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Animal models of human diseases are invaluable and inevitable elements in identifying and testing novel treatments for serious diseases, including severe infections. Planning and conducting investigator-initiated human trials are generally accepted as being enormously challenging. In contrast, it is often underestimated how much planning, including background and modifying experiments, is needed to establish a relevant infectious disease animal model. However, representative animal infectious models, well designed to test generated hypotheses, are useful to improve our understanding of pathogenesis, virulence factors and host response and to identify novel treatment candidates and therapeutic strategies. Such results can subsequently proceed to clinical testing if suitable. The present review aims at presenting all the pulmonary Pseudomonas aeruginosa infectious models we have knowledge of and the detailed descriptions of established animal models in our laboratory focusing on macrolide therapy are presented.

A 47-year-old man with ulcerative colitis was transferred to our hospital due to progressive dyspnea. Electrocardiography on admission showed ST elevation in leads II, III, aVF, and V5-V6. Coronary angiography revealed no remarkable coronary stenosis, and left ventriculography showed a depressed left ventricular ejection fraction (EF) of 23%. Although the patient received percutaneous cardiopulmonary support, his EF progressively decreased (7-15%), and both ventricular tachycardia (VT) and high-degree atrial-ventricular block occurred. An endomyocardial biopsy showed eosinophilic infiltration in the myocardium. Steroid therapy improved the patient's EF. However, his severe inferior wall hypokinesis and non-sustained VT remained after the abovementioned treatment.

Some patients with insulinoma present with neuropsychiatric symptoms and are often misdiagnosed with psychiatric disease. We present the case of a 72-year-old Japanese female who exhibited violent behavior while asleep and received a diagnosis of suspected rapid eye movement sleep behavior disorder (RBD). She was admitted to the psychiatry ward after receiving levomepromazine 25 mg intramuscularly. The patient's blood glucose level was 27 mg/dL at the time of hospitalization, and a biochemical examination revealed that her insulin level was 9.1 µU/mL and C-peptide level was 2.16 ng/mL. A contrast-enhanced computed tomography revealed a mass 8 mm in diameter in the pancreatic head. The diagnosis was changed from RBD to insulinoma. The sleep behavior disorder disappeared after continuous glucose administration. After enucleation of the insulinoma, the administration of glucose was discontinued, and her blood glucose levels recovered. This case suggests that insulinoma should be considered by physicians and psychiatrists in the differential diagnosis of patients with symptoms presenting as RBD.

Aim Pneumonia in older adults is increasingly recognized as a healthcare issue in countries with an aging population. Long-term macrolide therapy reduces exacerbations of chronic respiratory diseases, but its effects on the prevention of pneumonia have not been determined. Methods We carried out a randomized, controlled trial to test the effect of long-term clarithromycin therapy on the prevention of pneumonia among older adults. People aged ≥65 years who had recovered from pneumonia within the previous 3 months were recruited and randomly allocated to a long-term, low-dose clarithromycin (CAM) therapy group (n = 13) or a control group (n = 15). Results Both groups were followed up until recurrence of pneumonia. The median follow-up period was 251 days (95% CI 171-330) in the CAM group and 132 days (95% CI 67-196) in the control group (P = 0.627). The recurrence rate of pneumonia was two out of 13 (15%) in the CAM group and five out of 15 (33%) in the control group (P = 0.268). The median time to recurrence of pneumonia was 315 days (95% CI 249-382) in the CAM group and 260 days (95% CI 184-335) in the control group (P = 0.260). None of the differences between groups were statistically significant. Conclusions No statistically significant suppressive effects of long-term, low-dose macrolide therapy on the development of pneumonia among older people were found in this small sample. A large-scale, randomized, controlled study is required. Geriatr Gerontol Int 2019; 19: 1006-1009.

Pseudomonas aeruginosa exhibits the biofilm mode of growth and causes chronic as well as acute infections in humans. Several reports have shown that the treatments with sub-minimum inhibitory concentrations (sub-MICs) of antimicrobial agents influence biofilm formation by P. aeruginosa. The antibiotic ceftazidime (CAZ) is used to treat P. aeruginosa infections, but few studies have examined the effects of β-lactams on biofilm formation by P. aeruginosa. In this study, we investigated the role of sub-MICs of CAZ in the formation of P. aeruginosa biofilms. 1/4 × MIC CAZ reduced the biofilm volume of P. aeruginosa PAO1, as quantified by crystal violet staining. The formation of P. aeruginosa PAO1 biofilms treated with 1/4 × MIC CAZ were observed by confocal laser scanning microscopy. They were more heterogeneous than the PAO1 biofilms without CAZ treatment. Furthermore, sub-MICs of CAZ inhibited the twitching motility, which played an important role in mature biofilm formation. 1/4 × MIC CAZ also reduced the gene expressions of lecA, lecB, pel and psl, which mediate the adhesion and polysaccharide matrix synthesis of P. aeruginosa. These effects suggest that sub-MICs of CAZ may affect a number of stages of biofilm formation. Investigating the effects of sub-MIC antibiotics on targeted bacterial biofilm may lead to the development of future antibiotic treatment modalities.

In order to clarify the relationship between myelodysplastic morphologic features of marrow cells and prognoses and 1.0 define other prognostic factors, 124 patients with the FAB criteria of myelodysplastic syndrome (MIX) were analysed. These included 57 patients with refractory anemia (RA), 5 RA with ring sideroblasts (RARS), 25 RA with excess of blasts (RAEB), 14 chronic myelomonocytic: leukemia (CMML) and 23 with RAEB in transformation (RAEB in T). Univariate analysis of all MDS patients or those of RA demonstrated that the following factors, which were not reported or fully investigated previously, were significantly associated with prognosis. These included neutrophil alkaline phosphatase (NAP) score (significant only for all MDS), the percentage of marrow erythroblasts and lymphocytes present, the percentage of cells with morphological abnormalities in individual cell lineages and the number of cell lineages showing atypia (significant for all MDS and RA). Multiple regression analysis showed that (%) of marrow erythroblasts, NAP score, hemoglobin levels and number of marrow granulocytes with atypia were significant for predicting the prognosis of all MDS patients while the number of marrow megakaryocytes and granulocytes with atypia were significant for prognosis in the subgroup with RA.

