persisted even after recovery of the TG and TC levels, although lecithin:cholesterol acyltransferase (LCAT) and cholesterol-ester-transfer protein (CETP), two key enzymes of HDL metabolism, were normal throughout the course. The exact reasons why their post-heparin LPL activities were impaired for a certain period and why their HDL cholesterol levels have remained low are still unclear. CONCLUSION: Transiently impaired LPL activity with no defect in LPL enzyme induced severe hypertriglyceridemia in infants. The transient occurrence of inhibitor(s) for LPL was proposed.

No mutations of SAP/SH2D1A/DSHP and perforin genes in patients with Epstein-Barr virus-associated hemophagocytic syndrome in Japan.

Recently, mutations of two genes, SAP/SH2D1A/DSHP and perforin genes, have been identified in two fatal inherited lymphoproliferative diseases, X-linked lymphoproliferative disease and familial hemophagocytic lymphohistiocytosis, respectively. Epstein-Barr virus (EBV)-associated hemophagocytic syndrome, a fulminant non-inherited T-cell lymphoproliferative disease, is relatively common in Japan and is extremely difficult to distinguish from X-linked lymphoproliferative disease and familial hemophagocytic lymphohistiocytosis, especially in sporadic cases, because of similarities in clinical and laboratory features. Mutation analysis was carried out of samples obtained from 14 patients with EBV-associated hemophagocytic syndrome by sequencing the genomic SAP/SH2D1A/DSHP and perforin genes. However, a specific mutation was not identified in either of the genes, suggesting that mutations of the SAP/SH2D1A/DSHP and perforin genes are not responsible for the pathogenesis of EBV-associated hemophagocytic syndrome in Japan.

Nonpolarizing guided-mode resonant grating filter for oblique incidence.

A new type of guided-mode resonant grating filter is described. The filter is independent of polarization state for oblique incidence. The filter has a crossed grating structure, and the plane of incidence on the filter contains the symmetric axis of the grating structure. Theoretical considerations and numerical calculations using two-dimensional rigorous coupled-wave analysis show that a rhombic lattice structure is suitable to such filters. In this configuration an incident light wave is diffracted into the waveguide and is divided into two propagation modes whose directions are symmetric with respect to the plane of incidence. In particular, when the propagation directions of the two modes are perpendicular to each other, the fill factor of grating structure can be approximately 50%. The filter was designed for an incident angle of 45 degrees. Tolerances of setting errors and fabrication errors for this filter were estimated by numerical calculations.

Numerical feasibility study of the fabrication of subwavelength structure by mask lithography.

Electromagnetic diffraction of a light wave by a single aperture of subwavelength width and subsequent propagation in a lossy medium are numerically investigated. This diffraction problem simulates exposure of a resist with an amplitude mask. It is found that there is the possibility of fabricating a lambda/2 structure on a resist of lambda/4 thickness, where lambda is the wavelength of the exposing light in vacuum, by conventional contact or by proximity lithography. It is also found that an air gap between a mask and a resist of up to lambda/2 does not have a significant effect on resolution. This approach permits easy and cost-effective fabrication of subwavelength structures and leads to wide availability of diffractive optical elements in the nonscalar domain.

T-cell lines from 2 patients with adenosine deaminase (ADA) deficiency showed the restoration of ADA activity resulted from the reversion of an inherited mutation.

Inherited deficiency of adenosine deaminase (ADA) results in one of the autosomal recessive forms of severe combined immunodeficiency. This report discusses 2 patients with ADA deficiency from different families, in whom a possible reverse mutation had occurred. The novel mutations were identified in the ADA gene from the patients, and both their parents were revealed to be carriers. Unexpectedly, established patient T-cell lines, not B-cell lines, showed half-normal levels of ADA enzyme activity. Reevaluation of the mutations in these T-cell lines indicated that one of the inherited ADA gene mutations was reverted in both patients. At least one of the patients seemed to possess the revertant cells in vivo; however, the mutant cells might have overcome the revertant after receiving ADA enzyme replacement therapy. These findings may have significant implications regarding the prospects for stem cell gene therapy for ADA deficiency.

Successful living-donor liver transplantation from an asymptomatic carrier mother in ornithine transcarbamylase deficiency.

A liver transplantation from an asymptomatic mother, who was a carrier of ornithine transcarbamylase deficiency, to her daughter, who had severe manifestation, was successfully performed. One-year monitoring of plasma amino acid and urinary orotate/orotidine levels revealed no abnormality in the urea cycle in either subject.