Data on 225 Japanese patients with primary or essential thrombocythemia (ET) were analyzed in an attempt to characterize the clinical and laboratory features in subgroups with thrombosis (T), hemorrhages (H), thrombohemorrhagic events (TH) or a non-thrombohemorrhagic (O) group, and in order to examine survival and the incidence of blastic transformation in the entire group and in the different subgroups. Higher platelet and leukocyte counts were related to hemorrhage (H and TH), prolonged activated partial thromboplastin times and high LDH levels to H while elevated FDP levels were more frequently linked to T. Increased spontaneous platelet aggregation (SPA) was noted in 80.3% of the entire group, independent of whether there was a tendency for thrombohemorrhagic events or not. Bleeding time, as measured by the Duke method, and hemoglobin levels were not different in the various subgroups. Transformation occurred in 11 patients (1.9% per year); seven developed acute leukemia (myeloblastic 4, lymphoblastic 2, megakaryoblastic 1) at a rate of 1.2% per year; and 4 developed other types of chronic myeloproliferative disorders. Nineteen patients died (3.3% per year), six from leukemia (32%), 4 from bleeding (21%) and 9 from unrelated diseases (47%). Survival was estimated to be 65% at ten years, and was significantly longer in females, younger individuals, and the groups with lower leukocyte counts, but did not differ between the subgroups when platelet count and hemoglobin level were considered. Survival was similar in patients with platelet counts between 700-1000 × 10(9)/L and in those with an even higher platelet count. These findings suggest that (1) young female patients with low leukocyte counts may survive longer, (2) SPA is not indicative of either a thrombotic or an hemorrhagic tendency and (3) the limit of the platelet count for establishing the diagnosis of this disorder could perhaps be lowered to 700 × 10(9)/L.

Summary Statins are 3-hydroxy-3-methylglutaryl-co-enzyme A reductase inhibitors of cholesterol biosynthesis, and have been reported to exert pleiotropic effects on cellular signalling and cellular functions involved in inflammation. Recent reports have demonstrated that previous statin therapy reduced the risk of pneumonia or increased survival in patients with community-acquired pneumonia. However, the precise mechanisms responsible for these effects are unclear. In the present study, we examined the effects of statins on cytokine production from lipopolysaccharide (LPS)-stimulated human bronchial epithelial cells (BEAS-2B). Interleukin (IL)-6 and IL-8 mRNA expression and protein secretion in LPS-stimulated cells were inhibited significantly by the lipophilic statin pitavastatin and the hydrophilic statin pravastatin. As these inhibitory effects of statin were negated by adding mevalonate, the anti-inflammatory effects of statins appear to be exerted via the mevalonic cascade. In addition, the activation levels of Ras homologue gene family A (RhoA) in BEAS-2B cells cultured with pitavastatin were significantly lower than those without the statin. These results suggest that statins have anti-inflammatory effects by reducing cytokine production through inhibition of the mevalonic cascade followed by RhoA activation in the lung.

Background and objective: More than 100 000 Japanese die of pneumonia every year. The number of people residing in nursing homes is increasing with the ageing of the population. In 2005, the American Thoracic Society/Infectious Diseases Society of America (ATS/IDSA) published important guidelines for the management of healthcare-associated pneumonia (HCAP). In Japan, however, the optimum strategy for management of HCAP is still unclear. The purpose of this study was to clarify the clinical features of patients with HCAP. Methods: Patients (n = 202) who were consecutively admitted with a diagnosis of acute pneumonia between October 2007 and September 2009 were retrospectively evaluated. Using the ATS/IDSA guidelines, patients were divided into three groups: a community-acquired pneumonia (CAP) group (n = 123), a nursing home-acquired pneumonia (NHAP) group (n = 46) and a HCAP other than NHAP (O-HCAP) group (n = 33). These groups were then compared with respect to laboratory data, microbiological findings and mortality. Results: Thirty-day mortality in the NHAP group (10.9%) tended to be higher than that in the CAP group (3.3%) or the O-HCAP group (0%). The pathogens most frequently identified were Streptococcus pneumoniae and Haemophilus influenzae in the CAP group, methicillin-resistant Staphylococcus aureus and Klebsiella pneumoniae in the NHAP group, and S. pneumoniae and K. pneumoniae in the O-HCAP group. Conclusions: The NHAP group was clinically different from the O-HCAP group, based on bacteriological examination and mortality rates. In order to accurately diagnose, and formulate optimum treatment strategies for Japanese patients, the categories of HCAP, as specified in the ATS/IDSA guidelines, should not be applied directly either to patients with NHAP or those with O-HCAP.

This retrospective analysis investigated the prognostic value of monitoring the response to imatinib using peripheral blood (PB) samples and the impact of the response on outcome in 133 patients with chronic myeloid leukemia (CML). We divided the response into 3 categories according to the results of neutrophil (N)-FISH and BCR-ABL transcript levels in PB; more than a 3-log reduction [major molecular response (MMR)], between a 2-log and 3-log reduction or negative with N-FISH [complete cytogenetic response equivalent (CCyRe)], N-FISH positive or less than a 2-log reduction (non-CCyRe). The median follow-up was 5.46 years. At 5 years, the overall survival (OS) rate and progression-free survival (PFS) rate were 94.4 and 92.0%, respectively. The estimated rate of the CCyRe and MMR were 81.7 and 67.1%, respectively. 106 patients achieving the CCyRe had significantly better OS and PFS than 27 patients without achieving the CCyRe. Patients with MMR had significantly better survival free from death, progression, imatinib withdrawal and a loss of the CCyRe, than patients whose response level remained in the CCyRe without achieving MMR until 18 months. Our observation suggests that the response level of the CCyRe on PB serve as a prognostic indicator, and achieving MMR provides stable long-term survival.

The chemokinetic inhibitory factor (CIF) is a recently described B-cell derived lymphokine that mediates a chemokinetic inhibitory effect on human polymorphonuclear leukocyte (PMN) migration. In the present report the interaction of CIF with the neutrophil plasma membrane was studied. Normal human peripheral blood neutrophils and purified neutrophil plasma membranes selectively removed biologic activity from CIF-containing concentrates obtained during the purification procedure from conditioned medium. Removal was obtained at both 4 degrees C and 37 degrees C. Furthermore, HL-60 cells treated with dimethyl sulfoxide removed CIF activity (granulocyte-like cells) but HL-60 cells treated with 12-O-tetradecanoylphorbol-13-acetate (macrophage-like cells) did not. Purified human blood monocytes, cells from the macrophage-like U-937 cell line and cells from the basophilic leukemia cell line KU-812 did not remove CIF. These studies suggest that neutrophils express specific binding sites for CIF-activity.

Multidrug-resistant Pseudomonas aeruginosa, especially metallo-beta-lactamase (MBL)-producing P. aeruginosa, is an important pathogen in nosocomial infection and emergence of this pathogen has revived interest in polymyxin B (PMB) and colistin (COL). In this study, we evaluated the efficacies of PMB, COL and other antipseudomonal agents against IMP-type MBL-producing P. aeruginosa both in vitro and in vivo. A total of 75 isolates of bla(IMP)-positive P. aeruginosa obtained from clinical specimens (94.6% of isolates demonstrated resistance to beta-lactam, fluoroquinolone and aminoglycoside agents) were evaluated in the in vitro study. More than 90% of the examined isolates were susceptible to PMB (minimum inhibitory concentration for 50/90% of the isolates (MIC(50)/MIC(90)) 4/4 mg/L), although COL was less potent (MIC(50)/MIC(90) 8/16 mg/L). Cyclophosphamide-treated mice were intraperitoneally inoculated with bla(IMP)-positive P. aeruginosa. Treatment with PMB, but not COL, imipenem/cilastatin or aztreonam, significantly improved the survival rate and decreased the number of bacteria in the blood in a dose-dependent manner. Our results indicate that, among the agents studied, PMB is the most effective agent against bla(IMP)-positive P. aeruginosa.

Neutropenic mice with latent trichosporonemia were given various antifungal agents (amphotericin B, fluconazole, itraconazole) or saline to determine which antifungal agent could be useful for prophylaxis. The 3-week-survival rate was 80% in the fluconazole group, 50% in the amphotericin B group, 45% in the itraconazole group, and 30% in the saline group. Compared with the other antifungal agents, fluconazole offered superior prophylaxis against the progression of trichosporonosis fungemia to disseminated disease (P

We clarified experimentally that an EA-DFB laser in which the EAM and DFB parts are coupled with a passive waveguide provides a large bandwidth, even when a large reverse bias is applied to the EAM. This allows the chip to operate in a negative-chirp condition. We fabricated an EA-DFB module terminated with a 50 Omega resistor through a dc-block capacitor to suppress the increase in dc current as the reverse bias increases. A module equipped with our new EA-DFB chip successfully transmitted data over a distance of 3 km at a rate of 40 Gbit/s and at a driving voltage as low as 2 V.

Rationale and Objectives Nocardiosis is difficult to diagnose, and the diagnosis is thus frequently delayed. High-resolution computed tomography (HRCT) findings of patients with pulmonary nocardiosis have been documented in few reports. Our study objective was to assess HRCT findings of patients with pulmonary nocardiosis. Materials and Methods This was a retrospective study of 20 consecutive patients with pulmonary Nocardia infections who underwent HRCT of the chest at our institutions from January 2011 to August 2014. After the exclusion of two patients with concurrent infections, the study group comprised 18 patients (11 men, 7 women; age range, 39–83 years; mean, 67.9 years) with pulmonary Nocardia infections. Parenchymal abnormalities, enlarged lymph nodes, and pleural effusion were evaluated on HRCT. Results Underlying conditions included respiratory disease ( n = 6, 33.3%), collagen diseases ( n = 5, 27.8%), and diabetes mellitus ( n = 4, 22.2%). All patients showed abnormal HRCT findings, including the presence of a nodule/mass ( n = 17, 94.4%), ground-glass opacity ( n = 14, 77.8%), interlobular septal thickening ( n = 14, 77.8%), and cavitation ( n = 12, 66.7%). Pleural effusion was seen in two patients. There were no cases of lymph node enlargement. Conclusions Among the HRCT findings in patients with pneumonia, a nodule/mass with interlobular septal thickening and/or cavitation are suggestive of pulmonary nocardiosis.

SummaryDiffuse panbronchiolitis (DPB) is a distinctive form of small airway disease, which is characterized by chronic inflammation with lymphocyte infiltration around bronchioles. The aim of this study was to evaluate the importance of factors related to apoptosis in peribronchiolar lymphocytes of DPB. We employed immunohistochemical methods for the localization of Bax (a promoter of apoptosis), Bcl-2 (an inhibitor of apoptosis), and caspase-3 (a key executioner molecule of apoptosis) in lung tissues of five patients with DPB. In all patients, immunostaining for Bax was almost completely absent in accumulated lymphocytes around the bronchioles and in lymphocytes of the parafollicular area that correspond to a zone populated by T cells. In contrast to the reaction for Bax, Bcl-2 immunoreactivity was uniformly strong in all of the patients. The pattern of staining for caspase-3 was similar to that for Bax in all of the patients. In normal lung tissue, a few lymphocytes showed negative immunostaining for Bcl-2 and a positive reaction for caspase-3. Our results suggest that Bcl-2 protein may provide T-lymphocyte survival and hypercellularity in the bronchioles, thereby contributing to the progression of DPB.