Unusual presentation of measles giant cell pneumonia in a patient with acquired immunodeficiency syndrome.

The typical clinical presentation of measles in a normal immunocompetent host includes cough, coryza, conjunctivitis, Koplik's spots, and rash. However, in an immunocompromised host, measles may have an atypical clinical presentation and may be commonly associated with severe pneumonia or encephalitis. We report a fatal case of measles pneumonia without any clinical features that suggest measles in a patient with acquired immunodeficiency syndrome.

Antireflection microstructures fabricated upon fluorine-doped SiO(2) films.

Two-dimensional periodic structures were fabricated upon a fluorine-doped SiO(2) film in which the fluorine content changed gradually in the direction of film thickness. The films were deposited by plasma-enhanced chemical-vapor deposition. The film was periodically patterned into a 1-mum period and an ~1-mum -groove depth by inductive coupled plasma reactive-ion etching followed by chemical etching in a diluted HF solution. A surface reflectance of 0.7% was attained at 1.85-mum wavelength, a value that is one fifth as large as the 3.5% Fresnel reflection of a SiO(2) substrate with a flat surface.

Lipase production in two-step fed-batch culture of organic solvent-tolerant Pseudomonas aeruginosa LST-03.

Efficient lipase production by two-step fed-batch culture of an organic solvent-tolerant bacterium, Pseudomonas aeruginosa LST-03, was investigated. When FB synthetic medium was used in flask culture, no lipase activity was detected, whereas lipase was produced at 2.3 I.U./ml in C2 complex medium. However, lipase production was induced in FB medium when a fatty acid was added to the culture broth in the stationary phase. Among fatty acids tested, long chain saturated fatty acids, such as C18 (stearic acid) and C20 (arachidic acid), were found to function as effective inducers for the production of lipase, giving an activity level almost the same as that obtained in C2 medium in flask culture. Two-step lipase production, comprised of a growth phase in fed-batch mode and a production phase in which lipase was induced by the addition of 5% (v/v) stearic acid, was carried out in a jar-fermentor. In the growth phase, the maximum cell concentration at 16 h was only 20 in terms of the optical density at 660 nm (OD660), and a low level of lipase production (8 I.U./ml) was obtained after 167 h. This was considered to be due to the exhaustion of several medium components brought about by the use of an unsuitable medium or feeding solution. After analyzing the contents of the compounds in the culture broth by inductively coupled plasma spectrometry for metal ions and HPLC for anions, a modified FB medium was designed. When this modified FB medium was used in two-step fed-batch culture, the maximum cell concentration reached an OD660 of 55 (30.2 g-dry cells/l) at 16.5 h, and lipase was produced at 96 I.U./ml after 35 h, which is approximately 40 times higher than the production level obtained in flask culture using C2 medium.

Eyelid abnormalities in lamellar ichthyoses.

PURPOSE: To describe eyelid abnormalities in lamellar ichthyoses (LI). DESIGN: Retrospective observational case series. PARTICIPANTS: Eight patients with classic LI and two patients with congenital ichthyosiform erythroderma. METHODS: Results of eyelid and corneal examinations of 10 patients with LI were reviewed and analyzed with emphasis on the relationship between eyelid ectropion and corneal damage. RESULTS: All patients presented with cicatricial lagophthalmos. Of the eight patients with classic LI, five had ectropion of the four eyelids, one had only lower ectropion, and two had no degree of ectropion. Two patients with congenital ichthyosiform erythroderma had distinct eyelid abnormalities, including madarosis and eyelash retraction. Loss of vision caused by corneal damage was found in three patients with classic LI. Of these three patients, two did not have upper eyelid ectropion. CONCLUSIONS: Severe corneal damage can occur in LI even if there is no upper or lower eyelid ectropion.

Reconstitution of humoral immunity during pregnancy.

BACKGROUND: Recently, increasing attention has been paid to hormonal regulations of the immune system. MATERIALS AND METHODS: In this study, cord sera and the corresponding maternal sera were obtained at delivery. Sera from pregnant women were obtained at early, middle, and late stages of pregnancy. These sera were tested for titer and avidity of measles or mumps virus-specific immunoglobulin G (IgG) by means of a single-dilution, urea-denaturation enzyme-linked immunosorbent assay method. RESULTS: A positive and significant correlation was found between the titer and avidity of the virus-specific IgG, both in the cord sera and in the maternal sera. This correlation was established already at the early stage of pregnancy. There were no such correlations found in nonpregnant individuals. CONCLUSIONS: This is the first observation in human subjects that the avidity and concentration of the virus-specific IgG had a positive and significant correlation. Pregnancy must have some significant effects on the regulation of humoral immunity.