We examined the protective effect of intratracheal immunization with Pseudomonas aeruginosa pili protein against respiratory infection caused by P. aeruginosa. Mice were immunized intratracheally or subcutaneously with purified pili protein or bovine serum albumin as a control. Intratracheally but not subcutaneously pili protein-immunized mice showed significant improvement of survival after intratracheal challenge with the PAO1 strain. Furthermore, bacterial cell counts in pili protein-immunized murine lungs were significantly decreased compared to controls at 18 h after the challenge. Antipili protein antibody titers in bronchoalveolar lavage fluid of intratracheally pili protein-immunized mice were higher than in bovine serum albumin immunized mice. However, antipili antibody titers were not increased in bronchoalveolar lavage fluid of subcutaneously pili protein-immunized mice, despite the high serum antipili antibody titers. Inoculation of P. aeruginosa induced immediate increases in interleukin-12 and interferon-gamma in bronchoalveolar lavage fluid of pili protein-immunized mice, reflecting an adequate and rapid immune response against P. aeruginosa respiratory tract infection. Our findings suggest that intratracheal pili protein immunization is effective against respiratory tract infection caused by P. aeruginosa in mice.

Long-term administration of macrolide antibiotics reduced the number of lymphocytes in bronchoalveolar lavage fluid in patients with chronic airway inflammatory disease. To evaluate the inflammatory activity of macrolides, their effect on apoptosis of activated lymphocytes isolated from human peripheral blood was compared with that of other antibiotics. Macrolides, including clarithromycin and azithromycin, at a final concentration of 100 μg/ml accelerated apoptosis of activated lymphocytes, while other antibiotics such as fosfomycin sodium, β-lactams — ceftazidime, piperacillin sodium and biapenem, and a quinolone, ofloxacin, did not cause significant induction of apoptosis. Our results suggest that 14- or 15-membered ring macrolides are specifically involved in the augmentation of apoptosis of activated lymphocytes, and this may be of value therapeutically for chronic airway diseases.

Objective In the current studies, we investigated the clinical effects of long-term macrolide antibiotic therapy for patients with chronic small airway disease (CAD) that clinically and radiologically mimics but is pathologically distinct from diffuse panbronchiolitis (DPB).Patients and Methods Twenty-one Japanese patients were selected on the basis of clinical criteria for DPB and were categorized as DPB or CAD following histological evaluation of surgical lung biopsies. All patients received long-term macrolide therapy, and therapeutic results were compared for the DPB and CAD groups.Results Clinical, laboratory, radiological, and bacterial features, as well as neutrophilia in bronchoalveolar lavage fluid were strikingly similar in both groups. Long-term treatment with macrolides improved the clinical symptoms and PaO2 in both groups. There was a significant improvement in forced expiratory volume in one second (FEV1), vital capacity (VC), and %VC in patients with DPB but not in patients with CAD. Neutrophilia in bronchoalveolar lavage fluid was also reduced following therapy in DPB patients but was refractory in CAD patients.Conclusion Based on the different responses to macrolides, CAD might be associated with conditions distinct from those of DPB. Nevertheless, low-dose macrolide therapy may be applied in CAD to achieve clinical improvement, such as in respiratory symptoms and PaO2.

Solid solutions of BaLaIn2O5.5 and BaLaInMO5.5 that formed by partial replacement of In by Sc and Ga have a cubic perovskite structure. The Sc3+- and Ga3+-doped compounds, which had almost the same electrical conductivities, exhibited lower conductivities than the parent compound BaLaIn2O5.5 in the temperature range from 573 to 1273K. The Ga compound showed a transference number, ti, of unity at both high and low oxygen partial pressures in the temperature range from 973 to 1273K. As for the Sc compound kept at 973-1273K, a ti of nearly unity was observed at low oxygen partial pressure, while the ti was slightly lower than unity at high oxygen partial pressures at higher temperatures. The In compound had a ti of almost unity at high oxygen partial pressure, whereas the number decreased to a value less than 0.9 at low partial pressures of oxygen. The stepwise increase in the Sc content from 0.1 to 1.0 in BaLaIn2-xScxO5.5 caused a monotonic decrease in conductivity. The decrease in conductivity and the increase in activation energy caused by Sc3+ and Ga3+ doping were ascribed to the partial ordering of originally disordered oxygen vacancies and a decrease in the size of the triangle formed by two A-site cations and one B-site cation, through which the oxide ion proceeds, respectively.

Suppression of endotoxin release and subsequent production of inflammatory cytokines is crucial in the treatment of septic shock. We investigated the effect of clindamycin (CLI) on endotoxic shock induced in mice byEscherichia colilipopolysaccharide (LPS). Mice were treated with CLI (160 to 600 mg/kg) or saline and then injected withE. coliLPS andd-(+)-galactosamine intraperitoneally 0.5 h after CLI administration. Pretreatment with CLI significantly improved survival in a dose-dependent manner (CLI, at 160, 300, and 440 mg/kg) and significantly lowered the peak concentrations of tumor necrosis factor alpha and interleukin-1β(IL-1β) in serum. However, the peak concentrations of IL-6 in the sera of CLI-treated mice were higher than in control mice. Our findings suggest that CLI alters LPS-induced inflammatory cytokine production and suppresses endotoxin-induced mortality in this murine model.

In this paper, we present highly reliable high-temperature spot-size converter integrated laser diodes (LDs) fabricated using a full-wafer process with dry etching and MOVPE. The lasers lase at 150°C and operate stably for 10,000 h at 85°C and 10 mW. The median lifetime is estimated to be over 10 5 h. We also demonstrate a novel screening test procedure for a bar configuration of LDs, which can greatly reduce optical module cost.

We demonstrate 1.55-/spl mu/m buried-heterostructure (BH) vertical-cavity surface-emitting lasers (VCSELs) on a GaAs substrate. Thin-film wafer-fusion technology enables InP-based BH VCSELs to be fabricated on GaAs/AlAs distributed Bragg reflectors. Detailed investigations of the device resistance are also described. As a result of introducing BH and obtaining low device resistance, the threshold current density under CW operation shows the independence of mesa size due to a strong index guide and small noneffective current. A 5-/spl mu/m VCSEL exhibits a record threshold current of 380 /spl mu/A at 20/spl deg/C. This VCSEL also operates with single transverse mode up to the maximum optical output power.

We demonstrated a 25-Gb/s direct modulation up to 85degC with a 1.3- mum InGaAlAs ridge-waveguide multiple-quantum-well distributed-feedback laser. The dependence of the relaxation oscillation frequency on current was 3.3 GHz/mA1/2, and this is the highest value ever reported for 200-mum-long lasers in the 1.3-mum wavelength region. The alpha parameter was around 2.7 at 25degC, and an error-free operation after a 10-km single-mode fiber transmission was obtained up to 85degC.

Eosinophil granule major basic protein (MBP) is expressed exclusively in eosinophils and basophils in hematopoietic cells. In our previous study, we demonstrated a major positive regulatory role for GATA-1 and a negative regulatory role for GATA-2 in MBP gene transcription. Further analysis of the MBP promoter region identified a C/EBP (CCAAT/enhancer-binding protein) consensus binding site 6 bp upstream of the functional GATA-binding site in the MBP gene. In the cell line HT93A, which is capable of differentiating towards both the eosinophil and neutrophil lineages in response to retinoic acid (RA), C/EBP mRNA expression decreased significantly concomitant with eosinophilic and neutrophilic differentiation, whereas C/EBPβ expression was markedly increased. Electrophoretic mobility shift assays (EMSAs) showed that recombinant C/EBPβ protein could bind to the potential C/EBP-binding site (bp −90 to −82) in the MBP promoter. Furthermore, we have demonstrated that both C/EBPβ and GATA-1 can bind simultaneously to the C/EBP- and GATA-binding sites in the MBP promoter. To determine the functionality of both the C/EBP- and GATA-binding sites, we analyzed whether C/EBPβ and GATA-1 can stimulate the MBP promoter in the C/EBPβ and GATA-1 negative Jurkat T-cell line. Cotransfection with C/EBPβ and GATA-1 expression vectors produced a 5-fold increase compared with cotransfection with the C/EBPβ or GATA-1 expression vectors individually. In addition, GST pull-down experiments demonstrated a physical interaction between human GATA-1 and C/EBPβ. Expression of FOG (F̲riendo̲fG̲ATA), which binds to GATA-1 and acts as a cofactor for GATA-binding proteins, decreased transactivation activity of GATA-1 for the MBP promoter in a dose-dependent manner. Our results provide the first evidence that both GATA-1 and C/EBPβ synergistically transactivate the promoter of an eosinophil-specific granule protein gene and that FOG may act as a negative cofactor for the eosinophil lineage, unlike its positively regulatory function for the erythroid and megakaryocyte lineages.

Eosinophil granule major basic protein (MBP) is expressed exclusively in eosinophils and basophils in hematopoietic cells. In our previous study, we demonstrated a major positive regulatory role for GATA-1 and a negative regulatory role for GATA-2 in MBP gene transcription. Further analysis of the MBP promoter region identified a C/EBP (CCAAT/enhancer-binding protein) consensus binding site 6 bp upstream of the functional GATA-binding site in the MBP gene. In the cell line HT93A, which is capable of differentiating towards both the eosinophil and neutrophil lineages in response to retinoic acid (RA), C/EBPα mRNA expression decreased significantly concomitant with eosinophilic and neutrophilic differentiation, whereas C/EBPβ expression was markedly increased. Electrophoretic mobility shift assays (EMSAs) showed that recombinant C/EBPβ protein could bind to the potential C/EBP-binding site (bp −90 to −82) in the MBP promoter. Furthermore, we have demonstrated that both C/EBPβ and GATA-1 can bind simultaneously to the C/EBP- and GATA-binding sites in the MBP promoter. To determine the functionality of both the C/EBP- and GATA-binding sites, we analyzed whether C/EBPβ and GATA-1 can stimulate the MBP promoter in the C/EBPβ and GATA-1 negative Jurkat T-cell line. Cotransfection with C/EBPβ and GATA-1 expression vectors produced a 5-fold increase compared with cotransfection with the C/EBPβ or GATA-1 expression vectors individually. In addition, GST pull-down experiments demonstrated a physical interaction between human GATA-1 and C/EBPβ. Expression of FOG ( F riend o f G ATA), which binds to GATA-1 and acts as a cofactor for GATA-binding proteins, decreased transactivation activity of GATA-1 for the MBP promoter in a dose-dependent manner. Our results provide the first evidence that both GATA-1 and C/EBPβ synergistically transactivate the promoter of an eosinophil-specific granule protein gene and that FOG may act as a negative cofactor for the eosinophil lineage, unlike its positively regulatory function for the erythroid and megakaryocyte lineages.

Although peroxidase activity in basophils can be detected by optical and ultrastructural cytochemistry, its characteristics remain to be determined. We have demonstrated the characteristics of peroxidase activity induced in the immature basophil cell line, KU812-F. Ultrastructurally, peroxidase activity was detected in granules as well as in the perinuclear space and endoplasmic reticulum. Immunocytochemistry revealed that KU812-F cells were stained by anti-eosinophil peroxidase antibodies, and eosinophil peroxidase mRNA, not myeloperoxidase, was detected in the cells using Northern hybridization and reverse transcription-polymerase chain reaction. Eosinophil peroxidase can be one of the molecules shared with eosinophils and basophils. The biological function of eosinophil peroxidase detected in basophils remains uncertain.

Acute promyelocytic leukemia (APL) has a specific genetic rearrangement between the retinoic acid receptor (RAR)-α gene and the pml nuclear protein gene. All-trans retinoic acid (ATRA) induces granulocytic differentiation of APL-derived cells and is used to treat APL patients. However, ATRA interacts with normal cells with RAR throughout the entire body, and when used at high doses or over a long duration, it induces several adverse effects. The development of drugs that selectively act on APL cells may contribute to increasing the therapeutic efficacy of APL treatment as well as elucidating the mechanisms of response to ATRA. In this study, 9-cis retinoic acid α-tocopherol ester (9CTT) inhibited the proliferation of APL-derived NB4 and HT93 cells and induced differentiation markers, such as granulocytic maturation, nitroblue tetrazolium reduction, and CD11b expression, in these cells. The effects of 9CTT on non-APL cells, including HL-60 and U937 cells, were much weaker than those on APL cells, and tretinoin tocoferil (TT), which is an α-tocopherol ester of ATRA, did not induce the differentiation of APL cells as effectively as 9CTT. The differentiation-inducing effects of 9CTT were inhibited by RAR antagonists. 9CTT and TT similarly induced the transactivating activity of RARs, but were not effective on RXRs. 9CTT downregulated the expression of PML/RAR-α protein more effectively than TT, which suggests that it may be involved in the selectivity of 9CTT against APL cells. Interestingly, 9CTT enhanced the differentiation of APL cells induced by ATRA, 9-cis retinoic acid, and synthetic retinobenzoic acids. Combined with 1α,25-dihydroxyvitamin D3 (VD3), 9CTT also more than additively induced the differentiation of APL cells. Thus, 9CTT, alone or in combination with other retinoids or VD3, may be useful for the treatment of APL.