Detection of TT virus sequences in children with liver disease of unknown etiology.

DNA sequences of TT virus (TTV) in 55 serum samples taken from 20 children with liver disease of unknown etiology (16 with acute liver disease, 2 with fulminant hepatitis, and 2 with chronic liver disease) and from 35 healthy children as controls were examined by using the hemi-nested polymerase chain reaction (PCR). The PCR was carried out using the established primers (NG059, NG061, NG063) to amplify TTV DNA sequences. The sequences were detected in 6 of the 20 patients (30.0%) with liver disease and in 5 of the 35 healthy children (14.2%) by direct gel analysis. There was no significant difference between the prevalence of liver disease patients and controls. However, both patients with fulminant hepatitis and both patients with chronic hepatitis had TTV DNA sequences. Four of the six TTV isolates from liver disease patients were genotype 1a, whereas only one of the five TTV isolates from controls was genotype 1a. Although the study population was small, it is possible that genotype 1a of TTV might be more pathogenic than other genotypes in children.

Detection of a novel DNA virus (TTV) sequence in peripheral blood mononuclear cells.

DNA sequences of a novel DNA virus (TTV) were examined in 81 peripheral blood mononuclear cell (PBMC) DNA samples from 48 children and 33 adults, 22 cord blood mononuclear cells (CBMC) DNA samples, and 7 autopsy liver tissue DNA samples by a hemi-nested polymerase chain reaction (PCR). The PCR was carried out using the published primers (NG059, NG061, NG063) to amplify TTV DNA sequences. The sequences were detected in 4 of 81 (5%) PBMC DNA samples, in none of 22 (0%) CBMC DNA samples, and in 2 of 7 (29%) liver tissue DNA samples by direct gel analysis. The PCR-amplified products were confirmed by direct sequencing. The sequencing showed considerable diversities, with differences of 0-55% in 6 TTV isolates, compared with the prototype sequence of TTV. These results suggest that TTV is a ubiquitous virus that produces asymptomatic infection in a large proportion of the general population without transfusion of blood-derived products. To our knowledge, this is the first report describing the detection of TTV DNA sequences in PBMCs.

Nasal and nasal-type T/NK-cell lymphoma with cutaneous involvement.

Natural killer (NK) cells are a third lymphocyte lineage, in addition to B- and T-cells, that mediate cytotoxicity without prior sensitization. NK cells also have phenotypic and genotypic characteristics; they express the NK-related antigen CD56 and T-cell markers such as CD2 and CD3 epsilon, but their T-cell receptor (TCR) locus is not rearranged. Non-Hodgkin's lymphomas are divided into B- and T-cell neoplasms and NK-cell lymphomas. We describe 2 Japanese patients with nasal and nasal-type T/NK-cell lymphoma in which the skin, nasal/nasopharyngeal region, bone marrow, and lymph node were the sites of involvement. The clinical and histopathologic findings were recorded. In addition, immunophenotyping, TCR gene rearrangement, and the existence of Epstein-Barr virus (EBV) DNA by polymerase chain reaction amplification were determined. Clinically, the cutaneous eruptions were purplish, hard, multiple nodules. Histologically, angiocentric proliferation of small-to medium-sized, pleomorphic, lymphoid cells were observed. They revealed hand-mirror-shaped lymphocytes with azurophilic granules with the use of Giemsa staining by touch smear. These lymphocytes were found to be positive to immunophenotyping for CD2 (Leu5b), CD3 epsilon (DAKO), CD4 (Leu3a), and CD56 (Leu 19). No clonal rearrangement of TCR-beta, -gamma, and -delta genes and immunoglobulin gene markers were found, and no positive results of identification of EBV DNA were shown. The patients underwent cyclophosphamide, doxorubicin, vincristine, and prednisone chemotherapy with complete remission; however, both had recurrence of disease. Because NK-cell lymphomas express some T-cell markers, they may be mistakenly diagnosed as peripheral T-cell lymphomas if they are not investigated for the NK-cell-specific marker, CD56. Therefore the importance of immunophenotypic investigations of CD56 should be stressed. Also, the importance of clinical investigation of nasal/nasopharyngeal lymphomas should be stressed when NK-cell lymphoma is diagnosed involving the skin, because NK-cell lymphomas are often associated with the nasal and nasopharyngeal region.