We have demonstrated an optical transceiver diode integrated with a spot-size converter. We clarify that the design of a detection layer is essential for the temperature- and polarization-insensitive characteristics of the responsivity. Because the detection layer was introduced, 0.65 dB of responsivity deviation was obtained in the wide temperature range of -10/spl deg/C-85/spl deg/C with a high laser performance. A spot-size converter enables the coupling loss to be 1.8 dB for a flat-end optical fiber. The frequency response was also obtained up to 1 GHz in both laser and detector modes.

Competitive RT-PCR was used to determine the quantitative variation in the expression of deoxycytidine kinase (dCK) gene in childhood leukemic cells. The degree of dCK gene expression varied over a 50-fold range. In two cases in which both primary and relapsed leukemic cells were analysed, decreased expression of dCK gene was found in relapsed leukemic cells. The sequence variation analysis using bisbenzimide/polyethylene glycol electrophoresis demonstrated no sequence alteration of dCK cDNA in all cases. These results indicate that the expression of dCK gene varies in patients and suggests decreased expression of the dCK gene as one of the mechanisms responsible for clinical resistance to ara-C.

The device structure and performance of 1.3-/spl mu/m narrow beam superluminescent diodes (NB-SLD's), which consist of a spot-size converter and a new type rear absorbing region, are reported. A butt-jointed selectively grown spot-size converter (SSC) is employed to realize the narrow beam characteristics. The rear absorbing region is designed as a taper structure with a part of the region is inclined from the active-stripe axes. In order to investigate the effects of both SSC length and active-region length on device performance, two types of NB-SLDs, whose SSC and active-region lengths differ, are fabricated. An electrode to sweep out photoexcited carriers in the absorption region is formed on one device. By comparing the characteristics of these devices, we clarify that a 500-/spl mu/m-active-region device is suitable for high-output power operation, and a 400-/spl mu/m-active-region device is suitable for realizing short coherent length. The light-output power is 13.9 mW at 200-mA-injection current for the former device, and the full-width at half-maximum (FWHM) of the spectrum is 62.6 nm (calculated coherence length is 26.5 /spl mu/m) for the latter device. Very small spectral modulation index (0.015 at 5 mW-output power) is attained by grounding the absorption-region electrode. For the SSC length, a 300-/spl mu/m SSC device shows very narrow far-field patterns (FFPs) and very good fiber-coupling characteristics. The FWHM of horizontal and vertical FFPs are 8.9 and 10.6/spl deg/, respectively. Because of this narrow beam divergence, the coupling efficiency of -1.9 dB to a flat-end 4-/spl mu/m spot-size fiber is obtained without lenses. The alignment tolerance of this device to the fiber for both horizontal and vertical direction is more than 3 /spl mu/m at a loss of when -1 dB from the optimum coupling.

A double-heterostructure (DH) laser with TM mode lasing has been achieved with a narrow active-layer width, and a laser-diode optical switch (LDSW) module with less than a 0.35-dB gain difference between the TE and TM modes over a wide wavelength range has been constructed by introducing a square bulk active layer formed by dry etching and regrowth. The polarization-insensitive width of a 0.3-/spl mu/m-thick DH laser is clarified to be between 0.30 and 0.35 /spl mu/m, since the 0.30- and 0.35-/spl mu/m-wide DH lasers lase in the TM mode and TE mode, respectively. The polarization-insensitive width of the fabricated 0.3-/spl mu/m-thick LDSW is estimated to be about 0.32 /spl mu/m for the fabricated LDSW with a trapezoidal active layer by measuring the single-pass gain and the gain difference between the TE and TM modes. This must be to within 0.01 /spl mu/m. A 0.35-/spl mu/m-wide, 300-/spl mu/m-long LDSW module has lossless gain in the wavelength range of 1.31 to 1.36 /spl mu/m at 20 mA. The gain difference between the TE and TM modes is as low as 0.35 dB, The rise and fall times are 1.0 and 0.55 ns, respectively. The bulk active-layer LDSW module is promising for use as a polarization-insensitive optical-gate switch in optical information systems.

We experienced a case of Burkitt's lymphoma showing an unusual surface phenotype, CD5 expression, at an early stage of the disease. Initially, this patient showed massive abdominal para-aortic lymph node swelling which rapidly developed into leukemic change. Based on the clinical course and cytogenetic features of lymphoblasts in the bone marrow, which showed t(8;14) and c-myc gene rearrangement, the patient was diagnosed with Burkitt's lymphoma. Combination chemotherapy induced short-term remission, but central nervous system (CNS) involvement developed, followed by a regrowth of lymphoma cells in the bone marrow. The bone marrow at the end stage showed monotonous expansion of large cells with conspicuous vacuolation in the basophilic cytoplasm. The initial lymphoma cells showed pan-B markers and were CD5 positive but weakly CD10 positive; however, the lymphoma cells obtained from the bone marrow at the terminal stage did not express CD5. The chromosomal t(8;14) was seen, and identical rearrangement of immunoglobulin heavy chain joining gene and c-myc gene were detected by Southern blot analysis in the bone marrow lymphoblasts throughout the clinical course. This case is evidence that remarkable transformation of CD5-positive lymphoblasts to CD5-negative lymphoblasts occurred in an identical clone of Burkitt's lymphoma.

A patient presented with chronic large granular lymphocyte leukemia associated with chronic active Epstein-Barr virus infection (CAEBV). Cell cycle analysis revealed a minimal growth compatible with chronic lymphocytic leukemia After 5 months of treatment, the patient died from acute transformation of the leukemia. Cell harvested during chronic phase were analyzed for sensitivity to interleukin 2 (IL-2) and interferon alpha (IFN alpha) in vitro by means of surface phenotyping and cell cycle assay. IL-2 induced remarkable growth of the cells, whereas IFN alpha did not confer a growth advantage. Since IFN alpha was expected to have no growth induction effect on the leukemia cells, it was administered to the patient to treat the CAEBV.

Primary familial and congenital polycythaemia (PFCP) is a rare disease characterized by congenital erythrocytosis inherited in an autosomal dominant fashion. Recently, mutations in the erythropoietin receptor (EpoR) have been identified in PFCP families. We describe a Japanese family with an autosomal dominant inheritance of PFCP. An in vitro colony assay demonstrated hypersensitivity of erythroid progenitors to erythropoietin (Epo) in affected family members. Sequence analysis of RT-PCR products amplified from the C-terminal region of EpoR transcripts in affected family members revealed that they were all heterozygous for C and T bases at position 5986, which suggested a genetic mutation (C to T) on one allele of EpoR. This mutation gave rise to a translation termination codon TAG at amino acid 435. Thus, the resulting EpoR is a truncated protein product lacking all 74 amino acids downstream of the mutation. To date, all genetic mutations affecting a family with PFCP, including this one, have been located in the cytoplasmic negative regulatory region of the EpoR. All mutations gave rise to truncated Epo receptors between Tyrosine 427 and Tyrosine 455. The phosphotyrosines in this region of EpoR have been demonstrated to be binding sites for SHP-1 phosphatase. Therefore PFCP is presumably brought about as a result of genetic mutations which cause the loss of the SHP-1 binding site in the cytoplasmic region of EpoR.

Laser diodes monolithically integrated with spot-size converters operating at 1.3 /spl mu/m, and having an almost circular narrow emitted beam, have been successfully fabricated using 2 inch (50.8 mm) full wafer fabrication technology. The spot-size converters consist of a lateral taper, thin-film guiding layer and ridge structure. The epitaxial structure is designed to have wide fabrication tolerance with no deterioration of device characteristics. The investigation of the coupling-loss dependence on the dimension parameters in the spot-size converters confirms that this device has a wide fabrication tolerance for these parameters. Furthermore, butt-coupling loss to fibre as low as -1.3 dB is attained with alignment tolerance wide enough for passive alignment. The overall device is as short as 450 /spl mu/m, enabling high availability from one wafer.

To assess the consequence of second BMT (BMT2) for leukemia relapse after allogeneic BMT, we analyzed the clinical course of 66 recipients who were treated by BMT2 in Japan. Diagnoses included 29 ANLL, 27 ALL, six CML and four MDS. Durations between the first BMT (BMT1) to relapse and BMT1 to BMT2 were 13.5 +/- 13.7 months and 17.4 +/- 13.9 months, respectively. Donors for BMT2 were replaced in 11 cases. Thirty-one patients were in CR (or CP) at BMT2. Earlier deaths were observed in those who received BMT2 within 12 months after BMT1, mostly caused by regimen-related toxicity and infections. Overall leukemia-free survival rate was 28% at 2 years and 16% at 4 years. Factors influencing the poor prognosis after BMT2 were early (

Laser diodes integrated with spotsize converters by butt-joint technology combined with selective area metal organic vapor phase epitaxial (MOVPE) growth have been successfully fabricated. Satisfactory uniformity, reproducibility (>99%) and tolerance for low threshold current, a narrow emitted beam, and low optical coupling loss to fiber (

Several means of analyzing minimal residual disease (MRD) in leukemia involving the rearranged T cell receptor (TCR) gene have been described. We investigated MRD in leukemia with TCR beta rearrangement by examining TCR beta-chain RNA. A complementary DNA (cDNA) corresponding to the variable region of the TCR beta-chains originating from the peripheral blood or bone marrow from four patients was amplified. Single strand conformation polymorphism (SSCP) analysis of amplified cDNA showed that all four patients had monoclonal leukemia with TCR beta rearrangement; two patients had Vbeta2+ leukemia, another patient had Vbeta14+ leukemia and the other had Vbeta9+ leukemia. Flow cytometry supported this finding. Sequencing of the Vbeta2-complementarity determining region 3 (CDR3), Vbeta9-CDR3 and Vbeta14-CDR3 revealed monoclonality. To investigate MRD using TCR beta-chain RNA, cDNA from each patient was diluted with the cDNA of a healthy person and amplified using a specific CDR3 clonotype primer. A band in the ethidium bromide-stained agarose gel was detected from samples diluted 10,000-fold. SSCP analysis determined which V region gene was utilized in monoclonal leukemic cells. The leukemic cell specific TCR, determined in such a manner, may be a target for immunotherapy. Because the MRD of T cell malignancy can be easily examined once the CDR3 clonotype primer is made, this novel analysis is considered to be a useful method.

We clarified the degradation behaviors of semi-insulating buried heterostructure lasers in which mesa structures were fabricated by reactive ion etching (RIE) and then buried in semi-insulating Fe-doped InP grown by metal organic vapor phase epitaxy (MOVPE). The degradation rate and mode correlated with the quality of the buried heterostructure (BH) interface. Based on the correlation, a condition for highly stable semi-insulating Fe-doped InP buried heterostructure (SIBH) lasers was demonstrated and confirmed experimentally and statistically.

Purpose: Frequently recurrent leukemia is encountered in childhood and is usually resistant to multidrug therapy. We evaluated the efficacy of differentiation induction therapy in a child with leukemia. Patient and Methods: We administered all-trans retinoic acid (ATRA) alone, and then a combination of low-dose 1-β-D-arabinofuranosylcytosine (Ara-C), to a 7-year-old boy who had leukemia secondary to etoposide with chromosomal abnormality 11q23. Results: The patient exhibited a good response to ATRA, and had a complete remission after combination therapy of ATRA and low-dose Ara-C. Conclusion: Differentiation induction therapy with ATRA was effective in treating a patient with recurrence of leukemia without t(15;17) translocation, and which is resistant to conventional cytotoxic treatment.

Two patients with hormone-refractory advanced prostate cancer were treated with high-dose chemotherapy and peripheral blood stem-cell transplantation. A satisfactory number of stem cells were collected by appropriately timed leukapheresis in these patients. The stem-cell samples collected from the first patient were positive for prostate-specific antigen messenger ribonucleic acid expression; such expression was not detected in the samples from the second patient. The first patient has remained in complete remission as of December 1995. The second patient died of the disease.

A new monolithically integrated opto-electronic device has been developed for fast wavelength switching in photonic packet switching. The integrated device, called a tunable duplex integrated light source comprises two distributed Bragg reflector lasers (DBR-LDs), two electroabsorption optical switches, a Y-shaped waveguide coupler, an electroabsorption modulator, and two thermal drift compensators (TDCs). The tuning range of the DBR-LDs is 3 nm, and the wavelength switching time of the optical switches is estimated to be 60 ps including a 50 ps rise time for the electrical pulse generator. The wavelength of a 10 Gb/s NRZ-modulated optical signal is switched without bit loss. Thermal crosstalk between the DBR-LDs, and wavelength drift of the DBR-LDs, which is due to a thermal transient, are effectively suppressed by thermal-drift-compensation operation using the TDCs.

Ten patients with acute promyelocytic leukemia (APL) were treated with all-trans retinoic acid (ATRA). Eight of 10 patients achieved complete remission (CR), and among the 8 newly diagnosed cases, 7 achieved CR. Five of 8 CR cases remained in CR after 8 to 30 months. Except for hypotension and a large gastric ulcer resulting from hyperhistaminemia, the adverse effects of ATRA were generally mild. Severe thrombotic tendency occurred in a patient treated with ATRA combined with tranexamic acid. Intensive chemotherapy consisting of daunorubicin (DNR) and other agents was scheduled for the patients who achieved CR with ATRA.

We report on an InP-based optical multiplexer integrated with high-speed multiquantum-well (MQW) electroabsorption modulators for 100 Gb/s transmission. The optical multiplexer is based on a multimode interference (MMI) splitter, a waveguide array, and a taper-type combiner designed using the beam propagation method (BPM). The modulators are integrated on the top of the optical multiplexer. The integrated device is fabricated by three step epitaxy using metal organic vapor phase epitaxy (MOVPE) and by dry etching. The static and dynamic performances are measured, leading to the generation and the modulation of a 100 GHz optical pulse train. This optical pulse train is also coded using a pseudorandom nonreturn to zero (NRZ) pattern.

PURPOSE We evaluated the changes in stem cells in peripheral blood and the optimal timing for harvesting peripheral blood stem cells (PBSCs) following the administration of conventional cancer chemotherapy in patients with urological malignancies. PATIENTS AND METHODS We evaluated the changes in PBSCs after the administration of cancer chemotherapy in 12 patients with urological malignancies (6 testis cancer, 3 bladder cancer, 3 prostate cancer) using flowcytometry analysis of CD34, a marker of blood stem cell. We evaluated the optimal timing for PBSCs harvesting following the administration of conventional cancer chemotherapy in 6 patients with urological malignancies (2 patients with testis cancer, 3 patients with prostate cancer, 1 patient with bladder cancer) using flowcytometry analysis of CD34 and granulocyte-macrophage colony-forming units (CFU-GM) counts. PBSC samples from 2 patients with prostate cancer were analyzed by the reverse transcriptase-polymerase chain reaction (RT-PCR) method to detect prostate cancer cells. RESULTS The percentages of CD34-positive cells tended to increase in the peripheral blood during the period of convalescence that followed the induction of marked myelosuppression produced by the cancer chemotherapy. The peak in CD34-positive cells occurred between days 15 and 31 after beginning of cancer chemotherapy. The percentage of CD34-positive cells was increased in the mononuclear cells in harvested PBSCs. The percentages of CD34-positive cells in peripheral blood and in mononuclear cells harvested from PBSCs was correlated with the CFU-GM counts in harvested PBSCs. The CFU-GM counts peaked between days 16 to 31 following cancer chemotherapy. RT-PCR detected prostate-specific antigen (PSA) mRNA expression in PBSCs obtained from 1 of the 2 patients with prostate cancer. An amount of contamination with PSA mRNA reduced as chemotherapy was repeated. CONCLUSION A sufficient number of PBSCs can be harvested after conventional cancer chemotherapy without the need for special cytotoxic mobilization that may be delayed the patient's treatment. RT-PCR results suggested that the harvested PBSCs should be examined for tumor cell contamination in patients with metastasis in bone marrow. If a sufficient amount of PBSCs will be harvested, the PBSC samples from the patient with bone marrow metastasis after repeated chemotherapy should be used by priority for PBSC transplantation as of today.

The expression of the c-kit gene product has been examined in normal mast cells, mast cell neoplasms, and basophil/mast cell precursors obtained from patients with chronic myelogenous leukaemia (CML). Formalin-fixed, paraffin-embedded sections or smears fixed with formalin vapour were studied by immunohistochemical methods, using a polyclonal antibody against the c-kit gene product. Normal and neoplastic mast cells showed a positive immunoreaction for c-kit gene product, but neoplastic basophil/mast cell precursors from CML patients lacked c-kit gene product by immunohistochemical and flow cytometric methods, even in cells having mast cell granules, together with or without basophil granules. Mast cell tryptase was, however, expressed in normal and neoplastic mast cells and basophil/mast cell precursors containing mast cell granules. In addition, cells of monocyte/macrophage lineage lacked c-kit gene product. These findings indicate that the c-kit gene product may play an important role in the development and function of mast cell but not of cell of basophil and monocyte/macrophage lineage.